Determination of glucose deficiency-induced cell death by mitochondrial ATP generation-driven proton homeostasis.
Glucose is one of major nutrients and its catabolism provides energy and/or building bricks for cell proliferation. Glucose deficiency results in cell death. However, the underlying mechanism still remains elusive. By using our recently developed method to monitor real-time cellular apoptosis and necrosis, we show that glucose deprivation can directly elicit necrosis, which is promoted by mitochondrial impairment, depending on mitochondrial adenosine triphosphate (ATP) generation instead of ATP depletion. We demonstrate that glucose metabolism is the major source to produce protons. Glucose deficiency leads to lack of proton provision while mitochondrial electron transfer chain continues consuming protons to generate energy, which provokes a compensatory lysosomal proton efflux and resultant increased lysosomal pH. This lysosomal alkalinization can trigger apoptosis or necrosis depending on the extent of alkalinization. Taken together, our results build up a metabolic connection between glycolysis, mitochondrion, and lysosome, and reveal an essential role of glucose metabolism in maintaining proton homeostasis to support cell survival.


Graphene-silicon-graphene Schottky junction photodetector with field effect structure.
Graphene has unique advantages in ultrabroadband detection. However, nowadays graphene-based photodetectors cannot meet the requirements for practical applications due to their poor performance. Here, we report a graphene-silicon-graphene Schottky junction photodetector assisted by field effect. Two separate graphene sheets are located on both sides of the n-doped silicon to form two opposite lateral series heterojunctions with silicon, and a transparent top gate is designed to modulate the Schottky barrier. Low doping concentration of silicon and negative gate bias can significantly raise the barrier height. Under the combined action of these two measures, the barrier height increases from 0.39 eV to 0.77 eV. Accordingly, the performance of the photodetector has been greatly improved. The photoresponsivity of the optimized device is 2.6 A/W at 792 nm, 1.8 A/W at 1064 nm, and 0.42 A/W at 1550 nm, and the on/off photo-switching ratio reaches 104. Our work provides a feasible solution for the development of graphene-based optoelectronic devices.


IGF binding proteins (IGFBPs) and regulation of breast cancer biology.
The IGFBP family comprises six proteins with high affinity for the IGFs. Changes in the balance of the components of the IGF system may contribute to the progression of breast cancer. In tumours the abundance of IGFBPs relates to the estrogen receptor status and their production in the breast is controlled by hormones, principally estrogen and progesterone. Important interactions occur between IGFBPs and key growth regulators such as TGF-beta, PTEN and EGF which are reviewed. The conflicting observations between the effects of IGFBPs on the risk of breast cancer, in particular IGFBP-3, obtained from epidemiology studies in comparison to in vivo observations are highlighted and potential explanations provided. The functional activity of IGFBPs can also be affected by proteolysis, phosphorylation and glycosylation and the implications of these are described. The IGFs are generally present at levels far in excess of that required for maximal receptor stimulation, and the IGFBPs are critical regulators of their cellular actions. IGFBPs can affect cell function in an IGF-dependent or independent manner. The key mechanisms underlying the intrinsic actions of the IGFBPs are still in debate. IGF bioactivity locally in the breast is influenced not only by local tissue expression and regulation of IGFs, IGFBPs and IGFBP proteases, but also by these factors delivered from the circulation. Finally, the therapeutic potential of IGFBPs-2 and -3 are considered together with key questions that still need to be addressed.


Effects of disorder in two-dimensional photonic crystal waveguides.
The effects of randomness on the guiding properties of waveguides embedded in disordered two-dimensional photonic crystals composed of a finite cluster of circular cylinders of infinite length are investigated for TM-polarized radiation. Different degrees of disorder in the radius, filling fraction, refractive index, and position are considered for both straight and 90 degrees bent guides. The crystals exhibit similar sensitivity to refractive index and radius disorder, with a degree of disorder from 15%-20% yielding little substantial change in the guiding properties. A smaller range of position disorder is also considered. For strong disorder in radius and refractive index, the guide effectively closes. These results were obtained by a Monte Carlo simulation method, and the performance of this method is analyzed. The method requires at least ten realizations in some cases for convergence to commence; substantially more realizations are required for moderate and strong disorder to achieve accurate results.


The butyrophilin 3A1 intracellular domain undergoes a conformational change involving the juxtamembrane region.
Small isoprenoid diphosphates, such as (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate (HMBPP), are ligands of the internal domain of BTN3A1. Ligand binding in target cells promotes activation of Vγ9Vδ2 T cells. We demonstrate by small-angle X-ray scattering (SAXS) that HMBPP binding to the internal domain of BTN3A1 induces a conformational change in the position of the B30.2 domain relative to the juxtamembrane (JM) region. To better understand the molecular details of this conformational rearrangement, NMR spectroscopy was used to discover that the JM region interacts with HMBPP, specifically at the diphosphate. The spectral location of the affected amide peaks, partial NMR assignments, and JM mutants (ST296AA or T304A) investigated, confirm that the backbone amide of at least one Thr (Thr304), adjacent to conserved Ser, comes close to the HMBPP diphosphate, whereas double mutation of nonconserved residues (Ser/Thr296/297) may perturb the local fold. Cellular mutation of either of the identified Thr residues reduces the activation of Vγ9Vδ2 T cells by HMBPP, zoledronate, and POM2-C-HMBP, but not by a partial agonist BTN3 antibody. Taken together, our results show that ligand binding to BTN3A1 induces a conformational change within the intracellular domain that involves the JM region and is required for full activation.-Nguyen, K., Li, J., Puthenveetil, R., Lin, X., Poe, M. M., Hsiao, C.-H. C., Vinogradova, O., Wiemer, A. J. The butyrophilin 3A1 intracellular domain undergoes a conformational change involving the juxtamembrane region.


Effects of Individualized Gait Rehabilitation Robotics for Gait Training on Hemiplegic Patients: Before-After Study in the Same Person.
Lower-limb exoskeleton robots are being widely used in gait rehabilitation training for patients with stroke. However, most of the current rehabilitation robots are guided by predestined gait trajectories, which are often different from the actual gait trajectories of specific patients. One solution is to train patients using individualized gait trajectories generated from the physical parameters of patients. Hence, we aimed to explore the effect of individual gaits on energy consumption situations during gait rehabilitation training for hemiplegic patients with lower-limb exoskeleton robots. A total of 9 unilateral-hemiplegic patients were recruited for a 2-day experiment. On the first day of the experiment, the 9 patients were guided by a lower-limb exoskeleton robot, walking on flat ground for 15 min in general gait trajectory, which was gained by clinical gait analysis (CGA) method. On the other day, the same 9 patients wore the identical robot and walked on the same flat ground for 15 min in an individualized gait trajectory. The main physiological parameters including heart rate (HR) and peripheral capillary oxygen saturation (SpO2) were acquired via cardio tachometer and oximeter before and after the walking training. The energy consumption situation was indicated by the variation of the value of HR and SpO2 after walking training compared to before. Between-group comparison showed that the individualized gait trajectory training resulted in an increase in HR levels and a decrease in SpO2 levels compared to the general gait trajectory training. The resulting difference had a statistical significance of p < 0.05. Using individualized gait guidance in rehabilitation walking training can significantly improve energy efficiency for hemiplegic patients with stroke.


[Molecular cloning and analysis of a monocot mannose-binding agglutinin from Zephyranthes grandiflora (family Amaryllidaceae)].
The monocot mannose-binding lectin can inhibit HIV from infecting the target cells. The total RNA of Zephyranthes grandiflora was extracted and reversely transcribed into cDNA. Degenerate primers were designed based on the conserved regions of other monocot mannose-binding agglutinins by homology alignment. The 694bp full-length cDNA of Zephyranthes grandiflora agglutinin (ZGA) was cloned by RT-PCR, 3' and 5' RACE (rapid amplification of cDNA ends). The start codon and stop codon of ZGA were at 37-39bp and 529-531bp respectively. The NCBI Blast analysis result showed that ZGA gene encoded a protein precursor with signal peptide, mature protein and C-terminal cleavage sequence. The mature ZGA protein contained 106 amino acids residues and its molecular weight was 11.6KD. The percentages of identity of the deduced mature ZGA protein with those of Galanthus nivalis agglutinin, Narcissus hybrid cultivar agglutinin, Lycoris radiate agglutinin and Clivia miniata agglutinin were 71.8%, 81%, 81.8% and 84.5%, respectively. Blocks analysis revealed that ZGA had three functional domains and three mannose-binding boxes (QDNY).


Inhibited insulin signaling in mouse hepatocytes is associated with increased phosphatidic acid but not diacylglycerol.
Although an elevated triacylglycerol content in non-adipose tissues is often associated with insulin resistance, the mechanistic relationship remains unclear. The data support roles for intermediates in the glycerol-3-phosphate pathway of triacylglycerol synthesis: diacylglycerol (DAG), which may cause insulin resistance in liver by activating PKCϵ, and phosphatidic acid (PA), which inhibits insulin action in hepatocytes by disrupting the assembly of mTOR and rictor. To determine whether increases in DAG and PA impair insulin signaling when produced by pathways other than that of de novo synthesis, we examined primary mouse hepatocytes after enzymatically manipulating the cellular content of DAG or PA. Overexpressing phospholipase D1 or phospholipase D2 inhibited insulin signaling and was accompanied by an elevated cellular content of total PA, without a change in total DAG. Overexpression of diacylglycerol kinase-θ inhibited insulin signaling and was accompanied by an elevated cellular content of total PA and a decreased cellular content of total DAG. Overexpressing glycerol-3-phosphate acyltransferase-1 or -4 inhibited insulin signaling and increased the cellular content of both PA and DAG. Insulin signaling impairment caused by overexpression of phospholipase D1/D2 or diacylglycerol kinase-θ was always accompanied by disassociation of mTOR/rictor and reduction of mTORC2 kinase activity. However, although the protein ratio of membrane to cytosolic PKCϵ increased, PKC activity itself was unaltered. These data suggest that PA, but not DAG, is associated with impaired insulin action in mouse hepatocytes.


Ultrasonographic approach to diagnosing chest wall tumors.
The value of ultrasonography and UGAB for diagnosing chest wall tumors was investigated in 21 patients of whom 13 had metastatic and eight benign disease. Chest wall tumor showed a hypoechoic mass with tapered edges. The margin between the tumor and lung was curved and smooth in all cases. Movement of the tumor during breathing was synchronized with that of the chest wall. We believe that the ultrasonography is of value in identifying tumors that are localized to the chest wall by examination of their margin and movement with respiration, in addition to their shape. Using UGAB, we were able to diagnose correctly all eight metastatic tumors and two of four benign tumors. UGAB is a safe and simple procedure for diagnosing chest wall tumors.


Immunoprecipitation of in vitro-translated products of Nippostrongylus brasiliensis RNA.
RNA has been isolated from adult Nippostrongylus brasiliensis (NB) homogenized in guanidium isothiocyanate. The mRNA has been translated in vitro utilizing a rabbit reticulocyte system and translated products immunoprecipitated with rabbit anti-adult NB antiserum and rat infection serum.


Sonoporation, drug delivery, and gene therapy.
Ultrasound is a very effective modality for drug delivery and gene therapy because energy that is non-invasively transmitted through the skin can be focused deeply into the human body in a specific location and employed to release drugs at that site. Ultrasound cavitation, enhanced by injected microbubbles, perturbs cell membrane structures to cause sonoporation and increases the permeability to bioactive materials. Cavitation events also increase the rate of drug transport in general by augmenting the slow diffusion process with convective transport processes. Drugs and genes can be incorporated into microbubbles, which in turn can target a specific disease site using ligands such as the antibody. Drugs can be released ultrasonically from microbubbles that are sufficiently robust to circulate in the blood and retain their cargo of drugs until they enter an insonated volume of tissue. Local drug delivery ensures sufficient drug concentration at the diseased region while limiting toxicity for healthy tissues. Ultrasound-mediated gene delivery has been applied to heart, blood vessel, lung, kidney, muscle, brain, and tumour with enhanced gene transfection efficiency, which depends on the ultrasonic parameters such as acoustic pressure, pulse length, duty cycle, repetition rate, and exposure duration, as well as microbubble properties such as size, gas species, shell material, interfacial tension, and surface rigidity. Microbubble-augmented sonothrombolysis can be enhanced further by using targeting microbubbles.


Iridium-Complexed Dipyridyl-Pyridazine Organosilica as a Catalyst for Water Oxidation.
The heterogenization of metal-complex catalysts to be applied in water oxidation reactions is a currently growing field of great scientific impact for the development of energy conversion devices simulating the natural photosynthesis process. The attachment of IrCp*Cl complexes to the dipyridyl-pyridazine N-chelating sites on the surface of SBA-15 promotes the formation of metal bipyridine-like complexes, which can act as catalytic sites in the oxidation of water to dioxygen, the key half-reaction of artificial photosynthetic systems. The efficiency of the heterogeneous catalyst, Ir@NdppzSBA, in cerium(IV)-driven water oxidation was thoroughly evaluated, achieving high catalytic activity even at a long reaction time. The reusability and stability were also examined after three reaction cycles, with a slight loss of activity. A comparison with an analogous homogeneous iridium catalyst revealed the enhanced durability and performance of the heterogeneous system based on the Ir@NdppzSBA catalyst due to the stability of the SBA-15 structure as well as the isolated metal active sites. Thereby, this new versatile synthesis route for the preparation of water oxidation catalysts opens a new avenue for the construction of alternative heterogeneous catalytic systems with high surface area, ease of functionalization, and facile separation to improve the efficiency in the water oxidation reaction.


Long-term results of therapy with intracavernousal injections and penile venous surgery in chronic erectile dysfunction.
A follow up study of 603 patients with chronic erectile failure was carried out concerning the long term effects of intracavernous injection therapy with papaverine/phentolamine (n = 172 evaluable) and venous surgery (n = 134 evaluable). Injection therapy was effective in 95.8% of injections. Side effects included prolonged erections in 0.15% of injections and fibrotic palpable indurations in 3.4%. Acceptance of therapy was high among patients and their partners and improvement of self image or partnership was reported in 77.8% and 79.5% respectively. Results of venous surgery were not satisfactory. Only 19.4% of patients achieved sufficient spontaneous erections and 35.1% needed adjuvant injections. Spongiosolysis yielded better results. Patients with neurological impairment, diabetes mellitus, postoperative onset of erectile dysfunction and age above 40 years showed poorer surgical results. Venous surgery should only be offered to a highly selected group of patients.


Undifferentiated endometrial carcinoma: a diagnosis frequently overlooked.
Undifferentiated endometrial carcinoma (UEC) is a relatively uncommon neoplasm with only few studies published thus far. It has always been a diagnostic challenge because of the lack of proper definition cited in most of the standard textbooks. Recently however, a few studies have highlighted the clinicopathologic features of UEC. The distinctive morphology of UEC was noted by the group from MD Anderson Cancer Center, which enabled them to establish the defining criteria. It appears to be more aggressive than endometrial endometrioid adenocarcinoma, FIGO (International Federation of Gynecology and Obstetrics) grade 3, its main differential diagnosis. Proper recognition of this entity is important owing to its aggressive behavior.


Targeting restoration sites to improve connectivity in a tiger conservation landscape in India.
Maintaining and restoring connectivity between source populations is essential for the long term viability of wide-ranging species, many of which occur in landscapes that are under pressure to meet increasing infrastructure needs. Identifying barriers in corridors can help inform conservation and infrastructure development agencies so that development objectives can be achieved without compromising conservation goals. Here, we use the tiger landscape in central India as a case study to identify barriers, associate them with existing infrastructure, and quantify the potential improvement by restoring or mitigating barriers. Additionally, we propose an approach to categorize linkages based on their current status within and between Protected Areas (PAs). We generated a hybrid landuse-landcover map of our study area by merging datasets. We used least-cost methods and circuit theory to map corridors and generate linkage metrics. We mapped barriers and used the improvement score (IS) metric to quantify potential improvement by restoring or mitigating them. Based on criteria that represent the status of corridors between-PAs and populations within-PAs, we ranked linkages into one of four categories: Cat1-linkages that currently have high quality and potential for tiger connectivity and should be maintained, Cat2W-linkages where focus on habitat and tiger populations may improve connectivity, Cat2B-linkages where focus on reducing barriers between PAs may improve connectivity, and Cat3-linkages where effort is needed to both reduce barriers between PAs and improve tiger populations and habitat within PAs. We associated barriers with infrastructure and present maps to show where restoration or mitigation measures can be targeted to have the highest potential impact. We mapped 567 barriers within 30 linkages in this landscape, of which 265 barriers intersect with infrastructure (694 km of roads, 150 km of railway, 48 reservoirs, 10 mines) and 302 barriers are due to land-use or gaps in forest cover. Eighty-six barriers have both roads and railways. We identified 7 Cat1, 4 Cat2w, 9 Cat2b, and 10 Cat3 linkages. Eighty surface mines and thermal power plants are within 10 km of the least-cost paths, and more coal mines are closer to connectivity areas where linkages are narrow and rank poorly on both axes. We present spatial and quantitative results that can help conservation practitioners target mitigation and restoration efforts. India is on the path to rapid economic growth, with infrastructure development planned in biodiversity-rich areas. The mitigation hierarchy of avoiding, minimizing, and offsetting impacts due to proposed development projects can be applied to corridors in this landscape. Cross-sectoral cooperation at early stages of project life-cycles to site, design, and implement solutions can maintain connectivity while meeting infrastructure needs in this rapidly changing landscape.


Bleeding related to disturbed fibrinolysis.
The components and reactions of the fibrinolysis system are well understood. The pathway has fewer reactants and interactions than coagulation, but the generation of a complete quantitative model is complicated by the need to work at the solid-liquid interface of fibrin. Diagnostic tools to detect disease states due to malfunctions in the fibrinolysis pathway are also not so well developed as is the case with coagulation. However, there are clearly a number of inherited or acquired pathologies where hyperfibrinolysis is a serious, potentially life-threatening problem and a number of antifibrinolytc drugs are available to treat hyperfibrinolysis. These topics will be covered in the following review.


Effects of extracellular vesicles released by peritoneal B-1 cells on experimental Leishmania (Leishmania) amazonensis infection.
B-1 cells are a B-lymphocyte subtype whose roles in immunity are not completely defined. These cells can produce cytokines (mainly IL-10) and natural and specific antibodies. Currently, extracellular vesicles (EVs) released by immune cells have emerged as new important entities in cell-cell communication. Immune cells release EVs that can activate and/or modulate other immune cells. Here, we characterized the EVs released by peritoneal B-1 cells infected or not with Leishmania (Leishmania) amazonensis. This Leishmania species causes cutaneous leishmaniasis and can infect macrophages and B-1 cells. Our results showed that peritoneal B-1 cells spontaneously release EVs, but the parasite stimulated an increase in EVs production by peritoneal B-1 cells. The treatment of BALB/c and C57BL/6 bone marrow-derived macrophages (BMDM) with EVs from infected peritoneal B-1 cells led to differential expression of iNOS, IL-6, IL-10, and TNF-α. Additionally, BALB/c mice previous treated with EVs released by peritoneal B-1 cells showed a significant lower lesion size and parasite burden. Thus, this study demonstrated that peritoneal B-1 cells could release EVs that can alter the functions of macrophages in vitro and in vivo these EVs altered the course of L. amazonensis infection. These findings represent the first evidence that EVs from peritoneal B-1 cells can act as a new mechanism of cellular communication between macrophages and B-1 cells, contributing to immunity against experimental leishmaniasis.


Imaging the neonate in the incubator: an investigation of the technical, radiological and nursing issues.
Modern neonatal incubators incorporate an X-ray tray device into the mattress support structure to facilitate patient examination with minimal disturbance and distress. However, the usual method of examination is to place the image plate directly underneath the baby. Users often cite radiological reasons for not using X-ray trays but modern quantitative evidence is lacking. This work looks at the technical and clinical aspects of imaging neonates in incubators and the impact that these may have in determining the imaging protocol. A number of hospitals were surveyed to determine their current method of examination and the reasons for their preference. Experimental measurements of the radiological impact of using (or not using) the X-ray tray were performed for a range of neonatal incubators. The average dose to the image plate was 5.9 microGy (range 5.4-6.4 microGy) for the "plate on mattress" method and 3.0 microGy (2.0-3.8 microGy) when using the tray--a 49% reduction owing to the mattress support materials. However, when using a computed radiography (CR) imaging system, the image quality differences were marginal. Survey results indicated that nurses preferred to use the tray but that radiographers were reluctant. We conclude that incubator manufacturers could do much to improve the radiological performance of their equipment and we offer recommendations. We also conclude that, with appropriate nurse and radiographer training and the advent of CR imaging systems, use of X-ray tray facilities may optimize imaging of the neonate in the incubator.


[Modifications of cerebral amine metabolism in psychiatric illness].
Abnormalities in brain monoamines have been implicated in the pathogenesis of several psychiatric disorders. A brief revue of biochemical studies on indolamines and catecholamines in mental disorders with special emphasis on endogenous depression will be presented. Progress in neurochemistry makes it possible to investigate the biochemical disturbances present in the major psychoses. Recent knowledge in the neuropharmacological mechanism of action of psychotropic drugs have shed some light on the basic understanding of the pharmacological treatment of psychotic disorders. The implication of biogenic amine studies in psychiatry for endocrinological research in mental disorders will also be discussed.


Corticosterone-Induced Lipogenesis Activation and Lipophagy Inhibition in Chicken Liver Are Alleviated by Maternal Betaine Supplementation.
We have shown previously that in ovo betaine injection can prevent nonalcoholic fatty liver induced by glucocorticoid exposure in chickens; yet it remains unknown whether feeding betaine to laying hens may exert similar effects in their progeny. In this study, we fed laying hens a betaine-supplemented diet, and the progeny were later exposed chronically to corticosterone (CORT) to test hepatoprotective effects and further elucidate underlying mechanisms. Rugao yellow-feathered laying hens (n = 120) were fed a basal (control, C) diet or a 0.5% betaine-supplemented (B) diet for 28 d before their eggs were collected for incubation. At 49 d of age, male chickens selected from each group were daily injected subcutaneously with solvent (15% ethanol; vehicle, VEH) or CORT (4.0 mg/kg body mass) for 7 d to establish a fatty liver model. Chickens in the 4 groups (C-VEH, C-CORT, B-VEH, and B-CORT) were killed at day 57. Plasma and hepatic triglyceride (TG) concentrations, as well as the hepatic expression of genes involved in lipogenesis and lipophagy, were determined. CORT induced a 1.6-fold increase in the plasma TG concentration (P < 0.05) and a 1.8-fold increment in the hepatic TG concentration (P < 0.05), associated with activation of lipogenic genes (70-780%). In contrast, lipophagy and mitochondrial β-oxidation genes were inhibited by 30-60% (P < 0.05) in CORT-treated chickens. These CORT-induced changes were completely normalized by maternal betaine supplementation or were partially normalized to intermediate values that were significantly different from those in the C-VEH and C-CORT groups. These effects were accompanied by modifications in CpG methylation and glucocorticoid receptor binding to the promoters of major lipogenic and lipophagic genes (P < 0.05). These results indicate that maternal betaine supplementation protects male juvenile chickens from CORT-induced TG accumulation in the liver via epigenetic modulation of lipogenic and lipophagic genes.


Observations of rat fetuses after irradiation with 2450-MHz (CW) microwaves.
Female Sprague-Dawley (CD) rats were exposed to 2450-MHz (CW) microwave radiation at incident power densities of 0 or 28 mW/cm2 for 100 min daily on the 6th through 15th day of gestation. The whole-body specific absorption rate at 28 mW/cm2 is estimated to be 4.2 W/kg. These exposure conditions raised rats' average colonic temperatures to 40.3 degrees C at the end of irradiation. There were 67 sham-irradiated and 70 microwave-irradiated females. When these groups were compared, no significant differences were found in pregnancy rates; in the numbers of live, dead, or total fetuses; in the incidences of external, visceral, or skeletal anomalies or variations; or in the body weight of live fetuses. It is concluded that these conditions do not have an effect on the gross structure of the fetal rat when applied repetitively during post-implantation pregnancy. It is also strongly suspected that this lack of an effect may hold true at any exposure level less than that which will kill a significant number of the dams by hyperthermia (colonic temperature greater than 40 degrees C).


Imaging of surfaces by concurrent surface plasmon resonance and surface plasmon resonance-enhanced fluorescence.
Surface plasmon resonance imaging and surface plasmon induced fluorescent are sensitive tools for surface analysis. However, existing instruments in this area have provided limited capability for concurrent detection, and may be large and expensive. We demonstrate a highly cost-effective system capable of concurrent surface plasmon resonance microscopy (SPRM) and surface plasmon resonance-enhanced fluorescence (SPRF) imaging, allowing for simultaneous monitoring of reflectivity and fluorescence from discrete spatial regions. The instrument allows for high performance imaging and quantitative measurements with surface plasmon resonance, and surface plasmon induced fluorescence, with inexpensive off-the-shelf components.


Lactobionic acid-conjugated TPGS nanoparticles for enhancing therapeutic efficacy of etoposide against hepatocellular carcinoma.
Many effective anti-cancer drugs have limited use in hepatocellular carcinoma (HCC) therapy due to the drug resistance mechanisms in liver cells. In recent years, tumor-targeted drug delivery and the inhibition of drug-resistance-related mechanisms has become an integrated strategy for effectively combating chemo-resistant cancer. Herein, lactobionic acid-conjugated d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS-LA conjugate) has been developed as a potential asialoglycoprotein receptor (ASGPR)-targeted nanocarrier and an efficient inhibitor of P-glycoprotein (P-gp) to enhance etoposide (ETO) efficacy against HCC. The main properties of ETO-loaded TPGS-LA nanoparticles (NPs) were tested through in vitro and in vivo studies after being prepared using the nanoprecipitation method and characterized by dynamic light scattering (DLS). According to the results, smaller (∼141.43 nm), positively charged ETO-loaded TPGS-LA NPs were more suitable for providing efficient delivery to hepatoma cells by avoiding the clearance mechanisms. It was found that ETO-loaded TPGS-LA NPs were noticeably able to enhance the cytotoxicity of ETO in HepG2 cells. Besides this, markedly higher internalization by the ASGPR-overexpressed HepG2 cells and efficient accumulation at the tumor site in vivo were revealed in the TPGS-LA NP group. More importantly, animal studies confirmed that ETO-loaded TPGS-LA NPs achieved the highest therapeutic efficacy against HCC. Interestingly, ETO-loaded TPGS-LA NPs also exhibited a great inhibitory effect on P-gp compared to the ETO-loaded TPGS NPs. These results suggest that TPGS-LA NPs could be used as a potential ETO delivery system against HCC.


Intestinal twin-to-twin transplant for short gut: Review of the literature and discussion of a complex case.
Paediatric Intestinal Transplantation (IT) presents the highest mortality on the waiting-list due to anatomical disproportion. Living-Donor IT (LDIT) offers the best advantages and when performed among identical monozygotic twins, it also benefits from unique immunology. According to MEDLINE/Pubmed, twin-to-twin LDIT has been performed in seven cases (6:7 males, median age of 32 years). None of the patients received immunosuppression postoperatively. Only one paediatric twinto- twin LDIT was carried out with a 160-cm mid-ileum tract: an interposed 4/5-cm arterial graft was required to ensure a tensionfree anastomosis to the anterior wall of the infra-renal abdominal aorta. In contrast, venous anastomosis was done directly to the inferior cava vein. We present a case for debate of a 13- month-old SBS patient where a twin-LDIT was discussed with parents, who decided to wait after careful analysis and ethical considerations.


Extended architectures constructed from sandwich tetra-metal-substituted polyoxotungstates and transition-metal complexes.
Three unprecedented 2D architectures made up of sandwich-type tetra-metal-substituted polyoxotungstates and transition-metal complexes, [Cu(dien)(H2O)]2{[Cu(dien)(H2O)]2[Cu(dien)(H2O)2]2[Cu4(SiW9O34)2]} x 5 H2O (1; dien = diethylenetriamine), [Zn(enMe)2(H2O)]2{[Zn(enMe)2]2[Zn4(HenMe)2(PW9O34)2]} x 8 H2O (2; enMe = 1,2-diaminopropane), and [Zn(enMe)2(H2O)]4[Zn(enMe)2]2{(enMe)2{[Zn(enMe)2]2[Zn4(HSiW9O34)2]}{[Zn(enMe)2(H2O)]2[Zn4(HSiW9O34)2]}} x 13 H2O (3) were hydrothermally synthesized and structurally characterized by elemental analysis, IR spectroscopy, thermogravimetric analysis, and single-crystal X-ray diffraction. Compound 1 consists of anions [Cu4(SiW9O34)2]12- linked by copper complexes into a 2D structure, whereas 2 is constructed from novel inorganic-organic hybrid anions [Zn4(HenMe)2(PW9O34)2]8- linked by zinc complexes into a 2D structure. The most interesting is the unique 2D network 3, which consists of anions [Zn4(PW9O34)2]10- with two types of bridging groups: zinc complexes and enMe ligands.


Inventory of pediatric neurology "manpower" in Canada.
To review the demographics and workload characteristics of pediatric neurology in Canada. A standardized survey questionnaire was mailed out to practicing pediatric neurologists in Canada in 2001. Variables examined were age, gender, hours on call, regular hours worked per week, type of practice and projected changes in practice over next five to ten years. Results were compared to the 1994 Pediatric Neurology Manpower Survey which had used the same survey instrument. Fifty-six (70%) pediatric neurologists practicing in Canada returned the survey. As was the case in 1994, no significant differences in workload were found based on age or gender. The average age of the practicing pediatric neurologist in 2001 was 51 years compared to 45 years in 1994. The proportion of physicians over 55 years in 2001 was 35% compared to 25% in 1994. Pediatric neurology in Canada is an aging specialty needing a significant recruitment of new members


Prediction of the Ebola Virus Infection Related Human Genes Using Protein-Protein Interaction Network.
Ebola hemorrhagic fever (EHF) is caused by Ebola virus (EBOV). It is reported that human could be infected by EBOV with a high fatality rate. However, association factors between EBOV and host still tend to be ambiguous. According to the "guilt by association" (GBA) principle, proteins interacting with each other are very likely to function similarly or the same. Based on this assumption, we tried to obtain EBOV infection-related human genes in a protein-protein interaction network using Dijkstra algorithm. We hope it could contribute to the discovery of novel effective treatments. Finally, 15 genes were selected as potential EBOV infection-related human genes.


Effects of contextual relevance on pragmatic inference during conversation: An fMRI study.
Contextual relevance, which is vital for understanding conversational implicatures (CI), engages both the frontal-temporal language and theory-of-mind networks. Here we investigate how contextual relevance affects CI processing and regulates the connectivity between CI-processing-related brain regions. Participants listened to dialogues in which the level of contextual relevance to dialogue-final utterance (reply) was manipulated. This utterance was either direct, indirect but relevant, irrelevant with contextual hint, or irrelevant with no contextual hint. Results indicated that compared with direct replies, indirect replies showed increased activations in bilateral IFG, bilateral MTG, bilateral TPJ, dmPFC, and precuneus, and increased connectivity between rTPJ/dmPFC and both IFG and MTG. Moreover, irrelevant replies activated right MTG along an anterior-posterior gradient as a function of the level of irrelevance. Our study provides novel evidence concerning how the language and theory-of-mind networks interact for pragmatic inference and how the processing of CI is modulated by level of contextual relevance.


The incidence of dentinal cracks during root canal preparations with reciprocating single-file and rotary-file systems: A meta-analysis.
As a dangerous factor in vertical root fracture, dentinal crack formation is often associated with root canal instruments. We conducted this meta-analysis to compare the influence of two types of nickel titanium (NiTi) instruments that have different movements (reciprocating single-file versus full-sequence rotary file systems) on dentinal cracks formation during root canal preparation. Searches were conducted in PubMed, ISI Web of Knowledge, Embase and Cochrane Library using a combination of keywords. Titles and abstracts of all articles were independently assessed by two reviewers in accordance with the predefined inclusion criteria. Relevant studies were acquired in full-text form. Data in these articles were independently extracted. A meta-analysis was conducted using Review Manager 5.3. The results showed that the WaveOne and Reciproc files with a reciprocating motion produced significantly fewer dentinal cracks than the conventional rotational ProTaper technique.


Perinatal use of triptans and other drugs for migraine-A nationwide drug utilization study.
To characterize nationwide utilization patterns of migraine pharmacotherapy before, during, and after pregnancy in women with triptan use. Population-based data were obtained by linking the Medical Birth Registry of Norway and the Norwegian Prescription Database from 2006 to 2017. We included 22,940 pregnancies among 19,669 women with at least one filled triptan prescription, a proxy for migraine, in the year before pregnancy or during pregnancy. The population was classified into four groups: i) continuers; ii) discontinuers; iii) initiators, and vi) post-partum re-initiators. Participant characteristics and prescription fills for other drugs such as analgesics, antinauseants, and preventive drugs among the groups were examined, along with an array of triptan utilization parameters. In total, 20.0% of the women were classified as triptan continuers, 54.1% as discontinuers, 8.0% as initiators, and 17.6% as re-initiators. Extended use of triptans (≥15 daily drug doses/month) occurred among 6.9% of the continuers in the first trimester. The top 10% of triptan continuers and initiators accounted for 41% (95% CI: 39.2% - 42.5%) and 33% (95% CI: 30.3% - 35.8%) of the triptan volume, respectively. Triptan continuers and initiators had similar patterns of acute co-medication during pregnancy, but use of preventive drugs was more common among the continuers before, during, and after pregnancy. Among women using triptans before and during pregnancy, one in four continued triptan treatment during pregnancy, and extended triptan use was relatively low. Triptan discontinuation during and in the year after pregnancy was common. Use of other acute migraine treatments was higher among both continuers and initiators of triptans. Women using preventive migraine treatment were most commonly triptan continuers and re-initiators after pregnancy. Prescribing to and counseling of women with migraine should be tailored to the condition severity and their information needs to promote optimal migraine management in pregnancy.


Strategies for plasma proteomic profiling of cancers.
Despite a voluminous literature on potential protein biomarkers and a compelling need for diagnostic tests based on biomarkers to detect cancers at much earlier, more treatable stages, progress has been limited. New methods and new instruments for analysis of differences in gene expression, gene methylation, and proteomics are being employed to try to accelerate the discovery phase. Given the heterogeneity of tumor mechanisms and the limitations of analytical methods, it is likely that a variety of strategies will be needed and will be complementary. That is the basis of this review of proteomic approaches. This article adopts a systems biology view, starting with mRNA transcripts in tumors and cultured tumor cells to detect mRNA overexpression, some of which will be correlated with protein overexpression. Some of those proteins may be secreted or released into proximal biofluids and plasma. Detection of low-abundance tumor proteins in the complex and dynamic mixture that is plasma requires combinations of increasingly powerful technologies. The biological amplification of protein signals through the immune system offers autoantibodies as potential biomarkers. Higher abundance proteins, including acute-phase reactants, may have practical value, especially if the proteins are modified as part of the cancer processes. Low molecular weight proteins, fragments, and peptides may offer complementary biomarkers. Promising biomarker candidates must be confirmed in independent studies. Then they must be submitted to higher-throughput methods practical for large-scale validation studies and, hopefully, for clinical and epidemiological applications. Standardized operating procedures for specimen handling, design and use of various reference standards, care to avoid bias and confounding, and guidelines for reporting findings and contributing datasets should enhance the prospects for predictive proteomic profiling of people at risk for cancers.


Post-infectious persistent cough: pathogenesis and therapeutic options.
Post-infectious cough is a common symptom associated with common colds and/or upper respiratory tract infection. This cough is expected to last for only for few days and resolve spontaneously, whilst when persists for longer than three weeks is defined "persistent" and is associated tickling or an irritating sensation in the throat which often leads to paroxysms of coughing. Persistent post-infectious cough can cause morbidity since it may interfere with usual living. Despite the recent advances in understanding the mechanisms that regulate cough, in physiological and pathological conditions, current therapeutic options for post-infectious cough are little or only moderately effective.


Palm-Based Neopentyl Glycol Diester: A Potential Green Insulating Oil.
The transesterification of high oleic palm oil methyl ester (HOPME) with neopentyl glycol (NPG) has been investigated. The present study revealed the application of low-pressure technology as a new synthesis method to produce NPG diesters. Single variable optimization and response surface methodology (RSM) were implemented to optimize the experimental conditions to achieve the maximum composition (wt%) of NPG diesters. The main objective of this study was to optimize the production of NPG diesters and to characterize the optimized esters with typical chemical, physical and electrical properties to study its potential as insulating oil. The transesterification reaction between HOPME and NPG was conducted in a 1L three-neck flask reactor at specified temperature, pressure, molar ratio and catalyst concentration. For the optimization, four factors have been studied and the diester product was characterized by using gas chromatography (GC) analysis. The synthesized esters were then characterized with typical properties of transformer oil such as flash point, pour point, viscosity and breakdown voltage and were compared with mineral insulating oil and commercial NPG dioleate. For formulation, different samples of NPG diesters with different concentration of pour point depressant were prepared and each sample was tested for its pour point measurement. The optimum conditions inferred from the analyses were: molar ratio of HOPME to NPG of 2:1.3, temperature = 182°C, pressure = 0.6 mbar and catalyst concentration of 1.2%. The synthesized NPG diesters showed very important improvement in fire safety compared to mineral oil with flash point of 300°C and 155°C, respectively. NPG diesters also exhibit a relatively good viscosity of 21 cSt. The most striking observation to emerge from the data comparison with NPG diester was the breakdown voltage, which was higher than mineral oil and definitely in conformance to the IEC 61099 limit at 67.5 kV. The formulation of synthesized NPD diesters with VISCOPLEX® pour point depressant has successfully increased the pour point of NPG diester from -14°C to -48°C. The reaction time for the transesterification of HOPME with NPG to produce NPG diester was successfully reduced to 1 hour from the 14 hours required in the earlier synthesis method. The main highlight of this study was the excess reactant which is no longer methyl ester but the alcohol (NPG). The optimum reaction conditions for the synthesis were molar ratio of 2:1.13 for NPG:HOPME, 182°C, 0.6 mbar and catalyst concentration of 1.2 wt%. The maximum NPG diester yield of 87 wt% was consistent with the predicted yield of 87.7 wt% obtained from RSM. The synthesized diester exhibited better insulating properties than the commercial products especially with regards to the breakdown voltage, flash point and moisture content.


Expression of parathyroid hormone-related peptide in human osteoarthritis.
To evaluate the involvement of the expression of parathyroid hormone-related peptide gene in human articular cartilage pathology, we performed immunohistochemical staining and in situ hybridization on specimens of femoral head cartilage obtained from 15 patients with osteoarthritis, 11 with rheumatoid arthritis, and 12 control subjects. Parathyroid hormone-related peptide-positive chondrocytes were observed predominantly in degenerated lesions of osteoarthritic tissue and were less evident in rheumatoid arthritic samples, while the normal cartilage expressed little parathyroid hormone-related peptide. In addition, the level of parathyroid hormone-related peptide expression was clearly dependent on the degree of cartilage degeneration; cartilage tissues with moderate degenerative changes contained more positive chondrocytes compared with mildly or severely degenerated cartilage. In situ hybridization confirmed the localization of parathyroid hormone-related peptide protein and demonstrated intense expression of mRNA of the peptide in osteoarthritic samples. This is the first demonstration of parathyroid hormone-related peptide expression in chondrocytes from pathologic articular cartilage of humans. Our results suggest that parathyroid hormone-related peptide may be involved in the pathophysiology of osteoarthritis.


[Classification of bronchial asthma].
The bronchial asthma disease is divided into 5 groups: 1. bronchial asthma caused by allergy, 2. bronchial asthma caused by infection, 3. bronchial asthma caused by physicochemical irritants, 4. bronchial asthma caused by exercise, 5. combination between the groups 1--4. Etiological factors, immunologic phenomena so far as known and pathogenic mechanisms are listed for each group and discussed.


Swift adsorptive removal of Congo red from aqueous solution by K1.33Mn8O16 nanowires.
A swift and efficient approach to converting organic dye effluents into fresh water could be of substantial benefit. In this study, we presented facile hydrothermal synthesis of K1.33Mn8O16 nanowires in ammonium fluoride (NH4F) aqueous solution. The crystallization process of K1.33Mn8O16 nanowires was investigated. The as-obtained K1.33Mn8O16 nanowires were used for swift adsorptive removal of Congo red from aqueous solution without adjusting pH value at room temperature. Adsorption kinetic experimental data are well described by pseudo-second-order rate kinetic model, and the adsorption isotherm fits Langmuir isotherm model. The present investigation provides an efficient approach to designing and fabricating manganese-based nanomaterials for environmental remediation.


Radiologic anatomic variation of the carpus in horses with carpal lameness and control horses.
Our purpose was to describe the variation of the radiologic appearance of the carpus of horses of different breeds, discipline, and gender with lameness related to the carpus and control horses, with particular reference to the ulnar carpal bone. Two hundred and eighty-six sets of carpal radiographs from 222 horses were analyzed. Breed, gender, discipline, and cause of lameness were recorded. Chi square tests were used to test for associations between radiologic findings and gender, breed and discipline, to test for associations between different radiologic findings, and to test for associations between radiologic findings and causes of lameness. Bonferroni correction was applied when necessary. The shape of the ulnar carpal bone and most of the anatomic variants, with the exception of the first carpal bone and a radiolucent area in the second carpal bone, were not breed or gender related. Radiolucent areas and associated fragments on the palmaromedial aspect of the ulnar carpal bone can be an incidental finding in horses from all disciplines.


Emergence of deterministic Green's functions from noise generated by finite random sources.
Two-point correlation functions of sufficiently diffuse wave fields generated by uncorrelated random sources are known to approximate deterministic Green's functions between the two points. This property is utilized increasingly for passive imaging and remote sensing of the environment. Here we show that the relation between the Green's functions and the noise cross-correlation function holds under much less restrictive conditions than previously thought. It can even hold when ambient noise sources have correlation ranges large compared to the wavelength. Admissible correlation ranges are limited from above by the size of the Fresnel zone at wave propagation between the points where noise cross correlation is evaluated.


Specificity and retinotectal projections of quarter-eye fragments in Xenopus laevis.
Three quarters of the eye anlage in Xenopus embryos of stage 33/34 were eliminated in three different sets of experiments. The remaining quadrant originated from the nasoventral part of the retina, from its ventral portion, or from the temporo-ventral area of the retina. All the fragments developed into small eyes of normal shape. The retinotectal connections did not deviate from those found in the control groups, even though mirror-image duplication was fairly frequent. For all fragments the tectal projection fields were rather limited. There was some indication of fragments retaining their original specificity. Irrespective, however, of their different origins, the optic projections always occupied the rostrolateral area of the tectum.


[Morphological and biochemical characterization of the developmental stages of fertilized eggs inSphaerechinus granularis lam : I. Rearing, Morphology and determination of stages].
1. Rearing to the developmental stage. Eggs and embryos of the sea urchin speciesSphaerechinus granularis were kept at a concentration of 3000 per ml in 101 batches at 22° C. They were stirred under continuous aeration to counteract sedimentation for up to 72 h. The development in the population could be kept nearly synchronous all the time. 2. Morphology. Morphology has been followed up to the pluteus stage-72 h after fertilization. At the 64 cell stage no cell coronas of macromeres, micromeres and mesomeres could be discerned in this embryo. The time periods of the different stages, their corresponding volumes and their cell numbers have been determined. 3. Electronic characterization of the stages. Embryogenesis has been followed using the Coulter Counter with size distribution plotter, the volume distribution curves being recorded continuously. The sudden elevation of the fertilization membrane manifests itself as a volume increase of 50 per cent. In the course of further development the volume increases 300-400 per cent. Gastrulation events cause a drop in embryonic volume of about 150 per cent. In the course of further development two additional smaller volume peaks occur, followed by a steep rise in volume, at least up to 72 h. The results obtained from electronic methods when compared with results from microscopy prove their relevance, the data from the Coulter Counter being the more consistent ones. The electronic method makes it possible to characterize the embryos according to their developmental stage and to their pattern of population distribution in a fast and reliable fashion.


Extracellular matrix derived from Trypanosoma cruzi infected endothelial cells directs phenotypic expression.
Infection of confluent human umbilical vein endothelial cells by the parasite Trypanosoma cruzi results in the appearance of an altered heparan sulfate proteoglycan (HSPG) isolated from the extracellular matrix of infected endothelial cells (ECMi). HSPG from ECMi differed from HSPG obtained from the extracellular matrix of uninfected endothelial cells (ECMu) by virtue of an 8-10-fold increase in sulfation and a different elution pattern using DEAE Sepharose chromatography. Analysis of the HSPG that binds to acidic fibroblast growth factor (aFGF) revealed that infection increased the proportion of HSPG that binds to aFGF by 35%. Heparitinase and alkaline borohydride treatment of aFGF-binding HSPG and chromatographic resolution on Sepharose CL4B column revealed an infection-associated 10-fold increase in sulfation of the GAG side chain with no significant change in the migration of the core protein. In addition, the aFGF binding HSPG isolated from ECMi demonstrated a markedly attenuated synergistic mitogenic activity with aFGF in a cell proliferation assay. All of the infection associated changes in HSPG could be demonstrated in HSPG obtained from uninfected endothelial cell cultures grown on ECMi. Hence, the ECMi is associated with signals capable of modulating the ECM associated metabolism of uninfected endothelial cells. This facility of ECMi was also shown to extend to patterns of Gs protein synthesis as revealed by Western blot analysis. The observation that the ECM produced by infected endothelial cells can direct the synthetic patterns of uninfected endothelial cells in a manner uniquely observed in infected endothelial cells suggests a plausible pathway by which infection of only a few cells can ultimately result in the coordinate responses of neighboring uninfected cells.


Benign pigmented apocrine vulvar hamartomas.
Pigmented vulvar lesions were observed in a child during a sexual abuse evaluation. Gross examination of the lesions appeared most consistent with bowenoid papulosis; however, biopsy confirmed the lesions to be pigmented apocrine hamartomas. To our knowledge, these rare and benign tumors have never been described as pigmented, but should be added to the differential diagnosis of pigmented vulvar lesions.


Synthesis of notoamide J: a potentially pivotal intermediate in the biosynthesis of several prenylated indole alkaloids.
An efficient total synthesis of notoamide J, a new prenylated indole alkaloid and potential biosynthetic precursor, is described herein. Starting from L-proline and a substituted tryptophan derivative, this synthesis also employs an oxidation and pinacol rearrangement for the formation of the oxindole in the final step.


Five-year evaluation of two resin-retained ceramic systems: a retrospective study in a general practice setting.
This study performed a clinical evaluation of two ceramic systems retained with resin-bonded cement, observed clinical outcomes over time, and compared the two systems used (IPS Empress and Vitadur Alpha) and differences between inlays and onlays. Three general practitioners placed 317 restorations (215 IPS Empress and 102 Vitadur Alpha) in 153 patients. The first clinical registration was performed 6 to 36 months after placement of the restorations by calibrated investigators using the CDA criteria. A second follow-up was performed 36 months later. The mean time in function for all restorations at follow-up was 60 months. At the first examination, five restorations were not clinically acceptable because of fractures and caries. At follow-up, another 16 restorations were judged as failures. The most significant changes over time were an increase in rough surfaces, evidence of increasing marginal discrepancy, and slight mismatch in color between restored tooth and ceramic restoration. When comparing IPS Empress to Vitadur Alpha, there was no difference in clinical performance. When comparing inlay and onlay restorations, there was a tendency toward a higher number of failures in the inlay group. The overall success rate was 92%. Ceramic restorations performed well after 5 years in function and provide a good treatment alternative that can be successfully managed in general dental practice.


Depletion in LpA-I:A-II particles enhances HDL-mediated endothelial protection in familial LCAT deficiency.
The aim of this study was to evaluate the vasoprotective effects of HDL isolated from carriers of LCAT deficiency, which are characterized by a selective depletion of LpA-I:A-II particles and predominance of preβ migrating HDL. HDLs were isolated from LCAT-deficient carriers and tested in vitro for their capacity to promote NO production and to inhibit vascular cell adhesion molecule-1 (VCAM-1) expression in cultured endothelial cells. HDLs from carriers were more effective than control HDLs in promoting eNOS activation with a gene-dose-dependent effect (PTrend = 0.048). As a consequence, NO production induced by HDL from carriers was significantly higher than that promoted by control HDL (1.63 ± 0.24-fold vs. 1.34 ± 0.07-fold, P = 0.031). HDLs from carriers were also more effective than control HDLs in inhibiting the expression of VCAM-1 (homozygotes, 65.0 ± 8.6%; heterozygotes, 53.1 ± 7.2%; controls, 44.4 ± 4.1%; PTrend = 0.0003). The increased efficiency of carrier HDL was likely due to the depletion in LpA-I:A-II particles. The in vitro findings might explain why carriers of LCAT deficiency showed flow-mediated vasodilation and plasma-soluble cell adhesion molecule concentrations comparable to controls, despite low HDL-cholesterol levels. These results indicate that selective depletion of apoA-II-containing HDL, as observed in carriers of LCAT deficiency, leads to an increased capacity of HDL to stimulate endothelial NO production, suggesting that changes in HDL apolipoprotein composition may be the target of therapeutic interventions designed to improve HDL functionality.


Facile, Energy-Efficient Microscale Fibrillation of Polyacrylamides under Ambient Conditions.
Insight into fiber formation can provide new rationale for the design and preparation of fibers with programmed mechanical properties. While synthetic bioinspired fibers have shown impressive tensile properties, the fiber formation process remains poorly understood. Moreover, these systems are highly complex and their formation is environmentally and economically costly. Controlled fiber formation under ambient conditions from polyacrylamide solutions with properties comparable to natural fibers such as wool and coir is demonstrated. Photopolymerization and subsequent microscale fibrillation of different acrylamides in water/ethanol mixtures yield a simple and energy-efficient route to fiber formation. This strategy reduces required processing energy by two-to-three orders of magnitude. Through extensive experimental elucidation, insight into precise fiber forming conditions of polymeric solutions is achieved. Ethanol is utilized as a chain transfer agent to control the molecular weight of the polyacrylamides, and the entanglement regimes of the solutions are determined through rheological characterization showing fiber formation above the entanglement concentration. Unique from previously reported hydrogel microfibers, it is shown that fibers with good mechanical properties can be obtained without the need for composites or crosslinkers. The reported approach offers a platform for fiber formation under ambient conditions with molecular-level understanding of their assembly.


polyglutamine aggregation nucleation: thermodynamics of a highly unfavorable protein folding reaction.
Polyglutamine (polyGln) aggregation is implicated in the disease progression of Huntington's disease and other expanded CAG repeat diseases. PolyGln aggregation in vitro follows a simple nucleated growth polymerization pathway without apparent complications such as populated intermediates, alternative assembly pathways, or secondary nucleation phenomena. Previous analysis of the aggregation of simple polyGln peptides revealed that the critical nucleus (the number of monomeric units involved in the formation of an energetically unfavorable aggregation nucleus) is equal to one, suggesting that polyGln nucleation can be viewed as an unfavorable protein folding reaction. We provide here a method for experimentally determining the number of elongation growth sites per unit weight for any polyGln aggregate preparation, a key parameter required for completion of the nucleation kinetics analysis and determination of the thermodynamics of nucleation. We find that, for the polyGln peptide Q(47), the second-order rate constant for fibril elongation is 11,400 liters/mol per s, whereas K(n*)), the equilibrium constant for nucleation of aggregation, is remarkably small, equal to 2.6 x 10(-9). The latter value corresponds to a free energy of nucleus formation of +12.2 kcal/mol, a value consistent with a highly unfavorable folding reaction. The methods introduced here should allow further analysis of the energetics of polyGln nucleus formation and accurate comparisons of the seeding capabilities of different fibril preparations, a task of increasing importance in the amyloid field.


Evaluation of the Marginal and Internal Fit of Implant-Supported Metal Copings Fabricated with 3 Different Techniques: An In Vitro Study.
To compare the marginal and internal fit of cobalt-chromium (Co-Cr) alloy copings fabricated by with lost wax technique (LW), computer-aided design and computer-aided manufacturing (CAD/CAM), and direct metal laser sintering (DMLS). Thirty-six tissue level, straight titanium abutments were screwed onto implant replicas. All specimens were embedded in acrylic resin and randomly divided into 3 subgroups according to the fabrication of metal coping: LW, CAD/CAM, and DMLS. In total, 36 (n = 12/group) Co-Cr implant-supported metal copings were prepared. Marginal, intermarginal, axial, and occlusal fits of each coping were measured using the silicone replica technique. The data were evaluated statistically using one-way ANOVA and Bonferioni post-hoc test (α = 0.05). The CAD/CAM group showed significantly lower marginal fit than the LW group and DMLS groups (p < 0.001). The marginal fit of the LW group was not significantly different from the DMLS group (p = 0.721). No significant difference found among the fabrication methods in terms of intermarginal fit (p = 0.913). The CAD/CAM group showed lower axial fit than the LW group (p = 0.026), but there was no statistical difference between the DMLS group and the LW (p = 0.999) and CAD/CAM groups (p = 0.247). No significant differences found among the fabrication methods in terms of occlusal fit (p = 0.158). The LW and DMLS groups showed better marginal fit compared to the CAD/CAM group; however, the CAD/CAM group was better than the LW group in terms of axial fit. All fabrication methods demonstrated similar intermarginal and occlusal fit.


Targeted PI3K/AKT/mTOR therapy for metastatic carcinomas of the cervix: A phase I clinical experience.
Activated PI3K/AKT/mTOR pathway frequently occurs in metastatic or recurrent cervical carcinomas. However, the clinical benefits of matched therapy, a therapeutic approach targeting a specific mutational abnormality, have not yet been established. We analyzed the outcomes of patients with metastatic or recurrent cervical carcinomas who had a test for PIK3CA mutation and/or PTEN loss/mutation, and received ≥1 phase I therapeutic regimen between January 2006 and June 2013. Patients with adenocarcinoma had fewer PIK3CA mutations (14%), and survived longer (median, 14.2 months) than those with squamous cell carcinoma (48% and 7.2 months; p = 0.016, and 0.001, respectively). Matched therapy targeting the activated PI3K/AKT/mTOR pathway led to a favorable rate of SD ≥ 6 months/CR/PR (53%) and significantly longer progression-free survival (median, 6.0 months) than non-matched therapy (11% and 1.5 months; p = 0.08 and 0.026; respectively). In patients with squamous cell carcinoma of the cervix, the presence of PIK3CA mutations was associated with a significantly longer overall survival (median, 9.4 months) than the absence of PIK3CA mutations (median, 4.2 months; p = 0.019). Matched therapy targeting the activated PI3K/AKT/mTOR pathway provided meaningful clinical benefits. Thus, further evaluation of PI3K/AKT/mTOR pathway targeted therapy is warranted, especially in metastatic or recurrent squamous cell carcinoma.


Thrombocytopenia--an indicator of acute vivax malaria.
Thrombocytopenia is frequently observed in vivax malaria but the exact mechanism has not been elucidated. We studied 27 cases of acute vivax malaria out of which 24 cases had thrombocytopenia. This was the most common hematological finding. None had bleeding from any site. Anaemia and splenomegaly were not present in any of the cases. Platelet counts reverted to normal on treatment. Other causes of thrombocytopenia were ruled out by complete history and physical examination, dengue serology and blood culture. DIC was ruled out by peripheral smear examination and measurement of FDP levels. Our study stresses the importance of thrombocytopenia as an early indicator for acute malaria; a finding that is frequent and present even before anemia and splenomegaly set in. The possible mechanisms leading to thrombocytopenia in malaria have been discussed which include immune mechanisms, oxidative stress, alterations in splenic functions and a direct interaction between plasmodium and platelets.


Modeling Pathological Hemodynamic Responses to the Valsalva Maneuver.
The Valsalva maneuver (VM) consisting in a forced expiration against closed airways is one of the most popular clinical tests of the autonomic nervous system function. When properly performed by a healthy subject, it features four characteristic phases of arterial blood pressure (BP) and heart rate (HR) variations, based on the magnitude of which the autonomic function may be assessed qualitatively and quantitatively. In patients with some disorders or in healthy patients subject to specific conditions, the pattern of BP and HR changes during the execution of the Valsalva maneuver may, however, differ from the typical sinusoidal-like pattern. Several types of such abnormal responses are well known and correspond to specific physiological conditions. In this paper, we use our earlier mathematical model of the cardiovascular response to the Valsalva maneuver to show that such pathological responses may be simulated by changing individual model parameters with a clear physiological meaning. The simulation results confirm the adaptability of our model and its usefulness for diagnostic or educational purposes.


Evidence for Wg-independent tergite boundary formation in the millipede Glomeris marginata.
The correlation between dorsal and ventral segmental units in diplopod myriapods is complex and disputed. Recent results with engrailed (en), hedgehog (hh), wingless (wg), and cubitus-interruptus (ci) have shown that the dorsal segments are patterned differently from the ventral segments. Ventrally, gene expression is compatible with the classical autoregulatory loop known from Drosophila to specify the parasegment boundary. In the dorsal segments, however, this Wg/Hh autoregulatory loop cannot be present because the observed gene expression patterns argue against the involvement of Wg signalling. In this paper, we present further evidence against an involvement of Wg signalling in dorsal segmentation and propose a hypothesis about how dorsal segmental boundaries may be controlled in a wg-independent way. We find that (1) the Notum gene, a modulator of the Wg gradient in Drosophila, is not expressed in the dorsal segments. (2) The H15/midline gene, a repressor of Wg action in Drosophila, is not expressed in the dorsal segments, except for future heart tissue. (3) The patched (ptc) gene, which encodes a Hh receptor, is strongly expressed in the dorsal segments, which is incompatible with Wg-Hh autoregulation. The available data suggest that anterior-posterior (AP) boundary formation in dorsal segments could instead rely on Dpp signalling rather than Wg signalling. We present a hypothesis that relies on Hh-mediated activation of Dpp signalling and optomotor-blind (omb) expression to establish the dorsal AP boundary (the future tergite boundary). The proposed mechanism is similar to the mechanism used to establish the AP boundary in Drosophila wings and ventral pleura.


Multiple miRNA Detection through a Suspended Microbead Array Encoded by Triple-Color Upconversion Luminescent Nanotags via Bi-Beam Splitter Hybrid-Multitrap Optical Tweezers.
In recent years, optical tweezers have become a novel tool for biodetection, and to improve the inefficiency of a single trap, the development of multitraps is required. Herein, we constructed a set of hybrid multitrap optical tweezers with the balance of stability and flexibility by the combination of two different beam splitters, a diffraction optical element (DOE) and galvano mirrors (GMs), to capture polystyrene (PS) microbeads in aqueous solutions to create an 18-trap suspended array. A sandwich hybridization strategy of DNA-miRNA-DNA was adopted to detect three kinds of target miRNAs associated with triple negative breast cancer (TNBC), in which different upconversion nanoparticles (UCNPs) with red, green, and blue emissions were applied as luminescent tags to encode the carrier PS microbeads to further indicate the levels of the targets. With encoded luminescent microbeads imaged by a three-channel microscopic system, the biodetection displayed high sensitivity with low limits of detection (LODs) of 0.27, 0.32, and 0.33 fM and exceptional linear ranges of 0.5 fM to 1 nM, 0.7 fM to 1 nM, and 1 fM to 1 nM for miR-343-3p, miR-155, and miR-199a-5p, respectively. In addition, this bead-based assay method was demonstrated to have the potential for being applied in patients' serum by satisfactory standard addition recovery experiment results.


[Methods and applications of animal genomic mating].
Genomic selection (GS) is a powerful tool which can be used to estimate the breeding value of individual animals by using the molecular markers of the animal's entire genome. GS improves the accuracy and intensity of selection, reduces the interval of generation, and realizes the effects of early accuracy selection contributing to a significant evolution in animal breeding. In the past decade, GS was successfully applied in the genetic improvement of dairy animals with improved selection accuracy and genetic gain of breeding animals. However, GS focuses on the genetic gain of target traits while it ignores the genetic relationship between mating pairs such that it ignores long term genetic merits such as an increase in inbreeding coefficient of offspring population, a decrease of genetic diversity and the homozygous presentation of harmful genes. In 2016, genomic mating (GM) was proposed as a sustainable genetic selection method using genomic information of the breeding candidate individuals to optimize selection and mating with resultant control of the growth rate of population inbreeding coefficient and achieving long-term and sustainable genetic progress. Therefore, GM is more suitable for modern animal breeding than GS, especially for the genetic improvement of indigenous species. In this review, we summarize the basic concepts, methods, and applications of GM, and then present examples comparing the effects of six simulated mating schemes. This review serves as a valuable reference for the applications of animal breeding methods.


Metagenomic Next-Generation Sequencing vs. Traditional Microbiological Tests for Diagnosing Varicella-Zoster Virus Central Nervous System Infection.
Unbiased metagenomic next-generation sequencing (mNGS) detects pathogens in a target-independent manner. It is not well-understood whether mNGS has comparable sensitivity to target-dependent nucleic acid test for pathogen identification. This study included 31 patients with chickenpox and neurological symptoms for screening of possible varicella-zoster virus (VZV) central nervous system (CNS) infection. Microbiological diagnosing of VZV cerebrospinal fluid (CSF) infection was performed on stored CSF samples using mNGS, quantitative and qualitative VZV-specific PCR assays, and VZV IgM antibodies test. The median age was 30.0 [interquartile range (IQR), 24.3-33.3] years. 51.6% of the patients were men. About 80.6% of the patients had normal CSF white blood cell counts (≤ 5 × 106/L). VZV IgM antibodies presented in 16.1% of the CSF samples, and nucleic acids were detectable in 16.1 and 9.7% using two different VZV-specific real-time PCR protocols. Intriguingly, maximal identification of VZV elements was achieved by CSF mNGS (p = 0.001 and p = 007; compared with qualitative PCR and VZV IgM antibody test, respectively), with sequence reads of VZV being reported in 51.6% (16/31) of the CSF samples. All VZV PCR positive samples were positive when analyzed by mNGS. Of note, human betaherpesvirus 6A with clinical significance was unexpectedly detected in one CSF sample. Our study suggests that CSF mNGS may have higher sensitivity for VZV detection than CSF VZV PCR and antibody tests, and has the advantage of identifying unexpected pathogens.


Sphingosine-1-Phosphate Catabolizing Enzymes Predict Better Prognosis in Triple-Negative Breast Cancer Patients and Correlates With Tumor-Infiltrating Immune Cells.
Background: Triple-negative breast cancer (TNBC) is associated with a poor prognosis. Sphingosine-1-phosphate (S1P), a potent sphingolipid metabolite, has been implicated in many processes that are important for breast cancer (BC). S1P signaling regulates tumorigenesis, and response to chemotherapy and immunotherapy by affecting the trafficking, differentiation or effector function of tumor-infiltrating immune cells (TIICs). Objective: In this study, using bioinformatics tools and publicly available databases, we have analyzed the prognostic value of S1P metabolizing genes and their correlation with TIICs in BC patients. Methods: The expression of S1P metabolizing genes and receptors was evaluated by the UALCAN cancer database. The correlation between mRNA expression of S1P metabolizing genes and receptors and survival outcome of breast cancer patients was analyzed by the Kaplan-Meier plotter database. The association between the gene expression and infiltration of immune cells in the tumors was analyzed by "Tumor-Infiltrating Immune Estimation Resource (TIMER). In silico protein expression analysis was done using the Human Protein Atlas" database. Results: TNBC patients with lower expression of S1P phosphatase 1 (SGPP1) or lipid phosphate phosphatase 3 (PLPP3) have much shorter relapse-free survival than the patients with a higher expression of these genes. SGPP1 and PLPP3 expression show a strong positive correlation with tumor-infiltrating dendritic cells (DCs), CD4+ and CD8+ T cells, neutrophils, and macrophages in the TNBC subtypes. In addition, S1P receptor 4 (S1PR4), an S1P receptor exhibit a strong positive correlation with DCs, CD4+ and CD8+ T cells and neutrophils in TNBC. We, therefore, conclude that low expression of SGPP1 and PLPP3 may hinder the recruitment of immune cells to the tumor environment, resulting in the blockage of cancer cell clearance and a subsequent poor prognosis.


Risk factors and management for early and late intrahepatic recurrence of solitary hepatocellular carcinoma after curative resection.
Intrahepatic recurrence is a significant problem for patients who have undergone a hepatic resection for hepatocellular carcinoma (HCC). The objective of the present study was to identify risk factors and evaluate the management of early and late recurrence of solitary HCC after curative resection. Included in this study were 816 patients with solitary HCC who underwent a curative partial hepatectomy. Intrahepatic recurrence in these patients was followed up retrospectively. Prognosis and therapy for the recurrence were investigated and analysed. Early and late intrahepatic recurrence occurred in 423 patients and 199 patients, respectively. Multivariate analysis showed that a tumour diameter >5 cm, the absence of a tumour capsule and the presence of microvascular invasion were correlated with early recurrence, whereas cirrhosis and alpha-fetal protein >400 μg/l were independent risk factors contributing to late recurrence. The 5-year survival of HCC patients with early recurrence was significantly lower than that of patients with late recurrence. Further curative treatment for intrahepatic recurrence offered a 5-year overall survival of 56.0%, which was better than alternative management. Early and late recurrences of solitary HCC after curative resection are associated with different predictive factors. The time to recurrence and further curative treatment after recurrence were the best predictors of survival post recurrence.


Beta-adrenergic receptors on human tracheal epithelial cells in primary culture.
Human tracheal epithelial cells in suspension, whether obtained by brushing at bronchoscopy or from necropsy specimens by proteolytic digestion and EDTA treatment, increase adenosine 3',5'-cyclic monophosphate (cAMP) production in response to isoproterenol. These cells in primary culture also respond to beta-adrenergic agonists in order of potency isoproterenol greater than epinephrine greater than norepinephrine. The response is inhibited by propranolol or ICI 118551 (a beta 2-adrenergic selective blocker) but not by atenolol (a beta 1-adrenergic blocker). Binding of [125I]iodocyanopindolol (ICYP) to membranes was rapid, stereoselective, and saturable and displayed receptor density 8.0 +/- 4.6 fmol/mg protein (mean 228 receptors/cell) and a dissociation constant (KD) for ICYP of 35 +/- 14 pM for freshly isolated cells and a KD of 25 +/- 13 pM and receptor density of 17 +/- 17 fmol/mg protein for cells in culture. The 50% inhibitory concentration (IC50) for atenolol was 470 microM and for ICI 118551 was 0.012 microM. Analysis of the ICI 118551 displacement curve indicates that greater than 90% of the receptors are of the beta 2-adrenergic class. Prostaglandins E1 or E2, vasoactive intestinal peptide, carbachol, phenylephrine, or platelet-activating factor did not affect either the maximal cAMP response or the isoproterenol dose-response relationship. Neither clonidine nor epinephrine plus propranolol altered cellular cAMP content, and cyclooxygenase inhibition did not change the cAMP response to epinephrine. We conclude that in human tracheal epithelial cells in primary culture, adrenergic stimulation affects cAMP levels only through beta 2-adrenergic receptors and that modulation of this system by platelet-activating factor or muscarinic, alpha 1-, or alpha 2-adrenergic agents does not occur.


hPSC-derived lung and intestinal organoids as models of human fetal tissue.
In vitro human pluripotent stem cell (hPSC) derived tissues are excellent models to study certain aspects of normal human development. Current research in the field of hPSC derived tissues reveals these models to be inherently fetal-like on both a morphological and gene expression level. In this review we briefly discuss current methods for differentiating lung and intestinal tissue from hPSCs into individual 3-dimensional units called organoids. We discuss how these methods mirror what is known about in vivo signaling pathways of the developing embryo. Additionally, we will review how the inherent immaturity of these models lends them to be particularly valuable in the study of immature human tissues in the clinical setting of premature birth. Human lung organoids (HLOs) and human intestinal organoids (HIOs) not only model normal development, but can also be utilized to study several important diseases of prematurity such as respiratory distress syndrome (RDS), bronchopulmonary dysplasia (BPD), and necrotizing enterocolitis (NEC).


[Hypothermia after cardiac arrest--underused method that saves lives].
Cardiorespiratory arrest causes ischemia and lesion of all organ systems, but the central nervous system is the most vulnerable. It is known that only few minutes of hypoperfusion and ischemia can cause irreversible damage to the brain which is the major frustration of reanimatology. Results of clinical trials suggest positive effects of hypothermia on survival and neurological recovery which led to including this method to Guidelines for resuscitation as a recommended standard method in post-resuscitation period for patients who have not regained consciousness. Methods for induction and maintenance of hypothermia are numerous and various, basically divided into invasive and non-invasive, each with its own advantages and disadvantages which are described in this paper. Despite recognised positive effects of mild therapeutic hypothermia after resuscitation from cardiac arrest, the method is not fully implemented as a standard method in post-resuscitation period.


Whey protein rich in alpha-lactalbumin increases the ratio of plasma tryptophan to the sum of the other large neutral amino acids and improves cognitive performance in stress-vulnerable subjects.
Cognitive performance often declines under chronic stress exposure. The negative effect of chronic stress on performance may be mediated by reduced brain serotonin function. The uptake of the serotonin precursor tryptophan into the brain depends on nutrients that influence the availability of tryptophan by changing the ratio of plasma tryptophan to the sum of the other large neutral amino acids (Trp-LNAA ratio). In addition, a diet-induced increase in tryptophan may increase brain serotonergic activity levels and improve cognitive performance, particularly in high stress-vulnerable subjects. We tested whether alpha-lactalbumin, a whey protein with a high tryptophan content, would increase the plasma Trp-LNAA ratio and improve cognitive performance in high stress- vulnerable subjects. Twenty-three high stress-vulnerable subjects and 29 low stress-vulnerable subjects participated in a double-blind, placebo-controlled, crossover study. All subjects conducted a memory-scanning task after the intake of a diet enriched with either alpha-lactalbumin (alpha-lactalbumin diet) or sodium caseinate (control diet). Blood samples were taken to measure the effect of dietary manipulation on the plasma Trp-LNAA ratio. A significantly greater increase in the plasma Trp-LNAA ratio after consumption of the alpha-lactalbumin diet than after the control diet (P = 0.0001) was observed; memory scanning improved significantly only in the high stress-vulnerable subjects (P = 0.019). Because an increase in the plasma Trp-LNAA ratio is considered to be an indirect indication of increased brain serotonin function, the results suggest that dietary protein rich in alpha-lactalbumin improves cognitive performance in stress-vulnerable subjects via increased brain tryptophan and serotonin activities.


Vasculitis and vasculopathy in Lupus Nephritis: Clinical variability, outcome, and new insight into treatment.
More than 50% of patients with systemic lupus erythematosus (SLE) have renal involvement at presentation or during their illness. Lupus nephritis (LN) encompasses several patterns of renal disease, including glomerular, tubulointerstitial, and vascular pathologies. The presence and significance of renal vascular lesions (VLs) are often overlooked. VLs in LN are not rare with an incidence of 10%-40% on renal biopsies from various studies and their presence is often labeled as poor prognostic markers. The current treatment protocol for LN is mainly based on the glomerular pathology, and no guidelines/consensus exists for treatment of LN with VLs. We describe the clinical presentation, course, response to therapy, and outcomes in five patients with SLE with histological evidence of renal VLs.


Diabetes ketoacidosis and Recurrent Childhood Stroke-like Episodes.
A 13-year and 4-month-old girl was brought to the emergency department due to fever, dizziness,vomiting, and blurred vision. Laboratory data revealed hyperglycemia with an HbA1C of 7.3 percent, ketonuria, and lactic acidosis. The initial impression was diabetic ketoacidosis. During admission, recurrent focal impaired awareness seizures were noted, and magnetic resonance imaging of the brain revealed multiple brain infarctions in the bilateral cerebrum. Mitochondrial gene report showed A3243 G with 64 percent heteroplasmy, and mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes was diagnosed. At 16 years and 7 months old, recurrence of vomiting and onset of right hemianopia and mild right limb weakness were observed and follow-up T2 images showed massive edema in her left parieto-occipital region. At 16 years and 10 months old, she developed clonus in her left hand associated with an unsteady gait and blurred vision. MRI of the brain revealed recurrent brain infarction, and T2 images showed massive edema of the right parieto-occipital region. MELAS is a rare disease entity and occasionally comorbid with mitochondrial diabetes in childhood. Characteristic radiological features of MELAS include infarction-like lesions over the parieto-occipital or parieto-temporal areas, which help distinguish MELAS from childhood ischemic stroke.


Bidirectional epithelial-mesenchymal crosstalk provides self-sustaining profibrotic signals in pulmonary fibrosis.
Idiopathic pulmonary fibrosis (IPF) is the prototypic progressive fibrotic lung disease with a median survival of 2 to 4 years. Injury to and/or dysfunction of the alveolar epithelium is strongly implicated in IPF disease initiation, but the factors that determine whether fibrosis progresses rather than normal tissue repair occurs remain poorly understood. We previously demonstrated that zinc finger E-box-binding homeobox 1-mediated epithelial-mesenchymal transition in human alveolar epithelial type II (ATII) cells augments transforming growth factor-β-induced profibrogenic responses in underlying lung fibroblasts via paracrine signaling. Here, we investigated bidirectional epithelial-mesenchymal crosstalk and its potential to drive fibrosis progression. RNA-Seq of lung fibroblasts exposed to conditioned media from ATII cells undergoing RAS-induced epithelial-mesenchymal transition identified many differentially expressed genes including those involved in cell migration and extracellular matrix regulation. We confirmed that paracrine signaling between RAS-activated ATII cells and fibroblasts augmented fibroblast recruitment and demonstrated that this involved a zinc finger E-box-binding homeobox 1-tissue plasminogen activator axis. In a reciprocal fashion, paracrine signaling from transforming growth factor-β-activated lung fibroblasts or IPF fibroblasts induced RAS activation in ATII cells, at least partially through the secreted protein acidic and rich in cysteine, which may signal via the epithelial growth factor receptor via epithelial growth factor-like repeats. Together, these data identify that aberrant bidirectional epithelial-mesenchymal crosstalk in IPF drives a chronic feedback loop that maintains a wound-healing phenotype and provides self-sustaining profibrotic signals.


Reference Transcriptome for Deriving Marmoset Induced Pluripotent Stem Cells.
Limited access to primary tissue from various nonhuman primate (NHP) species represents a significant unmet need that hampers progress in understanding unique cellular diversity and gene regulation of specific tissues and organs in stem cell translational research. Most comparative biology studies have been limited to using postmortem tissue usually frozen specimens with limited utility for research. The generation of induced pluripotent stem cell (iPSC) lines from somatic cells, such as adult skin or blood cells, offers an alternative to invasive and ethically controversial interventions for acquiring tissue. Pluripotent iPSCs have virtually an unlimited capacity to proliferate and differentiate into all cell types of the body. We are generating high-quality validated NHP iPSC lines to offer to scientific community and facilitate their research programs. We use the non-integrative episomal vector system to generate iPSCs from NHP skin biopsies. In this chapter we describe the validation of NHP iPSC lines by confirming pluripotency and their propensity to differentiate into all three germ layers ectoderm, mesoderm, and endoderm according to established standards and measurable limits for a set of marker genes incorporated into a scorecard.


Bilateral atypical fractures of the femur: Ten years AFTER ten years of bisphosphonate therapy.
Atypical femur fracture (AFF) is a clinically important complication of bisphosphonate (BP) use in the treatment of osteoporosis. The benefits of long-term BP therapy in preventing osteoporotic fractures have been shown to outweigh the risks of treatment. Discontinuation of BPs or "drug holidays" have been implemented as a strategy to reduce the risk of rare complications such as AFF. We present the case of a 70-year-old postmenopausal woman who suffered bilateral AFF ten years after discontinuation of BP treatment. Management of this patient included fixation of the complete AFF with an intramedullary rod. A single dose of denosumab was administered prior to referral to endocrinology and seemed to contribute to callus formation. Denosumab was discontinued to prevent progression of the contralateral incomplete AFF. Teriparatide was indicated for the treatment of this patient's osteoporosis and also led to the resolution of the incomplete AFF. Patients receiving long-term BP therapy should be periodically reevaluated in order to maximize the benefit and minimize the risk of treatment. Current research supports the implementation of drug holidays to decrease the risk of AFF; however, this case report confirms the need for continued monitoring after discontinuation of BP therapy. Additionally, our review of current literature highlights the need for more specific research regarding duration of BP treatment and drug holidays.


Targeting GSK3 and Associated Signaling Pathways Involved in Cancer.
Glycogen synthase kinase 3 (GSK-3) is a serine/threonine (S/T) protein kinase. Although GSK-3 originally was identified to have functions in regulation of glycogen synthase, it was subsequently determined to have roles in multiple normal biochemical processes as well as various disease conditions. GSK-3 is sometimes referred to as a moonlighting protein due to the multiple substrates and processes which it controls. Frequently, when GSK-3 phosphorylates proteins, they are targeted for degradation. GSK-3 is often considered a component of the PI3K/PTEN/AKT/GSK-3/mTORC1 pathway as GSK-3 is frequently phosphorylated by AKT which regulates its inactivation. AKT is often active in human cancer and hence, GSK-3 is often inactivated. Moreover, GSK-3 also interacts with WNT/β-catenin signaling and β-catenin and other proteins in this pathway are targets of GSK-3. GSK-3 can modify NF-κB activity which is often expressed at high levels in cancer cells. Multiple pharmaceutical companies developed small molecule inhibitors to suppress GSK-3 activity. In addition, various natural products will modify GSK-3 activity. This review will focus on the effects of small molecule inhibitors and natural products on GSK-3 activity and provide examples where these compounds were effective in suppressing cancer growth.


Nanotoxoid Vaccines.
To improve innate defense against diseases, vaccine formulations are routinely administered to mount immune responses against disease-causing organisms or their associated toxins. These formulations are typically prepared with weakened forms of microbes, their surface proteins, or their virulence factors, which can train the immune system to recognize and neutralize similar infectious threats in later exposures. Owing to many unique properties of nanoparticles in enhancing vaccine potency, nanoscale carriers are drawing increasing interest as a platform for developing safer and more effective vaccine formulations. Notably, a nanoparticle-based strategy was recently demonstrated to safely deliver intact, non-denatured protein toxins to mount a potent anti-toxin immune response. A biomimetic nanoparticle cloaked in biological membranes was used to sequester membrane-active toxins. Upon interaction with the nanoparticles, the toxins become retrained and lose their toxicity as they are precluded from interacting with cellular targets. The resulting particle/toxin complex adopts a nanoparticulate morphology that facilitates the toxins' intracellular delivery. This sequestration approach has immense immunological implications owing to its ability in enabling structurally preserved toxins for immune processing. This technique offers opportunities in novel toxoid vaccine designs that promise more effective anti-toxin immune responses and contrasts the existing paradigm in toxoid preparation, in which toxins are antigenically altered to ensure virulence removal. The potent nanotoxoid formulations provide a viable anti-virulence measure in combating microbial infections that involve membrane-damaging toxins, including methicillin-resistant Staphylococcus aureus (MRSA) and Group A streptococcal infections.


Once weekly fluconazole for antifungal prophylaxis post-liver transplantation.
Invasive fungal infections (IFI) remain a significant cause of morbidity and mortality in orthotopic liver transplantation (OLT) recipients. In this retrospective study, the outcomes of a protocol using once weekly fluconazole for 3 months after OLT in low- and high-risk patients were reviewed. In total, 221 OLTs were evaluated in the 3-year period after institution of the new protocol to determine the incidence of IFI within 6 months post-OLT. In this cohort, 11 IFIs developed during the 6-month post-transplant period, with the majority being non-albicans Candida. High-risk patients had a greater rate of IFI (16.7% versus 3.4%, P = 0.038) and a significantly longer intensive unit care (ICU) and hospital lengths of stay compared with low-risk patients. Patient and graft survival were similar between the groups. Our patient population appeared to be at low risk for IFI, with 92% of the entire cohort considered low risk. Given the low incidence of IFI in the low-risk group and the possibility of such protocol selecting out for fluconazole-resistant fungi, the use of weekly fluconazole for 3 months may not be justifiable in low-risk OLT recipients. Given the increased resource utilization observed with IFI, further examination of a more intensive prophylactic strategy in high-risk patients may be warranted.


Intraductal Tubulopapillary Epithelial Proliferation Associated with Type 1 Autoimmune Pancreatitis.
A 70-year-old man was referred to our hospital with exacerbation of diabetes. His blood tests showed elevated levels of serum IgG4 and HbA1c. Computed tomography of the pancreatic body demonstrated a weakly enhanced mass, 2 cm in size, with indistinct borders. Magnetic resonance cholangiopancreatography revealed a narrowing of the main pancreatic duct (MPD) at the pancreatic body, a markedly dilated upstream duct, and a slightly dilated downstream duct. Endoscopic ultrasonography demonstrated an iso-hypoechoic heterogeneous mass, protruding and spreading in the pancreatic duct. The histology of a fine needle aspiration sample demonstrated fibrous tissue containing abundant IgG4-positive plasma cells and atypical epithelial cells. The imaging findings and histology were not typical for either pancreatic ductal adenocarcinoma or type 1 autoimmune pancreatitis (AIP), but these were not completely excluded, and a distal pancreatectomy was performed. Histological examination showed an intraductal tubulopapillary epithelial proliferation, which contained cytoplasmic mucin (MUC5AC and MUC6), and severe IgG4-positive lymphoplasmacytic infiltration in the interstitium around the MPD. Next-generation sequencing using DNA extracted from the tumor revealed no mutation of K-ras, GNAS, or TP53. The entire lesion was ultimately diagnosed as AIP with an intraductal tubular and papillary epithelial hyperplasia producing gastric-type mucin. Some recent reports have described AIP development in the background of intraductal papillary mucinous neoplasms, and some have hypothesized a paraneoplastic occurrence of IgG4-related disease. The current case indicates issues in the clinical diagnosis of rare variants of AIP, and raises questions about the relationship between AIP and pancreatic epithelial lesions.