Comparing the intracellular mobility of fluorescent proteins following in vitro expression or cell loading with streptolysin-O.
The application of fluorescent proteins in live cells has greatly improved our ability to study molecular mobility, which both reflects molecular function in live cells and reveals the properties of the local environment. Although measuring molecular mobility with fluorescent fusion proteins is powerful and convenient, certain experiments still require exogenous macromolecules to be loaded into cells. Cell viability provides a rough gauge of cellular damage following membrane permeabilization, but it is unknown how permeabilization will affect intracellular mobility. We have used fluorescence correlation spectroscopy to measure the intracellular dynamics of the enhanced green fluorescent protein (EGFP) in living human embryonic kidney (HEK) cells under conditions where the EGFP is either expressed or loaded using streptolysin O (SLO) permeabilization to determine how permeabilization effects mobility. We found that purified EGFP loaded with SLO has the same mobility as the expressed EGFP, while the mobility of the expressed EGFP after SLO permeabilization treatment becomes slightly slower. Our results indicate that SLO permeabilization is often accompanied by the loss of cellular soluble proteins to the surrounding medium, which explains the apparent decrease in diffusion rates following treatment. These measurements are also relevant to the role of molecular crowding in the intracellular mobility of proteins.


[Mass screening or early detection of cancer of the prostate by echography].
Mass screening of prostatic cancer by transrectal ultrasonography would result in between 600 and 2,500 negative biopsies for one positive biopsy, depending on the quality of the ultrasonographic image obtained. It is clear that such a procedure is not profitable. Early detection of prostatic cancer is profitable in a male population over the age of 50 with benign prostatic enlargement. For one positive biopsy, there would be 7 to 25 negative biopsies. In our opinion, a 4 to 16% yield justifies endorectal ultrasonography to look for a malignant prostatic tumor and is fully justified in men scheduled to undergo surgery on an adenoma of the prostate.


Mathematical spatio-temporal model of drug delivery from low temperature sensitive liposomes during radiofrequency tumour ablation.
Studies have demonstrated a synergistic effect between hyperthermia and chemotherapy, and clinical trials in image-guided drug delivery combine high-temperature thermal therapy (ablation) with chemotherapy agents released in the heating zone via low temperature sensitive liposomes (LTSL). The complex interplay between heat-based cancer treatments such as thermal ablation and chemotherapy may require computational models to identify the relationship between heat exposure and pharmacokinetics in order to optimise drug delivery. Spatio-temporal data on tissue temperature and perfusion from heat-transfer models of radiofrequency ablation were used as input data. A spatio-temporal multi-compartmental pharmacokinetic model was built to describe the release of doxorubicin (DOX) from LTSL into the tumour plasma space, and subsequent transport into the extracellular space, and the cells. Systemic plasma and tissue compartments were also included. We compared standard chemotherapy (free-DOX) to LTSL-DOX administered as bolus at a dose of 0.7 mg/kg body weight. Modelling LTSL-DOX treatment resulted in tumour tissue drug concentration of approximately 9.3 microg/g with highest values within 1 cm outside the ablation zone boundary. Free-DOX treatment produced comparably uniform tissue drug concentrations of approximately 3.0 microg/g. Administration of free-DOX resulted in a considerably higher peak level of drug concentration in the systemic plasma compartment (16.1 microg/g) compared to LTSL-DOX (4.4 microg/g). These results correlate well with a prior in vivo study. Combination of LTSL-DOX with thermal ablation allows localised drug delivery with higher tumour tissue concentrations than conventional chemotherapy. Our model may facilitate drug delivery optimisation via investigation of the interplays among liposome properties, tumour perfusion, and heating regimen.


Crumpled states of a wire in a two-dimensional cavity with pins.
In this paper, we report an extensive experimental study of the configurations of a plastic wire injected into a two-dimensional planar cavity populated with fixed pins. The wire is not allowed to cross any pin, but it can move in a wormlike manner within the cavity until to become jammed in a crumpled state. The jammed packing fraction depends heavily on the topology of the cavity, which depends on the number of pins. The experiment reveals nontrivial entanglement effects and scaling laws which are largely independent of the details of the distribution of pins, the symmetry of the cavity or the type of the wire. A mean-field model for the process is presented and analogies with some basic aspects of statistical thermodynamics are discussed.


A new modification of the technique of arthroscopy of the knee joint.
Experience from arthroscopy in 224 patients with knee complaints is reported. The common method for arthroscopy is compared with a new modification developed during the study. The new method includes introduction of a 5 mm arthroscope through the patellar tendon at the level of the joint line and the use of hooks to test the menisci and the ligaments. The experience from this method shows that it gives an expanded field of vision and that technical failures are less frequent. No complications occurred. It is stated that arthroscopy is of great value in the diagnosis of knee injuries. However, a high diagnostic accuracy can only be reached by an experienced operator using a strictly standardized method.


Discovery of 3-substituted aminocyclopentanes as potent and orally bioavailable NR2B subtype-selective NMDA antagonists.
A series of 3-substituted aminocyclopentanes has been identified as highly potent and selective NR2B receptor antagonists. Incorporation of a 1,2,4-oxadiazole linker and substitution of the pendant phenyl ring led to the discovery of orally bioavailable analogues that showed efficient NR2B receptor occupancy in rats. Unlike nonselective NMDA antagonists, the NR2B-selective antagonist 22 showed no adverse affects on motor coordination in the rotarod assay at high dose. Compound 22 was efficacious following oral administration in a spinal nerve ligation model of neuropathic pain and in an acute model of Parkinson's disease in a dose dependent manner.


An efficient, adaptive method of measuring loudness growth functions.
This paper presents a new categorical loudness scaling procedure that differs from previously published loudness scaling procedures by (i) adaptively selecting a new set of levels for each new sequence, (ii) deriving levels that are equispaced on the loudness scale, and (iii) using a continuous scale with few labels. A major advantage of the adaptive procedure is that the individual dynamic range need not be measured prior to loudness testing. The adaptive procedure proved to be time efficient and to produce complete loudness functions from Not heard to Uncomfortably loud for normal hearing and hearing impaired subjects. The pattern of short-term and long-term reliability was similar to that reported for non-adaptive loudness scaling procedures. Three presentations produced a stable loudness function. Normative curves for one octave babble-noise at six test frequencies are presented and compared to normative data obtained with a selection of published categorical scaling procedures.


[An autopsy case of cerebral calcification--with special reference to the morphogenesis of the calcified deposits].
The case was a 51 years old male who died of cervical spinal cord injury. On admission, X-ray disclosed distinct hyperostosis such as ossification of posterior longitudinal ligament, ankylosing spondylitis and callus luxurians of bilateral hip joints. He had no familial history. He normally developed into adult without any mental and neurological abnormalities. In the latter half of the fifth decade, he developed progressive spastic diplegia of his legs. Laboratory studies revealed evident hypocalcaemia (2.3 meq/l). Hormonal examination of the parathyroid gland was not performed. Postmortem examination of the brain (1,400 g) disclosed widely spreading numerous spherical deposits of various sizes stained deeply with hematoxilin. These deposits were positive with PAS, colloidal iron, prussian blue and Kossa's Method for calcium etc. as shown in Table 1. According to their histochemical properties, these deposits were considered to consist of both acid mucopolysaccharides and proteins to which calcium and iron have been bound later. These deposits were predominantly observed in the basal ganglia, dorso-lateral portion of the thalamus and the depth of the cerebellum. The dentate nucleus was mostly spared. To the lesser degree, these deposits were also seen around the capillaries and subadventitial space of the small vessels in the cerebral cortex, cerebral white matter, capsula interna and red nucleus. These deposits were shown to be adjacent to the capillary walls microscopically. Electron microscopy disclosed that many electron dense spherical bodies surrounding capillary were related to the basement membrane of the endothelium or pericyte.(ABSTRACT TRUNCATED AT 250 WORDS)


A unique episode of REM sleep behavior disorder triggered during surgery for Parkinson's disease.
REM sleep behavior disorder (RBD) is characterized by vigorous movements during REM sleep. Here, the authors report the case of a patient who presented such a disorder immediately after the implantation of the definitive electrode for left subthalamic stimulation. Interestingly, this was and has remained a unique episode in his medical history. It was found that a microlesion in or near the upper part of the pars compacta of the substantia nigra was very likely responsible for this phenomenon.


Virology in the Classroom: Current Approaches and Challenges to Undergraduate- and Graduate-Level Virology Education.
The pervasive effects of the current coronavirus disease 2019 pandemic are but one reason for educators to refocus their efforts on virology teaching. Additionally, it is critical to understand how viruses function and to elucidate the relationship between virus and host. An understanding of current virology education may improve pedagogical approaches for educating our students and trainees. Faculty who teach undergraduate microbiology indicate that approximately 10% of the course content features viruses; stand-alone virology courses are infrequently offered to undergraduates. Fortunately, virology taught to undergraduates includes foundational material; several approaches for delivery of lecture- and lab-based content exist. At the graduate education level, there is growing appreciation that an emphasis on logic, reasoning, inference, and statistics must be reintroduced into the curriculum to create a generation of scientists who have a greater capacity for creativity and innovation. Educators also need to remove barriers to student success, at all levels of education.


Local treatment for extensive deep dermal thickness burn and follow-up study.
For the local treatment of extensive burn patients including superficial dermal thickness burn and deep dermal thickness burn, it is often difficult to determine whether conservative treatment should be followed consistently or grafting be employed. The present study was made to determine when grafting should be provided if required and the treatment method to provide the best results. For the study, a total of 41 cases were taken to which the author rendered treatment from the initial stages, in the period from January 1981 to December 1985. In these cases, the burn area was more than 10 per cent of the body surface area (BSA) in infants and children, and more than 20 per cent BSA in adults. Both superficial dermal thickness burn and deep dermal thickness burn were coexisting. The treatments and results of examination after convalescence are as follows. 1. In conservative local treatment, impregnated ointment gauze, L(D)PS and silver sulfadiazine were used, with wet-to-dry dressing, and hydrotherapy or skeletal suspension used jointly in some cases. 2. Of 41 cases, 38 received grafting during the treatment process. Patch autograft was applied to all cases, and allograft was jointly applied to 5 cases. 3. From post-convalescence examination, most cases showed hypertrophic scar when grafting was provided at a later stage and with the younger age of the patient. From this, it can be emphasized that earlier grafting will cause reduction in treatment period, with better functional and esthetic results, even in deep dermal thickness burn.


Knowledge and awareness of age related eye diseases: a population-based survey.
To determine general awareness and knowledge about cataracts, glaucoma and diabetic retinopathy (DR), as common avoidable causes of blindness in an Iranian population. This cross-sectional population-based survey was performed on residents over 45 years of age in Tehran. The sampling frame was the list of all landline phone numbers registered by the Telecommunications Center of Iran, through which systematic random sampling was performed. Data was collected by phone-call interviews and completing a semi-structured questionnaire. Awareness was defined as whether the respondent had ever heard of the disease. Knowledge was assessed by realizing different aspects of each disease. Of a total of 1,084 eligible people including 574 (52.9%) women and 510 (47.1%) men were included and 957 subjects (response rate, 88.3%) completed the interview. Awareness regarding glaucoma, cataract and DR was 46.6% (95% confidence interval [CI]:43.4 -49.8%), 82.9% (95% CI: 80.5 -85.3%) and 86.2% (95% CI: 84-88.4%). In addition, 19.2% (95% CI: 16.7 -21.7%), 57.3% (95% CI: 54.2-60.4%) and 72% (95% CI: 69.2 -74.8%) of respondents could give at least a basic definition of the mentioned diseases, respectively. Only 22.6% (95% CI: 20-25.2%) and 41.6% (95% CI: 38.5-44.7%) realized glaucoma and DR as a treatable condition; in contrast, 77.2% (95% CI: 74.5-79.9%) categorized cataract as treatable. Only 19% and 7.1% knew that DR and glaucoma may commence without any apparent symptoms. Compared with cataract and DR, most participants had limited information about glaucoma. In addition, few of the respondents were familiar with the initial symptoms of DR and glaucoma.


Highly efficient continuous-wave and passively Q-switched Yb:YLuGdCOB compact lasers.
Highly efficient operation of an Yb:YLuGdCOB compact laser was demonstrated, producing an output power of 14.30 W around 1054 nm, with an optical-to-optical efficiency of 70.0% and a slope efficiency of 81%. In dual-polarization oscillation in an emission range of 1080 - 1085 nm, 11.37 W of output power was generated with a slope efficiency of 76%. Efficient passively Q-switched operation was also realized by incorporating a 2D MoTe2 saturable absorber into the compact resonator, producing a maximum pulsed output power of 1.56 W at 1030 nm at a repetition rate of 481 kHz; while the largest attainable pulse energy, minimum pulse duration, and highest peak power were, respectively, 3.87 μJ, 26.2 ns, and 147.7 W. The pulse width proves to be the shortest, while the peak power is the highest ever reported for solid-state lasers passively Q-switched with 2D saturable absorbers.


Artificial Chiral Probes and Bioapplications.
The development of artificial chiral architectures, especially chiral inorganic nanostructures, has greatly promoted research into chirality in nanoscience. The nanoscale chirality of artificial chiral nanostructures offers many new application opportunities, including chiral catalysis, asymmetric synthesis, chiral biosensing, and others that may not be allowed by natural chiral molecules. Herein, the progress achieved during the past decade in chirality-associated biological applications (biosensing, biolabeling, and bioimaging) combined with individual chiral nanostructures (such as chiral semiconductor nanoparticles and chiral metal nanoparticles) or chiral assemblies is discussed.


Human adenoviruses in water: occurrence and health implications: a critical review.
Adenoviruses are important human pathogens that are responsible for both enteric illnesses and respiratory and eye infections. Recently, these viruses have been found to be prevalent in rivers, coastal waters, swimming pool waters, and drinking water supplies worldwide. United Sates Environmental Protection Agency (USEPA) listed adenovirus as one of nine microorganisms on the Contamination Candidate List for drinking water because their survival characteristic during water treatment is not yet fully understood. Adenoviruses have been found to be significantly more stable than fecal indicator bacteria and other enteric viruses during UV treatment. Adenovirus infection may be caused by consumption of contaminated water or inhalation of aerosolized droplets during water recreation. The goal of this review is to summarize the state of technology for adenovirus detection in natural and drinking waters and the human health risk imposed by this emerging pathogen. The occurrence of these viruses in natural and treated waters is summarized from worldwide reports.


Thermal convection in monodisperse and bidisperse granular gases: a simulation study.
We present results of a simulation study of inelastic hard disks vibrated in a vertical container. An event-driven molecular dynamics method is developed for studying the onset of convection. Varying the relevant parameters (inelasticity, number of layers at rest, intensity of the gravity) we are able to obtain a qualitative agreement of our results with recent hydrodynamical predictions. Increasing the inelasticity, a first continuous transition from the absence of convection to one convective roll is observed, followed by a discontinuous transition to two convective rolls, with hysteretic behavior. At fixed inelasticity and increasing gravity, a transition from no convection to one roll can be evidenced. If the gravity is further increased, the roll is eventually suppressed. Increasing the number of monolayers the system eventually localizes mostly at the bottom of the box: in this case multiple convective rolls as well as surface waves appear. We analyze the density and temperature fields and study the existence of symmetry breaking in these fields in the direction perpendicular to the injection of energy. We also study a binary mixture of grains with different properties (inelasticity or diameters). The effect of changing the properties of one of the components is analyzed, together with density, temperature, and temperature ratio fields. Finally, the presence of a low fraction of quasielastic impurities is shown to determine a sharp transition between convective and nonconvective steady states.


Allyl compounds selectively killed human immunodeficiency virus (type 1)-infected cells.
Various allyl compounds were tested as anti-HIV reagents. Of all the allyl compounds tested, allyl disulfide (CH2 = CH-CH2-S-S-CH2-CH = CH2), allyl alcohol (CH2 = CH-CH2-OH), and its ester strongly depressed cell proliferation of HIV-1-infected cells. KC50 (50% killing concentration) of the allyl disulfide and the allyl alcohol were 34 microM and 10 microM, respectively. The allyl alcohol, in particular, completely inhibited cell growth of HIV-1-infected cells by a single injection at a concentration of 40 microM, ultimately killing the viable CEM/LAV-1 cells within 96 hr cultivation. No effect of the allyl alcohol and its esters on cell proliferation of CEM cells, which are a HIV-1 noninfected human T-cell cell line, was observed at the concentration range of 20 to 200 microM under the same experimental conditions. The results suggest that the effect of the allyl alcohol and its esters resulted in remarkable promotion of the cytopathic effect induced by HIV-1 virus.


Short communication: The effect of calcium supplementation at calving on neutrophil function.
Low total blood calcium concentration after calving has been demonstrated to be a risk factor for reduced neutrophil function. The objective of this study was to evaluate whether administration of an injectable calcium supplement product soon after calving increased neutrophil oxidative burst or phagocytosis capacity. Cows (n = 27) from 4 farms were blocked by parity and randomly assigned to receive either calcium gluconate (35% wt/vol) in combination with calcium glucoheptonate (10% wt/vol; Theracalcium, Vétoquinol Canada Inc., Lavaltrie, Quebec, Canada) or a placebo within 12 h after calving and again 24 h later. Each dose of 120 mL was injected subcutaneously over 2 sites. Total serum calcium concentration, neutrophil oxidative burst, and neutrophil phagocytosis capacity were measured from coccygeal blood samples before (time 0) and 72 h after first treatment. There was no difference between treatment groups in lactation number, total calcium concentration, oxidative burst, or phagocytosis at time of enrollment. There was no effect of treatment on oxidative burst or phagocytosis by neutrophils. This preliminary study does not support an effect of supplemental calcium to improve neutrophil oxidative burst or phagocytosis capacity of low-parity parturient cows.


Induction of regulatory T cells by high-dose gp96 suppresses murine liver immune hyperactivation.
Immunization with high-dose heat shock protein gp96, an endoplasmic reticulum counterpart of the Hsp90 family, significantly enhances regulatory T cell (Treg) frequency and suppressive function. Here, we examined the potential role and mechanism of gp96 in regulating immune-mediated hepatic injury in mice. High-dose gp96 immunization elicited rapid and long-lasting protection of mice against concanavalin A (Con A)-and anti-CD137-induced liver injury, as evidenced by decreased alanine aminotransaminase (ALT) levels, hepatic necrosis, serum pro-inflammatory cytokines (IFN-γ, TNF-α, and IL-6), and number of IFN-γ (+) CD4(+) and IFN-γ (+) CD8(+) T cells in the spleen and liver. In contrast, CD4(+)CD25(+)Foxp3(+) Treg frequency and suppressive function were both increased, and the protective effect of gp96 could be generated by adoptive transfer of Treg cells from gp96-immunized mice. In vitro co-culture experiments demonstrated that gp96 stimulation enhanced Treg proliferation and suppressive function, and up-regulation of Foxp3, IL-10, and TGF-β1 induced by gp96 was dependent on TLR2- and TLR4-mediated NF-κB activation. Our work shows that activation of Tregs by high-dose gp96 immunization protects against Con A- and anti-CD137-induced T cell-hepatitis and provides therapeutic potential for the development of a gp96-based anti-immune hyperactivation vaccine against immune-mediated liver destruction.


Binding of exogenous protein fragments to native proteins: possible explanation for the overestimation of uptake of extrinsically labelled macromolecules from the gut.
Investigators studying the transport of macromolecules across mucosal surfaces have noted that techniques involving exogenously labelled macromolecules result in estimation of transport which exceed estimates based on immunochemical methods. The present study tested the possibility that radio-labelled fragments of macromolecules might bind to host proteins resulting in an apparent increase in 'size' of the fragments. Fragments obtained by dialysis of bovine serum albumin (BSA) labelled with tritium gas by Wilzbach method, bound to normal rabbit serum in vitro and in vivo. Binding to BSA exceeded binding to bovine gamma globulin (BGG). Dinitrophenylated BGG was administered by gavage to adult rats; DNP-bearing fragments were identified in their serum and recovered by gel filtration. After radio-labelling, these serum fractions also bound to macromolecular constituents of normal rat serum. These findings suggest that several different fragments of proteins may bind to native serum components. These results may bear upon in vivo observations since it is likely that following intestinal instillation of extrinsically labelled protein, labelled amino acids or small peptides are generated by the digestive process, become absorbed, bind to serum macromolecules and thereby mimic the uptake of macromolecules.


Oral Delivery of Honokiol Microparticles for Nonrapid Eye Movement Sleep.
Honokiol (HNK) is a small-molecule lignin extracted from Magnolia Officinalis, demonstrating high potency in promoting nonrapid eye movement (NREM) sleep by modulating the benzodiazepine site of the GABAA receptor. However, the clinical use of HNK in the treatment of insomnia is restricted by its extremely low oral bioavailability. In the present work, enhanced oral bioavailability of HNK was achieved by loading it into poly lactide-glycolide acid microparticles (HNK-MP). After oral administration, HNK-MP demonstrated 15-fold increase of AUC0-12 h in comparison to free HNK. The maximum blood concentration ( Cmax) of HNK in HNK-MP-treated rats was 3.6 μg/mL at 2 h after oral administration, which was 6.5-fold of that in free HNK-treated rats. Oral administration of HNK-MP (20 mg/kg) efficiently increased NREM sleep by 60% by enhancing the transition from wakefulness to NREM sleep in rats. The biosafety of HNK-MP was assessed in vivo, and no damage occurred in the gastrointestinal tract. The present study provides a promising oral HNK formulation for the treatment of insomnia.


B-vitamins and heat processed fermented starchy and vegetable foods in sub-Saharan Africa: A review.
Micronutrient deficiency still occurs in sub-Saharan Africa (SSA) despite the availability of several food resources, particularly fermented foods and vegetables, with high nutritional potential. Fermentation enhances the quality of food in several aspects. Organoleptically, certain taste, aroma, and textures are developed. Health and safety are improved by inhibiting the growth of several foodborne pathogens and removing harmful toxic compounds. Furthermore, nutrition is enhanced by improving micronutrient contents and bioavailability from the food, especially vitamin B content. However, during processing and before final consumption, many fermented foods are heat treated (drying, pasteurization, cooking, etc.) to make the food digestible and safe for consumption. Heat treatment improves the bioavailability of B-vitamins in some foods. In other foods, heating decreases the nutritional value because some B-vitamins are degraded. In SSA, cooked starchy foods are often associated with vegetables in household meals. This paper reviews studies that have focused fermented starchy foods and vegetable foods in SSA with the potential to provide B-vitamins to consumers. The review also describes the process of the preparation of these foods for final consumption, and techniques that can prevent or lessen B-vitamin loss, or enrich B-vitamins prior to consumption.


Chromatin assembly factor 1 is essential and couples chromatin assembly to DNA replication in vivo.
De novo chromatin assembly maintains histone density on the daughter strands in the wake of the replication fork. The heterotrimer chromatin assembly factor 1 (CAF-1) couples DNA replication to histone deposition in vitro, but is not essential for yeast cell proliferation. Depletion of CAF-1 in human cell lines demonstrated that CAF-1 was required for efficient progression through S-phase. Cells lacking CAF-1 accumulated in early and mid S-phase and replicated DNA slowly. The checkpoint kinase Chk1, but not Chk2, was phosphorylated in response to CAF-1 depletion, consistent with a DNA replication defect. CAF-1-depleted cell extracts completely lacked DNA replication-coupled chromatin assembly activity, suggesting that CAF-1 is required for efficient S-phase progression in human cells. These results indicate that, in contrast to yeast, human CAF-1 is necessary for coupling chromatin assembly with DNA replication.


[Multiple brown tumors in a female hemodialyzed patient with severe secondary hyperparathyroidism].
Skeletal brown tumours are relatively uncommon, and brown tumours that involve multiple bones are considered very rare. We describe a 29-year-old woman with chronic renal failure (CRF) who had undergone hemodialysis for 21 years and developed multiple brown tumours associated with severe secondary hyperparathyroidism. Computed tomography (CT) revealed multiple brown tumours involving scapula, ribs, spine and sacroiliac bone. Microscopic analysis of the brown tumour showed dense infiltration of the marrow space by reactive fibroblastic tissue with irregularly distributed multinucleated osteoclastic giants cells and marked increase in hematopoietic elements.


Power gains by using external information in clinical trials are typically not possible when requiring strict type I error control.
In the era of precision medicine, novel designs are developed to deal with flexible clinical trials that incorporate many treatment strategies for multiple diseases in one trial setting. This situation often leads to small sample sizes in disease-treatment combinations and has fostered the discussion about the benefits of borrowing of external or historical information for decision-making in these trials. Several methods have been proposed that dynamically discount the amount of information borrowed from historical data based on the conformity between historical and current data. Specifically, Bayesian methods have been recommended and numerous investigations have been performed to characterize the properties of the various borrowing mechanisms with respect to the gain to be expected in the trials. However, there is common understanding that the risk of type I error inflation exists when information is borrowed and many simulation studies are carried out to quantify this effect. To add transparency to the debate, we show that if prior information is conditioned upon and a uniformly most powerful test exists, strict control of type I error implies that no power gain is possible under any mechanism of incorporation of prior information, including dynamic borrowing. The basis of the argument is to consider the test decision function as a function of the current data even when external information is included. We exemplify this finding in the case of a pediatric arm appended to an adult trial and dichotomous outcome for various methods of dynamic borrowing from adult information to the pediatric arm. In conclusion, if use of relevant external data is desired, the requirement of strict type I error control has to be replaced by more appropriate metrics.


Duodenal adenocarcinoma with neuroendocrine features in a patient with acromegaly and thyroid papillary adenocarcinoma: a unique combination of endocrine neoplasia.
A 67-year-old woman with familial clustering of thyroid papillary adenocarcinoma was diagnosed with acromegaly due to pituitary macroadenoma. She had multiple skin vegetations, but had no parathyroid and pancreas diseases. Before transsphenoidal surgery, she was further diagnosed as having a duodenal tumor and multiple hypervascular liver nodules. Biopsy specimens from the duodenal tumor and liver nodules were diagnosed histologically as moderately differentiated adenocarcinoma. Immunohistochemically, the tumor cells were positive for chromogranin, synaptophysin and somatostatin receptor 2a, suggestive for neuroendocrine features. After surgery, the patient was not in biochemical remission, and octreotide treatment was initiated. The duodenal cancer was treated with chemotherapy (neoadjuvant cisplatin and S-1). After 24 months, the patient's insulin-like growth factor I level had been normalized, and her liver tumors had not progressed macroscopically. This is a rare case of acromegaly associated with multiple endocrine tumors, not being categorized as conventional multiple endocrine neoplasia. Octreotide treatment might have had beneficial effects on our patient's duodenal adenocarcinoma and liver metastases, both directly via SSTR2a and indirectly via GH suppression, thereby contributing to their slow progression.


Use, option and externality values: are contingent valuation studies in health care mis-specified?
A general population sample of Australian respondents completed a contingent valuation (CV) survey that asked them to value six scenarios. These varied according to whether the scenario was seeking to elicit: (i) use value; (ii) externality value; (iii) option value; or (iv) a combination. Results indicate that use plus externality and/or option value was significantly greater than use value alone. As CV studies in health (care) overwhelmingly focus on use value alone - often implicitly through study design rather than explicitly - this raises the possibility of mis-specification in CV research in health (care). The implications for CV in health (care) are considered.


Genome-Wide Association Studies of Metabolites in Patients with CKD Identify Multiple Loci and Illuminate Tubular Transport Mechanisms.
Background The kidneys have a central role in the generation, turnover, transport, and excretion of metabolites, and these functions can be altered in CKD. Genetic studies of metabolite concentrations can identify proteins performing these functions.Methods We conducted genome-wide association studies and aggregate rare variant tests of the concentrations of 139 serum metabolites and 41 urine metabolites, as well as their pairwise ratios and fractional excretions in up to 1168 patients with CKD.Results After correction for multiple testing, genome-wide significant associations were detected for 25 serum metabolites, two urine metabolites, and 259 serum and 14 urinary metabolite ratios. These included associations already known from population-based studies. Additional findings included an association for the uremic toxin putrescine and variants upstream of an enzyme catalyzing the oxidative deamination of polyamines (AOC1, P-min=2.4×10-12), a relatively high carrier frequency (2%) for rare deleterious missense variants in ACADM that are collectively associated with serum ratios of medium-chain acylcarnitines (P-burden=6.6×10-16), and associations of a common variant in SLC7A9 with several ratios of lysine to neutral amino acids in urine, including the lysine/glutamine ratio (P=2.2×10-23). The associations of this SLC7A9 variant with ratios of lysine to specific neutral amino acids were much stronger than the association with lysine concentration alone. This finding is consistent with SLC7A9 functioning as an exchanger of urinary cationic amino acids against specific intracellular neutral amino acids at the apical membrane of proximal tubular cells.Conclusions Metabolomic indices of specific kidney functions in genetic studies may provide insight into human renal physiology.


Functional consequences of S1P receptor modulation in rat oligodendroglial lineage cells.
Fingolimod (FTY720) and its phosphorylated form FTY720P are modulators of sphingosine-1-phosphate (S1P) receptors, which are G-protein coupled receptors linked to cell migration and vascular maturation. The efficacy of FTY720 in autoimmune diseases such as multiple sclerosis and its animal models has been attributed to its inhibition of lymphocyte trafficking to target organs. In this study, we examined the role of S1P receptors in cultured rat oligodendrocytes (OLGs) and OLG progenitor cells (OPCs) using the active phosphorylated form of FTY720. We found that (1) FTY720P improves the survival of neonatal rat OLGs during serum withdrawal, which is associated with the phosphorylation of extracellular signal regulated kinases (ERK1/2) and Akt; (2) FTY720P regulates OPC differentiation into OLGs in a concentration-dependent manner; and (3) S1P receptors are differentially modulated by platelet-derived growth factor (PDGF) resulting in downregulation of S1P5 and upregulation of S1P1 in OPCs. In addition, siRNA studies revealed that S1P1 participates in PDGF-induced OPC mitogenesis. We conclude that S1P1 and S1P5 serve different functions during oligodendroglial development, and possibly during remyelination.


[Progress in the studies of prostate biopsy methods].
Prostate cancer (PCa) is one of the most common malignancies in aged males, ranking the second in the incidence of malignant tumors in men. Early diagnosis is essential, as advanced PCa is quite difficult to be managed, especially when it becomes castration-resistant or neuroendocrine PCa. Currently, the diagnosis of PCa is often based on pathology by prostate biopsy. Many recent studies focus on the impact of different biopsy methods on the diagnosis of the malignancy, but no consensus has been reached hitherto. This review summarizes various prostate biopsy methods and their latest studies.


Advancing Imbalanced Domain Adaptation: Cluster-Level Discrepancy Minimization With a Comprehensive Benchmark.
Unsupervised domain adaptation methods have been proposed to tackle the problem of covariate shift by minimizing the distribution discrepancy between the feature embeddings of source domain and target domain. However, the standard evaluation protocols assume that the conditional label distributions of the two domains are invariant, which is usually not consistent with the real-world scenarios such as long-tailed distribution of visual categories. In this article, the imbalanced domain adaptation (IDA) is formulated for a more realistic scenario where both label shift and covariate shift occur between the two domains. Theoretically, when label shift exists, aligning the marginal distributions may result in negative transfer. Therefore, a novel cluster-level discrepancy minimization (CDM) is developed. CDM proposes cross-domain similarity learning to learn tight and discriminative clusters, which are utilized for both feature-level and distribution-level discrepancy minimization, palliating the negative effect of label shift during domain transfer. Theoretical justifications further demonstrate that CDM minimizes the target risk in a progressive manner. To corroborate the effectiveness of CDM, we propose two evaluation protocols according to the real-world situation and benchmark existing domain adaptation approaches. Extensive experiments demonstrate that negative transfer does occur due to label shift, while our approach achieves significant improvement on imbalanced datasets, including Office-31, Image-CLEF, and Office-Home.


The Plasmodium falciparum exported J domain proteins fine-tune human and malarial Hsp70s: pathological exploitation of proteostasis machinery.
Cellular proteostasis requires a network of molecular chaperones and co-chaperones, which facilitate the correct folding and assembly of other proteins, or the degradation of proteins misfolded beyond repair. The function of the major chaperones, heat shock protein 70 (Hsp70) and heat shock protein 90 (Hsp90), is regulated by a cohort of co-chaperone proteins. The J domain protein (JDP) family is one of the most diverse co-chaperone families, playing an important role in functionalizing the Hsp70 chaperone system to form a powerful protein quality control network. The intracellular malaria parasite, Plasmodium falciparum, has evolved the capacity to invade and reboot mature human erythrocytes, turning them into a vehicles of pathology. This process appears to involve the harnessing of both the human and parasite chaperone machineries. It is well known that malaria parasite-infected erythrocytes are highly enriched in functional human Hsp70 (HsHsp70) and Hsp90 (HsHsp90), while recent proteomics studies have provided evidence that human JDPs (HsJDPs) may also be enriched, but at lower levels. Interestingly, P. falciparum JDPs (PfJDPs) are the most prominent and diverse family of proteins exported into the infected erythrocyte cytosol. We hypothesize that the exported PfJPDs may be an evolutionary consequence of the need to boost chaperone power for specific protein folding pathways that enable both survival and pathogenesis of the malaria parasite. The evidence suggests that there is an intricate network of PfJDP interactions with the exported malarial Hsp70 (PfHsp70-x) and HsHsp70, which appear to be important for the trafficking of key malarial virulence factors, and the proteostasis of protein complexes of human and parasite proteins associated with pathology. This review will critically evaluate the current understanding of the role of exported PfJDPs in pathological exploitation of the proteostasis machinery by fine-tuning the chaperone properties of both human and malarial Hsp70s.


Models for discovery of targeted therapy in genetic epileptic encephalopathies.
Epileptic encephalopathies are severe disorders emerging in the first days to years of life that commonly include refractory seizures, various types of movement disorders, and different levels of developmental delay. In recent years, many de novo occurring variants have been identified in individuals with these devastating disorders. To unravel disease mechanisms, the functional impact of detected variants associated with epileptic encephalopathies is investigated in a range of cellular and animal models. This review addresses efforts to advance and use such models to identify specific molecular and cellular targets for the development of novel therapies. We focus on ion channels as the best-studied group of epilepsy genes. Given the clinical and genetic heterogeneity of epileptic encephalopathy disorders, experimental models that can reflect this complexity are critical for the development of disease mechanisms-based targeted therapy. The convergence of technological advances in gene sequencing, stem cell biology, genome editing, and high throughput functional screening together with massive unmet clinical needs provides unprecedented opportunities and imperatives for precision medicine in epileptic encephalopathies.


Skin-Resident T Cells Drive Dermal Dendritic Cell Migration in Response to Tissue Self-Antigen.
Migratory dendritic cell (DC) subsets deliver tissue Ags to draining lymph nodes (DLNs) to either initiate or inhibit T cell-mediated immune responses. The signals mediating DC migration in response to tissue self-antigen are largely unknown. Using a mouse model of inducible skin-specific self-antigen expression, we demonstrate that CD103+ dermal DCs (DDCs) rapidly migrate from skin to skin DLN (SDLNs) within the first 48 h after Ag expression. This window of time was characterized by the preferential activation of tissue-resident Ag-specific effector T cells (Teffs), with no concurrent activation of Ag-specific Teffs in SDLNs. Using genetic deletion and adoptive transfer approaches, we show that activation of skin-resident Teffs is required to drive CD103+ DDC migration in response to tissue self-antigen and this Batf3-dependent DC population is necessary to mount a fulminant autoimmune response in skin. Conversely, activation of Ag-specific Teffs in SDLNs played no role in DDC migration. Our studies reveal a crucial role for skin-resident T cell-derived signals, originating at the site of self-antigen expression, to drive DDC migration during the elicitation phase of an autoimmune response.


Precarious employment, ill health, and lessons from history: the case of casual (temporary) dockworkers 1880-1945.
An international body of scientific research indicates that growth of job insecurity and precarious forms of employment over the past 35 years have had significant negative consequences for health and safety. Commonly overlooked in debates over the changing world of work is that widespread use of insecure and short-term work is not new, but represents a return to something resembling labor market arrangements found in rich countries in the 19th and early 20th centuries. Moreover, the adverse health effects of precarious employment were extensively documented in government inquiries and in health and medical journals. This article examines the case of a large group of casual dockworkers in Britain. It identifies the mechanisms by which precarious employment was seen to undermine workers and families' health and safety. The article also shows the British dockworker experience was not unique and there are important lessons to be drawn from history. First, historical evidence reinforces just how health-damaging precarious employment is and how these effects extend to the community, strengthening the case for social and economic policies that minimize precarious employment. Second, there are striking parallels between historical evidence and contemporary research that can inform future research on the health effects of precarious employment.


Hormonal influences on psychomotor reminiscence.
Psychomotor reminiscence was measured in contraceptively medicated and non-medicated subjects on Days 2, 8, and 14 of their menstrual cycles. As previously reported, mean reminiscence in non-medicated subjects was relatively low on Days 2 and 8, but significantly higher on Day 14. By contrast, reminiscence means in medicated subjects were relatively low and essentially alike across all testing days. These results are discussed within the context of reactive inhibition theory, wherein the adrenergic properties of mid-cycle concentrations of estrogenic hormones are thought responsible for more vigorous responding, hence more reactive inhibition and higher reminiscence. Such concentrations were, of course, precluded by contraceptive medication.


CTX-M extended-spectrum β-lactamase-producing Klebsiella spp, Salmonella spp, Shigella spp and Escherichia coli isolates in Iranian hospitals.
This study was conducted in Iran in order to assess the distribution of CTX-M type ESBLs producing Enterobacteriaceae. From January 2012 to December 2013, totally 198 E. coli, 139 Klebsiella spp, 54 Salmonella spp and 52 Shigella spp from seven hospitals of six provinces in Iran were screened for resistance to extended-spectrum cephalosporins. After identification and susceptibility testing, isolates presenting multiple-drug resistance (MDR) were evaluated for ESBL production by the disk combination method and by Etest using (cefotaxime and cefotaxime plus clavulanic acid). All isolates were also screened for blaCTX-M using conventional PCR. A total of 42.92%, 33.81%, 14.81% and 7.69% of the E. coli, Klebsiella spp, Salmonella spp and Shigella spp isolates were MDR, respectively. The presence of CTX-M enzyme among ESBL-producing isolates was 85.18%, 77.7%, 50%, and 66.7%, in E. coli, Klebsiella spp, Salmonella spp and Shigella spp respectively. The overall presence of CTX-M genes in Enterobacteriaceae was 15.4% and among the resistant isolates was 47.6%. This study indicated that resistance to β-lactams mediated by CTX-M enzymes in Iran had similar pattern as in other parts of the world. In order to control the spread of resistance, comprehensive studies and programs are needed.


Using token reinforcement to increase walking for adults with intellectual disabilities.
The purpose of this study was to evaluate the effectiveness of token reinforcement, using an ABAB reversal design, for increasing distance walked for adults with mild to moderate intellectual disabilities at an adult day-training center. Five participants earned tokens for walking 50-m laps and exchanged tokens for back-up reinforcers that had been identified through preference assessments. Token reinforcement resulted in a substantial increase from baseline in laps walked for 4 participants.


Underdiagnosed CKD in Geriatric Trauma Patients and Potent Prevention of Renal Impairment from Polypharmacy Risks through Individual Pharmacotherapy Management (IPM-III).
The aging global patient population with multimorbidity and concomitant polypharmacy is at increased risk for acute and chronic kidney disease, particularly with severe additional disease states or invasive surgical procedures. Because from the expertise of more than 58,600 self-reviewed medications, adverse drug reactions, drug interactions, inadequate dosing, and contraindications all proved to cause or exacerbate the worsening of renal function, we analyzed the association of an electronic patient record- and Summaries of Product Characteristics (SmPCs)-based comprehensive individual pharmacotherapy management (IPM) in the setting of 14 daily interdisciplinary patient visits with the outcome: further renal impairment with reduction of eGFR ≥ 20 mL/min (redGFR) in hospitalized trauma patients ≥ 70 years of age. The retrospective clinical study of 404 trauma patients comparing the historical control group (CG) before IPM with the IPM intervention group (IG) revealed a group-match in terms of potential confounders such as age, sex, BMI, arterial hypertension, diabetes mellitus, and injury patterns. Preexisting chronic kidney disease (CKD) > stage 2 diagnosed as eGFR < 60 mL/min/1.73 m2 on hospital admission was 42% in the CG versus 50% in the IG, although in each group only less than 50% of this was coded as an ICD diagnosis in the patients' discharge letters (19% in CG and 21% in IG). IPM revealed an absolute risk reduction in redGFR of 5.5% (11 of 199 CG patients) to 0% in the IPM visit IG, a relative risk reduction of 100%, NNT 18, indicating high efficacy of IPM and benefit in improving outcomes. There even remained an additive superimposed significant association that included patients in the IPM group before/beyond the 14 daily IPM interventions, with a relative redGFR risk reduction of 0.55 (55%) to 2.5% (5 of 204 patients), OR 0.48 [95% CI 0.438-0.538] (p < 0.001). Bacteriuria, loop diuretics, allopurinol, eGFR ≥ 60 mL/min/1.73 m2, eGFR < 60 mL/min/1.73 m2, and CKD 3b were significantly associated with redGFR; of the latter, 10.5% developed redGFR. Further multivariable regression analysis adjusting for these and established risk factors revealed an additive, superimposed IPM effect on redGFR with an OR 0.238 [95% CI 0.06-0.91], relative risk reduction of 76.2%, regression coefficient -1.437 including patients not yet visited in the IPM period. As consequences of the IPM procedure, the IG differed from the CG by a significant reduction of NSAIDs (p < 0.001), HCT (p = 0.028) and Würzburger pain drip (p < 0.001), and significantly increased prescription rate of antibiotics (p = 0.004). In conclusion, (1) more than 50% of CKD in geriatric patients was not pre-recognized and underdiagnosed, and (2) the electronic patient records-based IPM interdisciplinary networking strategy was associated with effective prevention of further periinterventional renal impairment and requires obligatory implementation in all elderly patients to urgently improve patient and drug safety.


DIAPH1 mediates progression of atherosclerosis and regulates hepatic lipid metabolism in mice.
Atherosclerosis evolves through dysregulated lipid metabolism interwoven with exaggerated inflammation. Previous work implicating the receptor for advanced glycation end products (RAGE) in atherosclerosis prompted us to explore if Diaphanous 1 (DIAPH1), which binds to the RAGE cytoplasmic domain and is important for RAGE signaling, contributes to these processes. We intercrossed atherosclerosis-prone Ldlr-/- mice with mice devoid of Diaph1 and fed them Western diet for 16 weeks. Compared to male Ldlr-/- mice, male Ldlr-/- Diaph1-/- mice displayed significantly less atherosclerosis, in parallel with lower plasma concentrations of cholesterol and triglycerides. Female Ldlr-/- Diaph1-/- mice displayed significantly less atherosclerosis compared to Ldlr-/- mice and demonstrated lower plasma concentrations of cholesterol, but not plasma triglycerides. Deletion of Diaph1 attenuated expression of genes regulating hepatic lipid metabolism, Acaca, Acacb, Gpat2, Lpin1, Lpin2 and Fasn, without effect on mRNA expression of upstream transcription factors Srebf1, Srebf2 or Mxlipl in male mice. We traced DIAPH1-dependent mechanisms to nuclear translocation of SREBP1 in a manner independent of carbohydrate- or insulin-regulated cues but, at least in part, through the actin cytoskeleton. This work unveils new regulators of atherosclerosis and lipid metabolism through DIAPH1.


Circadian Influences of Diet on the Microbiome and Immunity.
Host circadian rhythmicity and the timing of feeding are increasingly recognized to cross-regulate and entrain each other, and may play crucial roles in regulating multiple physiological functions including host immunity and metabolic health. Of relevance, these circadian diet-immune interactions may be modulated by the gut microbiota. We review current knowledge linking the circadian clock and dietary timing to host immune-microbiota interactions, exemplifying how this axis may impact on host immunity in health and in a variety of immune-mediated diseases. We also discuss current challenges in reaching mechanistic insights regarding the functions of the diurnally shifting diet-microbiome-host immune axis. We highlight the possible implications of circadian reprogramming by dietary timing patterns as a future intervention to modulate a variety of immune-related diseases.


Lung Surfactant Protein B Peptide Mimics Interact with the Human ACE2 Receptor.
Lung surfactant is a complex mixture of phospholipids and surfactant proteins that is produced in alveolar type 2 cells. It prevents lung collapse by reducing surface tension and is involved in innate immunity. Exogenous animal-derived and, more recently, synthetic lung surfactant has shown clinical efficacy in surfactant-deficient premature infants and in critically ill patients with acute respiratory distress syndrome (ARDS), such as those with severe COVID-19 disease. COVID-19 pneumonia is initiated by the binding of the viral receptor-binding domain (RBD) of SARS-CoV-2 to the cellular receptor angiotensin-converting enzyme 2 (ACE2). Inflammation and tissue damage then lead to loss and dysfunction of surface activity that can be relieved by treatment with an exogenous lung surfactant. Surfactant protein B (SP-B) is pivotal for surfactant activity and has anti-inflammatory effects. Here, we study the binding of two synthetic SP-B peptide mimics, Super Mini-B (SMB) and B-YL, to a recombinant human ACE2 receptor protein construct using molecular docking and surface plasmon resonance (SPR) to evaluate their potential as antiviral drugs. The SPR measurements confirmed that both the SMB and B-YL peptides bind to the rhACE2 receptor with affinities like that of the viral RBD-ACE2 complex. These findings suggest that synthetic lung surfactant peptide mimics can act as competitive inhibitors of the binding of viral RBD to the ACE2 receptor.


Pantothenic acid transport and metabolism in the central nervous system.
The mechanisms by which pantothenic acid (PA) enters and leaves brain, choroid plexus, and cerebrospinal fluid (CSF) were investigated by injecting [3H]PA either intravenously or intraventricularly into adult rabbits. [3H]PA, either alone or together with unlabeled PA, was infused at a constant rate into conscious rabbits. At 180 min, [3H]PA readily entered CSF, choroid plexus, and brain. In brain, CSF, and plasma, greater than 90% of the 3H was associated with [3H]PA. The addition of 200 mumol/kg PA to the infusion syringe decreased the penetration of [3H]PA into brain and CSF by approximately 70%. Two hours after the intraventricular injection of [3H]PA, [3H]PA was rapidly cleared from the CSF by a probenecid-sensitive mechanism. No metabolism of the [3H]PA occurred in brain. However, 18 h after the intraventricular injection of 37 microCi (34 nmol) of [3H]PA, approximately 40% of the 3H remaining in forebrain was converted to [3H]CoA. These results show that PA enters and leaves CSF and brain by saturable transport systems. However, [3H]PA is very slowly converted to [3H]CoA in brain in vivo.


Recommendations for management of diabetes during Ramadan: update 2020, applying the principles of the ADA/EASD consensus.
Fasting the Holy month of Ramadan constitutes one of the five pillars of the Muslim faith. Although there is some evidence that intermittent fasting during Ramadan may be of benefit in losing weight and cardiometabolic risk factors, there is no strong evidence these benefits apply to people with diabetes. The American Diabetes Association/European Association for the Study of Diabetes consensus recommendations emphasize the importance of patient factors and comorbidities when choosing diabetes medications including the presence of comorbidities, atherosclerotic cardiovascular disease, heart failure, chronic kidney disease, hypoglycemia risk, weight issues and costs. Structured education and pre-Ramadan counseing are key components to successful management of patients with diabetes. These should cover important aspects like glycemic targets, self-monitoring of blood glucose, diet, physical activity including Taraweeh prayers, medication and dose adjustment, side effects and when to break the fast. The decision cycle adapted for the specific situation of Ramadan provides an aid for such an assessment. Children with type 1 diabetes should strongly be advised not to fast due to the high risk of acute complications such as hypoglycemia and probably diabetic ketoacidosis (DKA), although there is very little evidence that DKA is increased in Ramadan. Pregnant women with diabetes or gestational diabetes should be advised to avoid fasting because of possible negative maternal and fetal outcomes. Hypoglycemia is a common concern during Ramadan fasting. To prevent hypoglycemic and hyperglycemic events, we recommend the adoption of diabetes self-management education and support principles. The use of the emerging technology and continuous glucose monitoring during Ramadan could help to recognize hypoglycemic and hyperglycemic complications related to omission and/or medication adjustment during fasting; however, the cost represents a significant barrier.


The Role of Non-genetic Therapies to Reduce the Incidence of Sickle Cell Crisis: A Systematic Review.
Sickle cell anemia is a hemoglobinopathy that causes complications such as Vaso-Occlusive Crisis (VOC), stroke, priapism, Acute Chest Syndromes (ACS), and bone infarcts due to blood vessel occlusion, resulting in hypoxia, ischemia, and inflammation. Preventing these incidents improves the quality of life and lowers mortality rates in Sickle Cell Disease (SCD) patients. This systematic review aims to describe the drugs, their mechanisms of action, dosages, changes in hemoglobin parameters, decrease in VOCs, delay the time for the next VOC, decrease in the length of hospital stay, and side effects associated with these drugs. This review adheres to the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) 2020 guidelines. For this review, we searched the PubMed, Google Scholar, and Cochrane databases and screened them for full free texts published in English and studied in humans in the last five years beginning in 2018. Randomized clinical trials (RCT), observational studies, meta-analyses, systemic reviews, and traditional reviews were all included in the search. According to the type of study, quality assessment tools are used, and eight papers are chosen. Full-text articles from these papers are studied, analyzed, and tabulated. We discussed seven interventions that are used to treat sickle cell disease. Voxelotor, crizanlizumab, L-glutamate, long-term blood transfusions, Zinc (Zn), Niprisan®, and Ciklavit* were found to reduce the number and severity of VOC. We discovered that VOCs containing L -glutamate reduced the length of hospitalization. Magnesium (Mg) did not affect the number and severity of VOCs. This review includes a few articles for the study. Future papers on this subject should include a large sample size and many papers. More clinical trials are required to evaluate the dosages and outcomes of using these drugs in combination to prevent VOCs.


A case of robotic assisted percutaneous coronary intervention of the left main coronary artery in a patient with very late baffle stenosis after surgical correction of anomalous left coronary artery from the pulmonary artery.
A 34-year-old woman with history of surgical correction (Takeuchi procedure) of anomalous left coronary artery from the pulmonary artery (ALCAPA) presented with reduced left ventricular ejection fraction of 48% and severe ischemia quantified as 21% by stress Positron Emission Tomography (PET) scan. A coronary angiogram revealed ostial 90% stenosis of the left main coronary artery (LMCA). A guidewire (Sion Blue, Asahi Intecc USA, Inc., Santa Ana, CA) was navigated robotically and after pre-dilation with 3.5 × 15 mm cutting balloon, the lesion length was measured by marking the distal end of the lesion with the balloon marker and withdrawing back robotically to the ostium of the LMCA. A 3.5 × 16 mm drug-eluting stent was deployed robotically after intravascular ultrasound (IVUS) with good results. The main advantage of robotic percutaneous coronary intervention includes the precise measurement and positioning of the stent. Since the guide catheter and balloon can be adjusted without guide catheter and device interaction, precise placement of stent is possible by advancing the device distal to the lesion, positioning the guide catheter just proximal to the proximal edge of the stent and pulling the guidecatheter and device back as a unit. Final IVUS after post-dilation with 4.0 noncompliant and 5.0 compliant balloon revealed precise placement at the ostium and full stent expansion.


Therapeutic mechanism of Yīn-Chén-Hāo decoction in hepatic diseases.
Yīn-Chén-Hāo decoction (YCHD) is a traditional Chinese medicine formula composed of capillaris (Artemisia capillaris), gardenia (Gardenia jasminoides), and rhubarb (Rheum rhabarbarum) that is used for the treatment of damp-heat jaundice. In modern clinics, YCHD is mostly used for hepatic diseases. This review summarizes the biological activities of YCHD and its medical applications. The main active compounds of YCHD are chlorogenic acid, rhein, geniposide, emodin, and scoparone. The pharmacological actions of YCHD include inhibition of hepatic steatosis, apoptosis, necrosis, anti-inflammation, and immune regulation. YCHD could be developed as a new therapeutic strategy for the treatment of hepatic diseases.


Optimization of extreme ultra-violet light emitted from the CO2 laser-irradiated tin plasmas using 2D radiation hydrodynamic simulations.
We studied Extreme Ultra-Violet (EUV) emission characteristics of the 13.5 nm wavelength from CO2 laser-irradiated pre-formed tin plasmas using 2D radiation hydrodynamic simulations. Our results indicate that when a CO2 laser irradiates pre-formed tin plasma, the heated plasma expands towards the surrounding plasma, steepening the density at the ablation front and lowering the density near the laser axis due to the transverse motion of the plasma. Consequently, the laser absorption fraction decreases, and the contribution to EUV output from the ablation front becomes dominant over that from the low-density plasmas. We estimated that an EUV conversion efficiency of 10% from laser to EUV emission could be achieved with a larger laser spot size, shortened laser pulse width, and longer pre-formed plasma density scale length. Our results offer one optimizing solution to achieve an efficient and powerful EUV light source for the next-generation semiconductors.


The atopen: a rehabilitation.
Consideration of the physicochemical and biological properties of the common inhalant, food and nematode allergens has led the author to propose reintroduction of the term atopen. Atopens are nonspecific activators of physiological enzyme systems in man and are indicators of the atopic status. Allergens are considered to be those substances, not necessarily immunogenic by themselves, capable of deliberately directing the immune response into IgE production. Ongoing IgE synthesis may eventually produce IgE antibodies reacting with nonallergenic antigens.


Histologic differentiation of desmoplastic melanoma from cicatrices.
Desmoplastic malignant melanoma (DMM) is a rare variant of melanoma that can be very difficult to diagnose correctly both clinically and histologically. The problem is compounded by the fact that many lesions persist at previous biopsy or excision sites so that scar tissue is often present admixed with or adjacent to the spindle cell neoplasm which may exhibit fibroblastic differentiation itself. In order to assess this problem, we compared and contrasted the histologic features of six DMM with 15 examples of cicatrices from various sources. Mature scars were readily differentiated from DMM by light microscopy. In contrast, immature scar and DMM had many features in common including hypercellularity, nodular lymphoid infiltrates, myxoid stroma, and atypical nuclei. The presence of a melanocytic proliferation within the epidermis above the dermal component, neurotropism, and S-100 and/or HMB-45 positivity of neoplastic cells were the only features that permitted reliable differentiation between the two. Clinical correlation and review of previous biopsy specimens are crucial in preventing a delayed diagnosis of DMM. Re-excision is advised in all questionable cases.


Modeling Solvation in Supercritical CO2.
In recent decades, a microscopic understanding of solute-solvent intermolecular interactions has been key to advances in technologies based on supercritical carbon dioxide. In many cases, computational work has provided the impetus for new discoveries, shedding new light on important concepts such as the local structure around the solute in the supercritical medium, the influence of the peculiar properties of the latter on the molecular behavior of dissolved substances and, importantly, CO2 -philicity. In this Review, the theoretical work that has been relevant to these developments is surveyed and, by presenting some crucial open questions, the possible routes to achieving further progress based on the interplay between theory and experiments is discussed.


MAMP and DAMP signaling contributes resistance to Fusarium graminearum in Arabidopsis.
Plants perceive externally produced microbe-associated molecular patterns (MAMPs) and endogenously produced danger-associated molecular patterns (DAMPs) to activate inducible immunity. While several inducible immune responses have been observed during Fusarium graminearum infection, the identity of the signaling pathways involved is only partly known. We screened 227 receptor kinase and innate immune response genes in Arabidopsis to identify nine genes with a role in F. graminearum resistance. Resistance-promoting genes included the chitin receptors LYK5 and CERK1, and the reactive oxygen species (ROS)-producing NADPH oxidase RbohF, which were required for full inducible immune responses during infection. Two of the genes identified in our screen, APEX and the PAMP-induced peptide 1 (PIP1) DAMP receptor RLK7, repressed F. graminearum resistance. Both RbohF and RLK7 were required for full chitin-induced immune responses and PIP1 precursor expression was induced by chitin and F. graminearum infection. Together, this indicates that F. graminearum resistance is mediated by MAMP and DAMP signaling pathways and that chitin-induced signaling is enhanced by PIP1 perception and ROS production.


Controlled release of pentoxifylline from porous chitosan-pectin scaffolds.
Measures to suppress inflammatory reactions are taken to prevent fibrous encapsulation of implants. It is proposed in this study that tissue engineered scaffolds that can slowly release anti-inflammatory drugs can help reduce inflammatory reactions around implants. Chitosan and chitosan cross-linked with different concentrations of pectin were made into films and porous scaffolds. Results seen from Fourier-transform infrared spectra and thermal gravimetric analysis showed that polyelectrolyte complexation took place between chitosan and pectin units. As the amounts of pectin added to chitosan increased (0%, 0.5%, 1%, and 2%) the scaffolds became more wettable (contact angle decreased from 81 degrees to 76 degrees ), less swellable (swelling ratio decreased from 35% to 30%), and less capable of releasing pentoxifylline (PTX) (release efficacies decreased from 93% to 83%). Higher degrees of pectin cross-linking made the scaffolds more resistant to compression (Young's modulus increased from 2.4 kPa to 3.7 kPa) and more favorable for initial cell attachment (percentage of attached cells increased from 55% to 67%). In vitro tests showed that, with the reduction of PTX release rates, PTX became more effective in inhibiting TNF-alpha and IL-6 production from activated macrophages. This investigation has demonstrated that the changes in the basic drug release properties of chitosan scaffolds were proportional to the amount of pectin added. The changes could help improve the effectiveness of PTX.


Active integral imaging system based on multiple structured light method.
In this paper, we simplify the equipments for integral imaging (II) pickup and implement an active II system based on multiple structured light (MSL) method. In the active II system, the complete three-dimensional (3D) shape of the 3D scene can be reconstructed, and the tunable parallaxes can be generated without occlusions. Therefore, the high-quality 3D images can be displayed efficiently by the II. We also use the Compute Unified Device Architecture implementing the processing algorithms in graphics processing unit. The experimental results demonstrate the effectiveness of the MSL method for the II pickup and the acceleration for the elemental image array generation. Especially, the proposed method is suitable for the real scene with high precision.


Effect of manual lymphdrainage massage on blood components and urinary neurohormones in chronic lymphedema.
In an earlier paper we have shown that manual lymph drainage massage of edematous limbs can result in the excretion of up to 1 liter urine derived from reabsorption and transport from the interstitial fluid, simultaneously with significant changes in the excretion of urinary neurohormones. These findings indicated that histamine and serotonin were released from the edematous tissue and that circulation improved through increased output of adrenaline and noradrenaline. The results achieved led us to assume that similar changes may have occurred in the blood during treatment, and induced us to study the effect of manual lymphdrainage on various blood constituents and urinary neurohormones.


Osteoclastogenesis and osteoimmunology.
Bone is continuously renewed through a dynamic balance between bone resorption and formation and is the fundamental basis for maintaining normal bone mass and architecture by osteoclasts (bone resorption) and osteoblasts (bone formation). Bone resorption is an elementary cellular activity in the modeling of the skeleton during growth and development. Later in life, bone remodeling occurs, and is locally initiated by resorption. During remodeling, bone resorption is coupled to new bone formation, that ensures bone renewal with only minor and temporary local bone loss. Osteoclasts play a crucial role in both physiological and pathological bone resorption, and receptor activator of nuclear factor-κB ligand is the key cytokine that induces osteoclastogenesis. At the same time, various factors produced during immune responses are capable of profoundly affecting bone regulation; bone and immune systems share an abundance of molecules and regulatory mechanisms. We summarize the new insights on the link between osteoclastogenesis and osteoimmunology.


Effect of geometric parameters in the design of customized hip implants.
In this paper, the effects of geometric parameters on the development of stress in custom cementless hip implant have been studied. Taguchi methods and analysis of variance (ANOVA) were used to find out which geometric parameters are statistically significant for the maximum von Mises stresses in the hip implant. The multiple regression analysis was also performed, to replace the FE solver and accurately and quickly predict the stress values for new patients. Three parameters horizontal offset (HO), vertical offset and neck shaft angle (NSA), with three levels have been selected. Based on L9 orthogonal array, nine hip implant shapes with three different geometric parameters were modelled. Stress analysis was carried out by using finite element analysis. ANOVA results showed that HO has the maximum impact on von Mises stresses followed by the NSA. It is shown that as the HO increases, the stress at the given region increases, however, as the neck shaft (NSA) increases the stress at the given region decreases. Good agreement is found between von Mises stresses, calculated from multiple regression analysis and obtained from FE analysis. To redesign customised hip implant for the minimum stresses, the results of this work will be important.


Thermal characteristics of neutralization therapy and water dilution for strong acid ingestion: an in-vivo canine model.
To determine whether in-vivo neutralization therapy for acid ingestions will superimpose a thermal injury upon tissue already damaged by acid. An in-vivo canine model was used with repeated measures of tissue and luminal temperatures. All dogs were placed under halothane general anesthesia. The stomach was exteriorized and temperature probes were placed in the lumen and mucosa. 25 mL of 0.5 N HCl (25 degrees C) was placed in the gastric lumen followed 5 minutes later by 75 mL of either 8% NaHCO3 neutralization (25 degrees C, n = 10) or water dilution (25 degrees C, n = 10). Temperature measurements were recorded at specified intervals for 5 minutes post HCl acid exposure and for 30 minutes post treatment. Temperature profiles were analyzed by repeated-measures ANOVA. Post-treatment changes were evaluated using signed-rank tests. In both treatment groups, treatment resulted in significant decreases in initial mucosa and intraluminal temperatures. Both the mucosa and intraluminal temperatures decreased immediately after treatment with HCO3 by an average of 1.6 degrees C (p = 0.05). In the water dilution treatment group, both temperatures decreased by 1.1 degrees C (p = 0.05). Ensuing post-treatment temperatures increased but did not reach baseline temperatures at any time up to 30 minutes post treatment. In the in-vivo setting, there is no evidence of hazardous temperature elevation when a weak alkali or dilution therapy is used to neutralize strong acid-induced injury. Contraindication of this form of emergency treatment should not be based on the preconceived idea that a resultant exothermic reaction will cause a superimposed thermal injury. Further clinical study is needed to determine the clinical utility of this emergency therapeutic modality.


Religious schema and values.
Past research has consistently shown marked differences between religious believers and nonbelievers. Such differences were suggested in this study to be due to the presence of religious schema in believers. It was hypothesized that non-believers would be less schematic, or aschematic with respect to religion. This hypothesis was examined by comparing the value system of college student believers (Protestants and Catholics) and non-believers (those indicating no religious faith). Extensive differences were found. Consistent with the hypothesis, the differences were found mainly on the schema-relevant values. In essence, believers showed greater preference for and possession of the moral and relational values, and lesser on the personal-extrinsic, competency, and egoistic values. As predicted, no difference was found on the social and intellectual types of values which are basically schema-irrelevant. The notion of schema availability was also supported. When subjects' religiosity (as indicated by their preference for the salvation value and their self-rated possession of the religiousness value) was controlled for, all the above differences were removed. When high and low religiosity believers were compared, great differences were found. The differences between the high and low religiosity non-believers were however much less extensive.


Effects of Light Quality on Morphology, Enzyme Activities, and Bioactive Compound Contents in Anoectochilus roxburghii.
The aim of this study was to investigate the effects of light quality on the morphological traits, leaf anatomical characteristics, antioxidant enzyme (superoxide dismutase, catalase, and peroxidase) activities, photosynthetic pigments content, and bioactive compounds (phenols, flavonoids, and polysaccharides) content in Anoectochilus roxburghii. Plants of A. roxburghii were grown under light filtered through four differently colored films for 8 months. The four treatments were red film (RF), blue film (BF), yellow film (YF), and colorless plastic film (control, CK). Compared with the A. roxburghii plants in CK, those in the BF treatment showed significantly greater stem diameter, fresh weight, leaf area, stomatal frequency, chlorophyll content (Chl a, Chl b, Chl a+b), antioxidant enzyme activities, and active compound (polysaccharides, flavones) content. The plants in the RF treatment showed the greatest plant height and phenolics contents. These results show that growing A. roxburghii plants under blue film is a useful technique to improve quality. This technique is conducive to achieving large-scale sustainable production of high-quality plant materials.


Novel bionic inspired nanosystem construction for precise delivery of mRNA.
The intracellular delivery of messenger (m)RNA holds great potential for the discovery and development of vaccines and therapeutics. Yet, in many applications, a major obstacle to clinical translation of mRNA therapy is the lack of efficient strategy to precisely deliver RNA sequence to liver tissues and cells. In this study, we synthesized virus-like mesoporous silica (V-SiO2) nanoparticles for effectively deliver the therapeutic RNA. Then, the cationic polymer polyethylenimine (PEI) was included for the further silica surface modification (V-SiO2-P). Negatively charged mRNA motifs were successfully linked on the surface of V-SiO2 through electrostatic interactions with PEI (m@V-SiO2-P). Finally, the supported lipid bilayer (LB) was completely wrapped on the bionic inspired surface of the nanoparticles (m@V-SiO2-P/LB). Importantly, we found that, compared with traditional liposomes with mRNA loading (m@LNPs), the V-SiO2-P/LB bionic-like morphology effectively enhanced mRNA delivery effect to hepatocytes both in vitro and in vivo, and PEI modification concurrently promoted mRNA binding and intracellular lysosomal escape. Furthermore, m@V-SiO2-P increased the blood circulation time (t1/2 = 7 h) to be much longer than that of the m@LNPs (4.2 h). Understanding intracellular delivery mediated by the V-SiO2-P/LB nanosystem will inspire the next-generation of highly efficient and effective mRNA therapies. In addition, the nanosystem can also be applied to the oral cavity, forehead, face and other orthotopic injections.


Science to Practice: What Light through Yonder Tumor Breaks: Noninvasive Staging of Cancer--Shining a Light on Disease.
Photoacoustic imaging imparts the ability to distinguish materials according to their differences in optical absorption (ie, their color) with the high spatial and temporal resolution of ultrasonography (US). Experiments in rats show the advantages this approach would have in the clinically important noninvasive determination of the presence of tumor in the lymph nodes draining a tumor, providing the ability to achieve the results of total axillary lymph node dissection without the risks of surgery or even percutaneous sentinel node biopsy. The coupling of this technique to a conventional US imaging system gives the promise of rapid translation to clinical use.


Growth factor delivery approaches in hydrogels.
The controlled delivery of growth factors is a very challenging task because many different issues have to be addressed to develop the best suited system. A wide range of approaches have been employed for the controlled delivery of growth factors by hydrogels. Direct loading, electrostatic interaction, covalent binding, and the use of carriers are the main strategies presented in the literature. They are all detailed in the first part of this review. Recent work emphasizing biologically inspired strategies is also included. Also, both natural and synthetic materials are discussed. The second part comprises the methods to evaluate such delivery approaches. Both in vivo and in vitro techniques are presented. Improvements based on the discussed approaches may illustrate future paths toward the development of an ideal growth factor delivery system.


A contrast-enhanced-CT-based classification tree model for classifying malignancy of solid lung tumors in a Chinese clinical population.
To develop and validate a contrast-enhanced CT based classification tree model for classifying solid lung tumors in clinical patients into malignant or benign. Between January 2015 and October 2017, 827 pathologically confirmed solid lung tumors (487 malignant, 340 benign; median size, 27.0 mm, IQR 18.0-39.0 mm) from 827 patients from a dedicated Chinese cancer hospital were identified. Nodules were divided randomly into two groups, a training group (575 cases) and a testing group (252 cases). CT characteristics were collected by two radiologists, and analyzed using a classification and regression tree (CART) model. For validation, we used the decision analysis threshold to evaluate the classification performance of the CART model and radiologist's diagnosis (benign; malignant) in the testing group. Three out of 19 characteristics [margin (smooth; slightly lobulated/lobulated/spiculated), and shape (round/oval; irregular), subjective enhancement (no/uniform enhancement; heterogeneous enhancement)] were automatically generated by the CART model for classifying solid lung tumors. The sensitivity, specificity, PPV, NPV, and diagnostic accuracy of the CART model is 98.5%, 58.1%, 80.6%, 98.6%, 79.8%, and 90.4%, 54.7%, 82.4% 98.5%, 74.2% for the radiologist's diagnosis by using three-threshold decision analysis. Tumor margin and shape, and subjective tumor enhancement were the most important CT characteristics in the CART model for classifying solid lung tumors as malignant. The CART model had higher discriminatory power than radiologist's diagnosis. The CART model could help radiologists making recommendations regarding follow-up or surgery in clinical patients with a solid lung tumor.


Hemodynamic findings in patients with brain stroke.
Standard procedures carried out at a stroke department in patients after a cerebral event may prove insufficient for monitoring hemodynamic indices. Impedance cardiography enables hemodynamic changes to be monitored non-invasively. The aim of the work was to describe hemodynamic parameters in patients with acute phase of ischemic and hemorrhagic stroke and to analyse the correlation between the type of hemodynamic response and long-term prognosis. The 45 consecutive subjects with ischemic stroke and 16 with a hemorrhagic stroke were examined additionally with impedance cardiography during the first day of hospitalization. The heart contractility, pump performance, afterload and preload indices were recorded and calculated automatically and the data analyzed in terms of 6-month mortality. We found a significant association between the systemic vascular resistance index, Heather index, stroke index, heart rate, systolic and diastolic and mean arterial blood pressure and mortality in patients with ischemic stroke (p = 0.002, p = 0.008, p = 0.012, p = 0.005, p = 0.007, p = 0.009, p = 0.002 respectively). Logistic regression analysis identified the thoracic fluid content as the most significant variable correlating with the non-survival of the patients with ischemic stroke and in the whole group (ischemic and hemorrhagic stroke). The significant parameters were also mean arterial pressure and stroke index in ischemic stroke (the correct answer ratio was 86.67%) and heart rate in the whole group (the correct answer ratio was 80.33%). There were no significant associations in hemorrhagic stroke. The hemodynamic parameters correlate with long term prognosis in patients with ischemic brain stroke.


An investigation into the validity of specifying 5 "O' levels in the General Certificate of Education as an entry requirement for the education and training of district nurses.
District nursing, like many other professionalizing occupations, has stipulated that possession of five ordinary level GCE passes might constitute one criterion of entry. This paper reports a longitudinal study over four intakes of district nurse students in one teaching centre and examines the relationship between selection criteria, in particular educational achievement prior to entry to the course, and the students' achievement during this. The research explored the relationship between educational achievement prior to commencing their course of study and the level of academic and practical performance during it. Additionally other factors, including the results of aptitude and personality tests, were examined to see if they would provide indication of the level of the students; grades during training. No factors were discovered that might provide an adequate basis for selection, which raises questions about trying to specify any pre-requisite for selection other than state registration and appropriate professional experience until further research has been undertaken.


Syncope in association with Prinzmetal variant angina.
A case of Prinzmetal variant angina with transient complete atrioventricular block and syncopal episodes following an anteroseptal myocardial infarction is described. The syncopal attacks were not prevented by demand cardiac pacing and were presumably caused by transient severe ischaemia of the left ventricle, with a consequent reduction in cardiac output. The left ventriculogram showed a large anterior dyskinetic area corresponding to the high grade proximal obstruction in the left anterior descending artery demonstrated by coronary angiography. All other coronary vessels appeared free of disease and it is suggested that the anginal episodes were caused by transient proximal segmental spasm of the right coronary artery. The anginal episodes were successfully prevented by a regimen of two-hourly coronary arterial vasodilator therapy.


Parietal Cortex Integrates Saccade and Object Orientation Signals to Update Grasp Plans.
Coordinated reach-to-grasp movements are often accompanied by rapid eye movements (saccades) that displace the desired object image relative to the retina. Parietal cortex compensates for this by updating reach goals relative to current gaze direction, but its role in the integration of oculomotor and visual orientation signals for updating grasp plans is unknown. Based on a recent perceptual experiment, we hypothesized that inferior parietal cortex (specifically supramarginal gyrus [SMG]) integrates saccade and visual signals to update grasp plans in additional intraparietal/superior parietal regions. To test this hypothesis in humans (7 females, 6 males), we used a functional magnetic resonance paradigm, where saccades sometimes interrupted grasp preparation toward a briefly presented object that later reappeared (with the same/different orientation) just before movement. Right SMG and several parietal grasp regions, namely, left anterior intraparietal sulcus and bilateral superior parietal lobule, met our criteria for transsaccadic orientation integration: they showed task-dependent saccade modulations and, during grasp execution, they were specifically sensitive to changes in object orientation that followed saccades. Finally, SMG showed enhanced functional connectivity with both prefrontal saccade regions (consistent with oculomotor input) and anterior intraparietal sulcus/superior parietal lobule (consistent with sensorimotor output). These results support the general role of parietal cortex for the integration of visuospatial perturbations, and provide specific cortical modules for the integration of oculomotor and visual signals for grasp updating.SIGNIFICANCE STATEMENT How does the brain simultaneously compensate for both external and internally driven changes in visual input? For example, how do we grasp an unstable object while eye movements are simultaneously changing its retinal location? Here, we used fMRI to identify a group of inferior parietal (supramarginal gyrus) and superior parietal (intraparietal and superior parietal) regions that show saccade-specific modulations during unexpected changes in object/grasp orientation, and functional connectivity with frontal cortex saccade centers. This provides a network, complementary to the reach goal updater, that integrates visuospatial updating into grasp plans, and may help to explain some of the more complex symptoms associated with parietal damage, such as constructional ataxia.


External scaffold of spherical immature poxvirus particles is made of protein trimers, forming a honeycomb lattice.
During morphogenesis, poxviruses undergo a remarkable transition from spherical immature forms to brick-shaped infectious particles lacking helical or icosahedral symmetry. In this study, we show that the transitory honeycomb lattice coating the lipoprotein membrane of immature vaccinia virus particles is formed from trimers of a 62-kD protein encoded by the viral D13L gene. Deep-etch electron microscopy demonstrated that anti-D13 antibodies bound to the external protein coat and that lattice fragments were in affinity-purified D13 preparations. Soluble D13 appeared mostly trimeric by gel electrophoresis and ultracentrifugation, which is consistent with structural requirements for a honeycomb. In the presence or absence of other virion proteins, a mutated D13 with one amino acid substitution formed stacks of membrane-unassociated flat sheets that closely resembled the curved honeycombs of immature virions except for the absence of pentagonal facets. A homologous domain that is present in D13 and capsid proteins of certain other lipid-containing viruses support the idea that the developmental stages of poxviruses reflect their evolution from an icosahedral ancestor.


Use of fluorescent protein to analyse recombination at three loci in Neurospora crassa.
We have inserted a histone H1-GFP fusion gene adjacent to three loci on different chromosomes of Neurospora crassa and made mating pairs in which a wild type version of GFP is crossed to one with a mutation in the 5' end of GFP. The loci are his-3, am and his-5, chosen because recombination mechanisms appear to differ between his-3 and am, and because crossing over adjacent to his-5, like his-3, is regulated by rec-2. At his-3, the frequencies of crossing over between GFP and the centromere and of conversion of 5'GFP to GFP(+) are comparable to those obtained by classical recombination assays, as is the effect of rec-2 on these frequencies, suggesting that our system does not alter the process of recombination. At each locus we have obtained sufficient data, on both gene conversion and crossing over, to be able to assess the effect of deletion of any gene involved in recombination. In addition, crosses between a GFP(+) strain and one with normal sequence at all three loci have been used to measure the interval to the centromere and to show that GFP experiences gene conversion with this system. Since any gene expressed in meiosis is silenced in Neurospora if hemizygous, any of our GFP(+) strains can be used as a quick screen to determine if a gene deleted by the Neurospora Genome Project is involved in crossing over or gene conversion.


Prediction of worsening heart failure in hypertrophic cardiomyopathy using plasma proteomics.
Heart failure (HF) is one of the most common and lifestyle-limiting complications of hypertrophic cardiomyopathy (HCM). Prediction of worsening HF using clinical measures alone remains limited. Moreover, the mechanisms by which patients with HCM develop worsening HF have not been elucidated. Therefore, the aim of this study was to develop a plasma proteomics-based model to predict worsening HF among patients with HCM and to identify signalling pathways that are differentially regulated in those who subsequently develop worsening HF. In this multi-centre, prospective cohort study of 389 patients with HCM, plasma proteomics profiling of 4986 proteins was performed at enrolment. A proteomics-based random forest model was developed to predict worsening HF using data from one institution (training set, n=268). This model was externally validated in patients from a different institution (test set, n=121). Pathway analysis of proteins significantly dysregulated in patients who subsequently developed worsening HF compared with those who did not was executed, using a false discovery rate (FDR) threshold of <0.001. Using the 11-protein proteomics-based model derived from the training set, the area under the receiver-operating characteristic curve to predict worsening HF was 0.87 (95% CI: 0.76 to 0.98) in the test set. Pathway analysis revealed that the Ras-MAPK pathway (FDR<0.00001) and related pathways were dysregulated in patients who subsequently developed worsening HF. The present study with comprehensive plasma proteomics profiling demonstrated a high accuracy to predict worsening HF in patients with HCM and identified the Ras-MAPK and related signalling pathways as potential underlying mechanisms.


[Local injection of alpha, beta-methylene ATP induces excitation of primary afferent fibers in rats].
To observe the excitation and sensitization of dorsal cutaneous primary afferent fibers induced by P2X agonist alpha, beta-methylene ATP (alphabetame-ATP) in rats. By means of single fiber electrophysiological recordings on the nerve filaments isolated from the dorsal cutaneous branches of the T(9)-T(13) spinal nerves, the effects of alphabetame-ATP (100 micromol/L, 10 microl) injection into the cutaneous receptive field on the mechanical threshold and spontaneous discharge of rat primary sensory afferent units were observed. The mean mechanical threshold of Adelta and C fibers prior to alphabetame-ATP injection were 0.384+/-0.018 and 0.943+/-0.102 mN, and lowered to 0.304+/-0.013 and 0.659+/-0.071 mN after the injection, respectively (P<0.05, P<0.01). The mechanical threshold of the Abeta fibers before alphabetame-ATP injection was 0.301+/-0.019 mN, and slightly lowered to 0.288+/-0.018 mN after the injection (P>0.05). Injection of alphabetame-ATP (10 microl) into the receptive fields evoked spontaneous discharge in 7.7% of the Abeta fibers, 66.7% of Adelta fibers and 75.0% of C fibres, respectively, and the proportions of Adelta and C fibers with spontaneous discharge evoked by alphabetame-ATP were significantly greater than that of Abeta fibers (P<0.05). In the control experiments, injection of saline did not significantly affect spontaneous discharge or excite the nerve fibers. The mean discharge frequency of Adelta and C fibers increased from 0.73+/-0.24 and 0.54+/-0.21 impulses/min before injection to 3.05+/-0.65 and 8.53+/-2.04 impulses/min during the injection, with subsequent reduction to 2.40+/-0.60 and 6.68+/-1.68 impulses/min in the following 5 min (P<0.05). In contrast to Adelta and C fibers, Abeta fibers exhibited no significant changes in the mean discharge frequency in response to alphabetame-ATP (0.23+/-0.09 impulses/min before injection, 0.28+/-0.09 impulses/min during injection and 0.22+/-0.14 impulses/min after injection, P>0.05). The excitatory effects of alphabetame-ATP on the discharge rate in Adelta and C primary afferent terminal could be observed in the entire course of experiment. Peripheral application of alphabetame-ATP, an ATP analogue, excites and sensitizes a subpopulation of Adelta and C fibers but not Abeta fibers of rat dorsal cutaneous primary afferent fibers.


Oral, perioral and systemic pathosis in HgCl2-induced autoimmunity in the BN rat.
Male BN rats were repeatedly skin-injected with HgCl2 solution and sacrificed after 6, 9, 14, 21, 28 or 24 days. Mononuclear cell infiltrates were observed in the oral mucosa and in lacrimal, salivary and thyroid glands from 6-9 days onwards, with a peak at 14-21 days. Immunohistochemistry identified these cells as predominantly T cells with some NK cells but very few B cells. Reversible parenchymal changes were observed but there was no obvious persistent tissue destruction. Serum titers of IgE, IgG1, anti-laminin and anti-DNP, but not IgG2a antibodies, were raised and peaked at 14-21 days. However, there was no correlation, within animals, between these titers and the extent of mononuclear cell infiltration. Mercury was histochemically detected within dendritic cells/macrophages in the connective tissue stroma of the glands and in the oral mucosa, but no correlation was found between the distribution of mercury and the degree of inflammation. We conclude that the accumulation of mononuclear cells in oral and perioral tissues of HgCl2-treated BN rats does not represent a local immune response to tissue-retained Hg. Instead, we propose that the extravasation represents an epiphenomenon that is not necessarily deleterious to the infiltrated organ.


Hypoxia induces miR-153 through the IRE1α-XBP1 pathway to fine tune the HIF1α/VEGFA axis in breast cancer angiogenesis.
It is well documented that hypoxia activates the hypoxia-inducible factor 1-alpha (HIF1α)/vascular endothelial growth factor A (VEGFA) axis to promote angiogenesis in breast cancer. However, it is unclear how this axis is negatively regulated. In this study, we demonstrated that miR-153 directly inhibits expression of HIF1α by binding to the 3'UTR of HIF1A mRNA, as well as suppresses tube formation of primary human umbilical vein endothelial cells (HUVECs) and breast cancer angiogenesis by decreasing the secretion of VEGFA. Importantly, expression of miR-153 was induced by hypoxia-stimulated ER stress, which activates IRE1α and its downstream transcription factor X-box binding protein 1 (XBP1). X-box binding protein 1 directly binds to the promoter of the miR-153 host gene PTPRN and activates transcription. These results indicate that hypoxia induces miR-153 to fine tune the HIF1α/VEGFA axis in breast cancer angiogenesis and miR-153 could be used for breast cancer anti-angiogenesis therapy.