Polysaccharides derived from natural sources regulate triglyceride and cholesterol metabolism: a review of the mechanisms.
With great changes in people's lifestyles, the incidence of hyperlipidaemia has dramatically increased in recent years. Numerous studies have demonstrated that natural polysaccharides have lipid lowering effects. In this review, the causes and mechanisms of hyperlipidaemia are discussed in order to better understand how polysaccharides alleviate hyperlipidaemia. Natural polysaccharides reduce triglyceride levels through ATGL-(PPAR-α)/(PGC-1α), (SREBP-1c)-ACC/FAS and ACC-CPT1 signal pathways, and exert cholesterol lowering effects via (SREBP-2)-HMGCR and bile acid biosynthesis pathways. Activation of adenosine monophosphate-activated protein kinase (AMPK) is the key factor that mediated the simultaneous regulation of both glucose and lipid metabolism by polysaccharides. The new discovery of polysaccharides increasing the production of endogenous H2S, an important physiological gaseous signaling molecule, is also discussed. Collectively, the current available data suggest that natural polysaccharides could be potentially developed as new and safe lipid-lowering drugs; yet further mechanistic and clinical studies are required during this long-term process.


Bartonella bacilliformis: dangerous pathogen slowly emerging from deep background.
Bartonella bacilliformis was perhaps the most lethal bacterial human pathogen in the pre-antibiotic era, but infections were and are limited to a specific geographical area, largely in Peru, corresponding to the range of its sand fly vector. B. bacilliformis targets both red cells and endothelial cells. Recent phylogenetic realignments have revealed a close genetic relationship to other bacteria which cause human diseases, including bacterial angiomatosis, to the former Grahamella species which infect red cells in other mammals, and to plant pathogens and symbionts including Agrobacterium tumefaciens and Rhizobium meliloti. Features of B. bacilliformis that contribute to its pathogenesis are slowly coming into view, and are here reviewed.


Venous thromboembolism prophylaxis in mental healthcare: do the benefits outweigh the risks?
Venous thromboembolism is an important cause of morbidity and mortality. In recent years, growing awareness has led to the development of strategies to prevent venous thromboembolism in individuals admitted to hospital who are deemed to be at high risk. However, there remains a considerable degree of uncertainty over whether these strategies are of overall benefit and there are few published studies on people who are admitted to psychiatric hospitals. In this editorial I review current clinical practice and areas of uncertainty with respect to venous thromboembolism prophylaxis and its implementation in mental healthcare settings.


Photoacoustic imaging for surgical guidance: Principles, applications, and outlook.
Minimally invasive surgeries often require complicated maneuvers and delicate hand-eye coordination and ideally would incorporate "x-ray vision" to see beyond tool tips and underneath tissues prior to making incisions. Photoacoustic imaging has the potential to offer this feature but not with ionizing x-rays. Instead, optical fibers and acoustic receivers enable photoacoustic sensing of major structures-such as blood vessels and nerves-that are otherwise hidden from view. This imaging process is initiated by transmitting laser pulses that illuminate regions of interest, causing thermal expansion and the generation of sound waves that are detectable with conventional ultrasound transducers. The recorded signals are then converted to images through the beamforming process. Photoacoustic imaging may be implemented to both target and avoid blood-rich surgical contents (and in some cases simultaneously or independently visualize optical fiber tips or metallic surgical tool tips) in order to prevent accidental injury and assist device operators during minimally invasive surgeries and interventional procedures. Novel light delivery systems, counterintuitive findings, and robotic integration methods introduced by the Photoacoustic & Ultrasonic Systems Engineering Lab are summarized in this invited Perspective, setting the foundation and rationale for the subsequent discussion of the author's views on possible future directions for this exciting frontier known as photoacoustic-guided surgery.


Clonal growth of mammalian cells in vitro; growth characteristics of colonies from single HeLa cells with and without a feeder layer.
Two methods for simple and rapid plating of single HeLa cells, human, carcinomatous cells, are described. These result in growth and formation of colonies from each single cell. One of these procedures uses irradiated, non-multiplying "feeder" cells to condition the medium. The second requires more gentle handling of the cells, but otherwise is virtually the same as that used in plating bacteria on semisolid, nutrient media. By extension of these methods, it is possible to isolate single mutant colonies and grow pure clonal stocks of animal cells. These genetically uniform strains are much more homogeneous in their behavior than the parental HeLa cell population. Growth curves obtained from developing colonies are highly reproducible. The most active mutant stocks so far isolated display a generation time of 18 to 20 hours. In pooled human serum HeLa cells assume a highly stretched, ameboid form, with marked motility; whereas growth of the same cells in a variety of non-human sera results in tightly packed, columnar, epithelial-like morphology. The two cell types possess volumes, nuclear cross-sections, plating efficiencies, and generation times which are identical within experimental error, but display widely different cross-sectional areas, suggesting that the basic change occurs in the cell surface. It is conceivable that this change may be related to that which enables the cells of a compact tumor to become invasive. Animal cells subjected to the standard trypsinization procedures which involve mechanical trauma and repeated washings in incomplete media leak large amounts of P and suffer impaired ability to reproduce as isolated cells. Application of the methods described in this paper as a tool for quantitative study of normal mammalian cell growth, physiology, genetics, and biochemistry, and the response of cells to drugs, viruses, high energy radiation, and other agents have been indicated.


Endometrial metastasis of colorectal cancer with coincident endometrial adenocarcinoma.
Metastasis to the uterine corpus is uncommon and secondary colorectal tumours of the endometrium are rare. We describe a uterine tumour with components of both primary endometrial and metastatic colorectal carcinomata. In this case, a 72-year-old obese woman presented with a 2-week history of postmenopausal bleeding per vaginum and weight loss. She had an abdominoperineal resection 3 years previously for a Dukes stage B rectal carcinoma. A transvaginal ultrasonography showed a thickened endometrium. Histology immunophenotyping showed a CK7+, CK20+, CA125- and CEA+ colorectal metastasis (a profile consistent with her previous cancer) associated with a primary CK7+, CK20-, CA125+ and CEA- endometroid endometrial adenocarcinoma. We conclude this represents endometrial metastasis of colorectal carcinoma with coincident primary endometrial adenocarcinoma. We speculate as to whether the endometrial carcinoma arose de novo or was induced by the colorectal metastasis, or whether the primary endometrial tumour provided a fertile site for the colorectal metastasis.


Modulation of heme redox potential in the cytochrome c6 family.
Cytochrome c6A is a unique dithio-cytochrome of green algae and plants. It has a very similar core structure to that of bacterial and algal cytochromes c6 but is unable to fulfill the same function of transferring electrons from cytochrome f to photosystem I. A key feature is that its heme midpoint potential is more than 200 mV below that of cytochrome c6 despite having His and Met as axial heme-iron ligands. To identify the molecular origins of the difference in potential, the structure of cytochrome c6 from the cyanobacterium Phormidium laminosum has been determined by X-ray crystallography and compared with the known structure of cytochrome c6A. One salient difference of the heme pockets is that a highly conserved Gln (Q51) in cytochrome c6 is replaced by Val (V52) in c6A. Using protein film voltammetry, we found that swapping these residues raised the c6A potential by +109 mV and decreased that of c6 by almost the same extent, -100 mV. X-ray crystallography of the V52Q protein showed that the Gln residue adopts the same configuration relative to the heme as in cytochrome c6 and we propose that this stereochemistry destabilizes the oxidized form of the heme. Consequently, replacement of Gln by Val was probably a key step in the evolution of cytochrome c6A from cytochrome c6, inhibiting reduction by the cytochrome b6f complex and facilitating establishment of a new function.


Vascular and blood-brain barrier-related changes underlie stress responses and resilience in female mice and depression in human tissue.
Prevalence, symptoms, and treatment of depression suggest that major depressive disorders (MDD) present sex differences. Social stress-induced neurovascular pathology is associated with depressive symptoms in male mice; however, this association is unclear in females. Here, we report that chronic social and subchronic variable stress promotes blood-brain barrier (BBB) alterations in mood-related brain regions of female mice. Targeted disruption of the BBB in the female prefrontal cortex (PFC) induces anxiety- and depression-like behaviours. By comparing the endothelium cell-specific transcriptomic profiling of the mouse male and female PFC, we identify several pathways and genes involved in maladaptive stress responses and resilience to stress. Furthermore, we confirm that the BBB in the PFC of stressed female mice is leaky. Then, we identify circulating vascular biomarkers of chronic stress, such as soluble E-selectin. Similar changes in circulating soluble E-selectin, BBB gene expression and morphology can be found in blood serum and postmortem brain samples from women diagnosed with MDD. Altogether, we propose that BBB dysfunction plays an important role in modulating stress responses in female mice and possibly MDD.


Uncertainties in deforestation emission baseline methodologies and implications for carbon markets.
Carbon credits generated through jurisdictional-scale avoided deforestation projects require accurate estimates of deforestation emission baselines, but there are serious challenges to their robustness. We assessed the variability, accuracy, and uncertainty of baselining methods by applying sensitivity and variable importance analysis on a range of typically-used methods and parameters for 2,794 jurisdictions worldwide. The median jurisdiction's deforestation emission baseline varied by 171% (90% range: 87%-440%) of its mean, with a median forecast error of 0.778 times (90% range: 0.548-3.56) the actual deforestation rate. Moreover, variable importance analysis emphasised the strong influence of the deforestation projection approach. For the median jurisdiction, 68.0% of possible methods (90% range: 61.1%-85.6%) exceeded 15% uncertainty. Tropical and polar biomes exhibited larger uncertainties in carbon estimations. The use of sensitivity analyses, multi-model, and multi-source ensemble approaches could reduce variabilities and biases. These findings provide a roadmap for improving baseline estimations to enhance carbon market integrity and trust.


Multiple neuronal populations control the eating behavior in Hydra and are responsive to microbial signals.
Although recent studies indicate the impact of microbes on the central nervous systems and behavior, it remains unclear how the relationship between the functionality of the nervous system, behavior, and the microbiota evolved. In this work, we analyzed the eating behavior of Hydra, a host that has a simple nervous system and a low-complexity microbiota. To identify the neuronal subpopulations involved, we used a subpopulation-specific cell ablation system and calcium imaging. The role of the microbiota was uncovered by manipulating the diversity of the natural microbiota. We show that different neuronal subpopulations are functioning together to control eating behavior. Animals with a drastically reduced microbiome had severe difficulties in mouth opening due to a significantly increased level of glutamate. This could be reversed by adding a full complement of the microbiota. In summary, we provide a mechanistic explanation of how Hydra's nervous system controls eating behavior and what role microbes play in this.


The risk of iridocyclitis in juvenile rheumatoid arthritis.
A retrospective study comparing 14 patients with juvenile rheumatoid arthritis (JRA) complicated by chronic iridocyclitis (Group I) with an equal number of JRA patients without this complication (Group II) was conducted to determine which factors might aid in the earlier diagnosis of chronic iridocyclitis. Eleven patients in Group I had oligoarticular disease compared to 5 in Group II. Antinuclear antibody was found in 13 patients in Group I compared to only 2 in Group II. Only one of nine patients tested in Group I for the histocompatibility antigen W-27 was positive. The patient with oligoarticular disease and a positive ANA test should be followed closely by the ophthalmologist for the development of chronic iridocyclitis.


A chemoenzymatic total synthesis of the structure assigned to the alkaloid (+)-montabuphine.
An enantioselective synthesis of the structure, 3, assigned to the alkaloid (+)-montabuphine has been achieved using the readily available metabolite 4 as starting material. A comparison of the physical and spectral data recorded on compound 3 with those reported for (+)-montabuphine suggests that they are different compounds.


Maternal care in wild toque macaques (Macaca sinica) involves prolonged lactation and interbirth intervals as adaptations to reduce maternal depletion and infant mortality in harsh environments.
Weaning age in primates has been challenging to measure and new methods, involving molecular biomarkers in feces, tissue, or teeth have contributed to a solution. Here, we used a direct approach by briefly anesthetizing 442 female toque macaques (Macaca sinica) of Sri Lanka (over a 17-year period) and manually testing their mammary tissue for the presence or absence of milk. Milk tests were related to known offspring ages and maternal care behaviors and indicated that older infants suckled milk well past the weaning age of 7 months that is often reported for food-provisioned primates. Mothers strongly rejected their infants' nursing attempts in two phases, the first at 7 months as an honest signal "giving notice" promoting a shift to greater independence from milk to solid food, and when "shutting down" at final weaning after 12-18 months. The shift to supplementary lactation coincided also with the cessation of mothers carrying their infants and a resumption of cycling. All infants up to 7.2 months suckled milk, 91% of them did up to 18 months, this continued for 42% of infants beyond 18 months, and normally none received milk after 22 months. Lactation extended into 2.2% of cycling and 10.7% of pregnant females (up to 50% of gestation). The interbirth interval was prolonged by factors predicted to draw on female metabolic energy reserves and included the duration of lactation, growth among primiparas, and dietary limitations. The last also increased menarche. Females offset the metabolic costs of lactation with increased foraging and catabolism, but infants died when lactation costs seemingly compromised maternal condition. The prolonged lactation and slowed reproduction are considered adaptations to promote infant survival and growth in an environment where the natural food supply limits population growth and competition for food and water impacts the mortality of the youngest the most.


Subungual trichogranuloma in a hairdresser.
Hairdressers may be subject to occupational skin diseases other than contact dermatitis. Interdigital trichogranuloma is a common disorder among those who cut men's hair but is much less common among those who cut women's hair. Interdigital trichogranulomas or interdigital sinuses are the result of penetration of the skin by short, sharp hair clippings. Subungual hair penetration appears to be much less common. This article reports the first case of subungual trichogranuloma in a hairdresser with psoriatic onycholysis. Onycholysis may be both a risk factor for and a consequence of cut hairs becoming imbedded subungually.


Cell type-specific β2-adrenergic receptor clusters identified using photoactivated localization microscopy are not lipid raft related, but depend on actin cytoskeleton integrity.
Recent developments in the field of optical super-resolution techniques allow both a 10-fold increase in resolution as well as an increased ability to quantify the number of labeled molecules visualized in the fluorescence measurement. By using photoactivated localization microscopy (PALM) and an experimental approach based on the systematic comparison with a nonclustering peptide as a negative control, we found that the prototypical G protein-coupled receptor β2-adrenergic receptor is partially preassociated in nanoscale-sized clusters only in the cardiomyocytes, such as H9C2 cells, but not in other cell lines, such as HeLa and Chinese hamster ovary (CHO). The addition of the agonist for very short times or the addition of the inverse agonist did not significantly affect the organization of receptor assembly. To investigate the mechanism governing cluster formation, we altered plasma membrane properties with cholesterol removal and actin microfilament disruption. Although cholesterol is an essential component of cell membranes and it is supposed to be enriched in the lipid rafts, its sequestration and removal did not affect receptor clustering, whereas the inhibition of actin polymerization did decrease the number of clusters. Our findings are therefore consistent with a model in which β2 receptor clustering is influenced by the actin cytoskeleton, but it does not rely on lipid raft integrity, thus ruling out the possibility that cell type-specific β2 receptor clustering is associated with the raft.


[Morphoimmunohistochemical differential diagnosis of B-cell lymphomas].
The paper considers the stages of B-cell differentiation at the level of central (bone marrow) and peripheral (lymph node) organs of the immune system. The immunophenotype of B-cell lymphoid tumors and their "normal" cell analogues was compared, by taking into account 4 stages of B-cell differentiation: bone marrow B-cell precursors, mature B-cell, B-cell with follicular differentiation, postfollicular B-cell. Algorithms of immunohistochemical differential diagnosis of B-cell lymphomas are proposed when their morphological patterns are similar.


Organization of brain networks governed by long-range connections index autistic traits in the general population.
The dimensional approach to autism spectrum disorder (ASD) considers ASD as the extreme of a dimension traversing through the entire population. We explored the potential utility of electroencephalography (EEG) functional connectivity as a biomarker. We hypothesized that individual differences in autistic traits of typical subjects would involve a long-range connectivity diminution within the delta band. Resting-state EEG functional connectivity was measured for 74 neurotypical subjects. All participants also provided a questionnaire (Social Responsiveness Scale, SRS) that was completed by an informant who knows the participant in social settings. We conducted multivariate regression between the SRS score and functional connectivity in all EEG frequency bands. We explored modulations of network graph metrics characterizing the optimality of a network using the SRS score. Our results show a decay in functional connectivity mainly within the delta and theta bands (the lower part of the EEG spectrum) associated with an increasing number of autistic traits. When inspecting the impact of autistic traits on the global organization of the functional network, we found that the optimal properties of the network are inversely related to the number of autistic traits, suggesting that the autistic dimension, throughout the entire population, modulates the efficiency of functional brain networks. EEG functional connectivity at low frequencies and its associated network properties may be associated with some autistic traits in the general population.


Regulation of the G1 phase of the cell cycle by periodic stabilization and degradation of the p25rum1 CDK inhibitor.
In fission yeast, the cyclin-dependent kinase (CDK) inhibitor p25(rum1) is a key regulator of progression through the G1 phase of the cell cycle. We show here that p25(rum1) protein levels are sharply periodic. p25(rum1) begins to accumulate at anaphase, persists in G1 and is destroyed during S phase. p25(rum1 )is stabilized and polyubiquitinated in a mutant defective in the 26S proteasome, suggesting that its degradation normally occurs through the ubiquitin-dependent 26S proteasome pathway. Phosphorylation of p25(rum1 )by cdc2-cyclin complexes at residues T58 and T62 is important to target the protein for degradation. Mutation of one or both of these residues to alanine causes stabilization of p25(rum1) and induces a cell cycle delay in G1 and polyploidization due to occasional re-initiation of DNA replication before mitosis. The CDK-cyclin complex cdc2-cig1, which is insensitive to p25(rum1 )inhibition, seems to be the main kinase that phosphorylates p25(rum1). Phosphorylation of p25(rum1) in S phase and G2 serves as the trigger for p25(rum1) proteolysis. Thus, periodic accumulation and degradation of the CDK inhibitor p25(rum1 )in G1 plays a role in setting a threshold of cyclin levels important in determining the length of the pre-Start G1 phase and in ensuring the correct order of cell cycle events.


Survival advantage from higher-dose radiation therapy for clinically localized prostate cancer treated on the Radiation Therapy Oncology Group trials.
We evaluated the effect of external-beam radiation therapy on disease-specific survival (death from causes related to prostate cancer) and overall survival in men with clinically localized prostate cancer. From 1975 to 1992, 1,465 men with clinically localized prostate cancer received radiation therapy on four Radiation Therapy Oncology Group phase III randomized trials and were pooled for this analysis. No one received androgen-deprivation therapy with his initial treatment. All original histology had central pathologic review for grading using the Gleason classification system. Total delivered radiation dose ranged from 60 to 78 Gy (median, 68.4 Gy). The median follow-up time was 8 years. A Cox regression model revealed that Gleason score was an independent predictor of disease-specific survival and overall survival. The 10-year disease-specific survival rates by Gleason score were as follows: score of 2 through 5, 85%; score of 6, 79%; score of 7, 62%; and score of 8 through 10, 43%. Stratifying outcome by this important prognostic factor revealed that higher radiation dose was a significant predictor for improved disease-specific survival and overall survival only for those patients whose cancers had Gleason scores of 8 through 10 (P <.05). After adjusting for clinical T stage, nodal status, and age, treating with a higher radiation dose was associated with a 29% lower relative risk of death from prostate cancer and 27% reduced mortality rate (P <.05). These data demonstrate that higher-dose radiation therapy can significantly reduce the risk of dying from prostate cancer in men with clinically localized disease. This survival benefit is restricted to men with poorly differentiated cancers.


Atorvastatin attenuation of ABCB1 expression is mediated by microRNA miR-491-3p in Caco-2 cells.
Atorvastatin, a HMG-CoA reductase inhibitor, used in the treatment of hypercholesterolemia, has been previously shown to regulate ABCB1 expression in vivo and in vitro. We hypothesized that the statin could regulate gene expression of ABCB1 transporter via microRNAs. Expression of microRNAs and ABCB1 mRNA was examined in atorvastatin-treated and control cells using real-time PCR. miR-491-3P mimic and inhibitor were transfected in Caco-2 and ABCB1 expression was monitored by western blot and real-time PCR. In HepG2 cells, none of the microRNAs predicted to target ABCB1 3'UTR was regulated by atorvastatin treatment. In agreement with this, ABCB1 3'UTR activity was not modulated in HepG-2 cells after 48h-treatment as measured by luciferase assay. In Caco-2 cells, atorvastatin treatment provoked a decrease in luciferase activity and, accordingly, miR-491-3p was upregulated about 2.7 times after 48h-statin treatment. Luciferase analysis of miR-491-3p with a mimetic or inhibitor of miR-491-3p revealed that this microRNA could target ABCB1 3'UTR, as after miR-491-3p inhibition, ABCB1 levels were increased by two-fold, and miR-491-3p superexpression decreased ABCB1 3'UTR activity. Finally, functional analysis revealed that treatment with miR-491-3p inhibitor could reverses atorvastatin attenuation of ABCB1 (Pg-p) protein levels. Our results suggest atorvastatin control ABCB1 expression via miR-491-3p in Caco-2 cells. This finding may be an important mechanism of statin drug-drug interaction, since common concomitant drugs used in the prevention of cardiovascular diseases are ABCB1 substrates.


Identification of protein targets in cerebral endothelial cells for brain arteriovenous malformation (AVMs) molecular therapies.
To develop a new molecular targeted treatment for brain (AVMs), identification of membrane proteins that are localised on the AVM endothelium is crucial. Current treatment methods are surgery and radiosurgery. However, complete occlusion post radiosurgery are achieved within 3 years, while patient remain at risk of haemorrhage. This study aims to identify potential protein targets in AVM endothelial cells that discriminate these vessels from normal vessels; these proteins targets will be investigated for the molecular therapy of brain AVMs to promote rapid thrombosis after radiosurgery. We employed in vitro biotinylation that we developed, and mass spectrometry to detect cell surface-exposed proteins in cultures of murine cerebral endothelial cells (bEnd.3). Two forms of mass spectrometry were applied (iTRAQ-MS and MSE) to identify and quantify membrane protein expression at various time-points following irradiation which simulates a radiosurgical treatment approach. Immunocytochemistry was used to confirm the expression of selected membrane proteins. ProteinPilot V4.0 software was used to analyse the iTRAQ-MS data and the MSE data was analysed using ProteinLynx Global Server version 2.5 software. The proteomics data revealed several differentially expressed membrane proteins between irradiated and non-irradiated cells at specific time points, e.g. PECAM-1, cadherin-5, PDI, EPCR and integrins. Immunocytochemistry data confirmed the expression of these proteins. Cell surface protein biotinylation and proteomics analysis successfully identified membrane proteins from murine brain endothelial cells in response to irradiation. This work suggests potential target protein molecules for evaluation in animal models of brain-AVM.


Facile core-shell nanoparticles with controllable antibacterial activity assembled by chemical and biological molecules.
Antibiotic residues can induce the production of bacterial variability and increase the variety of resistant bacterial strains. In order to cope up with the global crisis, there is an urgent need to develop alternative drugs and strategies to effectively kill the bacteria and inhibit the emergence of superbugs. Herein, we developed an antibacterial strategy based on the conjugated polymer nanoparticles (CPNs) and the biomolecule-mediated regulation for the controllable antibacterial treatment. Upon being modified with Hoechst 33258 (H33258), the biocompatible CPNs tend to be effective antibacterial agents, with the antibacterial activity being regulated through the binding and separation of H33258 and calf thymus double-stranded DNA (dsDNA) with the aid of deoxyribonuclease I (DNase I). This eco-friendly antibacterial system not only provides a simple and controllable antibacterial strategy but also inspires the design of biomolecule-mediated antibacterial activity regulation.


White matter injury in infants with intraventricular haemorrhage: mechanisms and therapies.
Intraventricular haemorrhage (IVH) continues to be a major complication of prematurity that can result in cerebral palsy and cognitive impairment in survivors. No optimal therapy exists to prevent IVH or to treat its consequences. IVH varies in severity and can present as a bleed confined to the germinal matrix, small-to-large IVH or periventricular haemorrhagic infarction. Moderate-to-severe haemorrhage dilates the ventricle and damages the periventricular white matter. This white matter injury results from a constellation of blood-induced pathological reactions, including oxidative stress, glutamate excitotoxicity, inflammation, perturbed signalling pathways and remodelling of the extracellular matrix. Potential therapies for IVH are currently undergoing investigation in preclinical models and evidence from clinical trials suggests that stem cell treatment and/or endoscopic removal of clots from the cerebral ventricles could transform the outcome of infants with IVH. This Review presents an integrated view of new insights into the mechanisms underlying white matter injury in premature infants with IVH and highlights the importance of early detection of disability and immediate intervention in optimizing the outcomes of IVH survivors.


Nanoparticle-Mediated Delivery of Neuroprotective Substances for the Treatment of Diabetic Retinopathy.
Diabetic retinopathy (DR) is a major complication of diabetes, characterized by extensive vascular pathology leading to vision loss. Neuronal suffering and death are also present in the diabetic retina as a result of different molecular mechanisms that are compromised or modified in response to high glucose. The aim of this paper is to highlight recent data indicating that neurodegeneration is likely to play a primary role in the development of DR and that strategies based on nanomedicine may be exploited to deliver neuroprotection to the retina. An extensive analysis of the publications dealing with the role of neuroprotection in DR and with nanoparticle-mediated drug delivery to the retina has been conducted using PubMed, with particular attention to the most recent papers. There are important limitations related to possible systemic side effects of neuroprotective substances and to drug bioavailability in the retina such as, for instance, the amount of drug reaching the retina, the need of keeping to a minimum the number of administrations (especially, for example, in the case of intraocular injections) and the need of assuring a long-lasting, graded intraocular drug delivery. In recent years, a variety of investigations have been aimed at the exploitation of approaches of nanomedicine to enhance the pharmacokinetics and pharmacodynamic activity of intraocularly delivered drugs. In particular, we provide some preliminary results that we have obtained about the feasibility of delivering magnetic nanoparticles functionalized with a neuroprotectant to mouse eyes through intraocular injections. We propose that nanoparticles functionalized with neuroprotective substances may be used to protect the diabetic retina, thus causing an impact in the design of future pharmacologic treatments for DR.


Radiation protection aspects of repairing a highly contaminated tritium cold box.
Radiation protection aspects of modifying a system exposed to tritium gas are discussed. This paper includes a brief discussion of the tritium gas and tritium surface contamination hazards associated with a facility designed to detritiate heavy water. Also described are the protective equipment, dosimetry, instrumentation, contamination control, and decontamination procedures used during welding of tubes that have been exposed to almost pure tritium gas. Absorption of the tritium gas on materials resulted in exposures to high peak concentrations of tritium oxide and high levels of tritium surface contamination (TSC).


The ontogeny of the androgen receptor in human foreskin.
To investigate the relationship between the androgen receptor content of human foreskin and age-dependent physiological changes in genital development, cytosolic and nuclear androgen receptors were measured in preputial skin specimens from male subjects of various ages. Optimum incubation conditions (4 C, 20 h) were established for measurement of androgen receptors by the exchange method with the synthetic androgen ligand methyltrienolone. The number of total androgen receptor sites, cytosolic plus nuclear, was fairly constant at all ages studied. However, the androgen receptor was predominantly localized to the nuclear compartment at those developmental stages characterized by higher levels of circulating blood androgens, i.e. newborns, pubertal males, and adults. By contrast, the androgen receptor in specimens from prepubertal boys was confined almost exclusively to the cytosolic compartment. This corresponds to a time of low plasma androgen levels and quiescence in genital maturation. It is suggested that changes in the intracellular distribution of androgen receptors may bear some relationship to the sequence of normal genital growth and development with nuclear androgen receptors having major physiological importance.


[Isolation of fibrin assembly inhibitor from plasma and tissue extracts].
Procedures for isolation of an inhibitor of the fibrin self association are described. The procedures involved dialysis across cuprophane or cellophane T-100, ion exchange chromatography (the first method) or gel filtration (the second method). The product containing amino acid impurities was obtained after dialysis across cuprophane and ion exchange chromatography; a homogeneous inhibitor was isolated by means of dialysis across cellophane T-100 and gel filtration through Sephadex G-50.


The fine specificity of human T cell lines towards myelin basic protein peptides in southern Italian multiple sclerosis patients.
We studied the relationship between the HLA specificities associated with multiple sclerosis (MS) susceptibility in southern Italy and the reactivity of the human myelin basic protein (hMBP) immunogenic peptides 84-98 and 143-168, using short-term T-cell lines established from 9 MS patients and from 8 healthy individuals. In our population, DR15 was significantly associated with MS (34.9% in MS versus 13.7% in healthy controls, P < 0.05). This result is in agreement with the association found in northern Europe, but not with data obtained in a population from the island of Sardinia (Italy). In MS patients the frequency of reactive T-cell lines (TCL), tested for fine specificity against the immunodominant hMBP peptides 84-98 and 143-168, was increased for the hMBP 143-168 peptide (P < 0.05) but not for the 84-98 peptide. Although this reactivity was higher in DR15+ MS patients than in DR 15- MS patients, it seemed not to be associated with DR15 specificity in the MS population. Furthermore, there were no significant differences in frequency of reactive TCL to hMBP peptide 84-98 in DR15-positive or DR15-negative MS patients. Consequently, it appears that peptide 84-98, considered as a relevant autoantigen, is not implicated in the pathogenesis of MS in our population from southern Italy.


Characterization of the outer-membrane proteins of Haemophilus parainfluenzae expressed under iron-sufficient and iron-restricted conditions.
SDS-PAGE of the outer-membrane (OM) proteins of Haemophilus parainfluenzae P205 grown under iron-sufficient conditions revealed three major proteins of 40, 37 and 13 kDa. In addition, growth under conditions of iron-restriction resulted in the expression of at least four iron-repressible OM proteins (IROMPs) of 72, 81, 88 and 90 kDa. OM proteins of 40 and 13 kDa were non-covalently associated with peptidoglycan and were resistant to digestion with trypsin. A 38 kDa peptidoglycan-associated protein, which was masked by the abundant 37 kDa protein, was also observed following tryptic digestion of whole cells or OMs. Neither the 37 kDa protein (which was heat-modifiable) nor the IROMPs were peptidoglycan-associated, and both were cleaved following treatment of whole cells with trypsin, indicating that they are exposed at the cell surface. A variety of IROMPs from five other H. parainfluenzae strains was also observed. In each strain, both the IROMPs and a major protein of 37 kDa were exposed at the cell surface.


Exploring the potential of clay catalysts in catalytic pyrolysis of mixed plastic waste for fuel and energy recovery.
Developing low-cost and high-activity catalysts is one of the keys to promoting the catalytic pyrolysis of waste plastics to fuels for plastic recycling. This work studied the effect of clay as the catalyst on mixed plastic pyrolysis for fuel and energy recovery. Four kinds of clay, including nanoclay, montmorillonite, kaolin, and hydrotalcite, were used as catalysts for the pyrolysis of mixed plastic consisted of polyethylene terephthalate, polystyrene, polypropylene, low-density polyethylene, and high-density polyethylene. The product yield and distribution varied with different clay in pyrolysis. The highest yield of oil was 71.0 % when using montmorillonite as the catalyst. While the highest contents of gasoline range hydrocarbons and diesel range hydrocarbons in the oil were achieved when using kaolin and nanoclay, respectively as catalysts. For the gas products, the CO, C2H4, C2H6, C3H6, and C4H10 increased with decreased CO2 in the gaseous products when using clay as catalysts. In general, the mild acidity of clay catalyst was essential to improve the oil yields and the proportion of the gasoline or diesel range fuels in the catalytic pyrolysis of mixed plastic waste.


Clinical manifestations, associated risk factors and treatment outcomes of Chronic Pulmonary Aspergillosis (CPA): Experiences from a tertiary care hospital in Lahore, Pakistan.
Chronic pulmonary aspergillosis (CPA) has a wide spectrum of illnesses depending on the progression of the disease and comorbid conditions. However, there is an inadequacy of investigations regarding clinical, laboratory, risk factor and prognostic data on CPA. The current study is aimed to consider the clinical manifestations, risk factors and outcomes of CPA. Retrospective records of all patients with a confirmed diagnosis of CPA who sought treatment at Gulab Devi Chest Hospital Lahore, Pakistan from January 2017 to December 2019 were evaluated. Data regarding demographics, clinical manifestations, comorbidities, radiographic and microbiological findings, length of hospital stay (LOS) and intensive care unit (ICU) admission was collected and analyzed to identify the factors associated with mortality. The independent factors associated with mortality were also identified by appropriate analyses. A total of 218 CPA patients were included in this study. The mean age was 45.75 ± 6.26 years. Of these, 160 (73.4%) were male, and 65 (29.8%) had diabetes. The mean LOS was 18.5 ± 10.9 days. The most common type of CPA was simple aspergilloma (56%) followed by chronic cavitary pulmonary aspergillosis (CCPA) (31.2%). About one half of the patients had a history of pulmonary tuberculosis (TB) and treatment response rates were low in patients with active TB. The overall mortality rate was 27.1%. ICU admission was required for 78 (35.8%) patients. Diabetes mellitus (DM), hematological malignancies and chronic kidney disease (CKD) were the common underlying conditions predicting a poor outcome. Mean LOS, hematological malignancies, consolidation and ICU admission were identified as the independent factors leading to mortality. CPA had a significant association with TB in the majority of cases. Treatment response rates in cases with active TB were comparatively low. Cases with high mean LOS, hematological malignancies, consolidation, ICU admission, CKD and DM experienced poor outcomes. High mean LOS, hematological malignancies, consolidation and ICU stay were identified as independent risk factors for mortality. Future large prospective studies, involving aspergillus specific immunoglobulin G (IgG) antibody testing, are required for a better understanding of CPA in Pakistan.


Preparative electrophoretic method for the purification of a hydrophobic membrane protein: subunit c of the mitochondrial ATP synthase from rat liver.
A method is described for the purification of subunit c of ATP synthase from rat liver mitochondria. After sample preparation and solvent extraction, the protein was purified to homogeneity by a single-step preparative electrophoretic procedure, using aqueous buffer and containing lithium dodecyl sulfate. The subunit is an extremely hydrophobic and insoluble protein and all solubilization attempts, using a variety of detergents, were unsuccessful except for lithium dodecyl sulfate. Buffer exchange and FPLC gel filtration removed the detergent from the purified sample, leaving the protein in a soluble form. The mammalian protein is composed of 75 amino acid residues, with a molecular mass of 7602 Da and is classified as a proteolipid. Subunit c accounts for 25 and 85% of the intralysosomal accumulation, within neurons, of storage material in juvenile and late-infantile forms of Batten's disease, respectively. This purification procedure allows access to a continuous supply of pure subunit c from a conventional source such as rat liver and preserves precious autopsy materials. The protein could be used as substrate in future proteolytic studies involving pepstatin-insensitive lysosomal proteases and for raising of more specific antibodies. The procedure could also be adapted/modified and used as a model for purifying other extremely insoluble proteins.


Influence of CYP2C19 genetic polymorphisms on clinical outcomes of intracranial aneurysms treated with stent-assisted coiling.
To investigate the influence of CYP2C19 genetic polymorphisms on clinical outcomes of intracranial aneurysms treated with stent-assisted coiling. Between September 2014 and October 2015, we prospectively recruited 215 patients with intracranial aneurysms who were treated with stent-assisted coiling. CYP2C19 genotypes were determined and clopidogrel response was tested. The primary endpoints included symptomatic or silent ischemic events, and bleeding events. The secondary endpoint was clinical outcome at 3 months. Of the 215 patients, 108 (50.2%) were classified as intermediate metabolizers (IMs, CYP2C19*1/*2, *1/*3), 76 (35.3%) as extensive metabolizers (EMs, CYP2C19*1/*1) and 31 (14.4%) as poor metabolizers (PMs, CYP2C19*2/*2, *2/*3, *3/*3). Carriers of CYP2C19 loss-of-function (LOF) alleles (*2 or *3, p=0.001), especially PMs (p=0.004), had an increased risk for clopidogrel resistance. After the procedures, cerebral ischemic events occurred in 69 patients (32.1%) and bleeding was seen in 20 patients (9.3%). In comparison with IMs and PMs, EMs had a lower risk for ischemic events (21.1% vs 37.0% and 41.9%, p=0.02 and 0.027, respectively) and a relatively higher risk for bleeding events (18.4% vs 5.6% and 0%, p=0.006 and 0.01, respectively). Based on multivariate analysis, the carriage of CYP2C19 LOF alleles (p=0.032) and clopidogrel resistance (p=0.047) were considered as predictors of cerebral ischemic events, and EMs were significantly associated with bleeding (p=0.002). Posterior circulation aneurysms (p=0.038), hemorrhagic history (p=0.001) and poor metabolic genotypes (p=0.001) could result in poor clinical outcomes (modified Rankin Scale >2). CYP2C19 genetic polymorphisms had significant influence on the antiplatelet effect of clopidogrel, and could be considered as risk factors of ischemic or bleeding events and even clinical outcomes of patients with intracranial aneurysms treated with stent-assisted coiling.


Non-steroidal anti-inflammatory drugs: assessment of risks.
The prescription-event monitoring procedure developed at the University of Southampton was used to evaluate five different non-steroidal anti-inflammatory drugs (NSAIDs), including piroxicam, in approximately 55,000 patients. The overall incidence of side effects was what would be expected. The risks of gastrointestinal haemorrhage and peptic ulceration were spread uniformly across the five drugs under study. No real difference was seen in the incidence of these conditions when patients discontinued the medication or switched to another NSAID. Overall, serious side effects were extremely rare. One provocative finding was the possibility that drugs of this class may exert a cardioprotective effect. There appeared to be a deficit of cases of myocardial infarction while the patients were taking an NSAID. Responsibility for the efficacy and safety of all drugs resides with many people and organizations: the manufacturers, national health departments, licensing departments, and physicians. The press, which has contributed to many unwarranted panics concerning drug safety, also must refrain from using power without responsibility.


The impact of interventions for youth experiencing homelessness on housing, mental health, substance use, and family cohesion: a systematic review.
Youth often experience unique pathways into homelessness, such as family conflict, child abuse and neglect. Most research has focused on adult homeless populations, yet youth have specific needs that require adapted interventions. This review aims to synthesize evidence on interventions for youth and assess their impacts on health, social, and equity outcomes. We systematically searched Medline, Embase, PsycINFO, and other databases from inception until February 9, 2018 for systematic reviews and randomized controlled trials on youth interventions conducted in high income countries. We screened title and abstract and full text for inclusion, and data extraction were completed in duplicate, following the PRISMA-E (equity) review approach. Our search identified 11,936 records. Four systematic reviews and 18 articles on randomized controlled trials met the inclusion criteria. Many studies reported on interventions including individual and family therapies, skill-building, case management, and structural interventions. Cognitive behavioural therapy led to improvements in depression and substance use, and studies of three family-based therapies reported decreases in substance use. Housing first, a structural intervention, led to improvements in housing stability. Many interventions showed inconsistent results compared to services as usual or other interventions, but often led to improvements over time in both the intervention and comparison group. The equity analysis showed that equity variables were inconsistently measured, but there was data to suggest differential outcomes based upon gender and ethnicity. This review identified a variety of interventions for youth experiencing homelessness. Promising interventions include cognitive behavioural therapy for addressing depression, family-based therapy for substance use outcomes, and housing programs for housing stability. Youth pathways are often unique and thus prevention and treatment may benefit from a tailored and flexible approach.


Direction of visual apparent motion driven solely by timing of a static sound.
In temporal ventriloquism, auditory events can illusorily attract perceived timing of a visual onset [1-3]. We investigated whether timing of a static sound can also influence spatio-temporal processing of visual apparent motion, induced here by visual bars alternating between opposite hemifields. Perceived direction typically depends on the relative interval in timing between visual left-right and right-left flashes (e.g., rightwards motion dominating when left-to-right interflash intervals are shortest [4]). In our new multisensory condition, interflash intervals were equal, but auditory beeps could slightly lag the right flash, yet slightly lead the left flash, or vice versa. This auditory timing strongly influenced perceived visual motion direction, despite providing no spatial auditory motion signal whatsoever. Moreover, prolonged adaptation to such auditorily driven apparent motion produced a robust visual motion aftereffect in the opposite direction, when measured in subsequent silence. Control experiments argued against accounts in terms of possible auditory grouping, or possible attention capture. We suggest that the motion arises because the sounds change perceived visual timing, as we separately confirmed. Our results provide a new demonstration of multisensory influences on sensory-specific perception [5], with timing of a static sound influencing spatio-temporal processing of visual motion direction.


Novel Machine Learning Approach for the Prediction of Hernia Recurrence, Surgical Complication, and 30-Day Readmission after Abdominal Wall Reconstruction.
Despite advancements in abdominal wall reconstruction (AWR) techniques, hernia recurrences (HRs), surgical site occurrences (SSOs), and unplanned hospital readmissions persist. We sought to develop, validate, and evaluate machine learning (ML) algorithms for predicting complications after AWR. We conducted a comprehensive review of patients who underwent AWR from March 2005 to June 2019. Nine supervised ML algorithms were developed to preoperatively predict HR, SSOs, and 30-day readmission. Patient data were partitioned into training (80%) and testing (20%) sets. We identified 725 patients (52% women), with a mean age of 60 ± 11.5 years, mean body mass index of 31 ± 7 kg/m2, and mean follow-up time of 42 ± 29 months. The HR rate was 12.8%, SSO rate was 30%, and 30-day readmission rate was 10.9%. ML models demonstrated good discriminatory performance for predicting HR (area under the receiver operating characteristic curve [AUC] 0.71), SSOs (AUC 0.75), and 30-day readmission (AUC 0.74). ML models achieved mean accuracy rates of 85% (95% CI 80% to 90%), 72% (95% CI 64% to 80%), and 84% (95% CI 77% to 90%) for predicting HR, SSOs, and 30-day readmission, respectively. ML identified and characterized 4 unique significant predictors of HR, 12 of SSOs, and 3 of 30-day readmission. Decision curve analysis demonstrated that ML models have a superior net benefit regardless of the probability threshold. ML algorithms trained on readily available preoperative clinical data accurately predicted complications of AWR. Our findings support incorporating ML models into the preoperative assessment of patients undergoing AWR to provide data-driven, patient-specific risk assessment.


Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine through 6 Months.
BNT162b2 is a lipid nanoparticle-formulated, nucleoside-modified RNA vaccine encoding a prefusion-stabilized, membrane-anchored severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) full-length spike protein. BNT162b2 is highly efficacious against coronavirus disease 2019 (Covid-19) and is currently approved, conditionally approved, or authorized for emergency use worldwide. At the time of initial authorization, data beyond 2 months after vaccination were unavailable. In an ongoing, placebo-controlled, observer-blinded, multinational, pivotal efficacy trial, we randomly assigned 44,165 participants 16 years of age or older and 2264 participants 12 to 15 years of age to receive two 30-μg doses, at 21 days apart, of BNT162b2 or placebo. The trial end points were vaccine efficacy against laboratory-confirmed Covid-19 and safety, which were both evaluated through 6 months after vaccination. BNT162b2 continued to be safe and have an acceptable adverse-event profile. Few participants had adverse events leading to withdrawal from the trial. Vaccine efficacy against Covid-19 was 91.3% (95% confidence interval [CI], 89.0 to 93.2) through 6 months of follow-up among the participants without evidence of previous SARS-CoV-2 infection who could be evaluated. There was a gradual decline in vaccine efficacy. Vaccine efficacy of 86 to 100% was seen across countries and in populations with diverse ages, sexes, race or ethnic groups, and risk factors for Covid-19 among participants without evidence of previous infection with SARS-CoV-2. Vaccine efficacy against severe disease was 96.7% (95% CI, 80.3 to 99.9). In South Africa, where the SARS-CoV-2 variant of concern B.1.351 (or beta) was predominant, a vaccine efficacy of 100% (95% CI, 53.5 to 100) was observed. Through 6 months of follow-up and despite a gradual decline in vaccine efficacy, BNT162b2 had a favorable safety profile and was highly efficacious in preventing Covid-19. (Funded by BioNTech and Pfizer; ClinicalTrials.gov number, NCT04368728.).


Regulation of the response regulator gene degU through the binding of SinR/SlrR and exclusion of SinR/SlrR by DegU in Bacillus subtilis.
Bacillus subtilis DegU is a response regulator of the DegS-DegU two-component regulatory system. Phosphorylated DegU (DegU-P) controls many genes and biological processes, such as exoprotease and γ-polyglutamic acid production, in addition to the degU gene, by binding to target gene promoters. Nonphosphorylated DegU and low levels of DegU-P are required for swarming motility and genetic competence. The DNA-binding repressors SinR and SlrR are part of a double-negative feedback loop and comprise the epigenetic switch governing biofilm formation. In this study, we found that SinR repressed degU. Furthermore, SlrR, which interacts with SinR through protein-protein interaction, seems to have an active role in degU expression in in vivo lacZ analysis. An in vitro transcription assay supported this observation. An electrophoretic mobility shift assay (EMSA) showed that SinR bound to the degU promoter and that SlrR formed a complex with SinR on the degU promoter. In EMSA, DegU-P excluded the SinR/SlrR complex but not SinR from the degU promoter in the presence of RNA polymerase. These findings suggest that DegU-P interacts with SlrR. In support of this hypothesis, disruption of the slrR gene resulted in decreased degU expression. This newly identified regulatory mechanism for degU is considered to be sequential transcription factor replacement.


[Mechanical vascular clips with an automatic stapler in reconstructive microsurgery. Apropos of 16 clinical cases].
From October 1995 through March 1997, we performed 30 microsurgical reconstructions and achieved microvascular anastomoses in 16 with an automatic stapler (VCS) which allows insertion of titanium clips in the everted vessel walls. The mean age of the patients (15 men, 1 woman) was 42 years. Mean vessel diameter of the donor and recipient sites was 2 mm for the arteries and 3 mm for the veins. Recipient vessels had suffered radiation damage in 3 cases. Micro-anastomoses were performed in 26 cases, with end-to-end sutures in 24 and end-to side sutures in 2. There were 11 arterial procedures including 3 bypasses and 15 venous procedures with 1 bypass. Peroperative thrombosis occurred in 2 cases and were treated by undoing the anastomoses and resuturing manually. There were no early or late postoperative vascular complications. This clinical experience confirms the advantages of this mechanical approach to microanastomotic procedures for small vessels as previously demonstrated in experimental work: a system avoiding transfixation, rapid procedure, reliability.


[Thyroid status of the body in leukemias].
Disordered thyroid status was detected in patients with various clinical and morphological forms of leukemia and with hypoplastic anemia. These disorders present as low triiodothyronine blood concentration and a deficit of this hormone in the body. A certain correlation has been detected between the degree of the thyroid status disorders and the leukemic process malignancy.


Incidental multifocal calcifying pseudoneoplasm of the neuraxis: case report and literature review.
Calcifying pseudoneoplasm of the neuraxis (CAPNON) is thought to be a rare tumefactive lesion with unknown pathogenesis. Its prevalence is questionable with few previously reported cases of incidental CAPNON, and likely underdiagnosis. We report a unique case of incidental multifocal CAPNON. A 64-year-old female was admitted with loss of consciousness due to a ruptured right middle cerebral artery aneurysm with subarachnoid and intraventricular hemorrhage. She has a craniotomy and clipping. At time of operation, numerous small dural-based nodules were found, and one was excised for biopsy and was diagnosed as CAPNON. Retrospective review of her CT images identified nodules that were all ipsilateral to the ruptured aneurysm. A literature review revealed that incidental and/or multifocal CAPNONs are rare but likely underreported. Our case suggests a reactive process in the pathogenesis of CAPNON.


Impact of Pharmacist-Led Drug Therapy Management Services on HbA1c Values in a Predominantly Hispanic Population Visiting an Outpatient Endocrinology Clinic.
To assess the impact pharmacists have on improving glycemic control among predominantly Hispanic diabetic patients visiting an endocrinology clinic in South Texas. Pharmacists were recently integrated into this clinic to be part of a collaborative team. This study follows a retrospective cohort design. All patients received diabetic care from endocrinologists, and some received pharmacist-led drug therapy management (PDTM). Patients with ≥1 PDTM were categorized as the intervention group and those without PDTM as the standard of care (SOC) group. The outcome variables were the mean absolute change in glycosylated hemoglobin (HbA1c) from baseline and the proportion of patients at goal HbA1c (<7%) postintervention. Data were collected from 222 patients (n = 120 SOC patients, n = 102 PDTM patients). The mean age was 61 ± 14 years, 136 (61%) were female, and 197 (89%) were Hispanic. The mean absolute change in HbA1c was -1.3%. In the adjusted model, the mean absolute change in HbA1c in the PDTM compared to the SOC group was not significant (-0.1% ± 0.2%; P < .74), and concurrent interventions from registered dieticians (RDs) and licensed professional counselors (LPC) were identified as effect modifiers of the association. The stratum specific analysis identified the greatest decrease in HbA1c when the three interventions (ie, PDTM, RD, and LPC) coincided (-1.0% ± 0.3%; P < .01). Postintervention, 25% of those who received PDTM achieved an HbA1c<7% as compared to 19% in the SOC group. The clinical importance of pharmacists is enhanced when integrated with behavioral modifying programs to achieve additional improvement in HbA1c.


Multimodal Machine Learning Workflows for Prediction of Psychosis in Patients With Clinical High-Risk Syndromes and Recent-Onset Depression.
Diverse models have been developed to predict psychosis in patients with clinical high-risk (CHR) states. Whether prediction can be improved by efficiently combining clinical and biological models and by broadening the risk spectrum to young patients with depressive syndromes remains unclear. To evaluate whether psychosis transition can be predicted in patients with CHR or recent-onset depression (ROD) using multimodal machine learning that optimally integrates clinical and neurocognitive data, structural magnetic resonance imaging (sMRI), and polygenic risk scores (PRS) for schizophrenia; to assess models' geographic generalizability; to test and integrate clinicians' predictions; and to maximize clinical utility by building a sequential prognostic system. This multisite, longitudinal prognostic study performed in 7 academic early recognition services in 5 European countries followed up patients with CHR syndromes or ROD and healthy volunteers. The referred sample of 167 patients with CHR syndromes and 167 with ROD was recruited from February 1, 2014, to May 31, 2017, of whom 26 (23 with CHR syndromes and 3 with ROD) developed psychosis. Patients with 18-month follow-up (n = 246) were used for model training and leave-one-site-out cross-validation. The remaining 88 patients with nontransition served as the validation of model specificity. Three hundred thirty-four healthy volunteers provided a normative sample for prognostic signature evaluation. Three independent Swiss projects contributed a further 45 cases with psychosis transition and 600 with nontransition for the external validation of clinical-neurocognitive, sMRI-based, and combined models. Data were analyzed from January 1, 2019, to March 31, 2020. Accuracy and generalizability of prognostic systems. A total of 668 individuals (334 patients and 334 controls) were included in the analysis (mean [SD] age, 25.1 [5.8] years; 354 [53.0%] female and 314 [47.0%] male). Clinicians attained a balanced accuracy of 73.2% by effectively ruling out (specificity, 84.9%) but ineffectively ruling in (sensitivity, 61.5%) psychosis transition. In contrast, algorithms showed high sensitivity (76.0%-88.0%) but low specificity (53.5%-66.8%). A cybernetic risk calculator combining all algorithmic and human components predicted psychosis with a balanced accuracy of 85.5% (sensitivity, 84.6%; specificity, 86.4%). In comparison, an optimal prognostic workflow produced a balanced accuracy of 85.9% (sensitivity, 84.6%; specificity, 87.3%) at a much lower diagnostic burden by sequentially integrating clinical-neurocognitive, expert-based, PRS-based, and sMRI-based risk estimates as needed for the given patient. Findings were supported by good external validation results. These findings suggest that psychosis transition can be predicted in a broader risk spectrum by sequentially integrating algorithms' and clinicians' risk estimates. For clinical translation, the proposed workflow should undergo large-scale international validation.


Folding of the hydrogen bond network of H(+)(CH3OH)7 with rare gas tagging.
A number of isomer structures can be formed in hydrogen-bonded clusters, reflecting the essential variety of structural motifs of hydrogen bond networks. Control of isomer distribution of a cluster is important not only in practical use for isomer-specific spectroscopy but also in understanding of isomerization processes of hydrogen bond networks. Protonated methanol clusters have relatively simple networks and they are model systems suitable to investigate isomer distribution changes. In this paper, isomer distribution of H(+)(CH(3)OH)(7) is studied by size-selective infrared spectroscopy in the OH and CH stretching vibrational region and density functional theory calculations. While the clusters produced by a supersonic jet expansion combined with electron ionization were predominantly isomers having open hydrogen bond networks such as a linear chain, the Ar or Ne attachment (so-called rare gas tagging) entirely switches the isomer structures to compactly folded ones, which are composed only of closed multiple rings. The origin of the isomer switching is discussed in terms of thermal effects and specific isomer preference.


Dietary lignan intakes and risk of pre- and postmenopausal breast cancer.
Lignans are plant compounds metabolized in the mammalian gut to produce the phytoestrogens enterolactone and enterodiol. Because estrogens have been linked to breast cancer etiology, lignans could affect breast cancer risk through modulation of endogenous estrogen metabolism or competitive inhibition with estrogen receptors. We examined breast cancer risk and dietary lignan intake in a population-based case-control study of 1,122 women with primary, incident, histologically confirmed breast cancer and 2,036 controls frequency matched to cases on age and county of residence as part of the Western New York Exposures and Breast Cancer (WEB) Study. Diet was assessed with a self-administered 104-item food frequency questionnaire and other relevant data were collected by detailed in-person interviews. Lignans were expressed as the sum of the dietary precursors secoisolariciresinol and matairesinol. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated by unconditional logistic regression, adjusting for age, total energy and other breast cancer risk factors. Premenopausal women in the highest quartile of dietary lignan intake had reduced breast cancer risk (OR = 0.66; 95% CI = 0.44-0.98). No association was observed between lignan intakes and postmenopausal breast cancer. Our results suggest that dietary lignans may be important in the etiology of breast cancer, particularly among premenopausal women.


The Texas Medication Algorithm Project Patient and Family Education Program: a consumer-guided initiative.
Educating patients with mental illness and their families about the illness and its treatment is essential to successful medication (disease) management. Specifically, education provides patients and families with the background they need to participate in treatment planning and implementation as full "partners" with clinicians. Thus, education increases the probability that appropriate and accurate treatment decisions will be made and that a treatment regimen will be followed. The Texas Medication Algorithm Project (TMAP) has incorporated these concepts into its philosophy of care and accordingly created a Patient and Family Education Program (PFEP) to complement the utilization of medication algorithms for the treatment of schizophrenic, bipolar, and major depressive disorders. This article describes how a team of mental health consumers, advocates, and professionals developed and implemented the PFEP. In keeping with the TMAP philosophy of care, consumers were true partners in the program's development and implementation. They not only created several components of the program and incorporated the consumer perspective, but they also served as program trainers and advocates. Initially, PFEP provides basic and subsequently more in-depth information about the illness and its treatment, including such topics as symptom monitoring and management and self-advocacy with one's treatment team. It includes written, pictorial, videotaped, and other media used in a phased manner by clinicians and consumer educators, in either individual or group formats.


Trophoblasts, invasion, and microRNA.
MicroRNAs (miRNAs) have recently become essential actors in various fields of physiology and medicine, especially as easily accessible circulating biomarkers, or as modulators of cell differentiation. To this respect, terminal differentiation of trophoblasts (the characteristic cells of the placenta in Therian mammals) into syncytiotrophoblast, villous trophoblast, or extravillous trophoblast constitutes a good example of such a choice, where miRNAs have recently been shown to play an important role. The aim of this review is to provide a snapshot of what is known today in placentation mechanisms that are mediated by miRNA, under the angles of materno-fetal immune dialog regulation, trophoblast differentiation, and angiogenesis at the materno-fetal interface. Also, two aspects of regulation of these issues will be highlighted: the part played by oxygen concentration and the specific function of imprinted genes in the developing placenta.


Lil3 Assembles with Proteins Regulating Chlorophyll Synthesis in Barley.
The light-harvesting-like (LIL) proteins are a family of membrane proteins that share a chlorophyll a/b-binding motif with the major light-harvesting antenna proteins of oxygenic photoautotrophs. LIL proteins have been associated with the regulation of tetrapyrrol biosynthesis, and plant responses to light-stress. Here, it was found in a native PAGE approach that chlorophyllide, and chlorophyllide plus geranylgeraniolpyrophosphate trigger assembly of Lil3 in three chlorine binding fluorescent protein bands, termed F1, F2, and F3. It is shown that light and chlorophyllide trigger accumulation of protochlorophyllide-oxidoreductase, and chlorophyll synthase in band F3. Chlorophyllide and chlorophyll esterified to geranylgeraniol were identified as basis of fluorescence recorded from band F3. A direct interaction between Lil3, CHS and POR was confirmed in a split ubiquitin assay. In the presence of light or chlorophyllide, geranylgeraniolpyrophosphate was shown to trigger a loss of the F3 band and accumulation of Lil3 and geranylgeranyl reductase in F1 and F2. No direct interaction between Lil3 and geranylgeraniolreductase was identified in a split ubiquitin assay; however, accumulation of chlorophyll esterified to phytol in F1 and F2 corroborated the enzymes assembly. Chlorophyll esterified to phytol and the reaction center protein psbD of photosystem II were identified to accumulate together with psb29, and APX in the fluorescent band F2. Data show that Lil3 assembles with proteins regulating chlorophyll synthesis in etioplasts from barley (Hordeum vulgare L.).


Quantitative Read-across structure-activity relationship (q-RASAR): A new approach methodology to model aquatic toxicity of organic pesticides against different fish species.
We have developed quantitative toxicity prediction models for organic pesticides of agricultural importance considering different fish species using a novel quantitative Read-across structure-activity relationship (q-RASAR) approach. The current study uses experimental (Log 1/LC50) data of organic pesticides to various fish species, including Rainbow trout (RT: Oncorhynchus mykiss: 715 data points), Lepomis (LP: Lepomis macrochirus: 136 data points), and Miscellaneous (Pimephales promelas, Brachydanio rerio: 226 data points). This study has also discussed the validation of the developed models and the analysis of structural features that are important for aquatic toxicity towards fishes. The read-across-derived similarity, error, and concordance measures (RASAR descriptors) have been extracted from the preliminary 0D-2D descriptors; the combined pool of RASAR and selected 0D-2D descriptors have been used to develop the final models by employing partial least squares algorithm. All the q-RASAR models are acceptable in terms of goodness of fit, robustness, and external predictivity, superseding the quality of the respective QSAR models, as seen from the computed validation metrics. The q-RASAR is an effective approach that has the potential to be used as a good alternative way to enhance external predictivity, interpretability, and transferability for aquatic toxicity prediction as well as ecotoxicity potential identification.


High-dose cyclophosphamide without stem cell rescue in scleroderma.
To investigate the safety and tolerability of high-dose cyclophosphamide without stem cell rescue in scleroderma. An open-label, single-site, uncontrolled study design entered patients with active diffuse cutaneous scleroderma. Patients were treated with cyclophosphamide (50 mg/kg) intravenously daily for 4 consecutive days (total 200 mg/kg) followed by granulocyte colony-stimulating factor (5 microg/kg/day). The primary clinical efficacy end point was the modified Rodnan skin score (mRSS). Secondary end points included the Health Assessment Questionnaire-Disability Index (HAQ-DI), physician global assessment (PGA) and pulmonary function tests. Six patients (4 men, 2 women) aged 19-60 years were entered into the study. One patients died early in the protocol, thus five patients had follow-up data. The percentage reduction of the mRSS in these five evaluable patients within 1 month of treatment was 60%, 55%, 41%, 31% and 0%. The patient with no decline in skin score at 1 month showed a decrease in skin score from 41 to 26 by the 3-month visit, a 37% improvement. Three patients sustained the improvement after treatment for 24, 12 and 12 months. Two patients relapsed at 12 and 6 months after treatment. The PGA and HAQ-DI scores improved in five of the six patients by 72% and 79% respectively at 3 months. The only serious adverse event was a death that occurred owing to infection after neutrophil count recovery. High-dose cyclophosphamide without stem cell rescue can lead to a clinically significant improvement in skin score and measures of disease severity in patients with diffuse cutaneous scleroderma.


Puromycin-peptide bond formation with reticulocyte initiation factors M1 and M2.
The ability to form a "peptide" bond between various forms of Met-tRNA or Phe-tRNA and puromycin has been studied in the reticulocyte cell-free system. When Met-tRNA(F), fMet-tRNA(F), or N-acetylPhe-tRNA are used as substrate at low Mg(++) concentration (3 mM), reticulocyte initiation factors M(1) and M(2) (M(2A) + M(2B)) are required for puromycin-peptide synthesis. In contrast to bacterial systems, this reaction is also stimulated by the elongation factor T(1). When Met-tRNA(M) or Phe-tRNA is used as substrate, there is no M-factor requirement for the puromycin reaction; T(1) is absolutely required, and the reaction is stimulated by T(2). These studies indicate that reticulocyte factors M(1) and M(2) may function in part by placing the initiator tRNA into the P site. The detailed mechanism for mammalian initiation, however, may be more complex than that for bacterial systems.


An alternative scoring system to predict risk for surgical site infection complicating coronary artery bypass graft surgery.
To analyze the risk factors for surgical site infection (SSI) complicating coronary artery bypass graft (CABG) surgery and to create an alternative SSI risk score based on the results of multivariate analysis. A prospective cohort study involving inpatient and laboratory-based surveillance of patients who underwent CABG surgery over a 27-month period from January 1, 2003 through March 31, 2005. Data were obtained from 6 acute care hospitals in Victoria, Australia, that contributed surveillance data for SSI complicating CABG surgery to the Victorian Hospital Acquired Infection Surveillance System Coordinating Centre and the Australasian Society of Cardiac and Thoracic Surgeons, also in Victoria. A total of 4,633 (93%) of the 4,987 patients who underwent CABG surgery during this period were matched in the 2 systems databases. There were 286 SSIs and 62 deep or organ space sternal SSIs (deep or organ space sternal SSI rate, 1.33%). Univariate analysis revealed that diabetes mellitus, body mass index (BMI) greater than 35, and receipt of blood transfusion were risk factors for all types of SSI complicating CABG surgery. Six multivariate analysis models were created to examine either preoperative factors alone or preoperative factors combined with operative factors. All models revealed diabetes and BMI of 30 or greater as risk factors for SSI complicating CABG surgery. A new preoperative scoring system was devised to predict sternal SSI, which assigned 1 point for diabetes, 1 point for BMI of 30 or greater but less than 35, and 2 points for BMI of 35 or greater. Each point in the scoring system represented approximately a doubling of risk of SSI. The new scoring system performed better than the National Nosocomial Infections Surveillance System (NNIS) risk index at predicting SSI. A new weighted scoring system based on preoperative risk factors was created to predict sternal SSI risk following CABG surgery. The new scoring system outperformed the NNIS risk index. Future studies are needed to validate this scoring system.


Crystal growth of new hexahydroxometallates using a hydroflux.
A series of seven compounds, Sr2Mn(OH)6, Ba2Mn(OH)6, Sr2Co(OH)6, Ba2Co(OH)6, Sr2Ni(OH)6, Ba2Ni(OH)6, and Ba2Cu(OH)6, were synthesized using a low-melting hydroflux, a hybrid approach between aqueous hydrothermal and molten hydroxide flux techniques. Crystals of the hexahydroxometallates were obtained by dissolving appropriate amounts of alkaline-earth nitrates or hydroxides and transition-metal oxides, acetates, or chlorides in the hydroflux and reacting at 180-230 °C. The isostructural compounds all crystallize in the monoclinic space group P2(1/n). The monoclinic structure consists of isolated transition-metal octahedra within a three-dimensional framework of corner- and edge-shared eight-coordinate, alkaline-earth polyhedra. Magnetic susceptibility data show that all compounds are simple paramagnets. Thermogravimetric analysis indicates that these hydroxides lose water between 215 and 350 °C and transform into oxide products, the identity of which depends on the metal cations present in the parent hexahydroxometallates.


Increased incidence of gastrointestinal cancers among patients with pyogenic liver abscess: a population-based cohort study.
The relationship between pyogenic liver abscess (PLA) and gastrointestinal (GI) cancer was first reported more than 20 years ago, yet little is known about this connection. We evaluated this association in a population-based, retrospective, cohort study. Using Taiwan National Health Insurance claims data, we collected data on a cohort of 14,690 patients with PLA diagnosed from 2000 to 2007. A reference cohort of 58,760 persons without PLA (controls) was selected from the same database, frequency matched by age, sex, and index year. Both cohorts were followed up until the end of 2009, and incidences of GI cancer were calculated. The incidence of GI cancer was 4.30-fold higher among patients with PLA compared with controls (10.8 vs 2.51/1000 person-years). Site-specific analysis showed that the highest incidence of colorectal cancer was among patients with PLA and diabetes mellitus, followed by patients with PLA without diabetes and controls with diabetes (9.58, 5.76, and 1.49/10,000 person-years, respectively). The PLA cohort also had a high risk of small intestine cancer (adjusted hazard ratio [aHR], 12.7; 95% confidence interval [CI], 5.79-27.7) and biliary tract cancer (aHR, 9.56; 95% CI, 6.68-13.7). Their risk of pancreatic cancer (aHR, 2.51; 95% CI, 1.68-3.76) was also significant. However, patients with PLA did not have an increased risk of gastric cancer compared with controls. In a population-based study, we found that the incidence of GI cancer is increased more than 4-fold among patients with PLA compared with controls. PLA might therefore be an indicator of GI cancer. Patients with PLA had the highest incidence of colorectal cancer, followed by cancers of the biliary tract, pancreas, and small intestine.


Protein quality control systems associated with no-go and nonstop mRNA surveillance in yeast.
Quality control systems eliminate aberrant proteins derived from aberrant mRNAs. Two E3 ubiquitin ligases, Ltn1 and Not4, are involved in proteasomal protein degradation coupled to translation arrest. Here, we evaluated nonstop and translation arrest products degraded in a poly(A) tail-independent manner. Ltn1 was found to degrade aberrant nonstop polypeptides derived from nonstop mRNA lacking a termination codon, but not peptidyl-tRNA, even in the absence of the ribosome dissociation complex Dom34:Hbs1. The receptor for activated C kinase (RACK1/ASC1) was identified as a factor required for nascent peptide-dependent translation arrest as well as Ltn1-dependent protein degradation. Both Not4 and Ltn1 were involved in the degradation of various arrest products in a poly(A) tail-independent manner. Furthermore, carboxyl terminus-truncated degradation intermediates of arrest products were stabilized in a cdc48-3 mutant defective in unfolding or the disassembly related to proteasomal degradation. Thus, we propose that stalled ribosomes may be dissociated into subunits and that peptidyl-tRNA on the 60S subunit is ubiquitinated by Ltn1 and Cdc48 is required for the degradation following release from tRNA.


Effect of probiotic-fermented milk administration on gastrointestinal survival of Lactobacillus casei ATCC 393 and modulation of intestinal microbial flora.
The aim of the present study was to assess the survival of free and immobilized Lactobacillus casei ATCC 393 on apple pieces, contained in probiotic-fermented milk, after gastrointestinal (GI) transit and to investigate the potential regulation of intestinal microbial flora in a rat model. In in vitro GI stress tolerance tests, immobilized L. casei ATCC 393 exhibited significantly higher survival rates compared to free cells. At a second stage, probiotic-fermented milk produced by either free or immobilized cells was administered orally at a single dose or daily for 9 days in Wistar rats. By 12 h after single-dose administration, both free and immobilized cells were detected by microbiological and molecular analysis at levels ≥6 logCFU/g of feces. Moreover, daily administration led to significant reduction of staphylococci, enterobacteria, coliforms and streptococci counts. In conclusion, L. casei ATCC 393 contained in fermented milk survived GI transit and modulated intestinal microbiota.


VISTA: computational tools for comparative genomics.
Comparison of DNA sequences from different species is a fundamental method for identifying functional elements in genomes. Here, we describe the VISTA family of tools created to assist biologists in carrying out this task. Our first VISTA server at http://www-gsd.lbl.gov/vista/ was launched in the summer of 2000 and was designed to align long genomic sequences and visualize these alignments with associated functional annotations. Currently the VISTA site includes multiple comparative genomics tools and provides users with rich capabilities to browse pre-computed whole-genome alignments of large vertebrate genomes and other groups of organisms with VISTA Browser, to submit their own sequences of interest to several VISTA servers for various types of comparative analysis and to obtain detailed comparative analysis results for a set of cardiovascular genes. We illustrate capabilities of the VISTA site by the analysis of a 180 kb interval on human chromosome 5 that encodes for the kinesin family member 3A (KIF3A) protein.


Medications in the workplace.
This article provides guidance and web-based resources regarding two frequent practice issues associated with medications in the workplace: dispensing of over-the-counter (OTC) medications in the occupational health setting, and the proper disposal of expired or otherwise unwanted prescription and nonprescription medications.


Phthalates and neurotoxic effects on hippocampal network plasticity.
Phthalates are synthetically derived chemicals used as plasticizers in a variety of common household products. They are not chemically bound to plastic polymers and over time, easily migrate out of these products and into the environment. Experimental investigations evaluating the biological impact of phthalate exposure on developing organisms are critical given that estimates of phthalate exposure are considerably higher in infants and children compared to adults. Extensive growth and re-organization of neurocircuitry occurs during development leaving the brain highly susceptible to environmental insults. This review summarizes the effects of phthalate exposure on brain structure and function with particular emphasis on developmental aspects of hippocampal structural and functional plasticity. In general, it appears that widespread disruptions in hippocampal functional and structural plasticity occur following developmental (pre-, peri- and post-natal) exposure to phthalates. Whether these changes occur as a direct neurotoxic effect of phthalates or an indirect effect through disruption of endogenous endocrine functions is not fully understood. Comprehensive investigations that simultaneously assess the neurodevelopmental, neurotoxic, neuroendocrine and behavioral correlates of phthalate exposure are needed to provide an opportunity to thoroughly evaluate the neurotoxic potential of phthalates throughout the lifespan.


HMO direct costs and health care resources use after implementation of a monthly limit on sumatriptan.
The health care costs and resource use of patients with migraine before and after a quantity limit on sumatriptan was introduced in an HMO were compared. A longitudinal, retrospective review of a medical claims database and a pharmacy claims database was conducted for two six-month periods before and after a monthly limit (four tablets or injections) on sumatriptan reimbursement was instituted at an independent practice association-model HMO in February 1997. Patients with at least one medical claim with a diagnosis code for migraine or at least two pharmacy claims for sumatriptan, methysergide, ergotamine, dihydroergotamine, or an ergotamine combination product in 1996 or 1997 were eligible for inclusion. A total of 557 patients were included in the analysis. Migraine-related medical costs and total medical costs increased 1.5% and 24.4%, respectively; neither change was statistically significant. Physician office visits related to migraine increased by 7.8%. The number of hospital admissions for the cohort increased from three to five, but hospital costs decreased by 55.0%. The overall costs of medications for migraine therapy decreased by 4.5%. There was an 8.2% increase in prescriptions for drugs to treat migraine but a 40.0% decrease in their cost, primarily because of decreased sumatriptan use. There was a 33.9% increase in prescriptions for medications that could be used as prophylaxis for migraine and a 49.6% increase in their cost. Implementation of a monthly limit on sumatriptan decreased an HMO's pharmacy costs but did not significantly alter migraine-related direct medical costs and health care resource use of patients with migraine.


Potential in vivo roles of nucleic acid triple-helices.
The ability of double-stranded DNA to form a triple-helical structure by hydrogen bonding with a third strand is well established, but the biological functions of these structures remain largely unknown. There is considerable albeit circumstantial evidence for the existence of nucleic triplexes in vivo and their potential participation in a variety of biological processes including chromatin organization, DNA repair, transcriptional regulation, and RNA processing has been investigated in a number of studies to date. There is also a range of possible mechanisms to regulate triplex formation through differential expression of triplex-forming RNAs, alteration of chromatin accessibility, sequence unwinding and nucleotide modifications. With the advent of next generation sequencing technology combined with targeted approaches to isolate triplexes, it is now possible to survey triplex formation with respect to their genomic context, abundance and dynamical changes during differentiation and development, which may open up new vistas in understanding genome biology and gene regulation.


MAGI2/S-SCAM outside brain.
Membrane-associated guanylate kinase with an inverted arrangement of protein-protein interaction domains (MAGI)2 (also called synaptic scaffolding molecule (S-SCAM), atrophin-1-interacting protein 1, activin receptor-interacting protein 1) is a scaffold protein that binds a wide variety of receptors, cell adhesion molecules and signalling molecules. It also interacts with other scaffold proteins and adaptors, and forms a protein network that supports cell junctions. As it is highly expressed in brain, the study on its roles in synaptic organization initially preceded. However, mounting evidence indicates that MAGI2/S-SCAM functions as a tumour suppressor and plays essential roles to maintain the integrity of cell structures in non-neuronal tissues. We review the articles regarding to MAGI2/S-SCAM outside brain and discuss future perspectives for the research of MAGI family proteins.


Congenital canine myasthenia gravis: I. Deficient junctional acetylcholine receptors.
The neuromuscular junction was studied during growth in two breeds of dog with a congenital familial form of myasthenia gravis (CMG): the Jack Russell terrier and the springer spaniel. Light microscopy revealed no difference in endplate size or nerve terminal morphology in age-matched CMG dogs and unaffected littermates. At all ages there was in individual muscle fibers of CMG dogs an approximately 75% lower postsynaptic membrane density of acetylcholine receptors (AChR). There was no evidence that this abnormality had an autoimmune basis. Measurements of miniature endplate potential amplitude in relation to fiber diameter revealed that the low density of AChR in the postsynaptic membrane of CMG dogs remained constant and did not change with the marked progression of muscle weakness during growth. Antigenic determinants of AChR were deficient in CMG muscle to the same extent as alpha-bungarotoxin (alpha BT) binding sites. Thus, a low density of AChR in the muscle's postsynaptic membrane appears to be the principal abnormality of CMG. The data suggest that the normal increase in number of acetylcholine (ACh) quanta released by nerve impulse may not fully compensate for the reduction in depolarization produced by a single quantum as the muscle fiber diameter increases. This could cause progression of weakness during growth. In addition to being a useful animal model of a rare form of CMG that resembles one form of human CMG, canine CMG offers a unique model for investigating events of synaptogenesis at the neuromuscular junction.


Old Mice Have Less Transcriptional Activation But Similar Periosteal Cell Proliferation Compared to Young-Adult Mice in Response to in vivo Mechanical Loading.
Mechanical loading is a potent strategy to induce bone formation, but with aging, the bone formation response to the same mechanical stimulus diminishes. Our main objectives were to (i) discover the potential transcriptional differences and (ii) compare the periosteal cell proliferation between tibias of young-adult and old mice in response to strain-matched mechanical loading. First, to discover potential age-related transcriptional differences, we performed RNA sequencing (RNA-seq) to compare the loading responses between tibias of young-adult (5-month) and old (22-month) C57BL/6N female mice following 1, 3, or 5 days of axial loading (loaded versus non-loaded). Compared to young-adult mice, old mice had less transcriptional activation following loading at each time point, as measured by the number of differentially expressed genes (DEGs) and the fold-changes of the DEGs. Old mice engaged fewer pathways and gene ontology (GO) processes, showing less activation of processes related to proliferation and differentiation. In tibias of young-adult mice, we observed prominent Wnt signaling, extracellular matrix (ECM), and neuronal responses, which were diminished with aging. Additionally, we identified several targets that may be effective in restoring the mechanoresponsiveness of aged bone, including nerve growth factor (NGF), Notum, prostaglandin signaling, Nell-1, and the AP-1 family. Second, to directly test the extent to which periosteal cell proliferation was diminished in old mice, we used bromodeoxyuridine (BrdU) in a separate cohort of mice to label cells that divided during the 5-day loading interval. Young-adult and old mice had an average of 15.5 and 16.7 BrdU+ surface cells/mm, respectively, suggesting that impaired proliferation in the first 5 days of loading does not explain the diminished bone formation response with aging. We conclude that old mice have diminished transcriptional activation following mechanical loading, but periosteal proliferation in the first 5 days of loading does not differ between tibias of young-adult and old mice. © 2020 American Society for Bone and Mineral Research.


Functional changes in the regulatory subunit of the type II cyclic adenosine 3':5'-monophosphate-dependent protein kinase isozyme during normal and neoplastic lung development.
The abilities of cyclic adenosine 3':5'-monophosphate (cAMP) and cyclic 8-azidoadenosine 3':5'-[32P]monophosphate (8-N3-[32P]cAMP) to bind to the regulatory subunit (RII) of the type II cAMP-dependent protein kinase isozyme and to cause subsequent dissociation of the holoenzyme were compared in extracts from adult and neonatal mouse lung and lung adenoma. RII in extracts from adult lung exhibits equal numbers of high- (Kd 15 nM) and low- (Kd 230 nM) affinity 8-N3-[32P]cAMP binding sites. In the neonate, the proportion of high-affinity sites is reduced to 20% while, in lung adenoma, only low-affinity RII binding is observed. Low-affinity RII binding is correlated with an inability of cAMP to dissociate the type II holoenzyme completely. Sucrose gradient sedimentation of adult lung cytosol in the presence of cAMP shows complete dissociation of the type I isozyme, while only some of the type II holoenzyme is dissociated. This is in contrast to the case with lung tumor cytosol, in which only low-affinity binding is observed and no apparent dissociation of the type II isozyme occurs. cAMP does promote RII dephosphorylation within the holoenzyme, however, suggesting that cAMP can bind to RII without dissociating the tetramer. Consistent with this interpretation, photoincorporation of 8-N3-[32P]cAMP prior to sucrose gradient sedimentation results in the formation of a photolabeled RII complex which sediments at the same rate as does the holoenzyme. Two-dimensional gel electrophoresis of RII photolabeled at low and high concentrations of 8-N3-[32P]cAMP suggests that these altered binding and dissociation characteristics of the type II isozyme are not due to the presence of a structurally altered RII molecule. After DEAE-cellulose chromatography of lung cytosol, only high-affinity RII binding is observed, and all of the RII can now be dissociated with cAMP. Low-affinity binding may thus reflect either an altered conformational state of RII or the interaction of the type II kinase with other cytosolic molecules which can affect RII binding and dissociation without altering the functional properties of the type I isozyme.


[Identification methods of crop and weeds based on Vis/NIR spectroscopy and RBF-NN model].
The automated recognition of crop and weed by using Vis/NIR spectral in field is one of hottest research branches of agriculture engineering. If the recognition is efficient and effective, then the variate operations of herbicide or fertilizer spraying in field could be realized. Many researches have pointed out that the reflectance rate of green plant leaves could be used to identify the varieties. As the colors and surface textures of crop and weed were change in different living phases, these changes may exert great influence on the reflectance spectral of plant leaves. Vis/NIR spectra of three weeds and one crop in two different terms were recorded by spectral meter ASD FieldSpec Pro FR. Its wave band is from 325 to 1 075 nm. The scan time was 270 ms. The scanning times of per sample was set to 30 times. Firstly, 23 days after the planting of soybean, some soybean leaves and weeds leave were picked from the field, and brought to lab to record spectral. The lighting condition was controlled by an artificial halogen bulb. Secondly, on the 45th day, the same experiment was done. The three weeds were goose grass, alligator alternanthera and emarginate amaranth. The crop was soybean seedling. Totally 378 samples were drawn for two terms. As one original reflectance spectrum contains 651 float numbers, the total data volume was huge. Using wavelet transform to compress data volume and extract characteristic spectral data was tried. The result was 114 float numbers per sample. Among them, 250 samples from two terms were used as input data to build artificial neural network model, and those left were used to check the validation. Radial basis function neural network model is widely used in pattern recognition problems. It is a nonlinear and self adaptive parallel. By assigning a 1 by 4 vector to each spectral samples, the source data could be used to build an RBF-NN model. All the samples were assigned these standard output data. Then, the left 128 samples were used to check the performance of the model. The result is that only 3 samples from the second term of goose grass were wrongly classified as alligator alternanthera, which showed that RBF neural network have strong ability to differentiate spectra of species of plant, and that there was no massive difference of NIR spectra of one plant in different life periods. This demonstrated that the NIR spectra could be used to identify crop from weed with no need to care about the living stages of these plants.


Interpenetrating Polymer Network HA/Alg-RGD Hydrogel: An Equilibrium of Macroscopic Stability and Microscopic Adaptability for 3D Cell Growth and Vascularization.
Two-dimensional (2D) cell culture methods dominate the current research. However, the inherent responsiveness of cells to their native three-dimensional (3D) microenvironment necessitates a paradigm shift toward the development of advanced hydrogels that faithfully mimic the intricacies of the extracellular matrix (ECM) and enable continuous cell-ECM interactions. To address the constraints of traditional static hydrogel networks that impede effective cell-matrix and cell-cell interactions, and to tackle the inherent stability issues associated with dynamically cross-linked hydrogels, which have become a pressing concern. Herein, we present an interpenetrating polymer network (IPN) hydrogel (HA/Alg-RGD hydrogel) that combines a physically cross-linked network between alginate and calcium ions (Alg-Ca2+) for the enhanced cell growth adaptability with a chemically cross-linked hyaluronic acid (HA) network to ensure macroscopic stability during cell culture. The incorporation of arginine-glycine-aspartic peptide modified alginate (Alg-RGD) further facilitates cell adhesion and improves the cell-hydrogel interaction. Notably, this IPN hydrogel demonstrates mechanical stability and enables cell spreading and growth within its structural framework. Leveraging the reversible characteristics of the ionically cross-linked Alg-Ca2+ network within IPN hydrogels, we demonstrate the feasibility of the gelatin sacrificial solution for 3D printing purposes within the hydrogel matrix. Subsequent UV-induced covalent cross-linking enables the fabrication of vascularized microfluidic channels within the resulting construct. Our results demonstrate endothelial cell spreading and spontaneous cell sprouting within the hydrogel matrix, thus highlighting the efficacy of this IPN hydrogel system in facilitating 3D cell growth. Additionally, our study emphasizes the potential of 3D printed constructs as a promising approach for vascularization in tissue engineering. The importance of RGD peptides in promoting favorable cell-hydrogel scaffold interactions is also highlighted, emphasizing their critical role in optimizing biomaterial-cell interfaces.