Implementation of the 2015 European Society of Cardiology guidelines for the management of infective endocarditis in the Netherlands.
Because the occurrence of infective endocarditis (IE) continues to be associated with high mortality, a working group was created by the Dutch Society of Cardiology to examine how the most recent European Society of Cardiology (ESC) guidelines for IE management could be implemented most effectively in the Netherlands. In order to investigate current Dutch IE practices, the working group conducted a country-wide survey. Based on the results obtained, it was concluded that most ESC recommendations could be endorsed, albeit with some adjustments. For instance, the suggested pre-operative screening and treatment of nasal carriers of Staphylococcus aureus as formulated in the ESC guideline was found to be dissimilar to current Dutch practice, and was therefore made less restrictive. The recently adapted ESC diagnostic criteria for IE were endorsed, while the practical employment of the relevant diagnostic techniques was simplified in an adapted flowchart. In addition, the presence of a multidisciplinary, so-called 'endocarditis team' in tertiary centres was proposed as a quality indicator. An adapted flowchart specifically tailored to Dutch practice for microbiological diagnostic purposes was constructed. Lastly, the working group recommended the Stichting Werkgroep Antibioticabeleid (SWAB; Dutch Working Party on Antibiotic Policy) guidelines for IE treatment instead of the antibiotic regimens proposed by the ESC.


The potential molecular targets of marine sulfated polymannuroguluronate interfering with HIV-1 entry. Interaction between SPMG and HIV-1 rgp120 and CD4 molecule.
The potential targets of marine sulfated polymannuroguluronate (SPMG) involved in inhibition of HIV-1 entry were investigated by surface plasmon resonance and flow cytometry. Results indicated that binding of SPMG either to soluble oligomeric rgp120 or to complexed rgp120-sCD4 mainly resided in V3 loop region. In addition, SPMG was shown to be less accessible for sCD4 when sCD4 had pre-interacted with rgp120, though SPMG per se multivalently bound to sCD4 with relatively low affinity. While the pre-incubation of SPMG with rgp120 caused a partial blockade of rgp120 binding to sCD4, suggesting that SPMG either shared common binding sites on gp120 with sCD4 or masked the docking sites of gp120 for sCD4. Taken together, V3 domain was demonstrated to be the major site mediating interaction of SPMG with complexed rgp120-sCD4. It seems likely that SPMG binds to both rgp120 and sCD4, but has less accessibility for sCD4 when sCD4 has already bound to rgp120. Nevertheless, addition of SPMG either prior to or after the interaction of rgp120 with sCD4 may suppress rgp120 binding to sCD4. The exact pattern of this trimolecular complex formation at the cell membrane-anchored virus level requires further clarification.


Coat protein expression strategy of maize rayado fino virus and evidence for requirement of CP1 for leafhopper transmission.
Marafiviruses, including maize rayado fino virus (MRFV) and oat blue dwarf virus (OBDV), encode two carboxy co-terminal coat proteins, CP1 and CP2, which encapsidate the genome to form icosahedral virions. While CP2 expression is expected to be solely driven from a second start codon of a subgenomic RNA under a marafibox promoter sequence, the larger CP1 with an in-frame N-terminal extension relative to CP2 could potentially be expressed either by proteolytic release from the MRFV polyprotein or from subgenomic RNA translation. We examined MRFV CP expression strategy with a series of mutations in the CP coding region and identified mutants viable and nonviable for systemic plant infection. Polyprotein expression of MRFV CP1 was minimal. Mutants blocking CP2 expression failed to establish systemic infection, while mutants depleted in CP1 exhibited systemic infection and formation of virus-like particles but lost leafhopper transmissibility, indicating that CP1 is required for leafhopper transmission.


Isopimpinellin extends antiangiogenic effect through overexpression of miR-15b-5p and downregulating angiogenic stimulators.
Angiogenesis is the formation of new blood vessels from an existing vasculature through a series of processes such as activation, proliferation, and directed migration of endothelial cells. Angiogenesis is instrumental in the metastatic spread of tumors. Isopimpinellin, a furanocoumarin group of phytochemicals, is an anticarcinogenic agent. However, no studies have proven its antiangiogenic effects. The current study thus aimed to screen the antiangiogenic effect of isopimpinellin. Human Umblical Vein Endothelial Cell (HUVEC) as an in vitro model and zebrafish embryos as an in vivo model was used in this study. The experimental results showed that isopimpinellin effectively inhibited HUVEC proliferation, invasion, migration, and tube formation, which are the key steps in angiogenesis by markedly suppressing the expression of pro-angiogenic genes VEGF, AKT, and HIF-1α. In addition, isopimpinellin exerts its anti-angiogenic effect through the regulation of miR-15b-5p and miR-542-3p. Furthermore, in zebrafish embryos, isopimpinellin inhibited the development of intersegmental vessels (ISVs) through the significant downregulation of all pro-angiogenic genes vegf, vegfr2, survivin, angpt-1, angpt-2, and tie-2. Collectively, these experimental findings offer novel insights into the antiangiogenic nature of isopimpinellin and open new avenues for therapeutic approaches.


Pediatric Considerations in Endoscopic Retrograde Cholangiopancreatography.
Endoscopic retrograde cholangiopancreatography is a technically challenging endoscopic technique that provides a minimally invasive way of evaluating and treating pathologic abnormality in the bile ducts and pancreas. Its utilization in children is increasing rapidly, broadening the understanding of its pediatric indications, clinical utility, and technical limitations. This article updates providers about specific considerations of endoscopic retrograde cholangiopancreatography in children as they relate to appropriate indications, patient preparation, available equipment, as well as expected technical and clinical outcomes following the procedure in pediatric populations.


2D-QSAR, docking, molecular dynamics, studies of PF-07321332 analogues to identify alternative inhibitors against 3CLpro enzyme in SARS-CoV disease.
Given the results of the Pfizer-developed inhibitor PF-07321332 in the treatment of the SARS-Covid-19 epidemic, we aimed to identify potential alternatives to this compound by utilizing various methods; we developed 2 D-QSAR models to predict the therapeutic activity of 78 analogues of PF-07321332, three statistical learning techniques including (MLP-ANN), (SVR), and (MLR) were exploited. Various validation approaches were applied to the three models developed following the use of five most relevant descriptors. The study of the characteristics of these descriptors proved that the inhibitory activity of PF-07321332 analogues is specifically affected by the structure of the molecule, its polarizability, and by the hydrogen bonds. The best model, named MLP-ANN (with a 5-3-1 architecture), was selected on the basis of the following statistical parameters: r2 = 0.922, Q2 = 0.921. In addition, we performed a molecular docking and a molecular dynamics analysis of these compounds. The obtained results confirm that compound 8 can be a good alternative for compound PF-07321332.Communicated by Ramaswamy H. Sarma.


A 2,000-year-old specimen with intraerythrocytic Bartonella quintana.
Photogrammetry and cascading microscopy investigations of dental pulp specimens collected from 2,000-year-old individuals buried in a Roman necropolis in Besançon, France, revealed unprecedented preserved tissular and cellular morphology. Photogrammetry yielded 3-D images of the smallest archaeological human remains ever recovered. Optical microscopy examinations after standard haematoxylin-phloxine-saffron staining and anti-glycophorin A immunohistochemistry exposed dental pulp cells, in addition erythrocytes were visualised by electron microscopy, which indicated the ancient dental pulp trapped a blood drop. Fluorescence in situ hybridisation applied on red blood cells revealed the louse-borne pathogen Bartonella quintana, a finding confirmed by polymerase chain reaction assays. Through paleohistology and paleocytology, we demonstrate that the ancient dental pulp preserved intact blood cells at the time of the individual's death, offering an unprecedented opportunity to engage in direct and indirect tests to diagnose pathogens in ancient buried individuals.


Integrating T cell metabolism in cancer immunotherapy.
Activation and maintenance of the T cell response occurs concurrently with metabolic reprogramming. This ensures the T cell response is supported by sufficient energy and substrates necessary for cell survival, growth and proliferation. Different metabolic programs are associated with differentiation into different cell subsets, effector function and development of long-lasting memory. This provides an opportunity to improve the T cell response through manipulation of metabolism, which is instrumental to ameliorate the current protocols for cancer immunotherapy. Using drugs and molecules targeting selective metabolic pathways it is now possible to generate T cells that can mount a durable and stable anti-tumor response. On the other hand, cancer cells can take advantage of the metabolic requirements of T cells to evade the immune response. In this brief review we discuss recent findings of T cell metabolism in quiescence and activation, how the tumor microenvironment can affect T cell metabolism, and how T cell metabolism can be manipulated to improve the T cell response to tumors.


Angiographic morphology of recurrent stenoses after percutaneous transluminal coronary angioplasty: are lesions longer at restenosis?
Angiographic morphology was analysed in 32 patients who developed restenosis after initially successful coronary angioplasty. The mean minimal luminal diameter of the dilated coronary segments increased from 0.9 mm to 2.3 mm after dilatation, but decreased to 0.9 mm at restenosis. The reference diameter was unchanged after dilatation and at restenosis. Mean stenosis length before the initial angioplasty was 7.0 mm but at the repeat procedure had increased to 8.7 mm (mean increase 1.7 mm, 95% confidence interval 0.6 to 2.8 mm, p less than 0.01). There were no significant differences in mean trans-stenotic pressure gradient and mean eccentricity ratio between the initial and repeat angioplasty procedures. In individual patients the changes in stenosis morphology were unpredictable, but overall stenoses tended to be longer at restenosis. In some patients stenosis length increased by several millimetres but the success rate of repeat angioplasty was high and the clinical importance of the changes in stenosis morphology are uncertain.


The feasibility of using micro concentrated multiple reflection ATR FTIR accessory in pesticide analysis.
Micro-Multiple Reflection ATR (CMRATR) spectroscopy is a technique, using specialized equipment, which allows the enhanced sensitivity of multiple reflection ATR analysis of small amounts of liquids in a confined area hitherto reserved for single reflection equipment. This technique has demonstrated a high level of sensitivity, especially when used in conjunction with an evaporative technique. In this work, the technique will be used with a miniature CMRATR accessory, which has the added advantage of compatibility with the smallest current FTIR spectrometers, to analyze pesticides. The results presented here indicate that the CMRATR/evaporative technique can serve as both qualitative and quantitative support to the existing standard methodology.


Effects of thrombin, acetyl salicylic acid and prostaglandins on spontaneous efflux of norepinephrine and beta-thromboglobulin from platelets in calcium/magnesium deficient environment.
The effect of different substances on the spontaneous efflux of 14C-norepinephrine (14C-NE) and beta-thromboglobulin (beta TG) from platelets in Ca2+ - and Mg2+ - free buffer was tested. Addition of increasing concentrations of thrombin (0.015-0.625 U/ml) resulted in an immediate increase of 14C-NE detectable in the supernatants, but the slopes of the efflux curves from 10 min after thrombin addition were almost identical and parallel with the slope before thrombin addition. Also beta-thromboglobulin was steadily released after thrombin addition. Prostaglandin E1 and prostacyclin in concentrations that gave a 30-fold increase in cAMP accumulation resulted in a moderate decrease in efflux of 14C-NE from 10 to 30 min but did not influence the efflux thereafter up to 80 min. Acetyl salicylic acid in vivo and in vitro (200-500 mumol/l) did not influence the efflux of 14C-NE or beta TG. The conclusions are that factors known to exert considerable effects on aggregation, desaggregation and acute release produce only transitory effects in a Ca2+ - and Mg2+ - free environment and that the basal efflux of alpha-granule components seems to be a stable process.


Bioalerts: a python library for the derivation of structural alerts from bioactivity and toxicity data sets.
Assessing compound toxicity at early stages of the drug discovery process is a crucial task to dismiss drug candidates likely to fail in clinical trials. Screening drug candidates against structural alerts, i.e. chemical fragments associated to a toxicological response prior or after being metabolized (bioactivation), has proved a valuable approach for this task. During the last decades, diverse algorithms have been proposed for the automatic derivation of structural alerts from categorical toxicity data sets. Here, the python library bioalerts is presented, which comprises functionalities for the automatic derivation of structural alerts from categorical (dichotomous), e.g. toxic/non-toxic, and continuous bioactivity data sets, e.g. [Formula: see text] or [Formula: see text] values. The library bioalerts relies on the RDKit implementation of the circular Morgan fingerprint algorithm to compute chemical substructures, which are derived by considering radial atom neighbourhoods of increasing bond radius. In addition to the derivation of structural alerts, bioalerts provides functionalities for the calculation of unhashed (keyed) Morgan fingerprints, which can be used in predictive bioactivity modelling with the advantage of allowing for a chemically meaningful deconvolution of the chemical space. Finally, bioalerts provides functionalities for the easy visualization of the derived structural alerts.


Histone acetylation of bile acid transporter genes plays a critical role in cirrhosis.
Owing to the lack of genetic animal models that adequately recreate key clinical characteristics of cirrhosis, the molecular pathogenesis of cirrhosis has been poorly characterized, and treatments remain limited. Hence, we aimed to better elucidate the pathological mechanisms of cirrhosis using a novel murine model. We report on the first murine genetic model mimicking human cirrhosis induced by hepatocyte-specific elimination of microspherule protein 1 (MCRS1), a member of non-specific lethal (NSL) and INO80 chromatin-modifier complexes. Using this genetic tool with other mouse models, cell culture and human samples, combined with quantitative proteomics, single nuclei/cell RNA sequencing and chromatin immunoprecipitation assays, we investigated mechanisms of cirrhosis. MCRS1 loss in mouse hepatocytes modulates the expression of bile acid (BA) transporters - with a pronounced downregulation of Na+-taurocholate cotransporting polypeptide (NTCP) - concentrating BAs in sinusoids and thereby activating hepatic stellate cells (HSCs) via the farnesoid X receptor (FXR), which is predominantly expressed in human and mouse HSCs. Consistently, re-expression of NTCP in mice reduces cirrhosis, and genetic ablation of FXR in HSCs suppresses fibrotic marks in mice and in vitro cell culture. Mechanistically, deletion of a putative SANT domain from MCRS1 evicts histone deacetylase 1 from its histone H3 anchoring sites, increasing histone acetylation of BA transporter genes, modulating their expression and perturbing BA flow. Accordingly, human cirrhosis displays decreased nuclear MCRS1 and NTCP expression. Our data reveal a previously unrecognized function of MCRS1 as a critical histone acetylation regulator, maintaining gene expression and liver homeostasis. MCRS1 loss induces acetylation of BA transporter genes, perturbation of BA flow, and consequently, FXR activation in HSCs. This axis represents a central and universal signaling event in cirrhosis, which has significant implications for cirrhosis treatment. By genetic ablation of MCRS1 in mouse hepatocytes, we generate the first genetic mouse model of cirrhosis that recapitulates human features. Herein, we demonstrate that the activation of the bile acid/FXR axis in liver fibroblasts is key in cirrhosis development.


Comparison between sodium cromoglycate (MDI: metered-dose inhaler) and beclomethasone dipropionate (MDI) in treatment of adult patients with mild to moderate bronchial asthma. A double-blind, double-dummy randomized, parallel-group study.
This study compared the efficacy and tolerability of sodium cromoglycate (SC) and beclomethasone dipropionate (BDP) in adult patients with bronchial asthma inadequately treated with bronchodilators alone. The study was a double-blind, randomized, double-dummy, parallel-group study. Patients with mild to moderate symptomatic asthma, inadequately treated with bronchodilators only, were, after a 2-week run-in (base-line) period, randomized to 8 weeks of treatment with either SC 10 mg four times daily or BDP 100 micrograms four times daily. Salbutamol metered-dose inhaler was given as relief medication. A total of 37 patients were randomized for treatment, 19 patients in the SC group and 18 patients in the BD group. Efficacy and safety were determined by daily record card data: morning and evening peak-expiratory-flow rates (PEFR), daytime and nighttime asthma symptom scores, and rescue salbutamol use. At clinic visits, FEV1 and FVC were measured, as were the physician's and the patient's assessment of the medication at the end of the study. The safety and tolerability of the trial medication were assessed by monitoring adverse events throughout the study. A clinically and statistically significant improvement of the asthma in FEV1, symptom scores, rescue medication, and global opinion of efficacy was observed, and both groups provided equivalent efficacy. The morning PEFR as well as the evening PEFR for both groups improved, but was statistically significant only for the BDP group (M-PEFR). Both drugs were well tolerated with only a few minor adverse events. This trial shows that SC and BDP are equally effective anti-inflammatory treatments for mild to moderate bronchial asthma in adults.


Histological quantification of decomposed human livers: a potential aid for estimation of the post-mortem interval?
The objective of this study was to determine if a novel scoring-based model for histological quantification of decomposed human livers could improve the precision of post-mortem interval (PMI) estimation for bodies from an indoor setting. The hepatic decomposition score (HDS) system created consists of five liver scores (HDS markers): cell nuclei and cell structure of hepatocytes, bile ducts, portal triad, and architecture. A total of 236 forensic autopsy cases were divided into a training dataset (n = 158) and a validation dataset (n = 78). All cases were also scored using the total body score (TBS) method. We specified a stochastic relationship between the log-transformed accumulated degree-days (log10ADD) and the taphonomic findings, using a multivariate regression model to compute the likelihood function. Three models were applied, based on (i) five HDS markers, (ii) three partial body scores (head, trunk, limbs), or (iii) a combination of the two. The predicted log10ADD was compared with the true log10ADD for each case. The fitted models performed equally well in the training dataset and the validation dataset. The model comprising both scoring methods had somewhat better precision than either method separately. Our results indicated that the HDS system was statistically robust. Combining the HDS markers with the partial body scores resulted in a better representation of the decomposition process and might improve PMI estimation of decomposed human remains.


Discovery of Turn-On Fluorescent Probes for Detecting PDEδ Protein in Living Cells and Tumor Slices.
The first small-molecule fluorescent turn-on probes for detecting PDEδ protein were rationally designed, showing reasonable fluorescent properties and the fluorescent ability has been applied for visualization of the PDEδ protein in living cells and at tissue levels. The qPCR results showed that the mRNA expression of KRAS, PDEδ, AKT1, MAPK1, MEK7, RAF1, and mTOR were downregulated by probes 1-3 through PI3K/AKT/mTOR and MAPK signal pathways. The probes also can downregulate the protein level of pErk and tErk. Therefore, these small-molecule fluorescent probes are expected to be used in the screening of antipancreatic cancer drugs targeting the PDEδ protein, as well as in obtaining a better understanding of the pathological and physiological roles of PDEδ protein.


Dissipation and residue of S-metolachlor in maize and soil.
The purpose of this article was to establish a simple residue analysis method for S-metolachlor in maize and to study its dissipation and residue in maize field eco-system. The results showed that S-metolachlor declined rapidly in maize seedling and soil after application. The half-lives of S-metolachlor in maize seedlings in Beijing and Changchun were 6.68 and 4.85 days, respectively, and in soil were 12.81 and 14.81 days, respectively. The terminal residues of S-metolachlor in soil samples were very low (around 0.005-0.045 mg/kg), and the residues in maize seeds were not detectable. The use of S-metolachlor according to the recommended dosages in maize could be considered safe.


Absence of detectable effect of radiotherapy and chemotherapy for breast cancer on the presence of low voltage areas in patients receiving left atrial catheter ablation.
Radiation and chemotherapy for breast cancer are known to cause side effects to the heart. However, it still remains unclear whether those therapies affect left atrium fibrosis. We sought to examine the effects of radiation and chemotherapy on the electroanatomic features of the left atrium (LA) in patients who received catheter ablation for LA arrhythmias and underwent radiation and/or chemotherapy prior to the procedure. We compared 38 patients who underwent catheter ablation for LA arrhythmias and had a previous diagnosis of breast cancer with 38 patients without breast cancer. LA low voltage zones (LVZ) were analysed during the electrophysiological (EP) study. The existence of LA LVZ did not differ significantly between both groups (71.1% vs. 76.3%, p = .602; 13.7cm2 (IQR 0;20.6cm2) vs. 7.0cm2 (IQR 1.6;21.1cm2), p = .690). Also scar distribution revealed no difference between both groups. However, an involvement of the anterior wall was common in both groups (65.8% vs. 73.7%, p = .454). Patients with breast cancer and persistent AF showed a trend towards greater LA scar areas 14.5% vs. 6.9%, p = .383) compared to the control group. Age and LA volume index were the only independent predictors for greater LA scarring. Thoracic irradiation and chemotherapy for breast cancer do not lead to an increase in LA scar area or a changed distribution of LA scarring. However, patient with breast cancer showed a tendency towards greater LA scar areas. Patient's age and LA volume index were identified as independent predictors for LA scar development.


Brentuximab vedotin in relapsed/refractory Hodgkin lymphoma: An updated review of published data from the named patient program.
Brentuximab vedotin was available via named patient program (NPP) to patients with relapsed/refractory (R/R) Hodgkin lymphoma (HL) or systemic anaplastic large-cell lymphoma in ∼60 non-US/Canadian countries, before local approval. Published results were examined recently; through systematic literature review, we identified 12 new NPP publications. Most (10/12) publications included new NPP data describing 8 unique cohorts (N=480; all R/R HL) and new participating countries. Overall response rates were 58-80%, and complete remission rates were 10-40%. With median follow-up of 9.5-26 months, median progression-free survival was 5-10.5 months and median overall survival (OS) had not been reached in most cohorts; 1- and 2-year OS was 67-76% and 58-67%, respectively. Tolerability was as expected from previous reports. Despite intrinsic bias and heterogeneous cohorts, this update supports previous findings showing comparable efficacy and tolerability of brentuximab vedotin between real-world practice and phase 2 trial results in R/R HL.


Characteristics of genital herpes in Bilbao (Northern Spain): 12-year retrospective study.
Herpes simplex virus type 2 (HSV-2) is the most common cause of genital herpes (GH), but genital infection by herpes simplex virus type 1 (HSV-1) is increasing. The aim of this study was to analyze and compare epidemiological characteristics of patients with GH. Retrospective study conducted from January 2004 to December 2015 in patients with GH attended at two Sexually Transmitted Diseases (STDs) medical consultation of Bilbao-Basurto Integrated Health Organisation in Northern Spain. Patient's medical history was reviewed and data of interest was analyzed. One thousand three patients (524 male and 479 female) were reviewed. HSV-2 was detected in 74%. The proportion of HSV-1 increased during the study period, significantly in men (28% in 2004-2007 vs. 50% in 2012-2015). More female than male had HSV-1 infection (56% vs. 44%). The proportion of primary infection was higher among HSV-1 compared to HSV-2 (79% vs. 21%). Among the patients with HSV-1, primary infection was higher among men (86%) and in younger than 30 years. Recurrent GH was higher among HSV-2 infections (63%). In a multivariate model older age, geographic origin outside Spain, recurrent infection, prior contact with a partner's genital herpetic lesions, previous N. gonorrhoeae infection and prostitution were significantly associated with HSV-2 infection. HSV-2 was the most common causative agent of GH, but the proportion of HSV-1 increased. Overall, antecedent of STD and sexual risk behaviors were more frequent in patients with genital HSV-2 infection.


The treatment of erythropoietic protoporphyria. Experience with beta-carotene.
In a group of 19 subjects suffering from erythropoietic protoporphyria the administration of beta-carotene appeared to produce improvement based on a subjective assessment of alterations in exposure times required to produce the symptoms and signs of the condition. This clinical improvement however failed to show a direct correlation with the prophyrin levels in blood and faeces, or with the estimation by phototesting of the minimal response dose within the action spectrum of 400-600 nm.


A programmed surface on dental implants sequentially initiates bacteriostasis and osseointegration.
Osteogenesis surrounding dental implants is initiated by a series of early physiological events, including the inflammatory response. However, the persistence of an anti-infection surface often results in compromised histocompatibility and osseointegration. Here, we presented a programmed surface containing both silver nanoparticles (AgNPs) and silver ions (Ag+) with a heterogeneous structure and time-dependent functionalities. The AgNPs were located at the surface of the heparin-chitosan polyelectrolyte coating (PEM), whereas Ag+ was distributed at both the surface and inside of the coating under optimized conditions (pH=4). The optimized coating (Ag-4) exhibited potent bactericidal activity at the early stage (12 and 24 h after inoculation) and a sustained antibacterial efficacy in the subsequent stage (one or two weeks), as it gradually depleted. Furthermore, compared to coatings with sustained high silver concentrations in bacteria-cell coculture experiments, the degradable Ag-4 coating demonstrated improved cytocompatibility, better cell viability, and morphology over time. At a later stage (within one month), the in vivo test revealed that Ag-4-coated titanium had superior histocompatibility and osteogenesis outcomes compared to bare titanium in a bacteria-exposed environment. The programmed surface of dental implants presented in this study offers innovative ideas for sequential antibacterial effects and osseointegration.


Decreased baroreflex sensitivity is linked to sympathovagal imbalance, body fat mass and altered cardiometabolic profile in pre-obesity and obesity.
Though decreased baroreflex sensitivity (BRS), the predictor of cardiac morbidities and mortality has been reported in obesity, the mechanisms and metabolic biomarkers influencing BRS have not been studied. We aimed to assess the difference in cardiovascular (CV) risk profile in pre-obesity and obesity, and the contribution of body composition and cardiometabolic factors to CV risks in these two conditions. Obesity indices, body composition, blood pressure variability and autonomic function test parameters were recorded in 223 subjects divided into controls (n=72), pre-obese (n=77) and obese (n=74) groups. Insulin resistance (HOMA-IR), atherogenic index (AI), leptin, adiponectin, inflammatory and oxidative stress parameters were measured. Association and independent contribution of altered cardiometabolic parameters with BRS were performed by Pearson's correlation and multiple regression analysis, respectively. BRS was significantly decreased in pre-obese and obese group compared to controls. Sympathovagal imbalance (SVI) in the form of increased sympathetic and decreased parasympathetic cardiac drives was observed in pre-obesity and obesity. There was significant difference in general markers of obesity (body mass index, and waist-to-hip ratio), between pre-obese and obese group, however no such difference was observed in body composition and cardiometabolic parameters between the two groups. AI, high sensitive C-reactive protein (hs-CRP) and ratio of basal metabolism to body fat (BM/BF) in pre-obese group, and AI, HOMA-IR, leptin, adiponectin, ratio of basal metabolism to body weight (BM/BW), BM/BF, inflammatory and oxidative stress markers in obese group had independent contribution to BRS. Among these metabolic biomarkers, BRS had maximum association with leptin (β=0.532, p=0.000) in the obese group and hs-CRP (β=0.445, p=0.022) in the pre-obese group. The present study demonstrates decreased BRS, an important marker of increased CV risk in pre-obesity and obesity. The intensity of cardiometabolic derangements and CV risk was comparable between pre-obese and obese subjects. BM/BF ratio appears to be a better marker of metabolic activity in pre-obesity and obesity. SVI and increased basal metabolism appear to be the physiological link between metabolic derangements and CV risks in both pre-obesity and obesity.


Branched-Chain Amino Acids Enhance Retinal Ganglion Cell Survival and Axon Regeneration after Optic Nerve Transection in Rats.
This study's aim was to investigate the beneficial effects of branched-chain amino acids (BCAAs) on the neuronal survival and axon regeneration of retinal ganglion cells (RGCs) after optic nerve (ON) transection. The experimental rats received daily BCAA injections through the caudal vein after left intra-orbital ON transection. Neuroprotection was evaluated by counting Fluorogold-labeled RGCs. The role of mammalian target of rapamycin (mTOR) pathway activation in promoting RGC survival was studied after rapamycin administration. Moreover, a peripheral nerve (PN) graft was transplanted onto the transected ON to study the effects of BCAAs on axon regeneration of injured RGCs. Our results showed that BCAAs alleviated the death of RGCs 7 and 14 days after ON transection, accompanied by an activation of mTOR pathway in RGCs. Blocking mTOR pathway with rapamycin eliminated such neuroprotective effects of BCAAs. Moreover, BCAAs also promoted axon regeneration of injured RGCs into a PN graft. Our results suggest a neuroprotection of BCAAs through the activation of mTOR pathway. BCAAs also have a beneficial effect on axon regeneration of injured RGCs. Therefore, BCAAs could be considered for the clinical treatment of ON injury.


Three-dimensional spheroid formation of adipose-derived stem cells improves the survival of fat transplantation by enhance their therapeutic effect.
Adipose-derived stem cells (ADSCs) have important applications in basic research, especially in fat transplantation. Some studies have found that three-dimensional (3D) spheroids formed by mesenchymal stem cells have enhanced therapeutic potential. However, the fundamental basics of this effect are still being discussed. ADSCs were harvested from subcutaneous adipose tissues and 3D spheroids were formed by the automatic aggregation of ADSCs in a non-adhesive 6-well plate. Oxygen glucose deprivation (OGD) was used to simulate the transplantation microenvironment. We found that 3D culture of ADSCs triggered cell autophagy. After inhibiting autophagy by Chloroquine, the rates of apoptosis were increased. When the 3D ADSC-spheroids were re-planked, the number of senescent ADSCs decreased, and the proliferation ability was promoted. In addition, there were more cytokines secreted by 3D ADSC-spheroids including VEGF, IGF-1, and TGF-β. After adding the conditioned medium with human umbilical vein endothelial cells (HUVECs), 3D ADSC-spheroids were more likely to promote migration, and tube formation, stimulating the formation of new blood vessels. Fat grafting experiments in nude mice also showed that 3D ADSC-spheroids enhanced survival and neovascularization of fat grafts. These results suggested that 3D spheroids culturing of ADSCs can increase the therapeutic potential in fat transplantation.


Micropropagation of Andrographis producta through axillary and adventitious shoot regeneration.
Andrographis producta (C. B. Clarke) Gamble is a valuable medicinal plant that yields several therapeutic compounds. In addition, this species is endemic to the Western Ghats regions of South India. Natural populations of Andrographis producta have dwindled due to the overexploitation of this species. The objective of the present study was to develop a reliable In vitro propagation protocol for this plant species. In vitro plant regeneration protocol has been developed in Andrographis producta using nodal and shoot tip explants. The highest axillary shoots (14.60) were regenerated from nodal explants on MS medium amended with 10 μM 2-iP. Similarly, on MS media amended with 5 μM BAP, 17.50 shoots were regenerated from shoot tip explants. Optimal of 27.66 adventitious shoots were regenerated from the cut end of shoot tip explants on MS medium amended with 10 μM 2-iP. Medium amended with 10 μM 2-iP was optimum for regeneration of multiple axillary shoots from nodal explants and for the adventitious shoots regeneration from shoot tip explants. Shoot tips were ideal explants for the micropropagation of A. producta. Qarter sthrength MS media supplemented with 10 μM IBA has resulted in maximum rooting of the shoots. A reliable In vitro micropropagation method was developed in Andrographis producta through direct organogenesis, and this method is helpful for the multiplication, conservation, and utilization of this plant.


Hybrid spheroids containing mesenchymal stem cells promote therapeutic angiogenesis by increasing engraftment of co-transplanted endothelial colony-forming cells in vivo.
Peripheral artery disease is an ischemic vascular disease caused by the blockage of blood vessels supplying blood to the lower extremities. Mesenchymal stem cells (MSCs) and endothelial colony-forming cells (ECFCs) have been reported to alleviate peripheral artery disease by forming new blood vessels. However, the clinical application of MSCs and ECFCs has been impeded by their poor in vivo engraftment after cell transplantation. To augment in vivo engraftment of transplanted MSCs and ECFCs, we investigated the effects of hybrid cell spheroids, which mimic a tissue-like environment, on the therapeutic efficacy and survival of transplanted cells. The in vivo survival and angiogenic activities of the spheroids or cell suspension composed of MSCs and ECFCs were measured in a murine hindlimb ischemia model and Matrigel plug assay. In the hindlimb ischemia model, the hybrid spheroids showed enhanced therapeutic effects compared with the control groups, such as adherent cultured cells or spheroids containing either MSCs or ECFCs. Spheroids from MSCs, but not from ECFCs, exhibited prolonged in vivo survival compared with adherent cultured cells, whereas hybrid spheroids composed of MSCs and ECFCs substantially increased the survival of ECFCs. Moreover, single spheroids of either MSCs or ECFCs secreted greater levels of pro-angiogenic factors than adherent cultured cells, and the hybrid spheroids of MSCs and ECFCs promoted the secretion of several pro-angiogenic factors, such as angiopoietin-2 and platelet-derived growth factor. These results suggest that hybrid spheroids containing MSCs can serve as carriers for cell transplantation of ECFCs which have poor in vivo engraftment efficiency.


[Differential diagnosis of accident and resuscitation traumas].
The purpose of this paper is to define the criteria for the differential diagnosis of trauma following resuscitation and road accidents. To this end, 311 cases of thoracic and epigastric trauma were selected from the 2893 medico-legal autopsies carried out between 1979 and 1982 at the Institute of Forensic Medicine of the University of Heidelberg. Cardiopulmonary resuscitation had to be considered as the cause of trauma in 140 of these, but 45 of this group were excluded from further evaluation as they had been the victims of blunt trauma and no clear-cut distinction was possible between trauma resulting from an accident and trauma resulting from resuscitation. Thus, we were left with 95 cases of internal injury that presented as emergencies and in whom death followed resuscitation, as a group for comparison with 171 road accident victims who had not received cardiopulmonary resuscitation. Rib fractures, predominantly on the left side, were established in half the cases resuscitated, sternal fractures also being found in one-third of these victims. Bleeding at various sites, including hemato-thorax, was rare, with an incidence of 15%, thus making it highly unlikely that serious traumas caused by resuscitation were a major factor in the cause of death. This paper encompasses an extensive discussion on serious injuries, such as aortic and gastric ruptures, in this connection.


Bioassay-guided isolation of a low molecular weight PHB from Burkholderia sp. with phytotoxic activity.
This work reports on the bioassay-guided isolation and identification of the macrocyclic pentolide 1, a cyclic polyhydroxybutyrate (PHB) with low molecular weight. This metabolite is produced by Burkholderia sp. and it exhibited phytotoxic activity in a Lemna minor bioassay. Its structure was determined by (1)H and (13)C NMR, heteronuclear multiple quantum correlation, heteronuclear multiple bond correlation, IR, and electrospray ionization tandem mass spectrometry analyses. The period for maximum production of the pentolide was optimized and determined on the basis of multiple reaction monitoring experiments at 15 days. The potential of Burkholderia sp. as a producer of higher biopolymers of PHB was also investigated. The methodology employed here accelerated the isolation and characterization of a phytotoxic metabolite whose structure can serve as a model for the synthesis of new classes of herbicides.


Microvascular reactivity in rehabilitating cardiac patients based on measurements of retinal blood vessel diameters.
Exercise-based rehabilitation improves general cardiovascular fitness. The impact on the microvascular system has been studied in less detail. We measured changes in retinal blood vessel diameters, as a proxy for microvascular reactivity, in cardiac patients and we assessed the impact of a rehabilitation program on retinal vessel diameters. Cardiac patients (n = 78) and age-matched healthy controls (n = 32) performed an initial maximal endurance cycling test. Patients then participated in a 12-week rehabilitation program with additional endurance tests being performed six and twelve weeks after the initial test. Fundus images were collected immediately before and 0, 5, 10, 15 and 30 min after the endurance test. Widths of retinal blood vessels, represented as Central Retinal Arteriolar/Venular Equivalent (CRAE/CRVE) were calculated from the images. At the start of the rehabilitation program, CRAE and CRVE values of the patients changed immediately after the endurance test with respectively -1.90 μm (95% CI: -3.58; -0.22) and -5.32 μm (95% CI: -7.33; -3.30) compared to baseline values. In contrast, CRAE and CRVE values of healthy controls were respectively increased [3.52 μm (95% CI: 2.34; 4.69)] and decreased [-3.17 μm (95% CI: -5.27; -1.07)]. After six and twelve weeks, CRAE responses of patients immediately after endurance test increased respectively with 5.98 μm (95% CI: 4.25; 7.71) and 4.44 μm (95% CI: 3.18; 5.71). These responses were similar to the microvascular reactions observed in the control group. Arteriolar and venular retinal microvascular responses in cardiac patients were different from the ones of healthy controls. Retinal microvascular response of cardiac patients improved during rehabilitation.


In vivo reflectance confocal microscopy of the skin: a noninvasive means of assessing body cystine accumulation in infantile cystinosis.
Patients with infantile nephropathic cystinosis have progressive accumulation of cystine in tissues leading to delayed extrarenal complications. No simple tool is available to evaluate the level of body cystine accumulation. We sought to determine the value of in vivo reflectance confocal microscopy of the skin in patients with infantile nephrogenic cystinosis. Nine patients and control subjects were recruited for this study. Images were acquired by means of a near-infrared reflectance confocal laser scanning microscope. Scattered bright particles within the papillary dermis were observed in all patients but not in control subjects. The density of particles ranged from numerous (+++) to very few (+/-) and their distribution was heterogeneous. Electron microscopy confirmed that these particles corresponded to cystine crystal deposits within dermal fibroblasts. The density of cystine crystals within the dermis was greater in older patients, in patients with a high leukocyte cystine concentration, and with delayed cysteamine therapy. There was no correlation between the density of cystine deposits and renal disease or hypopigmentation but high levels of deposition occurred in association with extrarenal manifestations. This is a preliminary study on a small sample of patients. Repeated examination and longer follow-up is necessary. In vivo reflectance confocal microscopy of the skin appears to be a noninvasive means of assessing body cystine accumulation in infantile cystinosis and could be used as a complementary marker of treatment response in addition to leukocyte cystine measurement.


Knockdown of PIK3R1 by shRNA inhibits the activity of the splenic macrophages associated with hypersplenism due to portal hypertension.
Phosphatidylinositol 3-kinase (PI3K) plays a central role in the metabolic actions of insulin. One 85 kDa regulatory subunit of PIK3 is encoded by phosphoinositide-3-kinase, the regulatory subunit 1 (PIK3R1). Our previous study has demonstrated that PIK3R1 was up-regulated significantly in the splenic macrophage (MΦ) of portal hypertensive spleen. In the present study, RNA interference specific to PIK3R1 was employed to investigate its inhibitive effects on the activity of MΦ associated with hypersplenism due to portal hypertension (HS-PHT). The expression of PIK3R1 in the spleen was detected by immunohistochemical staining. Plasmid vector pGenesil-1 expressing specific small hairpin RNA (shRNA) against PIK3R1 and the scrambled shRNA control was constructed. MΦ were isolated and purified by anchored cultivation from patients with HS-PHT (HS-PHT-MΦ) and traumatic rupture of the spleen (Con-MΦ). After transfection into MΦ, PIK3R1 expression at both the mRNA and the protein level was examined by real-time polymerase chain reaction and Western blot. The activities of MΦ were determined, and the expression and activity of NF-κB were also detected. Immunohistochemistry revealed expression and cellular distribution of PIK3R1 in the spleen. The PIK3R1-shRNA was successfully synthesized and cloned into the plasmid vector pGenesil-1, and specifically suppressed PIK3R1 expression at both the mRNA and the protein level. After transfection into HS-PHT-MΦ and Con-MΦ, PIK3R1 knockdown inhibited the viability of MΦ, reduced the phagocytic rate, the rate of antigen-presenting positive cells, the metabolic rate, and the secretion of IL-1β and TNF-α (all p<0.05), and decreased the expression and activity of NF-κB. Our data showed that the knocking down of PIK3R1 with shRNA produced by pGenesil-1 led to inhibition of viability and to decreased activity of MΦ associated with HS-PHT in vitro. Therefore, it is tempting to speculate that PIK3R1 might play a considerable role in the pathogenesis of HS-PHT, and inhibition of PIK3R1 expression might be a novel therapeutic strategy for HS-PHT.


Understanding the context and appreciating the complexity of evaluating the Diversity Program Consortium.
The National Institutes of Health (NIH) made a sizeable investment in developing a scientific approach to understanding how to best increase diversity in the NIH-funded workforce by fostering inclusive excellence at a national scale through the Diversity Program Consortium (DPC). This chapter provides an overview of the context in which the consortium-wide evaluation study has taken place to provide readers with an understanding of its level of complexity. This evaluation effort is the first large-scale, national, systemic, longitudinal evaluation of harmonized interventions focused on undergraduate biomedical research training programs in the history of the NIH and the National Institute of General Medical Sciences.


Population ecology of Pardia tripunctana Schiff. and Notocelia roborana Den. and Schiff. (Lepidoptera, Tortricidae) -an example of "Equilibrium species".
The population ecology of Pardia tripunctana and Notocelia roborana, a bud- and a leaf-feeding tortricid respectively, on Rosa spp. was studied.Mortality factors of the egg and larval stages are low and density-independent.Pupal mortality in the soil is a regulatory mechanism: it compensates for density fluctuations, as independent of the initial pupal densities the densities of emerging adults are fairly constant.These mechanisms lead to low density levels, low resource utilization rates, and low density fluctuations. A steady state model is developed to describe the population dynamics.Both species therefore can be considered as "equilibrium species".


Cholesterol, Inflammasomes, and Atherogenesis.
Plasma cholesterol levels have been strongly associated with atherogenesis, underscoring the role of lipid metabolism in defining cardiovascular disease risk. However, atherosclerotic plaque is highly dynamic and contains elements of both the innate and adaptive immune system that respond to the aberrant accumulation of lipids in the subendothelial space. Previous research has focused on defining how proinflammatory cytokines synthesized by macrophages, such as interleukin-1β (IL-1β), modulate the progression of atherosclerosis, supporting the notion that chronic inflammation accelerates atherogenesis. More recently, emphasis has been placed on the elucidation of the mechanisms that contribute to pro-IL-1β production and finally its processing via multiprotein complexes termed the inflammasomes, a family of cytosolic multiprotein complexes that serve as sensors of either pathogen invasion or cellular stress (ie, cholesterol crystals) and work via triggering caspase-1-mediated processing of pro-IL-1β to IL-1β. Based on this link between cholesterol metabolism, NLRP3 inflammasome activation, and IL-1β release, it is important to re-evaluate how the atherogenic environment stimulates immune cells to produce IL-1β.


Motion-deblurring in human vision.
If photographs are taken of moving objects at slow shutter speeds the images of the objects are blurred. In human vision, however, we are not normally conscious of blur from moving objects despite the fact that the temporal response of the photoreceptors is sluggish. It has been suggested that there are motion-deblurring mechanisms specifically to aid the visual system in the analysis of the shape of retinally moving targets. Models of motion deblurring have been influenced by the finding that certain very precise spatial pattern discriminations are unaffected by motion. An example is vernier hyperacuity, in which the observer must detect the direction of offset between two lines with abutting ends. With a stationary stimulus, observers can detect a vernier cue of less than 10 arcsec and acuity is unaffected by retinal-image motion of up to 3 deg s-1 We confirm this finding, but provide evidence against any general deblurring mechanism by showing that another kind of hyperacuity, discrimination of the distance between two parallel lines (spatial interval acuity), is interfered with by motion. This argues against a general deblurring mechanism, such as a neural network 'shifter circuit', and we point out that the high level of vernier acuity for moving stimuli is susceptible to an alternative explanation.


Targeting Oncogenic Pathways in the Era of Personalized Oncology: A Systemic Analysis Reveals Highly Mutated Signaling Pathways in Cancer Patients and Potential Therapeutic Targets.
Cancer is the second leading cause of death globally. One of the main hallmarks in cancer is the functional deregulation of crucial molecular pathways via driver genetic events that lead to abnormal gene expression, giving cells a selective growth advantage. Driver events are defined as mutations, fusions and copy number alterations that are causally implicated in oncogenesis. Molecular analysis on tissues that have originated from a wide range of anatomical areas has shown that mutations in different members of several pathways are implicated in different cancer types. In recent decades, significant efforts have been made to incorporate this knowledge into daily medical practice, providing substantial insight towards clinical diagnosis and personalized therapies. However, since there is still a strong need for more effective drug development, a deep understanding of the involved signaling mechanisms and the interconnections between these pathways is highly anticipated. Here, we perform a systemic analysis on cancer patients included in the Pan-Cancer Atlas project, with the aim to select the ten most highly mutated signaling pathways (p53, RTK-RAS, lipids metabolism, PI-3-Kinase/Akt, ubiquitination, b-catenin/Wnt, Notch, cell cycle, homology directed repair (HDR) and splicing) and to provide a detailed description of each pathway, along with the corresponding therapeutic applications currently being developed or applied. The ultimate scope is to review the current knowledge on highly mutated pathways and to address the attractive perspectives arising from ongoing experimental studies for the clinical implementation of personalized medicine.


Factors restricting diffusion of water-soluble spin labels.
Line broadening of spin label signals is treated in terms of concentration, viscosity, charge and temperature dependencies. Line broadening of spin label signals may be caused either by spin label interactions or by the interaction between a spin label and a second paramagnetic species. Line broadening has been related to collision frequency in the literature and is treated in that way here. Collision frequency is related to diffusion processes in a way that allows information to be obtained about the diffusion environment. Several potential spin label line-broadening agents are compared as to their effectiveness. Small polymer beads with graduated pore sizes are used to show that collisional broadening has a marked dependence on the long-range structure of the diffusion environment. Application of these results to biological diffusion processes is considered.


Two-stage hepatectomy in resection of colorectal liver metastases - a single-institution experience with case-control matching and review of the literature.
Two-stage hepatectomy (TSH) has been proposed for patients with bilateral liver tumours who have a high risk of posthepatectomy liver failure after one-stage hepatectomy (OSH). This study aimed to determine the outcomes of TSH for extensive bilateral colorectal liver metastases. A retrospective review of a prospectively maintained database of liver resections for colorectal liver metastases was conducted. The TSH group was compared to the OSH group in terms of perioperative outcomes and survival. Case-control matching was performed. A total of 632 consecutive liver resections for colorectal liver metastases were performed between 2000 and 2020. The study group (TSH group) consisted of 15 patients who completed TSH. The control group included 151 patients who underwent OSH. The case-control matching-OSH group consisted of 14 patients. The major morbidity and 90-day mortality rates were 40% and 13.3% in the TSH group, 20.5% and 4.6% in the OSH group and 28.6% and 7.1% in the case-control matching-OSH group, respectively. The recurrence-free survival, median overall survival, and 3- and 5-year survival rates were 5 months, 21 months, 33% and 13% in the TSH group; 11 months, 35 months, 49% and 27% in the OSH group; and 8 months, 23 months, 36% and 21%, respectively, in the case-control matching-OSH group, respectively. TSH used to be a favourable therapeutic choice in a select population of patients. Now, OSH should be preferred whenever feasible because it has lower morbidity and equivalent oncological outcomes to those of completed TSH.


How rats process spatiotemporal information in the face of distraction.
How rats process spatiotemporal information in the face of distraction was assessed. Rats were trained on a time-place learning task in which the location of food availability depended on the amount of time elapsed since the beginning of the training session. In each training session each of four levers provided food pellets for 5 min on an intermittent schedule. In probe sessions interspersed with the final training sessions, the rats were presented with a second highly preferred food source-a piece of cheese-at various times into the session. Rats choose the correct lever after the cheese distraction, but it appeared that their internal clock had stopped during the cheese consumption period. Thus rats' internal clock, like that of pigeons, displays the properties of 'stop', 'reset', and 'restart'. Rat-pigeon differences in timing processes may be restricted to circadian or time of day timing. Present results also suggest that rats process spatial and temporal information separately.


Electrochemical growth of gold nanoparticles on horizontally aligned carbon nanotubes: a new platform for ultrasensitive DNA sensing.
A new platform based on electrochemical growth of Au nanoparticles on horizontally aligned single walled carbon nanotube (SWCNT) array was developed for ultrasensitive DNA detection. The as-prepared DNA-functionalized SWCNT-Au platform, in which every gold-coated SWCNT acts as an isolated micro electrode, could detect lower than 10 zmol complimentary 10-base DNA, which corresponded to having 6 DNA molecules in a 1 mL sample solution. For a 1-base mismatched DNA, the experimental detection limit was 100 amol. A linear relationship between the change of charge transfer resistance and target DNA concentration was achieved at low concentration range. Over the extended DNA concentration range, the change of charge transfer resistance was found to have a linear relationship with respect to the logarithm of the target DNA concentration. The sensor also showed great stability and could be conveniently regenerated via dehybridization in hot water. The significant improvement in sensitivity illustrates that combining Au nanoparticles with the on-site fabricated SWCNT array represents a promising platform for achieving ultrasensitive biosensor.


Enzymatic fingerprints of polysaccharides of Dendrobium officinale and their application in identification of Dendrobium species.
Enzymatic fingerprinting of polysaccharides from Dendrobium officinale was studied and applied to authenticate Dendrobium species. Results showed that Dendrobium officinale species from Anhui province, Fujian province, Yunnan province, Guangdong province and Guangxi province of China, could be identified by polysaccharide analysis using carbohydrate gel electrophoresis (PACE). However, the fingerprints of Dendrobium officinale from Jiangxi province, Hu'nan province and Wenzhou, Yandangshan and Fuyang in Zhejiang province were very similar. As far as the fingerprints of different Dendrobium species were concerned, the differences between Dendrobium officinale, Dendrobium huoshanense, Dendrobium moniliforme, Dendrobium devonianum, Dendrobium aphyllum, Dendrobium wilsonii and Dendrobium crystallinum were obvious. Moreover, the genetic relationships between different samples were analyzed by using principal component analysis and unweighted pair group method with arithmetic mean cluster analysis. Results suggested that polysaccharide fingerprint analysis by PACE has the potential to become a valuable new method for the identification and control of quality of herbal medicines in future.


Gene transfer to salivary glands.
This article provides a review of the application of gene transfer technology to studies of salivary glands. Salivary glands provide an uncommon target site for gene transfer but offer many experimental situations likely of interest to the cell biologist. The reader is provided with a concise overview of salivary biology, along with a general discussion of the strategies available for gene transfer to any tissue. In particular, adenoviral vectors have been useful for proof of concept studies with salivary glands. Several examples are given, using adenoviral-mediated gene transfer, for addressing both biological and clinical questions. Additionally, benefits and shortcomings affecting the utility of this technology are discussed.


Hemodynamic Significance of Internal Carotid or Middle Cerebral Artery Stenosis Detected on Magnetic Resonance Angiography.
We evaluated hemodynamic significance of stenosis on magnetic resonance angiography (MRA) using acetazolamide perfusion single photon emission computed tomography (SPECT). Of 171 patients, stenosis in internal carotid artery (ICA) and middle cerebral artery (MCA) (ICA-MCA) on MRA and cerebrovascular reserve (CVR) of MCA territory on SPECT was measured using quantification and a 3-grade system. Stenosis and CVR grades were compared with each other, and their prognostic value for subsequent stroke was evaluated. Of 342 ICA-MCA, 151 (44%) presented stenosis on MRA; grade 1 in 69 (20%) and grade 2 in 82 (24%) cases. Decreased CVR was observed in 9% of grade 0 stenosis, 25% of grade 1, and 35% of grade 2. The average CVR of grade 0 was significantly different from grade 1 (p<0.001) and grade 2 stenosis (p=0.007). In quantitative analysis, average CVR index was -0.56±7.91 in grade 0, -1.81±6.66 in grade 1 and -1.18±5.88 in grade 2 stenosis. Agreement between stenosis and CVR grades was fair in patients with lateralizing and non-lateralizing symptoms (κ=0.230 and 0.346). Of the factors tested, both MRA and CVR were not significant prognostic factors (p=0.104 and 0.988, respectively), whereas hypertension and renal disease were significant factors (p<0.05, respectively). A considerable proportion of ICA-MCA stenosis detected on MRA does not cause CVR impairment despite a fair correlation between them. Thus, hemodynamic state needs to be assessed for evaluating significance of stenosis, particularly in asymptomatic patients.


Morphology and water flux of produced cellulose acetate membranes reinforced by the design of experiments (DOE).
Cellulose acetate (CA) ultrafiltration membranes were successfully prepared using the non-solvent induced phase separation (NIPS) methodology. This technique is used to produce porous membranes for a large variety of applications. However, the formation of a dense skin during the process reduces membrane pure water flux (PWF). To overcome this issue, three parameters were investigated: CA/NMP (N-methyl-2-pyrrolidone) ratio in the casting solution, acetone (Ac)/water (W) ratio in the precipitation bath composition (PBC) and support material (glass/polyethylene). The effect of each factor on the mean pore size, water contact angle, porosity and PWF was supported by Taguchi design. The increase in the CA/NMP ratio reduced mean pore size and porosity. In contrast, there was an increase in porosity and hydrophilicity with increasing Ac/W ratio. The maximum value of PWF was obtained for membranes prepared using a PE support. ANOVA showed that most, but not all, factors had significant effects on the parameters measured.


"Forever young at the table": metabolic effects of eating speed in obesity.
Cardiometabolic diseases (CMD) are recognized as the main causes of morbidity and mortality in developed countries. In recent years eating speed (ES) has been of particular interest since some studies have associated it with the development of obesity and CMD. However, the different impact of the ES at which main meals are eaten on the risk of developing these diseases has not yet been identified. Thus, we aimed to investigate the effect of ES at the main meals (breakfast, lunch, and dinner) on the risk of developing cardiometabolic diseases (type 2 diabetes mellitus, dyslipidaemia and hypertension) in middle-aged Caucasian subjects with obesity. For this purpose we carried out a cross-sectional, observational study. One hundred and eighty-seven middle-aged subjects aged 43.6 ± 16 years were enrolled of which anthropometric parameters and lifestyle habits were studied. A dietary interview was performed to collect information about meal duration and eating habits at the main meals. According to median value of meal duration, meals were classified in two groups: fast eating group (FEG) and slow eating group (SEG). The prevalence of dyslipidaemia was more than twice in FEG compared to SEG at lunch and dinner. For all main meals, FEG had a significantly higher risk of dyslipidaemia than SEG (p < 0.05) in unadjusted model. However, when the model was adjusted for age, BMI, physical activity, smoking and alcohol use and medication, the result remained significant for lunch and dinner (p < 0.05). The results of our study suggest that fast eating increases at lunch and dinner increase the risk of developing dyslipidaemia in obesity.


D(5h) C50X10: saturn-like fullerene derivatives (X=F, Cl, Br).
Based on the recently reported D(5h) C(50), the geometries and stabilities of its Saturn-like derivatives C(50)X(10) (X=F, Cl, Br) have been investigated by DFT method. Compared with C(50), the equatorial carbon atoms in C(50)X(10) have been saturated by halogens and change to sp(3) hybridization to release the large angle strain. Because the equatorial carbon atoms have been taken out of the pi system by the halogens "ring," the C(50)X(10) system has been split into two well-delocalized conjugated annulene subunits, and then the electronic stabilization has been enhanced.


Cardiomyocyte--specific expression of the nuclear matrix protein, CIZ1, stimulates production of mono-nucleated cells with an extended window of proliferation in the postnatal mouse heart.
Myocardial injury in mammals leads to heart failure through pathological cardiac remodelling that includes hypertrophy, fibrosis and ventricular dilatation. Central to this is inability of the mammalian cardiomyocyte to self-renew due to entering a quiescent state after birth. Modulation of the cardiomyocyte cell-cycle after injury is therefore a target mechanism to limit damage and potentiate repair and regeneration. Here, we show that cardiomyocyte-specific over-expression of the nuclear-matrix--associated DNA replication protein, CIZ1, extends their window of proliferation during cardiac development, delaying onset of terminal differentiation without compromising function. CIZ1-expressing hearts are enlarged, but the cardiomyocytes are smaller with an overall increase in number, correlating with increased DNA replication after birth and retention of an increased proportion of mono-nucleated cardiomyocytes into adulthood. Furthermore, these CIZ1 induced changes in the heart reduce the impact of myocardial injury, identifying CIZ1 as a putative therapeutic target for cardiac repair.


Mutant pfcrt "SVMNT" haplotype and wild type pfmdr1 "N86" are endemic in Plasmodium vivax dominated areas of India under high chloroquine exposure.
Chloroquine resistance (CQR) phenotype in Plasmodium falciparum is associated with mutations in pfcrt and pfmdr-1 genes. Mutations at amino acid position 72-76 of pfcrt gene, here defined as pfcrt haplotype are associated with the geographic origin of chloroquine resistant parasite. Here, mutations at 72-76 and codon 220 of pfcrt gene and N86Y pfmdr-1 mutation were studied in blood samples collected across 11 field sites, inclusive of high and low P. falciparum prevalent areas in India. Any probable correlation between these mutations and clinical outcome of CQ treatment was also investigated. Finger pricked blood spotted on Whatman No.3 papers were collected from falciparum malaria patients of high and low P. falciparum prevalent areas. For pfcrt haplotype investigation, the parasite DNA was extracted from blood samples and used for PCR amplification, followed by partial sequencing of the pfcrt gene. For pfmdr-1 N86Y mutation, the PCR product was subjected to restriction digestion with AflIII endonuclease enzyme. In 240 P. falciparum isolates with reported in vivo CQ therapeutic efficacy, the analysis of mutations in pfcrt gene shows that mutant SVMNT-S (67.50%) and CVIET-S (23.75%) occurred irrespective of clinical outcome and wild type CVMNK-A (7.91%) occurred only in adequate clinical and parasitological response samples. Of 287 P. falciparum isolates, SVMNTS 192 (66.89%) prevailed in all study sites and showed almost monomorphic existence (98.42% isolates) in low P. falciparum prevalent areas. However, CVIETS-S (19.51%) and CVMNK-A (11.84%) occurrence was limited to high P. falciparum prevalent areas. Investigation of pfmdr-1 N86Y mutation shows no correlation with clinical outcomes. The wild type N86 was prevalent in all the low P. falciparum prevalent areas (94.48%). However, mutant N86Y was comparably higher in numbers at the high P. falciparum prevalent areas (42.76%). The wild type pfcrt gene is linked to chloroquine sensitivity; however, presence of mutation cannot explain the therapeutic efficacy of CQ in the current scenario of chloroquine resistance. The monomorphic existence of mutant SVMNT haplotype, infer inbreeding and faster spread of CQR parasite in areas with higher P. vivax prevalance and chloroquine exposure, whereas, diversity is maintained in pfcrt gene at high P. falciparum prevalent areas.


Activation of sterol ester hydrolase of bovine corpus luteum by N6,O2'-dibutyryl cyclic adenosine 3':5'-phosphate.
The direct activation of sterol ester hydrolase (E.C. 22.214.171.124) in homogenates of bovine corpus luteum by N6O2'-dibutyryl cyclic adenosine 3':5'-phosphate, (dibutyryl cAMP), adenosine triphosphate (ATP), and Mg2+ has been demonstrated. Variability in the extent of activation by the additions was minimized by homogenization of the tissue in 5 mM Mg2+'. Baseline sterol ester hydrolase activity was primarily associated with the 105,000 X g soluble fraction, and significant activation of the enzyme preparation preincubated with dibutyryl cAMP, ATP and Mg2+ occurred within the first 15 min, prior to addition of substrate. A requirement for protein kinase in the system was demonstrated by blocking the cofactor-dependent enzyme activation with commercial protein kinase inhibitor.


Thyrocyte specific killer cell activity is decreased in patients with thyroid carcinoma.
Natural killer cells (NK) are important in immune surveillance, and they have found to be decreased in the serum of patients with various types of malignancies. In patients with thyroid carcinoma, we investigated NK activity of peripheral lymphocytes directed against either a standard tumor cell target, K562, or against the clinically relevant target cell, the thyrocyte. Although NK-cell activity was normal when directed against K562 cell targets, NK-cell activity was reduced when labeled thyrocytes were used as target cells. At effector to target cell ratios of 100:1 and 20:1, there was 43 +/- 3% (n = 7) and 29 +/- 3% (n = 7) lyses of thyrocytes using normal lymphocytes, and 17 +/- 4% (n = 5) and 13 +/- 2% (n = 3) using lymphocytes from patients with thyroid carcinoma. These findings suggest that NK-cells have distinct subpopulations and that immune surveillance against specific thyrocyte target cells may be important in thyroid cancer.


Protocol to analyze the inhibitory effect of free organic carbon generated by chemoautotrophic bacteria on their CO2 fixation.
Here, we present a protocol to analyze the inhibition of self-generated extracellular free organic carbon (EFOC) on CO2 fixation by chemoautotrophic bacteria. We detail the construction and operation of membrane reactor, followed by a simulation experiment to verify the inhibition of EFOC on CO2 fixation. We further describe the analysis of main inhibitory components in EFOC and measurement of abundance and transcription level of ribulose bisphosphate carboxylase/oxygenase (RuBisCO) gene to clarify the mechanism of the main inhibition components on CO2 fixation. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2022).1.


A randomized placebo-controlled pilot study of Cat saliva 9cH and Histaminum 9cH in cat allergic adults.
Cat allergy is an abnormal immune response by the body to cat dander or saliva, leading to the development of a complex of symptoms which can negatively influence health. Cat saliva 9cH and Histaminum 9cH are indicated, according to isopathic principles, for the treatment of cat allergy, however no research has been done to date. To determine the effect of Cat saliva 9cH and Histaminum 9cH (combined) on cat allergic adults. 30 Participants with a positive test result for a cat allergy skin prick test (SPT) were recruited to a double-blind, randomised, placebo controlled clinical trial. Participants took two tablets twice daily for 4 weeks, and attended a follow-up consultation at the end of weeks 2 and 4. The measurement tool used was the SPT, conducted at the beginning and at the end of the study. Cat saliva 9cH and Histaminum 9cH produced a highly statistically significant reduction in the wheal diameter of the cat allergen SPT at the end of week 4. The placebo group showed no statistically significant change. The homeopathic medicine reduced the sensitivity reaction of cat allergic adults to cat allergen, according to the SPT. Future studies are warranted to further investigate the effect of Cat saliva and Histaminum and their role as a potential therapeutic option for this condition.


Envenomation by Tegenaria agrestis (Walckenaer) spiders in rabbits.
Preliminary studies in rabbits demonstrate that envenomation by the male Tegenaria agrestis spider produces a slowly developing dermal lesion, which exhibits gravitational drift and extensive subcutaneous hemorrhaging. Petechial hemorrhages were present on the lungs, liver and kidneys of rabbits bitten by male Tegenaria agrestis. Female Tegenaria agrestis trials in rabbits produced no significant effects in New Zealand white rabbits, but produced dermal lesions in the California giant strain.


Plant identification applications do not reliably identify toxic and edible plants in the American Midwest.
Exposure to potentially toxic plants is a global problem, resulting in thousands of calls to poison centers and emergency department visits annually and occasional deaths. Persons with limited botanical knowledge may be tempted to rely on smartphone applications to determine if plants are safe to forage. This study evaluated the reliability of several popular smartphone applications to identify foraged foods and distinguish them from potentially toxic plants in the Midwestern United States. Sixteen plant species were selected based on local availability, attractiveness as foraged food, and potential for misidentification. Of the 16 species, five are edible, three are potentially toxic if improperly harvested or prepared, and eight are considered to be toxic. Plant specimens were identified by graduate-level botanists and photographed during multiple stages of their growth cycles. LeafSnap, PictureThis, Pl@ntNet and PlantSnap were used to identify the plants. Overall accuracy of the applications in identifying plant genus was 76% (95% confidence interval: 73-79, range 96% for PictureThis to 53% for PlantSnap). Accuracy for identification of plant species was 58% (95% confidence interval 55-62%, range 94% for PictureThis to 34% for PlantSnap). Five of eleven potentially toxic species were identified as an edible species by at least one application. Accuracy of the smartphone applications varies, with PictureThis outperforming other apps. At this time, apps cannot be used to safely identify edible plants. Foragers must have adequate botanical knowledge to ensure safe harvesting of wild plants.


Benzoxaborole antimalarial agents. Part 2: Discovery of fluoro-substituted 7-(2-carboxyethyl)-1,3-dihydro-1-hydroxy-2,1-benzoxaboroles.
A series of new boron-containing benzoxaborole compounds was designed and synthesized for a continuing structure-activity relationship (SAR) investigation to assess the antimalarial activity changes derived from side-chain structural variation, substituent modification on the benzene ring and removal of boron from five-membered oxaborole ring. This SAR study demonstrated that boron is required for the antimalarial activity, and discovered that three fluoro-substituted 7-(2-carboxyethyl)-1,3-dihydro-1-hydroxy-2,1-benzoxaboroles (9, 14 and 20) have excellent potencies (IC(50) 0.026-0.209 μM) against Plasmodium falciparum.


Effects of raloxifene on Ki-67 and CD34 antigen expression in breast cancer.
The aim of this study was to evaluate the effect of raloxifene on CD34 and Ki-67 antigen expression in breast cancer specimens from postmenopausal women. Sixteen postmenopausal patients with operable, stage II (>or=3 cm), estrogen receptor-positive breast cancer, who took 60 mg of raloxifene daily for 28 days, participated in this study. Immunohistochemistry was carried out in tumor samples prior to and following raloxifene treatment to evaluate CD34 and Ki-67 protein expression. Angiogenesis was quantified in 10 randomly selected fields per slide, and Ki-67-stained nuclei were counted in 1,000 cells per slide using an image capture and analysis system with 400x magnification. Student's t test for paired samples was used for the statistical analysis of data. Statistical significance was established at p < 0.05. The mean number of microvessels was 44.44 +/- 3.54 prior to raloxifene therapy and 22.63 +/- 1.61 following therapy (p < 0.001), and the mean percentage of Ki-67-stained nuclei was 19.28 +/- 1.61 and 12.13 +/- 1.48 prior to and following raloxifene treatment, respectively (p < 0.001). Raloxifene significantly reduces CD34 and Ki-67 protein expression in breast carcinoma in postmenopausal women.


Hospitalized opioid-dependent patients: Exploring predictors of buprenorphine treatment entry and retention after discharge.
Few studies have explored predictors of entry into and retention in buprenorphine treatment following linkage from an acute medical hospitalization. This secondary analysis of a completed clinical trial focuses on medically hospitalized, opioid-dependent patients (n = 72) who were randomized to an intervention including buprenorphine induction and dose stabilization during hospitalization followed by post-discharge transition to office-based buprenorphine treatment (OBOT). Predictors included demographics, days hospitalized, prior buprenorphine/methadone treatment, PTSD symptoms, social support, and readiness for drug use cessation. Outcome variables were treatment entry and retention (number of days in OBOT). Previous buprenorphine treatment, more days hospitalized, and higher PTSD symptoms predicted OBOT entry. Prior treatment, older age, and non-minority status were associated with a higher mean number of days in OBOT. OBOT may appeal to patients who have tried buprenorphine in other settings. Linking hospitalized patients to OBOT may improve utilization of addiction treatment. Prior substance treatment, longer hospital stay, and mental health should be examined in future linkage studies. (Am J Addict 2017;26:667-672).


Comparative proteome analysis reveals proteins involved in salt adaptation in Photobacterium damselae subsp. piscicida.
Proteomic approaches were applied to investigate whether Photobacterium damselae subsp. piscicida (Phdp) can directly sense and respond to growth conditions under different salinities, 0.85% and 3.5% NaCl concentrations, mimicking the osmotic conditions in host and marine water bodies, respectively. Proteins significantly altered were analyzed by two-dimensional gel electrophoresis (2-DE), liquid chromatography-electrospray ionization-quadrupole-time-of-flight tandem mass spectrometry (LC-ESI-Q-TOF MS/MS) and bioinformatics analysis, thus resulting in 16 outer membrane proteins (OMPs), 12 inner membrane proteins (IMPs), and 20 cytoplasmic proteins (CPs). Quantitative real-time PCR was also applied to monitor the mRNA expression level of these target proteins. Cluster of orthologous groups of protein (COG) analysis revealed that when shifting from 3.5% to 0.85% salinity, the majority of the up-regulated proteins were involved in posttranslational modification, protein turnover, and chaperones, while the down-regulated proteins were mainly related to energy production and conversion, compatible solutes (carbohydrates, amino acids and their derivatives) biogenesis and transport. Differentially expressed proteins identified in the current study could be used to elucidate the salt adaptation mechanisms of Phdp during their transition between host cells and the marine habitats.


Modulation of hyperpolarization-activated cation current Ih by volatile anesthetic sevoflurane in the mouse striatum during postnatal development.
Volatile anesthetics have been reported to inhibit hyperpolarization-activated cyclic-nucleotide gated channels underlying the hyperpolarization-activated cation current (Ih) that contributes to generation of synchronized oscillatory neural rhythms. Meanwhile, the developmental change of Ih has been speculated to play a pivotal role during maturation. In this study, we examined the effect of the volatile anesthetic sevoflurane, which is widely used in pediatric surgery, on Ih and on functional Ih activation kinetics of cholinergic interneurons in developing striatum. Our analyses showed that the changes in Ih of cholinergic interneurons occurred in conjunction with maturation. Sevoflurane application (1-4%) caused significant inhibition of Ih in a dose-dependent manner, and apparently slowed Ih activation. In current-clamp recordings, sevoflurane significantly decreased spike firing during the rebound activation, which is essential for responses to the sensory inputs from the cortex and thalamus. The sevoflurane-induced inhibition of Ih in striatal cholinergic interneurons may lead to alterations of the acetylcholine-dopamine balance in the neural circuits during the early postnatal period.


ERα/LKB1 complex upregulates E-cadherin expression and stimulates breast cancer growth and progression upon adiponectin exposure.
Adiponectin is the major adipocytes-secreted protein involved in obesity-related breast cancer growth and progression. We proved that adiponectin promotes proliferation in ERα-positive breast cancer cells, through ERα transactivation and the recruitment of LKB1 as ERα-coactivator. Here, we showed that adiponectin-mediated ERα transactivation enhances E-cadherin expression. Thus, we investigated the molecular mechanism through which ERα/LKB1 complex may modulate the expression of E-cadherin, influencing tumor growth, progression and distant metastasis. We demonstrated that adiponectin increases E-cadherin expression in ERα-positive 2D and higher extent in 3D cultures. This occurs through a direct activation of E-cadherin gene promoter by ERα/LKB1-complex. The impact of E-cadherin on ERα-positive breast cancer cell proliferation comes from the evidence that in the presence of E-cadherin siRNA the proliferative effects of adiponectin is no longer noticeable. Since E-cadherin connects cell polarity and growth, we investigated if the adiponectin-enhanced E-cadherin expression could influence the localization of proteins cooperating in cell polarity, such as LKB1 and Cdc42. Surprisingly, immunofluorescence showed that, in adiponectin-treated MCF-7 cells, LKB1 and Cdc42 mostly colocalize in the nucleus, impairing their cytosolic cooperation in maintaining cell polarity. The orthotopic implantation of MCF-7 cells revealed an enhanced E-cadherin-mediated breast cancer growth induced by adiponectin. Moreover, tail vein injection of MCF-7 cells showed a higher metastatic burden in the lungs of mice receiving adiponectin-treated cells compared to control. From these findings it emerges that adiponectin treatment enhances E-cadherin expression, alters cell polarity and stimulates ERα-positive breast cancer cell growth in vitro and in vivo, sustaining higher distant metastatic burden.


B-MIC: An Ultrafast Three-Level Parallel Sequence Aligner Using MIC.
Sequence alignment is the central process for sequence analysis, where mapping raw sequencing data to reference genome. The large amount of data generated by NGS is far beyond the process capabilities of existing alignment tools. Consequently, sequence alignment becomes the bottleneck of sequence analysis. Intensive computing power is required to address this challenge. Intel recently announced the MIC coprocessor, which can provide massive computing power. The Tianhe-2 is the world's fastest supercomputer now equipped with three MIC coprocessors each compute node. A key feature of sequence alignment is that different reads are independent. Considering this property, we proposed a MIC-oriented three-level parallelization strategy to speed up BWA, a widely used sequence alignment tool, and developed our ultrafast parallel sequence aligner: B-MIC. B-MIC contains three levels of parallelization: firstly, parallelization of data IO and reads alignment by a three-stage parallel pipeline; secondly, parallelization enabled by MIC coprocessor technology; thirdly, inter-node parallelization implemented by MPI. In this paper, we demonstrate that B-MIC outperforms BWA by a combination of those techniques using Inspur NF5280M server and the Tianhe-2 supercomputer. To the best of our knowledge, B-MIC is the first sequence alignment tool to run on Intel MIC and it can achieve more than fivefold speedup over the original BWA while maintaining the alignment precision.


Mre11 nuclease and C-terminal tail-mediated DDR functions are required for initiating yeast telomere healing.
Mre11 is a central factor in creating an optimal substrate for telomerase loading and elongation. We have used a G2/M synchronized telomere-healing assay as a tool to separate different functions of Mre11 that are not apparent in null alleles. An analysis of healing efficiencies of several mre11 alleles revealed that both nuclease and C-terminal mutations led to a loss of healing. Interestingly, trans-complementation of the 49 amino acid C-terminal deletion (DeltaC49) and the D16A mutant, deficient in nuclease activity and partially defective in MRX complex formation, restores healing. DeltaC49 provokes Rad53 phosphorylation after treatment with the radiomimetic agent MMS exclusively through the Tel1 pathway, suggesting that a Tel1-mediated function is initiated through the C-terminal tail.


Unstable DNA in a patient with a severe form of congenital myotonic dystrophy.
Recently, an unstable DNA (expanded CTG repeat) was identified as the mutation that causes myotonic dystrophy (DM). By Southern blot analysis of DNA derived from peripheral blood, patients with congenital form of DM are shown to have greater expansion of DM specific band than is seen in the analyses of adult form of DM. We present here tissue expression of unstable DNA in an autopsied case of severe congenital form of DM. Our study revealed the largest expanding band of 19 kb was uniformly seen in various tissues from an autopsied infant with congenital form of DM.


Steady cone-jet electrosprays in liquid insulator baths.
This study shows that conducting liquids can be electrosprayed in steady cone-jet mode inside liquid insulator baths. Experimental results show that the current emitted from the meniscus fits well the scaling laws given in the literature for electrosprays in air at atmospheric pressure or vacuum. The technique may be of interest in obtaining fine liquid-liquid emulsions of uniformly sized droplets in the nanometric range.


Seizures and epilepsy of autoimmune origin: A long-term prospective study.
To follow prospectively a group of patients with seizures or epilepsy and suggestive clinical features of autoimmune aetiology and find out how many are finally diagnosed with acute symptomatic seizures (ASS) secondary to autoimmune encephalitis or autoimmune-related epilepsy, and how many develop epilepsy. Consecutive patients meeting the inclusion criteria from 2010 to 2018 were identified. Patients were classified as confirmed, probable autoimmune, non-autoimmune, or unknown. One-hundred and nine patients were included, 64 (48.7 %) women, mean age 55.2 years (SD 17.9). ASS were reported by 61 patients (56 %), while 48 presented epilepsy (44 %). During follow-up 18 patients died (16.5 %). Final diagnosis was autoimmune-relatedepilepsy (confirmed + probable) in 22 cases and ASS secondary to autoimmune encephalitis (confirmed or probable) in 27, non-autoimmune aetiologies or other diagnosis in 49 (44 %), and unknown aetiology in 11 (10.2 %). Neuronal antibodies (ab) were found in 27 patients (24.7 %). T-lymphocyte infiltration in temporal lobes was observed in 2/8 patients (20 %). Neuronal ab were more frequent in the autoimmune groups: 17 patients (29.8 %) vs 1(2.3 %), p:0.001, and they suffered more autoimmune diseases: 37 (75.5 %) vs 12 (24.48 %), p:0.0001, and 34 (69 %) vs 22 (44.9 %) p:0.027, respectively. All patients with GAD ab 17/17 (100 %) evolved to chronic disease. Four patients (29 %) with ASS secondary to autoimmune encephalitis developed epilepsy. ASS secondary to autoimmune encephalitis or autoimmune-related epilepsy will be diagnosed in nearly half of patients who have been suspected of it. The only diagnostic clue is neuronal ab. Patients who have suffered ASS secondary to autoimmune encephalitis may develop epilepsy over time.


Distribution and genome structures of temperate phages in acetic acid bacteria.
Acetic acid bacteria (AAB) are industrial microorganisms used for vinegar fermentation. Herein, we investigated the distribution and genome structures of mitomycin C-inducible temperate phages in AAB. Transmission electron microscopy analysis revealed phage-like particles in 15 out of a total 177 acetic acid bacterial strains, all of which showed morphology similar to myoviridae-type phage. The complete genome sequences of the six phages derived from three strains each of Acetobacter and Komagataeibacter strains were determined, harboring a genome size ranging from 34,100 to 53,798 bp. A phage AP1 from A. pasteurianus NBRC 109446 was predicted as an active phage based on the genomic information, and actually had the ability to infect its phiAP1-cured strain. The attachment sites for phiAP1 were located in the 3'-end region of the tRNAser gene. We also developed a chromosome-integrative vector, p2096int, based on the integrase function of phiAP1, and it was successfully integrated into the attachment site of the phiAP1-cured strain, which may be used as a valuable tool for the genetic engineering. Overall, this study showed the distribution of mitomycin C-inducible temperate phages in AAB, and identified the active temperate phage o f A. pasteurianus.


A comparative study of in-vitro and in-silico anti-candidal activity and GC-MS profiles of snow mountain garlic vs. normal garlic.
The study aimed to profile the volatile phytocomposition of snow mountain garlic (SMG) compared to normal garlic and investigate the anti-Candida efficacy against clinically relevant multi-drug resistant isolates of Candida species. Herein, SMG has shown significantly superior fungicidal power at 2x-MIC dose against C. albicans and C. glabrata in killing kinetic evaluation unlike the fungistatic effect of normal garlic. GC-MS headspace-based profiling of SMG showed 5 unique volatile compounds and a 5-fold higher content of saponins than normal garlic. In an in-silico analysis, cholesta-4,6-dien-3-ol,(3-beta) was uniquely identified in SMG as a potential inhibitor with high binding affinity to the active site of exo-1,3-betaglucan synthase, an established anti-candida drug target crucial for the biofilm matrix formation, thus suggesting a plausible anti-Candida mechanism. The in-vitro and in-silico studies have demonstrated the Candida-cidal and anti-biofilm activities of SMG, distinguishing it from the Candida-static efficacy of normal garlic. This is the first report that identifies several phytochemical signatures of SMG along with a potential anti-Candida compound, that is cholesta-4,6-dien-3-ol,(3-beta)-, which appears worthy of detailed studies in the future to explore the utility of SMG as a fungal phytotherapy agent, especially against drug-resistant Candida sp.


Marrow Fat Content and Composition in β-Thalassemia: A Study using 1 H-MRS.
β-thalassemia is a genetic disease that causes abnormal production of red blood cells (ineffective erythropoiesis, IE). IE is a condition known to change bone marrow composition. To evaluate the effect of IE on the marrow fat content and fat unsaturation levels in the proximal femur using 1 H-MRS. Prospective. Twenty-three subjects were included in this study, seven control and 16 β-thalassemia subjects. 3.0T; stimulated echo acquisition Mode (STEAM); magnetic resonance spectroscopy (MRS) sequence. Multiecho MRS scans were performed in four regions of the proximal left femur of each subject, that is, diaphysis, femoral neck, femoral head, and greater trochanter. The examined regions were grouped into red (diaphysis and femoral neck) and yellow marrow regions (femoral head and greater trochanter). The Jonckheere-Terpstra test was used to evaluate the impact of increasing disease severity on bone marrow fat fraction (BMFF), marrow conversion index, and fat unsaturation index (UI). Pairwise comparison analysis was performed when a significant trend (P < 0.05) was found. K-means clustering analysis was used to examine the clusters observed when BMFF in the red and yellow regions were studied (diaphysis against greater trochanter). BMFF showed a significant decreasing trend with increasing disease severity in both red (TJT = 109.00, z = -4.414, P < 0.05) and yellow marrow regions (TJT = 108.00, z = -4.438, P < 0.05). The opposite trend was observed in UI in both bone marrow regions (red marrow: TJT = 180.5, z = 3.515, P < 0.05; yellow marrow: TJT = 155.0, z = 2.282, P = 0.05). Three distinct forms of marrow adipogenesis were found when plotting BMFF diaphysis against BMFF greater trochanter: 1) normal (centroid: 80.4%, 66.6%), 2) partial disruption (centroid: 51.1%, 16.6%), and 3) total disruption (centroid: 2.6%, 1.6%). β-thalassemia is associated with decreased marrow fat, and increased marrow fat unsaturation level. 2 TECHNICAL EFFICACY STAGE: 3.


What data collection methods work best for COVID19 outbreak surveillance for people with end stage kidney disease? An observational cohort study using the UK Renal Registry.
Patients on kidney replacement therapy (KRT) are vulnerable to severe illness from COVID-19. Timely, accurate surveillance is essential for planning and implementing infection control at local, regional and national levels. Our aim was to compare two methods of data collection for COVID-19 infections amongst KRT patients in England. Adults receiving KRT in England were linked to two sources of data on positive COVID-19 tests recorded March-August 2020: (1) submissions from renal centres to the UK Renal Registry (UKRR) and (2) Public Health England (PHE) laboratory data. Patient characteristics, cumulative incidence by modality (in-centre haemodialysis (ICHD), home HD, peritoneal dialysis (PD) and transplant), and 28-day survival were compared between the two sources. 2,783/54,795 patients (5.1%) had a positive test in the combined UKRR-PHE dataset. Of these 2,783, 87% had positive tests in both datasets. Capture was consistently high for PHE (> 95% across modalities) but varied for UKRR (ranging from ICHD 95% to transplant 78%, p < 0.0001). Patients captured only by PHE were more likely to be on transplant or home therapies (OR 3.5 95% CI [2.3-5.2] vs. ICHD) and to be infected in later months (OR 3.3 95%CI [2.4-4.6] for May-June, OR 6.5 95%CI [3.8-11.3] for July-August, vs. March-April), compared to patients in both datasets. Stratified by modality, patient characteristics and 28-day survival were similar between datasets. For patients undergoing ICHD treatment the collection of data submitted directly by renal centres allows constant monitoring in real time. For other KRT modalities, using a national swab test dataset through frequent linkage may be the most effective method. Optimising central surveillance can improve patient care by informing interventions and assisting planning at local, regional and national levels.


The anti-emetic properties of 1-sulpiride in a ground-based model of space motion sickness.
L-sulpiride, at a dose of 4 mg/kg, essentially abolished motion-induced emesis in a group of 6 squirrel monkeys undergoing horizontal rotation at 25 rpm, a terrestrial model of space motion sickness (SMS). Extrapyramidal side effects were not observed. In the absence of the drug, the usual emetic response returned. In comparison while typical neuroleptics were also strongly anti-emetic, they produced a considerable degree of rigidity and akinesia.


Droperidol inhibits GABA(A) and neuronal nicotinic receptor activation.
Droperidol is used in neuroleptanesthesia and as an antiemetic. Although its antiemetic effect is thought to be caused by dopaminergic inhibition, the mechanism of droperidol's anesthetic action is unknown. Because gamma-aminobutyric acid type A (GABAA) and neuronal nicotinic acetylcholine receptors (nAChRs) have been implicated as putative targets of other general anesthetic drugs, the authors tested the ability of droperidol to modulate these receptors. gamma-Aminobutyric acid type A alpha1beta1gamma2 receptor, alpha7 and alpha4beta2 nAChRs were expressed in Xenopus oocytes and studied with two-electrode voltage clamp recording. The authors tested the ability of droperidol at concentrations from 1 nm to 100 microm to modulate activation of these receptors by their native agonists. Droperidol inhibited the GABA response by a maximum of 24.7 +/- 3.0%. The IC50 for inhibition was 12.6 +/- 0.47 nm droperidol. At high concentrations, droperidol (100 microm) activates the GABAA receptor in the absence of GABA. Inhibition of the GABA response is significantly greater at hyperpolarized membrane potentials. The activation of the alpha7 nAChR is also inhibited by droperidol, with an IC50 of 5.8 +/- 0.53 microm. The Hill coefficient is 0.95 +/- 0.1. Inhibition is noncompetitive, and membrane voltage dependence is insignificant. Droperidol inhibits activation of both the GABAA alpha1beta1gamma2 and alpha7 nAChR. The submaximal GABA inhibition occurs within a concentration range such that it might be responsible for the anxiety, dysphoria, and restlessness that limit the clinical utility of high-dose droperidol anesthesia. Inhibition of the alpha7 nAChR might be responsible for the anesthetic action of droperidol.