Engineering endoglucanase II from Trichoderma reesei to improve the catalytic efficiency at a higher pH optimum.
The catalytic efficiency and pH optimum of Trichoderma reesei endo-beta-1,4-glucanase II were improved by protein engineering. We subjected residue 342 to saturation mutagenesis, and further changed the enzyme by random mutagenesis and two rounds of DNA shuffling. Enzyme variants were purified and characterized. Variant N342V exhibited an optimal activity at pH 5.8, corresponding to a basic shift of 1 pH unit compared with the wild-type enzyme, and had improved catalytic efficiency (1.5-fold of k(cat)/K(m)) for the main substrates at pH 6.2. Variants N342R and N39R/L218H/W276R/N342T both had a pH optimum of 6.2 and the latter had improved catalytic efficiency (1.4-fold of k(cat)/K(m)) at pH 6.2. Variants L218H, Q139R/N342T and Q139R/L218H/W276R/N342T all had more than 4.5-fold higher activity in reactions compared with the wild-type at pH 7.0. The relationship between the structures and the activities of the variants were analyzed by modeling the structures of the endoglucanase II variants. More stable helixes and changed electrostatic interactions between the catalytic residues and substrates may explain the higher activities and higher pH optima of the variants.


Mycotoxicological control on raw material and tablets of cascara sagrada (Rhamnus purshiana).
Among phytotherapic medicines, tablets of cascara sagrada (Rhamnus purshiana) dried bark, usually used as laxative, are commercially widespread in our market. Taking into account natural origin and/or inappropriate procedures that may allow the occurrence of toxinogenic Aspergillus flavus group, a study on susceptibility to aflatoxin contamination and natural aflatoxin incidence was performed by TLC and HPLC methods. This survey allows one to conclude that bark of Cascara Sagrada is a good substrate for the growth of A. parasiticus NRRL 2999 and for aflatoxins production. Natural anatoxins presence was detected on 2 from 9 raw material samples. One of them (irradiated sample) had only aflatoxin B1 (10 μg/kg) and the other (pasteurized) was positive for aflatoxin B1 (19 μg/kg); G1 (6 μg/kg) and B2 (1.46 μg/kg). Only one from 10 lots of tablets analyzed was positive for aflatoxin B1 (5.42 μg/kg) and B2 (0.32 μg/kg).Therefore, adequate quality control including an aflatoxins assay must be performed to guarantee the harmlessness of natural drugs.


Drug repurposing to overcome microbial resistance.
Infections are a growing global threat, and the number of resistant species of microbial pathogens is alarming. However, the rapid development of cross-resistant or multidrug-resistant strains and the development of so-called 'superbugs' are in stark contrast to the number of newly launched anti-infectives on the market. In this review, I summarize the causes of antimicrobial resistance, briefly discuss different approaches to the discovery and development of new anti-infective drugs, and focus on drug repurposing strategy, which is discussed from all possible perspectives. A comprehensive overview of drugs of other indications tested for their in vitro antimicrobial activity to support existing anti-infective therapeutics is provided, including several critical remarks on this strategy of repurposing non-antibiotics to antibacterial drugs.


Phthalate exposure and odds of bacterial vaginosis among U.S. reproductive-aged women, NHANES 2001-2004.
Endocrine-disrupting chemicals, such as phthalates, are an unexamined potential risk factor for bacterial vaginosis (BV) and warrant investigation because hormones affect BV. We examined the association between phthalate exposure and BV in the National Health and Nutrition Examination Survey, 2001-2004. BV outcomes were defined as intermediate (Nugent score of 4-6) and positive (7-10). Phthalate metabolites, including monoethyl phthalate (MEP), mono-n-butyl phthalate (MnBP), and di(2-ethylhexyl) phthalate (DEHP) metabolites, were measured in urine. Among 854 women with complete data, multinomial logistic regression revealed that concentrations of MnBP (Q4 vs. Q1 OR = 3.01, 95% CI 1.76-5.15, p-trend <0.001) and ΣDEHP metabolites (Q4 OR = 2.55, 95% CI 1.45-4.47, p-trend = 0.03) were associated with Nugent-score BV, although only MnBP was significant after adjustment for confounders. Associations were null after adjustment for urinary creatinine (MnBP Q4 OR = 1.11, 95% CI 0.63-1.96; ΣDEHP Q4 OR = 0.72, 95% CI 0.37-1.39). Future work should further examine these relationships using direct measurements of intravaginal phthalates exposures.


Perception of opposite-moving dots in 3- to 5-month-old infants.
We conducted four experiments on the development of motion perception in a total of 109 3- to 5-month-old infants using motion stimuli consisting of opposite-moving dots. A psychophysical study showed that adult subjects perceived two global planes with opposite-moving dots, but this global perception collapsed when paired opposite-moving dots were located within 0.4 deg of one another (Qian, Andersen, & Adelson, 1994). We used this paired-dot stimulus as a non-target and the opponent motion stimulus as a target and examined target preference using methods based on forced-choice-preferential looking (Teller, 1979). In Experiment 1, we used 90 moving dots as stimuli. The results showed that 5-month-old infants had a significant preference for the targets but 4- and 3-month-olds did not. In Experiment 2, we used a small number of dots, and the results showed that 5-month-old infants did not prefer the target significantly. These results suggest that the preference for a target decreases according to the number of dots. In Experiment 3, we used opponent motion with long traveling length of the dots, and the results showed that all age groups, including 3-month-olds, had a preference for the moving targets. We showed that the preference observed in Experiment 3 was dependent not on local traveling length but on the global opponency. These results suggest that the perception of motion opponency based on a global motion cue emerges at 5 months of age (Experiments 1 and 2) and that the traveling length of the dots promote this perception (Experiments 3 and 4).


Randomized, Controlled, Multi-center Trial: Comparing the Safety and Efficacy of DA-9701 and Itopride Hydrochloride in Patients With Functional Dyspepsia.
Therapies of functional dyspepsia (FD) are limited. DA-9701 is a novel prokinetic agent formulated with Pharbitis semen and Corydalis Tuber. We aimed to assess the efficacy of DA-9701 compared with itopride in FD patients. Patients with FD randomly received either itopride 50 mg or DA-9701 30 mg t.i.d after a 2-week baseline period. After 4 weeks of treatment, 2 primary efficacy endpoints were analyzed: the change from baseline in composite score of the 8 dyspep-tic symptoms and the overall treatment effect. Impact on patients' quality of life was assessed using the Nepean Dyspepsia Index (NDI) questionnaire. We randomly assigned 464 patients with 455 having outcome data. The difference of the composite score change of the 8 symptoms between the 2 groups was 0.62, indicating that DA-9701 was not inferior to itopride. The overall treatment effect response rate was not different between the groups. When responder was defined as ≥ 5 of the 7 Likert scale, responder rates were 37% of DA-9701 and 36% of itopride group. Patients receiving DA-9701 experienced similar mean percentage of days with adequate relief during the 4-week treatment period compared with those receiving itopride (56.8% vs 59.1%). Both drugs increased the NDI score of 5 domains without any difference in change of the NDI score between the groups. The safety profile of both drugs was comparable. DA-9701 significantly improves symptoms in patients with FD. DA-9701 showed non-inferior efficacy to itopride with com-parable safety.


Effect of subinhibitory concentrations of josamycin on the expression of M protein by group A streptococci.
In an attempt to inhibit the biosynthesis of the type-specific M protein usually expressed on surface fimbriae group A Streptococcus pyogenes delta 2305 was cultivated in Todd-Hewitt broth containing 10% human serum and subinhibitory concentrations of either josamycin, erythromycin or clindamycin. Electron microscopy revealed that the antibiotic-pretreatment had little visible effect on the surface structures of the streptococci. However, josamycin and clindamycin-pretreated bacteria adhered less to hydrophobic gels than erythromycin-pretreated or untreated control cultures. Due to the decrease in surface hydrophobicity, the drug-pretreated bacteria also activated complement more readily and fixed more C3 on their surface. Consequently the killing of josamycin and clindamycin-pretreated bacteria by polymorphonuclear leucocytes was significantly enhanced. Similar findings were obtained when the M protein was removed from the bacteria by digestion with trypsin. These results suggest that josamycin, like clindamycin, reverses the capacity of group A streptococci to resist opsonization by normal human serum and interferes with the adhesion of the organisms to host epithelial cell surfaces.


Expanding scope of Kirkpatrick model from training effectiveness review to evidence-informed prioritization management for cricothyroidotomy simulation.
Modified Kirkpatrick model has been adopted to evaluate training effectiveness by 6 categories, including activity accounting (training objectives/success in organization change) at Level-0, reaction (satisfaction) at Level-1, learning (acquisition of surgical airway skills) at Level-2, behavior (post-training change in personal strengths) at Level-3, result (organizational or clinical outcomes) at Level-4, and Return on Investment (ROI) or Expectation (ROE) (monetary and societal values following training and other quality and safety related measures) at Level-5. The purpose of this hospital-based prospective observational study was twofold: i) To evaluate potential impacts on monetary and societal values and successful organization change following implementation of advanced Cricothyroidotomy simulator and standardized curriculum in healthcare simulation training, ii) To inform decisions of resource allocation by reviewing overall values and prioritization strategies for i) general surgeon/emergency physician ii) with seniority >5 years and iii) prior porcine training experience based on findings at Kirkpatrick Level-0, Level-4, and Level-5. Seventy doctors and 10 nurses completed Cricothyroidotomy training and follow-up questionnaires within 2021/22. All training usability scoring measured by Scales of Emergency Surgical Airway Simulator (SESAS-17) achieved over 4 out of 5 (Level-4) with effects in favor of emergency physicians or general surgeons (p < .5), regardless of seniority and prior training experience. Success in organization change (Level-0) and cost-effectiveness (Level-5) were hypothetically established using theoretical framework of Gleicher's formula and Roger's Diffusion of Innovation Theory. Overall training effectiveness, in terms of advantage in usability, cost-benefits and successful organizational changes, provided sound evidence to support continuous investment of new curriculum and innovative simulator and "Surgeon-and-emergency-physician-first" policy when it comes to resources allocation strategies for Cricothyroidotomy training. [ACGME competencies: Practice Based Learning and Improvement, Systems Based Practice.].


Regression models for censored time-to-event data using infinitesimal jack-knife pseudo-observations, with applications to left-truncation.
Jack-knife pseudo-observations have in recent decades gained popularity in regression analysis for various aspects of time-to-event data. A limitation of the jack-knife pseudo-observations is that their computation is time consuming, as the base estimate needs to be recalculated when leaving out each observation. We show that jack-knife pseudo-observations can be closely approximated using the idea of the infinitesimal jack-knife residuals. The infinitesimal jack-knife pseudo-observations are much faster to compute than jack-knife pseudo-observations. A key assumption of the unbiasedness of the jack-knife pseudo-observation approach is on the influence function of the base estimate. We reiterate why the condition on the influence function is needed for unbiased inference and show that the condition is not satisfied for the Kaplan-Meier base estimate in a left-truncated cohort. We present a modification of the infinitesimal jack-knife pseudo-observations that provide unbiased estimates in a left-truncated cohort. The computational speed and medium and large sample properties of the jack-knife pseudo-observations and infinitesimal jack-knife pseudo-observation are compared and we present an application of the modified infinitesimal jack-knife pseudo-observations in a left-truncated cohort of Danish patients with diabetes.


[C. Controversial technical and methodological problems in abdominal surgery. 1. Bile: a) Procedures in intraoperative actualization of papillary stenosis. Papillotomy in principle].
A critical comment on the question of papillotomy en principe is made, based on results from more than 7000 gallbladder operations and over 500 papillotomies, performed at the surgical departments of the municipal hospitals in Braunschweig and Lüneburg. In principle, too few papillotomies are carried out in many hospitals. The duration and seriousness of the cholecystitis is to a large degree responsible for the frequency of the formation of a papilla stenosis. Out of 232 multiple gallbladder operations performed at the Braunschweiger Hospital, 130 were necessary because of an incorrect opinion about the indication at the time of the first operation, in most cases due to an overlooked stenosis of the papilla.


Suburethral sling release.
The suburethral sling is an effective surgical procedure for curing genuine stress incontinence. However, a well-known complication is urinary retention. This report describes a method of modifying sling placement 6 weeks after the initial surgery to ameliorate the complication of urinary retention. A 68-year-old woman had undergone suburethral sling placement as part of her pelvic reconstructive surgery. Preoperatively, she had objective, urodynamic evidence of severe genuine stress incontinence, consistent with intrinsic sphincter deficiency. She was unable to void for 6 weeks postoperatively, at which time this procedure was offered. After an uneventful suburethral sling release, the patient had immediate return of voiding function, and symptoms resolved over the next 2 weeks. This procedure may be a useful alternative to sling removal in cases of persistent urinary retention after placement of a suburethral sling.


Acute and chronic role of 5-HT3 neuronal system on behavioral and neuroendocrine changes induced by intravenous cholecystokinin tetrapeptide administration in humans.
The influence of single and multiple oral doses of ondansetron, a selective 5-HT3 receptor antagonist, was evaluated against placebo on cholecystokinin tetrapeptide (CCK-4)-induced behavioral and neuroendocrine changes in humans. As compared to placebo, subjects receiving acute ondansetron treatment showed a significant decrease in the sum intensity of CCK-4-induced-panic symptoms (iPSS). Pre-CCK-4 neuropeptide Y (NPY) plasma levels were significantly higher and maximal changes in cortisol, growth hormone, and prolactin secretion from baseline (delta max) were significantly lower in the ondansetron group. After ondansetron and placebo chronic administration, there were no statistical differences in the iPSS between groups. Pre-CCK-4 NPY plasma levels were significantly higher; whereas, delta max for NPY significantly lower in the ondansetron group as compared to placebo. These results suggest a role for the 5-HT3 receptor in the neurobiology of panic disorder through a possible interaction with CCK and NPY systems. Ondansetron chronic effect on CCK-4-induced behavioral changes needs further exploration.


Circulating matrix metalloproteinase-9 is associated with cardiovascular risk factors in a middle-aged normal population.
Elevated levels of circulating matrix metalloproteinase-9 (MMP-9) have been demonstrated in patients with established coronary artery disease (CAD). The aim of this study was to analyse levels of MMP-9 in a population free from symptomatic CAD and investigate their associations with cardiovascular (CV) risk factors, including C-reactive protein (CRP). A cross-sectional study was performed in a population based random sample aged 45-69 (n = 345, 50% women). MMP-9 levels were measured in EDTA-plasma using an ELISA-method. CV risk factors were measured using questionnaires and standard laboratory methods. Plasma MMP-9 was detectable in all participants, mean 38.9 ng/mL (SD 22.1 ng/mL). Among individuals without reported symptomatic CAD a positive association (p<0.001) was seen, for both men and women, of MMP-9 levels regarding total risk load of eight CV risk factors i.e. blood pressure, dyslipidemia, diabetes, obesity, smoking, alcohol intake, physical activity and fruit and vegetable intake. The association was significant also after adjustment for CRP, and was not driven by a single risk factor alone. In regression models adjusted for age, sex, smoking, alcohol intake and CRP, elevated MMP-9 levels were independently positively associated with systolic blood pressure (p = 0.037), smoking (p<0.001), alcohol intake (p = 0.003) and CRP (p<0.001). The correlation coefficient between MMP-9 and CRP was r = 0.24 (p<0.001). In a population without reported symptomatic CAD, MMP-9 levels were associated with total CV risk load as well as with single risk factors. This was found also after adjustment for CRP.


Hyperprolactinemia and male sexual function: focus on erectile dysfunction and sexual desire.
The present paper aims to analyze and discuss the available evidence supporting the relationship between male sexual function and elevated prolactin (PRL) levels (HPRL). Two different sources of data were analyzed. Clinical data were derived from a series of patients seeking medical care for sexual dysfunction at our Unit. Out of 418 studies, 25 papers were used with a meta-analytic approach to evaluate the overall prevalence of HPRL in patients with erectile dysfunction (ED) and to study the influence of HPRL and its treatment on male sexual function. Among 4215 patients (mean age 51.6 ± 13.1 years) consulting for sexual dysfunction at our Unit, 176 (4.2%) showed PRL levels above the normal range. Meta-analytic data showed that HPRL is a rare condition among patients with ED (2 [1;3]%). Either clinical and meta-analytic data confirm a stepwise negative influence of PRL on male sexual desire (S = 0.00004 [0.00003; 0.00006]; I = -0.58915 [-0.78438; -0.39392]; both p < 0.0001 from meta-regression analysis). Normalization of PRL levels is able to improve libido. The role of HPRL in ED remains inconclusive. Data from a meta-analytic approach showed that either HPRL or reduced T levels were independently associated with ED rates. The normalization of PRL levels only partially restored ED. HPRL did not significantly contribute to ED severity, in our clinical setting. In conclusion, treating HPRL can restore normal sexual desire, whereas its effect on erection is limited.


[How to define chronically ill children].
The current overview article highlights specific features of defining chronic illnesses in children with a view on recent methodological developments. Methodological advantages and potential pitfalls of these approaches are discussed. Epidemiological trends are reported taking these definitions and methodological approaches into account. The conceptual interrelationships with other constructs in the area of health service research, for instance quality of life, are demonstrated.


Presynaptic action of neurotensin on cultured ventral tegmental area dopaminergic neurones.
Dopamine-containing neurones of the ventral tegmental area express neurotensin receptors which are involved in regulating cell firing and dopamine release. Although indirect evidence suggests that some neurotensin receptors may be localised on the nerve terminals of dopaminergic neurones in the striatum and thus locally regulate dopamine release, a clear demonstration of such a mechanism is lacking and a number of indirect sites of action are possible. We have taken advantage of a simplified preparation in which cultured rat ventral tegmental area dopaminergic neurones establish nerve terminals that co-release glutamate to determine whether neurotensin can act at presynaptic sites. We recorded glutamate-mediated synaptic currents that were generated by dopaminergic nerve terminals as an index of presynaptic function. The neurotensin receptor agonist NT(8-13) caused an inward current and an enhancement of the firing rate of dopaminergic neurones together with an increase in the frequency of spontaneous glutamate receptor-mediated excitatory postsynaptic currents (EPSCs). Incompatible with a direct excitatory action on nerve terminals, NT(8-13) failed to change the amplitude of individual action potential-evoked EPSCs or the frequency of miniature EPSCs recorded in the presence of tetrodotoxin. However, NT(8-13) reduced the ability of terminal D2 dopamine receptors to inhibit action potential-evoked EPSCs in isolated dopaminergic neurones. Taken together, our results suggest that in addition to its well-known somatodendritic excitatory effect leading to an increase in firing rate, neurotensin also acts on nerve terminals. The main effect of neurotensin on nerve terminals is not to produce a direct excitation, but rather to decrease the effectiveness of D2 receptor-mediated presynaptic inhibition.


Purinergic neuromuscular transmission is absent in the colon of P2Y(1) knocked out mice.
Purinergic and nitrergic co-transmission is the dominant mechanism responsible for neural-mediated smooth muscle relaxation in the gastrointestinal tract. The aim of the present paper was to test whether or not P2Y(1) receptors are involved in purinergic neurotransmission using P2Y(1)(−/−) knock-out mice. Tension and microelectrode recordings were performed on colonic strips. In wild type (WT) animals, electrical field stimulation (EFS) caused an inhibitory junction potential (IJP) that consisted of a fast IJP (MRS2500 sensitive, 1 μm) followed by a sustained IJP (N(ω)-nitro-L-arginine (L-NNA) sensitive, 1 mm). The fast component of the IJP was absent in P2Y(1)(−/−) mice whereas the sustained IJP (L-NNA sensitive) was recorded. In WT animals, EFS-induced inhibition of spontaneous motility was blocked by the consecutive addition of L-NNA and MRS2500. In P2Y(1)(−/−) mice, EFS responses were completely blocked by L-NNA. In WT and P2Y(1)(−/−) animals, L-NNA induced a smooth muscle depolarization but 'spontaneous' IJP (MRS2500 sensitive) could be recorded in WT but not in P2Y(1)(−/−) animals. Finally, in WT animals, 1 μm MRS2365 caused a smooth muscle hyperpolarization that was blocked by 1 μm MRS2500. In contrast, 1 μm MRS2365 did not modify smooth muscle resting membrane potential in P2Y(1)(−/−) mice. β-Nicotinamide adenine dinucleotide (β-NAD, 1 mm) partially mimicked the effect of MRS2365. We conclude that P2Y(1) receptors mediate purinergic neurotransmission in the gastrointestinal tract and β-NAD partially fulfils the criteria to participate in rodent purinergic neurotransmission. The P2Y(1)(−/−) mouse is a useful animal model to study the selective loss of purinergic neurotransmission.


Occurrence of two germline-related rheumatoid factor idiotypes in rheumatoid arthritis and in non-rheumatoid seropositive individuals.
Human rheumatoid factor (RF) paraproteins express two distinct light chain cross-reactive idiotypes defined by the monoclonal antibodies 17.109 and 6B6.6. These germline gene-related cross-reactive idiotypes are both carried on VK3 light chains and are each present on about one-third of IgM RF paraproteins. We assessed the degree to which these idiotypes are represented in polyclonal RFs. We used rheumatoid arthritis (RA) and non-RA RF-positive sera selected from a large cross-sectional population study (the Mini-Finland Health Survey), and sera from a community-based follow-up study of recent-onset RA patients from Heinola, Finland. In the Mini-Finland Health Survey, elevated levels of the 17.109 RF idiotype were seen in sera of 13% of the RA and 19% of the non-RA group; 6B6.6 RF was seen in 26% of the RA and 28% of the non-RA group. In sera of the Heinola follow-up study, 17.109 RF was seen in 12% initially, but in only 3% at 8 years. Similarly, 6B6.6 RF was detected in 25% initially, but in only 7% at 8 years. Ten sera positive for RF prior to the onset of clinical RA were identified from individuals of a second large population study from Finland (North Karelia project); two of these sera exhibited the 6B6.6 idiotype; none exhibited the 17.109 idiotype. The data are consistent with the concept that these germline gene-related cross-reactive RF idiotypes occur frequently in the polyclonal RF of non-RA as well as RA sera, and that in RA the idiotypes may sometimes be reduced or lost as a consequence of somatic diversification of the RF through somatic mutation, usage of new germline genes, or both.


Induced sputum supernatant bioactive lipid mediators can identify subtypes of asthma.
Induced sputum (IS) allows to measure mediators of asthmatic inflammation in bronchial secretions. The specific role of induced sputum supernatant (ISS) endogenous bioactive lipid mediators in subtypes of asthma is not well understood. To investigate the interactions between airway inflammation and clinical phenotypes of asthma, we integrated induced sputum supernatant (ISS) eicosanoids and quantitative assessment of infiltrating cells into new subtypes with the means of latent class analysis (LCA). One hundred and thirty-nine asthmatics with and without aspirin hypersensitivity underwent sputum induction. High-performance liquid chromatography or gas chromatography coupled with mass spectrometry was used to profile eicosanoids. Nineteen variables covering clinical characteristics, IS inflammatory cells and eicosanoids were considered in the LCA. Four phenotypic asthma classes were distinguished. Class 1 with mild-to-moderate asthma, chronic rhinosinusitis (CRS), high PGA2 in ISS and almost equal distribution of inflammation cell patterns. Class 3 subjects also had mild-to-moderate asthma but without upper airway symptoms. Induced sputum was often paucigranulocytic with low levels of lipid mediators. Classes 2 and 4 represented severe asthma with CRS and impaired lung function despite high doses of steroids. High blood and sputum eosinophilia was in line with high cysteinyl leukotrienes and PGD2 in ISS only in class 2. Class 4 subjects tended to have increased sputum neutrophilia and PGE2 in ISS. Aspirin hypersensitivity was most frequent among class 2 subjects. The LCA revealed four distinct asthma classes differing in eicosanoid pathways.


Transcatheter Cardiac Interventions in the Newborn: JACC Focus Seminar.
For neonates with critical congenital heart disease requiring intervention, transcatheter approaches for many conditions have been established over the past decades. These interventions may serve to stabilize or palliate to surgical next steps or effectively primarily treat the condition. Many transcatheter interventions have evidence-based records of effectiveness and safety, which have led to widespread acceptance as first-line therapies. Other techniques continue to innovatively push the envelope and challenge the optimal strategies for high-risk neonates with right ventricular outflow tract obstruction or ductal-dependent pulmonary blood flow. In this review, the most commonly performed neonatal transcatheter interventions will be described to illustrate the current state of the field and highlight areas of future opportunity.


[Pure red cell aplasia: Diagnosis, classification and treatment].
Pure red cell aplasia (PRCA) is a rare anemia characterised by profound reticulocytopenia caused by a marked reduction in bone marrow erythroblasts, without abnormalities in other blood lineages. Blackfan-Diamond anemia is an inherited ribosomopathy responsible for a hereditary form of PRCA. Acquired PRCA are separated in primary and secondary forms, including Parvovirus B19 infection, thymoma, lymphoproliferative disorders, autoimmune diseases (lupus) and drug-induced PRCA. The pathophysiology of PRCA is not fully understood and involves both humoral and T lymphocyte autoreactive cells. In Parvovirus B19-related PRCA, treatment is based on polyvalent immunoglobulins. Thymectomy for thymoma is mandatory but results in prolonged remission in a limited number of cases. The therapeutic strategy is based on expert opinion: corticosteroids in monotherapy provide few sustained responses. The choice of an additional immunosuppressant drug is guided by the presence of an underlying disease. In most cases, cyclosporine A is the first choice providing the best response rate but requires a concentration monitoring (150 to 250 ng/mL). The second choice is cyclophosphamide in large granular lymphocyte leukaemia. Sirolimus (mTOR inhibitor) seems to be a promising option especially in refractory cases. Transfusion independence is the main objective. If the patient receives numerous red blood cell transfusions (> 20 packs), iron overload assessment is crucial to initiate an iron chelation. A retrospective and prospective national cohort (EPIC-F) has been set up and is now available to include each case of PRCA to improve the knowledge of this disease and to optimize the therapeutic strategy.


Application of Sargassum biomass to remove heavy metal ions from synthetic multi-metal solutions and urban storm water runoff.
The ability of Sargassum sp. to biosorb four metal ions, namely lead, copper, zinc, and manganese from a synthetic multi-solute system and real storm water runoff has been investigated for the first time. Experiments on synthetic multi-solute systems revealed that Sargassum performed well in the biosorption of all four metal ions, with preference towards Pb, followed by Cu, Zn, and Mn. The solution pH strongly affected the metal biosorption, with pH 6 being identified as the optimal condition for achieving maximum biosorption. Experiments at different biosorbent dosages revealed that good biosorption capacity as well as high metal removal efficiency was observed at 3g/L. The biosorption kinetics was found to be fast with equilibrium being attained within 50 min. According to the Langmuir isotherm model, Sargassum exhibited maximum uptakes of 214, 67.5, 24.2 and 20.2mg/g for lead, copper, zinc, and manganese, respectively in single-solute systems. In multi-metal systems, strong competition between four metal ions in terms of occupancy binding sites was observed, and Sargassum showed preference in the order of Pb>Cu>Zn>Mn. The application of Sargassum to remove four heavy metal ions in real storm water runoff revealed that the biomass was capable of removing the heavy metal ions. However, the biosorption performance was slightly lower compared to that of synthetic metal solutions. Several factors were responsible for this difference, and the most important factor is the presence of other contaminants such as anions, organics, and other trace metals in the runoff.


Element distribution and morphology of spotted golden goatfish fish scales as affected by demineralisation.
Scales of spotted golden goatfish were subjected to non-collagenous protein removal followed by demineralisation with hydrochloric acid at different concentrations (0.25, 0.5, 0.75 and 1 M) for various times (30, 60 and 90 min). The morphology and element composition/distribution of scales from spotted golden goatfish as influenced by demineralisation conditions were determined. The appropriate demineralisation was pertained using 0.75 M hydrochloric acid for 30 min, in which the ash content was 0.62% (dry weight basis). The scales having non-collagenous protein removal with, and without, subsequent demineralisation were analysed for element content using X-ray fluorescence spectrometer. Images of different scales were determined using scanning electron microscopy (SEM) and scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX). Based on the images, an external layer rich in inorganic elements was removed. Most of Ca and P were eliminated with the coincidental increases in organic substances (C, N and O) after demineralisation. Demineralisation therefore mainly removed the external layer of scales, which facilitated the further extraction of collagen or gelatin.


Orientational order in the choline headgroup of sphingomyelin: A 14N-NMR study.
An aqueous dispersion of fully hydrated bovine sphingomyelin was studied using 14N-NMR spectroscopy. Spectra were obtained as a function of temperature over the range 15-80 degrees C, in both the liquid crystal and gel phases. In the liquid crystal phase, powder pattern lineshapes were obtained, whose quadrupolar splitting slowly decreases with increasing temperature. The spectra are increasingly broadened as the temperature is lowered through the phase transition into the gel phase. The linewidths and the second moments of these spectra indicate that the onset of a broad phase transition occurs at approx. 35 degrees C, in agreement with previous calorimetric and 31 P-NMR measurements. There is no evidence from the lineshapes for an hexagonal phase in this system, and this conclusion is supported by X-ray diffraction measurements carried out on aqueous dispersions of sphingomyelin in both phases. Assuming that the static nitrogen quadrupole coupling constant is the same for both sphingomyelin and dipalmitoyl-L-alpha-phosphatidylcholine (DPPC), the decrease observed in the quadrupolar splitting of sphingomyelin compared to that of DPPC indicates that the orientational order of the choline headgroup in liquid crystalline sphingomyelin is not the same as that of its counterpart in DPPC. Preliminary relaxation time measurements of T1 and T2 are presented which suggest that there are also dynamic differences between sphingomyelin and DPPC in the choline headgroup.


Mucoadhesive buccal patches based on interpolymer complexes of chitosan-pectin for delivery of carvedilol.
The study was designed to develop bioadhesive patches of carvedilol hydrochloride using chitosan (CH) and pectin (PE) interpolymer complexes and to systematically evaluate their in vitro and in vivo performances. Mucoadhesive buccal patches of carvedilol were prepared using solvent casting method. The physicochemical interaction between CH and PE was investigated by FTIR and DSC studies. The patches were evaluated for their physical characteristics like mass variation, content uniformity, folding endurance, ex vivo mucoadhesion strength, ex vivo mucoadhesion time, surface pH, in vitro drug release, in situ release study, and in vivo bioavailability study. The swelling index of the patches was found to be proportional to the PE concentration. The surface pH of all the formulated bioadhesive patches was found to lie between 6.2 and 7.2. The optimized bioadhesive patch (C1, CH:PE 20:80) showed bioadhesive strength of 22.10 ± 0.20 g, in vitro release of 98.73% and ex vivo mucoadhesion time of 451 min with in a period of 8 h. The optimized patch demonstrated good in vitro and in vivo results. The buccal delivery of carvedilol in rabbits showed a significant improvement in bioavailability of carvedilol from patches when compared to oral route.


Patient adherence to antiviral treatment for chronic hepatitis B and C: a systematic review.
Poor adherence to treatment for various chronic diseases is a frequent phenomenon. Current guidelines for the treatment of chronic hepatitis B (HBV) and hepatitis C (HCV) recommend optimal adherence, since it has been suggested that poor adherence is associated with an increased risk of virological failure. We aimed to give an overview of studies exploring adherence to combination treatment (PEG-interferon plus ribavirin) for HCV and nucleos(t)ide analogues for HBV. Material and methods. A systematic review was conducted using the databases PubMed, Embase, Cochrane Library and Web of Knowledge. Search terms included "adherence" or "compliance" combined with "hepatitis B", "hepatitis C" or "viral hepatitis". The final selection included 19 studies (13 HCV, 6 HBV). Large differences in patient numbers and adherence assessment methods were found between the various studies. For HCV mean adherence varied from 27 to 97%, whereas the proportion of patients with ≥ 80% adherence varied from 27 to 96%. Mean adherence reported in HBV studies ranged from 81 to 99%, with 66 to 92% of patients being 100% adherent. For both HCV and HBV studies, the highest adherence rates were reported in studies using self-report whereas lower adherence rates were reported in studies using pharmacy claims. Poor adherence to treatment was associated with an increased risk of virological failure. Non-adherence to treatment in chronic viral hepatitis is not a frequent phenomenon. However, given the increased risk of virological failure in poorly adherent patients, clinicians should routinely address adherence issues in all patients treated for chronic viral hepatitis.


Seven-year trajectories of body weight, quality of life and comorbidities following Roux-en-Y gastric bypass and sleeve gastrectomy.
There is limited long-term data comparing the outcomes of sleeve gastrectomy (SG) and Roux-en-Y gastric bypass (RYGB) for severe obesity, both with respect to body weight, quality of life (QOL) and comorbidities. We aimed to determine 7-year trajectories of body mass index (BMI), QOL, obesity-related comorbidities, biomarkers of glucose and lipid metabolism, and early major complications after SG and RYGB. Patients scheduled for bariatric surgery at two Norwegian hospitals, preferentially performing either SG or RYGB, were included consecutively from September 2011 to February 2015. Data was collected prospectively before and up to 7 years after surgery. Obesity-specific, generic and overall QOL were measured by the Impact of Weight on Quality of Life-Lite, Short-Form 36 and Cantril's ladder, respectively. Comorbidities were assessed by clinical examination, registration of medication and analysis of glucose and lipid biomarkers. Outcomes were examined with linear mixed effect models and relative risk estimates. Of 580 included patients, 543 (75% women, mean age 42.3 years, mean baseline BMI 43.0 kg/m2) were operated (376 SG and 167 RYGB). With 84.2% of participants evaluable after 5-7 years, model-based percent total weight-loss (%TWL) at 7 years was 23.4 after SG versus 27.3 after RYGB (difference 3.9%, p = 0.001). All levels of QOL improved similarly after the two surgical procedures but remained below reference data from the general population at all timepoints. Remission rates for type 2 diabetes, dyslipidemia, obstructive sleep-apnea and gastroesophageal reflux disease (GERD) as well as the rate of de novo GERD significantly favored RYGB. SG had fewer major early complications, but more minor and major late complications combined over follow-up. In routine health care, both SG and RYGB are safe procedures with significant long-term weight-loss, improvement of QOL and amelioration of comorbidities. Long-term weight-loss and remission rates of main obesity-related comorbidities were higher after RYGB.


Clinical investigation and management of Brucella suis seropositive dogs: A longitudinal case series.
Brucellosis in dogs caused by Brucella suis is an emerging zoonotic disease. To document clinical characteristics, serology, microbiology, and clinical response to treatment in B. suis-seropositive dogs. Longitudinal study of 27 privately-owned dogs. Dogs that tested positive by serology, culture, or real-time polymerase chain reaction (qPCR) were included in the study. Clinical (physical examination and imaging) and laboratory (serology, hematology, serum biochemistry, and qPCR or culture) assessments were made at baseline and after approximately 3, 6, 12, and 18 months. Dogs were followed for 10 895 dog days, with 17/27 dogs completing the 18-month follow-up. Ten dogs had signs consistent with brucellosis before enrollment (n = 4), at baseline (n = 2) or during follow-up (n = 6), with 2 dogs experiencing relapse of historical signs. Antibody titers persisted for the duration of follow-up in 15/17 dogs (88%). Radiographic (n = 5) and ultrasound (n = 11) findings, of variable clinical relevance, were observed. Brucella DNA and organisms were detected in 3 dogs, all of which had clinical signs, including in the milk of a bitch around the time of whelping. Brucella DNA was not detected in blood (n = 92 samples), urine (n = 80), saliva (n = 95) or preputial swabs (n = 78) at any time during follow-up. Six dogs underwent treatment, all of which achieved clinical remission although remission was not reflected by decreasing antibody titers. Most dogs with B. suis infections have subclinical infections. Serology is poorly associated with clinical disease. Excretion of organisms appears rare except in whelping bitches. Clinical management using antibiotics with or without surgery is recommended.


A systematic review of topical negative pressure therapy for acute and chronic wounds.
Topical negative pressure (TNP) therapy is becoming increasingly popular for all kinds of wounds. Its clinical and cost effectiveness is unclear. A search of randomized controlled trials (RCTs) on TNP in adult patients with all kinds of wounds in all settings was undertaken in Medline, Embase, Cinahl (to October 2007) and the Cochrane Library (to issue 4, 2007). Information was also sought from manufacturer of the VAC device. Selection of trials for analysis, quality assessment, data abstraction and data synthesis were conducted by two authors independently. The primary endpoint was any measure of wound healing. Secondary endpoints were infection, pain, quality of life, oedema, microcirculation, bacterial load, adverse events, duration of hospital stay and cost. The search identified 15 publications on 13 RCTs. These reported on patients with chronic wounds, diabetic wounds, pressure ulcers, skin grafts and acute wounds. In chronic and diabetic wounds, TNP did not allow earlier complete wound healing. It was, however, associated with a 1-10 day reduction in the time needed to prepare the wound for secondary closure surgery. In one trial on acute wounds, 17 (95 per cent confidence interval (c.i.) 0.02 to 0.32) per cent more wounds appeared to heal with TNP; the number needed to treat was six. This was, however, at the cost of an 11 (95 per cent c.i. 0.01 to 0.21) per cent higher complication rate; the number needed to harm was nine. There is little evidence to support the use of TNP in the treatment of wounds.


Remdesivir: Effectiveness and Safety in Hospitalized Patients with COVID-19 (ReEs-COVID-19)-Analysis of Data from Daily Practice.
Remdesivir was the first antiviral approved for treating COVID-19. We investigated its patterns of use, effectiveness and safety in clinical practice in Greece. This is a retrospective observational study of hospitalized adults who received remdesivir for COVID-19 in September 2020-February 2021. The main endpoints were the time to recovery (hospital discharge within 30 days from admission) and safety. The "early" (remdesivir initiation within 24 h since hospitalization) and "deferred" (remdesivir initiation later on) groups were compared. One thousand and four patients (60.6% male, mean age 61 years, 74.3% with severe disease, 70.9% with ≥1 comorbidities) were included, and 75.9% of them were on a 5-day regimen, and 86.8% were in the early group. Among those with a baseline mild/moderate disease, the median (95% CI) time to recovery was 8 (7-9) and 12 (11-14) days for the early and deferred groups, respectively (p < 0.001). The corresponding estimates for those with a severe disease were 10 (9-10) and 13 (11-15) days, respectively (p = 0.028). After remdesivir initiation, increased serum transaminases and an acute kidney injury were observed in 6.9% and 2.1%, respectively. Nine (0.9%) patients discontinued the treatment due to adverse events. The effectiveness of remdesivir was increased when it was taken within 24 h since admission regardless of the disease severity. Remdesivir's safety profile is similar to that described in clinical trials and other real-world cohorts.


Contribution to the knowledge of infectivity of Fasciola hepatica adolescariae after their passage through the digestive tract of white mice.
The study of infective elements of Fasciola hepatica which passed through the digestive tract of white mice revealed that those adolescariae (transitive adolescariae) (Table 1), which were not visibly damaged or possessed a more or less digested outer cyst wall, remained infective. The mice infected with transitive adolescariae harboured higher number of trematodes in the livers than those infected with infective elements which had not previously passed through the digestive tract of the definitive host.


Isolation of Cancer Stem Cells by Side Population Method.
The Hoechst side population (SP) method is a flow cytometry technique used to obtain stem cells based on the dye efflux properties of the ATP-binding cassette (ABC) transporters. The SP cells are characterized by their capability to efflux the fluorescent DNA-binding dye Hoechst 33342 through their ABC transporters and are enriched in stem cells, which are endowed with a self-renewal capacity and multilineage differentiation potential and express the stemness genes including ABC multidrug transporters. The protocols outlined in this book chapter describe the isolation method of the SP cells from human lung carcinoma cell lines by using Hoechst 33342. In addition, we refer to the propagation method of SP cells by successive rounds of fluorescence-activated cell sorting analysis for SP cells. These approaches will be helpful for the establishment of novel in vitro and in vivo models using cancer stem cells, which may play a key role during carcinogenesis and/or tumor progression.


3-(p-tolyl)-4-aminobutanoic acid synthesis, resolution into enantiomers and pharmacological activity.
Racemic 3-(p-totyl)-4-aminobutyric acid was obtained, resolved into enantiomers and their absolute configuration was determined. Both the racemic acid and two enantiomers were screened for CNS activity. R (+) enantiomers is 14--27-fold more effective than the S(-) one and 1.2--2.1 fold more effective than the racemate.


Gene expression profiles for the prediction of progression-free survival in diffuse large B cell lymphoma: results of a DASL assay.
We performed the whole genome cDNA-mediated annealing, selection and ligation assay with 164 formalin-fixed paraffin-embedded (FFPE) tumor samples to develop robust prognostic gene expression profiles in patients with diffuse large B cell lymphoma. The prognostic gene expression profiles were developed and validated by a gradient lasso and leave-one-out cross-validation process. We identified a set of genes whose expression provided prognostic indicators from whole data set (PRKCDBP, CASP10, FAM3C, KCNK12, MAN1A2, PRND, RAB1A, TMEM39B, SLC6A6, MMP12, FEM1B, C3orh37, RBP1, HK1, LOC400464, KIAA0746, and SLC25A23). This gene expression profile-based risk model could classify patients into two cross-validated risk groups with a significant difference in 5-year progression-free survival rates (71.1 vs. 45.5 %) and with a hazard ratio for recurrence of 2.45 (95 % CI, 1.44-4.16, P = 0.001). This model provided prognostic information independent of the International Prognostic Index (IPI), and discriminated high-risk group from patients belong to high/high-intermediate risk of IPI and activated B cell-like type. Thus, gene expression profiling from FFPE could provide additional prognostic information for diffuse large B cell lymphoma and our data underscore the need for development of risk-adapted treatment strategies based on gene expression profiles.


Lupus-prone NZBWF1/J mice, defective in cytokine signaling, are resistant to fumonisin hepatotoxicity despite accumulation of liver sphinganine.
Fumonisin B1 (FB1) is a mycotoxin produced by Fusarium verticillioides, commonly present in corn and other cereals. Exposure to FB1 causes organ-specific diseases in various species, e.g., equine leukoencephalomalacia and porcine pulmonary edema; in mice the response is hepatotoxicity. We earlier reported that ceramide synthase inhibition by FB1, the initial biochemical effect of this mycotoxin, results in modulation of cytokine network in response to accumulated free sphingoid bases. In the current study we used NZB/NZW-F1 (NZBW) mice that have modified cytokine expression and develop lupus beginning at 5 months of age. The NZBW and C57BL/6J (CBL) mice (appropriate control) were given five daily subcutaneous injections of either saline or 2.25 mg FB1/kg/day and euthanized 24 h after the last treatment. Peripheral leukocyte counts were higher after exposure to FB1 in CBL but not in NZBW. FB1 treatment caused increases of plasma alanine aminotransferase and aspartate aminotransferase activity in CBL mice indicating hepatotoxicity; no elevation of circulating liver enzymes was recorded in NZBW mice. Hepatotoxic responses were confirmed by microscopic evaluation of apoptotic cells. The FB1-induced proliferation of cells observed in CBL strain was abolished in NZBW animals. The sphinganine accumulation in liver after FB1 was equal in both strains of mice. The NZBW strain lacked the FB1-induced increases in the expression of liver tumor necrosis factor alpha, interferon gamma, receptor interacting protein (RIP), and tumor necrosis factor alpha-related apoptosis-inducing ligand (TRAIL), observed in CBL. Results confirmed our hypothesis that initial altered sphingolipid metabolism caused by FB1 leads to perturbation of liver cytokine network and ultimate cellular injury; the mice deficient in cytokine signaling are refractory to FB1 hepatotoxicity.


Probing the stress and depression circuits with a disease gene.
Selectively deleting a gene that has been linked to depression from specific neurons in mice sheds new light on a neural circuit that controls stress-induced depressive behaviors.


Decreased expression of BRAF-activated long non-coding RNA is associated with the proliferation of clear cell renal cell carcinoma.
BRAF-activated long non-coding RNA (BANCR) has been associated with various types of cancer. Nevertheless, the role of BANCR in clear cell renal cell carcinoma (ccRCC) is still not fully understood. This study aims to investigate the relationship between ccRCC and BANCR. Expression of BANCR in TCGA renal cancer data sets was analyzed. The expression pattern of BANCR in Immortalized normal human proximal tubule epithelial cell line HK-2 and ccRCC cell lines (ACHN, CAKI-1, A498 and 786-O) was detected by real-time quantitative RT-PCR (qRT-PCR). ccRCC tissues with adjacent normal renal tissues diagnosed by pathological methods from 62 patients were used to detect the expression of BANCR, and its correlation with prognosis of ccRCC patients was assessed by Kaplan-Meier method. The LV-BANCR vector was used to examine the influence of BANCR on the proliferation, migration, invasion, apoptosis and cell cycle distribution of ccRCC cells in vitro. BANCR was downregulated in renal cancer according to TCGA data sets. Compared with adjacent normal renal tissues and normal human proximal tubule epithelial cell line HK-2, BANCR expression was significantly decreased in ccRCC tissues and ccRCC cell lines, and its low expression was associated with poor prognosis. Moreover, in the condition of BANCR overexpression by LV-BANCR vector, the proliferation, migration, invasion capacity of ccRCC cells was inhibited, while the apoptosis was increased and the G1 cell cycle arrest was induced in vitro. BANCR is downregulated in ccRCC tissues and cell lines, and is associated with ccRCC progression. Thus, BANCR may represent a novel prognostic biomarker and a potential therapeutic target for ccRCC patients.


Editorial: Sex-Specific Developmental Trajectories of Psychopathology: The Case of Substance Misuse.
Childhood psychopathology can cause an array of adverse outcomes later in later life, including substance use disorders (SUDs). Perhaps because substance abuse is thought to be on the externalizing continuum, the prospective association between childhood externalizing disorders and SUDs is more frequently studied than the association with internalizing disorder. Indeed, research on attention-deficit/hyperactivity disorder (ADHD), oppositional defiant disorder, and conduct disorder (CD) quite consistently demonstrate that these disorders are associated with increased SUDs risk in later life.1 However, although it is often thought that only externalizing disorders in childhood increase the risk of SUDs in later life, there is evidence, albeit more fragmented, for a similar association between internalizing disorders in childhood and later substance use.1 Because internalizing and externalizing disorders are often comorbid, it is vital to investigate their joint effect on the developmental trajectory toward SUDs. So far, this trajectory has mainly been studied from an externalizing perspective, but Virtanen et al.2 studied this prospective association with internalizing disorders as the start point, while considering the possibility of biological sex-specific patterns.


[Effect of social factors on the oral health status and utilization of stomatologic care by elderly citizens].
Different social items (marital status, social contacts, indigence, level of education) and in addition the social area have consequences to the dental health behaviour and the health consciousness until old age, and influence the success of dental care for the prosthesis wearer.


Correlations between ultrastructural findings and electrocochleographic recordings in experimental aminoside intoxication.
Correlations are made between the electrocochleographic aspects and the ultrastructural findings in 60 guinea pigs intoxicated with either Gentamycin or Tobramycin. Small cochlear lesions are studied in order to identify the electrical aspect corresponding to the earliest anatomical cell alterations. The wave shape seems to be more important than the classical input-output functions as first signs of the inability of the organ of Corti to clear the aminoside antibiotics. The reversal of N1 and N2 and later the accentuation of P1, frequently correspond to early signs of cell intoxication such as disruption of the regular arrangement of the endoplasmic reticulum and proliferation of lysosomes. A similar situation may obtain in humans intoxicated with aminosides.


Effects of discontinuation of chronic feeding of diethylnitrosamine on the development of hepatomas in adult rats.
Diethylnitrosamine (DENA) at 10 mg/kg was fed to adult rats either continuously or for periods ranging from 1 to 10 weeks. Survival correlated inversely with the duration of carcinogen feeding. Less than 4 weeks of DENA feeding produced only preneoplastic foci that persisted indefinitely; 4 weeks were found to be necessary for the transformation of preneoplastic lesions into liver cancers; after 6 weeks, the incidence of hepatomas was 100%. The process of liver cancerization appeared to be identical whether DENA was fed for 8 weeks or continuously up to the time of death. These results are discussed in the light of the evolution of the homoeostatic control of liver-cell division during DENA feeding, in order to distinguish the different successive roles played by the carcinogen.


Alpha and delta ganglion cells in the rat retina.
In the rat retina a distinctive class of large ganglion cell was demonstrated by intracellular staining with Lucifer Yellow and with reduced silver staining. They are referred to as alpha cells because they resemble the alpha cells of other mammalian retinae. A second class, called delta cells, is also described. Both classes belong to the type I group defined by Perry (Proc. R. Soc. Lond. [Biol.] 204:363-375, '79). The dendritic trees of both classes stratify in either an inner or outer lamina of the inner plexiform layer which presumably corresponds to an on/off dichotomy in the response to light. Rat alpha cells constitute 2-4% of all ganglion cells, and their density, size, and detailed morphological appearance change with retinal location. Inner and outer stratifying alpha cells of the rat show significant differences compared to those of other mammals. In central retina (at the large cell density maximum) the densities and dendritic field sizes of inner and outer alpha cells are approximately equal. However, in peripheral retina outer alpha cells are up to three times more numerous and have dendritic field areas only one-third the size of those of the inner alpha cells. The maximal density is about 110 alpha cells/mm2; peripheral densities are about 30/mm2. The smallest central dendritic field diameters are 220 microns. Peripheral dendritic field diameters are 350-550 microns for outer and 570-790 microns for inner alpha cells. Each subpopulation is distributed in a regular mosaic, and the territorial arrangement of the dendritic fields provides a homogeneous coverage of the retina. The dendritic coverage is three- to 3.6-fold for each subpopulation, irrespective of their other quantitative differences. Eccentricity-dependent receptive field sizes of the alpha cells are predicted from the morphological data.


Evaluation of in-stent stenosis by magnetic resonance phase-velocity mapping in nickel-titanium stents.
To evaluate different grades of in-stent stenosis in a nickel-titanium stent with MRI. Magnetic resonance phase velocity mapping (MR-PVM) was used to measure flow velocity through a 9-mm NiTi stent with three different degrees of stenosis in a phantom study. The tested stenotic geometries were 1) axisymmetric 75%, 2) axisymmetric 90%, and 3) asymmetric 50%. The MR-PVM data were subsequently compared with the velocities from computational fluid dynamic (CFD) simulations of identical conditions. Good quantitative agreement in velocity distribution for the 50% and 75% stenoses was observed. The agreement was poor for the 90% stenosis, most likely due to turbulence and the high-velocity gradients found in the small luminal area relative to the pixel resolution in our imaging settings. The accuracy of the MRI velocities inside the stented area renders MRI a modality that may be used to assess moderate to severe in-stent restenosis (ISR) in medium-sized vascular stents in peripheral vessels, such as the iliac, carotid, and femoral arteries. Advances in MR instrumentation may provide sufficient resolution to obtain adequate velocity information from smaller vessels, such as the coronary arteries, and allow MRI to substitute for invasive and expensive catheterization procedures currently in clinical use.


Variations in the Regulatory Region of Alpha S1-Casein Milk Protein Gene among Tropically Adapted Indian Native (Bos Indicus) Cattle.
Regulatory region of milk protein alpha S1-casein (αS1-CN) gene was sequenced, characterized, and analyzed to detect variations among 13 Indian cattle (Bos indicus) breeds. Comparative analysis of 1,587 bp region comprising promoter (1,418 bp), exon-I (53 bp), and partial intron-I (116 bp) revealed 35 nucleotide substitutions (32 within promoter region, 1 in exon-I, and 2 in partial intron-I region) and 4 Indels. Within promoter, 15 variations at positions -1399 (A > G), -1288 (G > A), -1259 (T > C), -1158 (T > C), -1016 (A > T), -941 (T > G), -778 (C > T), -610 (G > A), -536 (A > G), -521 (A > G), -330 (A > C), -214 (A > G), -205 (A > T), -206 (C > A), and -175 (A > G) were located within the potential transcription factor binding sites (TFBSs), namely, NF-κE1/c-Myc, GATA-1, GATA-1/NF-E, Oct-1/POU3F2, MEF-2/YY1, GATA-1, AP-1, POU1F1a/GR, TMF, GAL4, YY1/Oct-1, HNF-1, GRalpha/AR, GRalpha/AR, and AP-1, respectively. Seventy-four percent (26/35) of the observed SNPs were novel to Indian cattle and 11 of these novel SNPs were located within one or more TFBSs. Collectively, these might influence the binding affinity towards their respective nuclear TFs thus modulating the level of transcripts in milk and affecting overall protein composition. The study provides information on several distinct variations across indicine and taurine αS1-CN regulatory domains.


Improved prediction of immune checkpoint blockade efficacy across multiple cancer types.
Only a fraction of patients with cancer respond to immune checkpoint blockade (ICB) treatment, but current decision-making procedures have limited accuracy. In this study, we developed a machine learning model to predict ICB response by integrating genomic, molecular, demographic and clinical data from a comprehensively curated cohort (MSK-IMPACT) with 1,479 patients treated with ICB across 16 different cancer types. In a retrospective analysis, the model achieved high sensitivity and specificity in predicting clinical response to immunotherapy and predicted both overall survival and progression-free survival in the test data across different cancer types. Our model significantly outperformed predictions based on tumor mutational burden, which was recently approved by the U.S. Food and Drug Administration for this purpose1. Additionally, the model provides quantitative assessments of the model features that are most salient for the predictions. We anticipate that this approach will substantially improve clinical decision-making in immunotherapy and inform future interventions.


Diagnostic accuracy of the BioFire® FilmArray® pneumonia panel in COVID-19 patients with ventilator-associated pneumonia.
Ventilator-Associated pneumonia (VAP) is one of the leading causes of morbidity and mortality in critically ill COVID-19 patients in lower-and-middle-income settings, where timely access to emergency care and accurate diagnostic testing is not widely available. Therefore, rapid microbiological diagnosis is essential to improve effective therapy delivery to affected individuals, preventing adverse outcomes and reducing antimicrobial resistance. We conducted a cross-sectional study of patients with suspected VAP and COVID-19, evaluating the diagnostic performance of the BioFire® FilmArray® Pneumonia Panel (FA-PP). Respiratory secretion samples underwent standard microbiological culture and FA-PP assays, and the results were compared. We included 252 samples. The traditional culture method detected 141 microorganisms, and FA-PP detected 277, resulting in a sensitivity of 95% and specificity of 60%, with a positive predictive value of 68% and negative predictive value of 93%. In samples with high levels of genetic material (> 10^5 copies/mL), the panel had a sensitivity of 94% and specificity of 86%. In addition, 40% of the culture-negative samples had positive FA-PP® results, of which 35% had > 10^5 copies/mL of genetic material. The most prevalent bacteria were Gram-negative bacilli, followed by Gram-positive cocci. The panel identified 98 genes associated with antimicrobial resistance, predominantly extended-spectrum beta-lactamases (28%). The FA-PP is a sensitive assay for identifying bacteria causing VAP in patients with COVID-19, with a greater capacity to detect bacteria than the conventional method. The timely microbiological recognition offered by this panel could lead to optimized decision-making processes, earlier tailored treatment initiation, and improved antibiotic stewardship practices.


Determinants of Perceived Physical Environment Barriers among Community-Dwelling Elderly in Taiwan.
To test the hypothesis that mobility, activities of daily living, and the interaction between them can play a key role in determining perceived physical environment barriers among community-dwelling elderly. Cross-sectional. Community. One hundred and ninety-seven community-dwelling elderly with more than 7 points on the Short Portable Mental State Questionnaire and less than 7 points on the Geriatric Depression Scale (15 items). None. Time Get-up and Go test (TUG), the subscales of basic activity of daily living (BADL)/instrumental activities of daily living (IADL) of the Hierarchy of Care Required (HCR), and the physical/structural subscale of the Craig Hospital Inventory of Environmental Factors in Community-dwelling Elderly in Taiwan were used to measure mobility, activities of daily living and perceived physical environment barriers, respectively. Hierarchical linear regression analyses were used to test the study hypothesis. Significant and positive relations were found to exist between perceived physical environment barriers and (1) the TUG time (β=.300, p<.05), and (2) the IADL score for the HCR (β=.322, p<.05), respectively. A significant and negative relation existed between perceived physical environment barriers and the interaction term (the TUG time and the IADL score for the HCR) (β=-.211, p<.05). Mobility, IADL and the interaction between them are found to be significant determinants of perceived physical environment barriers in the community-dwelling elderly under consideration. Strategies targeting the enhancement of mobility among community-dwelling elderly are suggested to lead to improvements in the degree to which physical environment barriers are perceived. This beneficial effect could be greater in the case of elderly individuals with better IADL function.


Localization of the intracellular activity domain of Pasteurella multocida toxin to the N terminus.
We have shown that Pasteurella multocida toxin (PMT) directly causes transient activation of Gqalpha protein that is coupled to phosphatidylinositol-specific phospholipase Cbeta1 in Xenopus oocytes (B. A. Wilson, X. Zhu, M. Ho, and L. Lu, J. Biol. Chem. 272:1268-1275, 1997). We found that antibodies directed against an N-terminal peptide of PMT inhibited the toxin-induced response in Xenopus oocytes, but antibodies against a C-terminal peptide did not. To test whether the intracellular activity domain of PMT is localized to the N terminus, we conducted a deletion mutational analysis of the PMT protein, using the Xenopus oocyte system as a means of screening for toxin activity. Using PCR and conventional cloning techniques, we cloned from a toxinogenic strain of P. multocida the entire toxA gene, encoding the 1,285-amino-acid PMT protein, and expressed the recombinant toxin as a His-tagged fusion protein in Escherichia coli. We subsequently generated a series of N-terminal and C-terminal deletion mutants and expressed the His-tagged PMT fragments in E. coli. These proteins were screened for cytotoxic activity on cultured Vero cells and for intracellular activity in the Xenopus oocyte system. Only the full-length protein without the His tag exhibited activity on Vero cells. The full-length PMT and N-terminal fragments containing the first 500 residues elicited responses in oocytes, but the C-terminal 780 amino acid fragment did not. Our results confirm that the intracellular activity domain of PMT is localized to the N-terminal 500 amino acids of the protein and that the C terminus is required for entry into cells.


High-resolution genetic and physical mapping of the cauliflower high-beta-carotene gene Or ( Orange).
Mutation in the cauliflower gene Or causes high levels of beta-carotene to accumulate in various tissues of the plant that are normally devoid of carotenoids. To decipher the molecular basis by which Or regulates carotenoid accumulation, we have undertaken the isolation of Or by a map-based cloning strategy. Two previously isolated, locus-specific, sequence-characterized amplified region (SCAR) markers that flank Or were employed for the analysis of a large segregating population consisting of 1632 F(2) individuals, and a high-resolution genetic linkage map of the Or locus region was developed. To facilitate positional cloning, we constructed a cauliflower genomic library in a bacterial artificial chromosome (BAC) vector, using high molecular weight DNA from Or homozygotes. The BAC library comprises 60,288 clones with an average insert size of 110 kb, and represents an estimated 10-fold coverage of the genome. A BAC contig encompassing the Or locus was established by screening the library with a marker that is closely linked to Or and by identifying overlapping BAC clones by chromosome walking. Physical mapping delimited the Or locus to a 50-kb DNA fragment within a single BAC clone, which corresponds to a genetic interval of 0.3 cM.


Why surgery won the SYNTAX trial and why it matters.
The Synergy Between Percutaneous Coronary Intervention With Taxus and Cardiac Surgery (SYNTAX) trial cause of death analysis shows that cardiac death due to myocardial infarction is 10 times higher with percutaneous coronary intervention than coronary artery bypass grafting in the higher-risk patients. There was a clear advantage for surgery in the prevention of death in both the intermediate and high SYNTAX score groups with 3-vessel disease and in the high SYNTAX score group with left main disease, and that incremental advantage is statistically significant and widening with time, which should be transparently communicated to patients.


Rooting pattern and water relations of three pasture grasses growing in drying soil.
Seedlings of perennial ryegrass (Lolium perenne L. S23), cocksfoot (Dactylis glomerata L. S37) and Timothy (Phleum pratense L. S48) were rooted into tubes of soil and plants were either watered well or remained unwatered for a period of seven days. Measurements were made of root water relations and of growth of roots and shoots. Root turgor of Dactylis was larger than that of Lolium and Phleum. As a result of accumulation of solutes, turgor of Dactylis and Lolium was maintained as the soil dried. Phleum roots lost turgor in drying soil and this characteristic correlated well with a water stress-induced reduction in root and shoot growth. Soil drying had marked effects on the rooting patterns of two of the three species. Phleum seedlings which, in wet soil are deep rooting were restricted in their depth of rooting, while roots of water-stressed Dactylis seedlings grew deeper into the profile than did roots of well-watered plants. When water was withheld from plants, deeper rooting apparently resulted in a more favourable shoot water balance which had a beneficial effect on shoot growth.


Targeting small-cell lung cancer with novel fluorescent analogs of somatostatin.
Early, accurate detection of small-cell lung cancer (SCLC), before it becomes systemic, is essential for successful treatment. Fluorescence-based imaging provides safe, sensitive detection of malignancies. Targeted delivery of fluorophores increases sensitivity of endoscopic imaging. We synthesized novel somatostatin analogs, based on backbone cyclic peptides, and conjugated them with fluorescent agents. Nineteen conjugates differing in core peptide, length of alkyl linker and fluorescence moiety (rhodamine and fluorescein) were tested in vitro, using a receptor binding assay, and nine of the more promising conjugates were tested in vivo by fiber-optic spectrofluorimetry and quantitative spectral imaging, on an H69 human SCLC tumor mouse xenograft model. The lead compound showed exceptional tumor/normal tissue ratios, ranging from 9 to 90, and has potential for targeting SCLC overexpressing somatostatin receptors.


Retroauricular skin: a flaps bank for ear reconstruction.
The retroauricular skin has always been given much attention by the reconstructive surgeon for ear and face reconstruction because it is richly vascularised, as many anatomical investigations show, it is hidden behind the ear, its skin is very similar to that of ear and face. All these reasons make it an ideal donor site for ear reconstruction. The authors propose their own algorithm for reconstruction of every kind of anterior defects of the auricle with different Retroauricular Island Flaps (RIFs) based on the location and size of the defect developed over a 16 years single institution's experience with a series of 216 consecutive cases. 216 patients have undergone ear reconstruction with RIFs from 1999 to 2006. In 52 a Superior Pedicle RIF (SP-RIF) was used for defects of the upper half of the auricle. In 68 cases a Perforator RIF (P-RIF) was used for conchal reconstruction. In 96 cases an Inferior Pedicle RIF (IP-RIF) was used for reconstruction of nonmarginal and superficial marginal defects of the auricle. No flap failure was recorded. Excellent morphological reconstruction was obtained with these flaps with no sequealae at the donor site in terms of form and function. Only in the case of P-RIFs the sulcus becomes flat in its central part, but this has never affected the possibility of wearing spectacles. The SP-RIFs may sometimes show some signs of venous stasis that invariably resolve in the first two postoperative days. The retroauricular skin may be considered a flaps bank for ear reconstruction. It offers in fact a great variety of island flaps that are suitable for every kind of loss of substance of the ear, have a safe vascularisation, skin of similar colour and texture, are easy to harvest under local anaesthesia on an outpatient basis and cause no relevant morbidity at the donor site. Location and size of the defects lead the choice between the different types of RIFs.


Delta hepatitis in Lebanon. Prevalence studies and a report on six siblings with chronic delta-positive active hepatitis.
The prevalence of delta infection in Lebanon is reported for the first time. Delta antigen and antibody were screened for in serum of 43 patients consecutively seen in hospital, 22 with acute hepatitis B (anti-HBc IgM-positive), and 21 with histological evidence of chronic active hepatitis, 6 of whom were brothers in a sibship of eleven. Twenty asymptomatic HBsAg carriers without clinical or biochemical evidence of liver disease were similarly studied. None of the asymptomatic carriers or acute hepatitis B patients had markers of delta infection. In contrast, delta antibody in high titre (greater than 1:5000) was found in 12 (57%) of the patients with chronic active hepatitis, including all six brothers. Five sisters in the sibship were anti-HBs-positive without evidence of liver disease, suggesting a horizontal mode of transmission of the delta virus, as a superinfection on a hepatitis B carrier state. Excluding the sibship, delta infection was present in 6 of 15 (40%) chronic active hepatitis patients. This prevalence is similar to other Middle Eastern countries. Delta infection was associated with severe liver disease.


The year(s) of the contrast agent - micro-MRI in the new millennium.
The beginning of the new millennium has been a dynamic time for the field of magnetic resonance imaging (MRI). Exciting recent advances have been made at all levels of imaging, ranging from the visualization of single cells to rodents, birds and the human brain. Many of these techniques employ contrast agents to visualize the movement or activity of cells or organs in vivo; examples of this include the observation of stem cell migration, the tracking of labeled T cells, and the visualization of the events of gastrulation in developing Xenopus embryos. Other advances include improved techniques for elucidating white matter tracts in brain by either monitoring the diffusion of water along the tracts or tracing active neuronal tracts in vivo with Mn(2+) ions. Imaging of the immune system presents two dramatically different challenges: imaging most if not all of the body to follow cell trafficking, and imaging at cellular resolution to follow key intercellular and intracellular events.


Primary left atrial tumor showing mitral valve stenosis and left upper lobe pulmonary edema: report of a case.
Myxoma and other neoplasms in the left atrium often show signs and symptoms similar to those of mitral valve stenosis. Malignant fibrous histiocytoma, one of the most common sarcomas, occurs in the deep soft tissue of the limbs and retroperitoneum in adults. With the exception of primary lung cancer, however, sarcomatous intravascular extension in the lung and infiltration into the left atrium are rare. We report a case of overt left upper lobe pulmonary edema observed on chest radiograms and a mass occupying much of the left atrium and exhibiting hemodynamics similar to those of mitral valve stenosis on transthoracic echocardiography. Intraoperative and pathologic examination showed the mass to be a primary malignant fibrous histiocytoma extending from the left upper pulmonary vein into the left atrium.


Visual feedback improves bimanual force control performances at planning and execution levels.
The purpose of this study was to determine the effect of different visual conditions and targeted force levels on bilateral motor synergies and bimanual force control performances. Fourteen healthy young participants performed bimanual isometric force control tasks by extending their wrists and fingers under two visual feedback conditions (i.e., vision and no-vision) and three targeted force levels (i.e., 5%, 25%, and 50% of maximum voluntary contraction: MVC). To estimate bilateral motor synergies across multiple trials, we calculated the proportion of good variability relative to bad variability using an uncontrolled manifold analysis. To assess bimanual force control performances within a trial, we used the accuracy, variability, and regularity of total forces produced by two hands. Further, analysis included correlation coefficients between forces from the left and right hands. In addition, we examined the correlations between altered bilateral motor synergies and force control performances from no-vision to vision conditions for each targeted force level. Importantly, our findings revealed that the presence of visual feedback increased bilateral motor synergies across multiple trials significantly with a reduction of bad variability as well as improved bimanual force control performances within a trial based on higher force accuracy, lower force variability, less force regularity, and decreased correlation coefficients between hands. Further, we found two significant correlations in (a) increased bilateral motor synergy versus higher force accuracy at 5% of MVC and (b) increased bilateral motor synergy versus lower force variability at 50% of MVC. Together, these results suggested that visual feedback effectively improved both synergetic coordination behaviors across multiple trials and stability of task performance within a trial across various submaximal force levels.


Gut-muscle axis and sepsis-induced myopathy: The potential role of gut microbiota.
Sepsis is described as an immune response disorder of the host to infection in which microorganisms play a non-negligible role. Most survivors of sepsis experience ICU-acquired weakness, also known as septic myopathy, characterized by skeletal muscle atrophy, weakness, and irreparable damage/regenerated or dysfunctional. The mechanism of sepsis-induced myopathy is currently unclear. It has been believed that this state is triggered by circulating pathogens and their related harmful factors, leading to impaired muscle metabolism. Sepsis and its resulting alterations in the intestinal microbiota are associated with sepsis-related organ dysfunction, including skeletal muscle wasting. There are also some studies on interventions targeting the flora, including fecal microbiota transplants, the addition of dietary fiber and probiotics in enteral feeding products, etc., aiming to improve sepsis-related myopathy. In this review, we critically assess the potential mechanisms and therapeutic prospects of intestinal flora in the development of septic myopathy.


Aquaporin-4 gene deletion in mice increases focal edema associated with staphylococcal brain abscess.
Brain abscess is associated with local vasogenic edema, which leads to increased intracranial pressure and significant morbidity. Aquaporin-4 (AQP4) is a water channel expressed in astroglia at the blood-brain and brain-CSF barriers. To investigate the role of AQP4 in brain abscess-associated edema, live Staphylococcus aureus (10(5) colony-forming units) was injected into the striatum to create a focal abscess. Wild-type and AQP4-deficient mice had comparable immune responses as measured by brain abscess volume (approximately 3.7 mm3 at 3 days), bacterial count and cytokine levels in brain homogenates. Blood-brain barrier permeability was increased comparably in both groups as assessed by extravasation of Evans blue dye. However, at 3 days the AQP4 null mice had significantly higher intracranial pressure (mean +/- SEM 27 +/- 2 vs. 17 +/- 2 mmHg; p < 0.001) and brain water content (81.0 +/- 0.3 vs. 79.3 +/- 0.5 % water by weight in the abscess-containing hemisphere; p < 0.01) than wild-type mice. Reactive astrogliosis was found throughout the abscess-containing hemisphere; however, only a subset of astrocytes in the peri-abscess region of wild-type mice had increased AQP4 immunoreactivity. Our findings demonstrate a protective effect of AQP4 on brain swelling in bacterial abscess, suggesting that AQP4 induction may reduce vasogenic edema associated with cerebral infection.


A Novel Approach to Functionalization of Aryl Azides through the Generation and Reaction of Organolithium Species Bearing Masked Azides in Flow Microreactors.
A novel straightforward method for aryl azides having functional groups based on generation and reactions of aryllithiums bearing a triazene group from polybromoarenes using flow microreactor systems was achieved. The present approach will serve as a powerful method in organolithium chemistry and open a new possibility in the synthesis of polyfunctional organic azides.


Inadvertent P-hacking among trials and systematic reviews of the effect of progestogens in pregnancy? A systematic review and meta-analysis.
Progestogens have been evaluated in numerous trials and meta-analyses, many of which concluded they were effective. However, two large trials PROMISE and OPPTIMUM have recently concluded that progesterone was ineffective. This raises the possibility that earlier studies and reviews had been biased by either selective publication or selective choice of outcomes, so called "P-hacking". To compare the findings all progestogen trials and systematic reviews with those of trials with pre-registered primary outcomes which avoided selective outcome reporting. Search of PubMed, the Cochrane Library and trial registries. Registration PROSPERO CRD42016035303. Systematic reviews of randomised trials comparing progestogen with placebo in pregnancy and the individual trials included in those reviews. The subset of trials reporting a pre-registered primary outcome were compared with the totality of trials and reviews. For reviews all outcomes were included. For individual trials all outcomes reported in the systematic reviews were included. For the comparison group we recorded the registered primary outcome from trials that were either registered before they started, or registered during the recruitment phase and also double blind. Nineteen of twenty-nine meta-analyses concluded that progestogens were effective. Twenty-two trials reported their pre-registered primary outcomes. There was no effect of progesterone on primary registered dichotomous outcome RR 1.00 (95% CI 0.94-1.07). Only one of the 22 showed a nominally statistically significant benefit. When evaluated in registered double-blind trials with analysis restricted to predefined primary outcomes, progestational agents in pregnancy are ineffective. Progestogens to prevent pregnancy loss, an example of P-hacking.


Actinomadura jiaoheensis sp. nov. and Actinomadura sporangiiformans sp. nov., two novel actinomycetes isolated from muddy soil and emended description of the genus Actinomadura.
Two novel actinomycetes, designated strains NEAU-Jh1-3(T) and NEAU-Jh2-5(T), were isolated from muddy soil collected from a riverbank in Jiaohe, Jilin Province, north China. A polyphasic study was carried out to establish the taxonomic positions of these strains. The 16S rRNA gene sequence analysis showed that the two novel isolates exhibited 99.9 % 16S rRNA gene sequence similarity with each other and that they are closely related to Actinomadura viridis IFO 15238(T) (99.6, 99.6 %) and Actinomadura vinacea IFO 14688(T) (99.3, 99.3 %). Phylogenetic analysis based on the 16S rRNA gene sequences indicated that the two cultures clustered together and formed a cluster with A. viridis IFO 15238(T), A. vinacea IFO 14688(T) and Actinomadura rugatobispora IFO 14382(T). However, the DNA-DNA hybridization value between strains NEAU-Jh1-3(T) and NEAU-Jh2-5(T) was 63.6 %, and the values between the two strains and their close phylogenetic relatives were also below 70 %. With reference to phenotypic characteristics, phylogenetic data and DNA-DNA hybridization results, the two strains can be distinguished from each other and their close phylogenetic relatives. Thus, strains NEAU-Jh1-3(T) and NEAU-Jh2-5(T) represent two novel species of the genus Actinomadura, for which the names Actinomadura jiaoheensis sp. nov. and Actinomadura sporangiiformans sp. nov. are proposed. The type strains are NEAU-Jh1-3(T) (=CGMCC 4.7197(T) = JCM 30341(T)) and NEAU-Jh2-5(T) (=CGMCC 4.7211(T) = JCM 30342(T)), respectively.


Combined arthroscopy and "mini-reconstruction" techniques in the acutely torn anterior cruciate ligament.
A concept of the "knee at risk" has been discussed. Such knees with acute anterolateral rotatory instability require stabilization. The combination procedure of arthroscopy for diagnosis and treatment of internal derangement with an extra-articular reconstruction has been described. In a follow-up study of 69 cases so treated, objective results were 94 per cent acceptable, and subjective results were 93 per cent acceptable. There was no apparent difference either objectively or subjectively between 12 to 24 months and 24 to 65 months postoperatively in this series. The high incidence (80 per cent) of meniscal lesions in this series re-emphasizes the need for a high index of suspicion of internal derangement in acute knee ligament injuries.


Thiol functionalised gold nanoparticles loaded with methotrexate for cancer treatment: From synthesis to in vitro studies on neuroblastoma cell lines.
Colloidal gold nanoparticles (AuNPs) functionalised with hydrophilic thiols can be used as drug delivery probes, thanks to their small size and hydrophilic character. AuNPs possess unique properties for their use in nanomedicine, especially in cancer treatment, as diagnostics and therapeutic tools. Thiol functionalised AuNPs were synthesised and loaded with methotrexate (MTX). Spectroscopic and morphostructural characterisations evidenced the stability of the colloids upon interaction with MTX. Solid state (GISAXS, GIWAXS, FESEM, TEM, FTIR-ATR, XPS) and dispersed phase (UV-Vis, DLS, ζ-potential, NMR, SAXS) experiments allowed to understand structure-properties correlations. The nanoconjugate was tested in vitro (MTT assays) against two neuroblastoma cell lines: SNJKP and IMR5 with overexpressed n-Myc. Molar drug encapsulation efficiency was optimised to be >70%. A non-covalent interaction between the π system and the carboxylate moiety belonging to MTX and the charged aminic group of one of the thiols was found. The MTX loading slightly decreased the structural order of the system and increased the distance between the AuNPs. Free AuNPs showed no cytotoxicity whereas the AuNPs-MTX nanoconjugate had a more potent effect when compared to free MTX. The active role of AuNPs was evidenced by permeation studies: an improvement on penetration of the drug inside cells was evidenced.


Intensity modulated radiation therapy with field rotation--a time-varying fractionation study.
This paper proposes a novel mathematical approach to the beam selection problem in intensity modulated radiation therapy (IMRT) planning. The approach allows more beams to be used over the course of therapy while limiting the number of beams required in any one session. In the proposed field rotation method, several sets of beams are interchanged throughout the treatment to allow a wider selection of beam angles than would be possible with fixed beam orientations. The choice of beamlet intensities and the number of identical fractions for each set are determined by a mixed integer linear program that controls jointly for the distribution per fraction and the cumulative dose distribution delivered to targets and critical structures. Trials showed the method allowed substantial increases in the dose objective and/or sparing of normal tissues while maintaining cumulative and fraction size limits. Trials for a head and neck site showed gains of 25%-35% in the objective (average tumor dose) and for a thoracic site gains were 7%-13%, depending on how strict the fraction size limits were set. The objective did not rise for a prostate site significantly, but the tolerance limits on normal tissues could be strengthened with the use of multiple beam sets.


Associations of renal function with diabetic retinopathy and visual impairment in type 2 diabetes: A multicenter nationwide cross-sectional study.
Diabetic retinopathy (DR) separately has been noted as a major public health problem worldwide as well. Currently, many studies have demonstrated an association between diabetic nephropathy and DR in type 1 diabetes mellitus (T1DM) patients, but this association is less strong in T2DM. The evidence for an association between renal function and DR and visual impairment among T2DM patients is limited, particularly in the Asian population. To assess the association between glomerular filtration rate (GFR) and DR, severe DR, and severe visual impairment among T2DM patients in Thailand. We conducted a nationwide cross-sectional study based on the DM/HT study of the Medical Research Network of the Consortium of Thai Medical Schools. This study evaluated adult T2DM patients from 831 public hospitals in Thailand in the year 2013. GFR was categorized into ≥ 90, 60-89, 30-59 and < 30 mL/min/1.73 m2. The association between GFR and DR, severe DR, and severe visual impairment were assessed using multivariate logistic regression. A total of 13192 T2DM patients with available GFR were included in the analysis. The mean GFR was 66.9 ± 25.8 mL/min/1.73 m2. The prevalence of DR, proliferative DR, diabetic macular edema, and severe visual impairment were 12.4%, 1.8%, 0.2%, and 2.1%, respectively. Patients with GFR of 60-89, 30-59 and < 30 mL/min/1.73 m2 were significantly associated with increased DR and severe DR when compared with patients with GFR of ≥ 90 mL/min/1.73 m2. In addition, increased severe visual impairment was associated with GFR 30-59 and < 30 mL/min/1.73 m2. Decreased GFR was independently associated with increased DR, severe DR, and severe visual impairment. GFR should be monitored in diabetic patients for DR awareness and prevention.


Simultaneous detection of antibody responses to multiple SARS-CoV-2 antigens by a Western blot serological assay.
The nucleic acid test is still the standard assessment for the diagnosis of coronavirus disease 2019 (COVID-19), which is caused by human infection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In addition to supporting the confirmation of disease cases, serological assays are used for the analysis of antibody status and epidemiological surveys. In this study, a single Western blot strip (WBS) coated with multiple Escherichia coli (E. coli)-expressed SARS-CoV-2 antigens was developed for comprehensive studies of antibody profiles in COVID-19 patient sera. The levels of specific antibodies directed to SARS-CoV-2 spike (S), S2, and nucleocapsid (N) proteins were gradually increased with the same tendency as the disease progressed after hospitalization. The signal readouts of S, S2, and N revealed by the multi-antigen-coated WBS (mWBS)-based serological assay (mWBS assay) also demonstrated a positive correlation with the SARS-CoV-2 neutralizing potency of the sera measured by the plaque reduction neutralization test (PRNT) assays. Surprisingly, the detection signals against the unstructured receptor-binding domain (RBD) purified from E. coli inclusion bodies were not observed, although the COVID-19 patient sera exhibited strong neutralizing potency in the PRNT assays, suggesting that the RBD-specific antibodies in patient sera mostly recognize the conformational epitopes. Furthermore, the mWBS assay identified a unique and major antigenic epitope at the residues 1148, 1149, 1152, 1155, and 1156 located within the 1127-1167 fragment of the S2 subunit, which was specifically recognized by the COVID-19 patient serum. The mWBS assay can be finished within 14-16 min by using the automatic platform of Western blotting by thin-film direct coating with suction (TDCS WB). Collectively, the mWBS assay can be applied for the analysis of antibody responses, prediction of the protective antibody status, and identification of the specific epitope. KEY POINTS: • A Western blot strip (WBS) coated with multiple SARS-CoV-2 antigens was developed for the serological assay. • The multi-antigen-coated WBS (mWBS) can be utilized for the simultaneous detection of antibody responses to multiple SARS-CoV-2 antigens. • The mWBS-based serological assay (mWBS assay) identified a unique epitope recognized by the COVID-19 patient serum.


External validation of the 2008 Framingham cardiovascular risk equation for CHD and stroke events in a European population of middle-aged men. The PRIME study.
To test the applicability of the sex-specific 2008 Framingham general cardiovascular risk equation for coronary heart disease (CHD) and stroke in European middle-aged men from Ireland and France. In the PRIME study, 9638 healthy middle-aged men recruited in France and Ireland were surveyed for 10 years for the occurrence of first CHD and stroke events. The original Framingham equation, the partially calibrated Framingham equation (using the PRIME baseline survival at 10 years), and the completely calibrated Framingham equation (additionally using risk factor means calculated in PRIME) were assessed. Model fit (expected versus observed events) and discrimination ability were assessed using a modified Hosmer-Lemeshow Chi-square statistic and Harrell's c-index respectively. The original (uncalibrated) Framingham equation overestimated by 1.94-fold the risk of CHD and stroke combined in PRIME, and by 2.23 and 1.42-fold in PRIME-France and PRIME-Ireland respectively. Adequate fit was found after complete calibration. However, discrimination ability of the Framingham equation was poor as shown by Harrell's c-index lower than 0.70. The (completely) calibrated 2008 Framingham equation predicted accurate number of CHD and stroke events but discriminated poorly individuals at higher from those at lower event risk in a European population of middle-aged men.


Birefringent Glass-Engraved Tilted Pillar Metasurfaces for High Power Laser Applications.
Birefringent materials-which are highly needed in high power laser systems-may be limited in usage due to the laser-induced damage threshold of traditional birefringent materials. This work reports here on all-glass metasurfaces, fabricated by angled etching through sacrificial metal nanoparticle (NP) etching masks, for generation of effective birefringence in the formed layer. As a result, a fused silica metasurface, monolithic to the underlying substrate, is demonstrated to exhibit a birefringence of 6.57° under 375 nm illumination. Full-wave analysis shows a good agreement with the measurement and presents potential paths forward to increasing the effective metasurface birefringence. This is the first demonstration, to the best of knowledge, of an etching technique to obtain the resulting tilted pillar-like nanofeatures. The anisotropy of the metasurface nanoelements along the two window in-plane major axes presents different effective paths for the two polarizations and thus generates birefringence in a nonbirefringent material. Additionally, the imparted anisotropy lends itself to manipulation of physical properties of the surface as well, with metasurface feature orientation suppressing water flow along one principal axis and giving rise to water flow steering capabilities.


Intrauterine transfusion--intraperitoneal versus intravascular approach: a case-control comparison.
Intravascular fetal transfusion has gained widespread acceptance and has supplanted the use of intraperitoneal fetal transfusion in management of severe alloimmune disease in many centers. This study compares the two methods with regard to multiple objective end points of performance, therapy, and outcome in a highly matched case-control fashion. The intravascular approach is better on almost every level. More surviving infants who are in better condition at a mature gestation and whose mothers have fewer complications and sequelae are the result. Whereas intraperitoneal transfusion should not be abandoned altogether, it is a second-line procedure used only in very limited circumstances. Intravascular fetal transfusion offers realistic prognosis for intact survival at virtually any extreme of alloimmune disease.