Elastography for hepatic fibrosis severity in chronic hepatitis B or C.
To assess the value of transient elastography for predicting significant fibrosis or cirrhosis in chronic hepatitis B or C (CHB or CHC) patients. 75 patients (CHB: 45, CHC: 32) were included. All underwent elastography and liver biopsy concurrently. Biopsies were evaluated using Ishak's classification. Fibrosis was mild, moderate or severe/cirrhosis when scores were 0-1 (n = 30), 2-3 (n = 20), 4-6 (n = 25), respectively. Median liver stiffness values were higher in patients with severe fibrosis or cirrhosis than in those with moderate or mild fibrosis (14.8 vs. 6.4 vs. 5.3 kPa, p < 0.001). The diagnostic accuracy of elastography for severe fibrosis and cirrhosis was excellent [area under the receiver operating characteristic (AUROC) curve 0.938 vs. 0.948], but it was not optimal for mild fibrosis (AUROC 0.78). Values of 7.5, 9.0 and 12 kPa had a sensitivity and specificity for severe fibrosis/cirrhosis of 96, 84 and 60%, and 76, 90 and 94%, respectively. The median stiffness value in cirrhotic patients (score 5-6) was 16.6 kPa (7.7-48). No differences in accuracy of elastography between CHB or CHC patients were found. Cutoff was 12.5 kPa for cirrhosis; 10/75 patients (13%) were misclassified. Transient elastography has an excellent diagnostic accuracy for severe fibrosis and cirrhosis in CHB and CHC, but the cutoffs need further evaluation.


Single versus multiple deficiencies of methionine, zinc, riboflavin, vitamin B-6 and choline elicit surprising growth responses in young chicks.
A soy-protein isolate diet that was deficient in methionine (Met), zinc (Zn), riboflavin, vitamin B-6 and choline for chick growth (Assay 1) was used to study individual or multiple deficiencies of several of these nutrients. In all cases, adding all three deficient nutrients together resulted in growth responses that were superior to those resulting from supplementation with any pairs of deficient nutrients. In Assay 2, single addition of Zn but not of methionine or riboflavin produced a growth response, but the combination of either Zn and Met or Zn and riboflavin resulted in growth responses that were greater than the response elicited by Zn alone. Assay 3 involved individual or multiple deficiencies of choline, riboflavin and vitamin B-6, and individual additions suggested that choline was first limiting. Choline + riboflavin supplementation, however, produced marked growth and gain:food responses that were far greater than those resulting from supplemental choline or riboflavin alone. Moreover, the growth response to a combination of choline + pyridoxine (PN) was also greater than that obtained from any of the three nutrients fed alone; even PN + riboflavin (in the absence of choline) produced responses greater than those observed with the unsupplemented negative-control diet. In Assay 4, chicks responded to individual additions of riboflavin, PN or Met, and in Assay 5, to either riboflavin or PN; all two-way combinations resulted in growth rates that were far greater than those occurring with any single addition. The data from these experiments show that unlike the situation with three deficient amino acids, the expected responses to first-, second- and third-limiting B-vitamins or deficient vitamins combined with deficient levels of Zn or Met do not follow the expected pattern of response to first-, further response to first- and second- and an even further response to first-, second- and third-limiting nutrients.


The LIM-only protein FHL2 attenuates lung inflammation during bleomycin-induced fibrosis.
Fibrogenesis is usually initiated when regenerative processes have failed and/or chronic inflammation occurs. It is characterised by the activation of tissue fibroblasts and dysregulated synthesis of extracellular matrix proteins. FHL2 (four-and-a-half LIM domain protein 2) is a scaffolding protein that interacts with numerous cellular proteins, regulating signalling cascades and gene transcription. It is involved in tissue remodelling and tumour progression. Recent data suggest that FHL2 might support fibrogenesis by maintaining the transcriptional expression of alpha smooth muscle actin and the excessive synthesis and assembly of matrix proteins in activated fibroblasts. Here, we present evidence that FHL2 does not promote bleomycin-induced lung fibrosis, but rather suppresses this process by attenuating lung inflammation. Loss of FHL2 results in increased expression of the pro-inflammatory matrix protein tenascin C and downregulation of the macrophage activating C-type lectin receptor DC-SIGN. Consequently, FHL2 knockout mice developed a severe and long-lasting lung pathology following bleomycin administration due to enhanced expression of tenascin C and impaired activation of inflammation-resolving macrophages.


Left ventricular performance after intravenous infusion of captopril in patients with congestive heart failure.
Although oral administration of captopril, an angiotensin-converting enzyme inhibitor, is effective for the treatment of congestive heart failure (CHF), the effect of its intravenous (iv) administration is not well known. Ten patients (age range 48-72 years), with CHF belonging to the second and third NYHA class, were given an iv bolus of 25 mg of captopril. Before and 30 minutes after the infusion of captopril, a number of parameters of the left ventricular function were evaluated by echocardiography IREX 3 M-B Mode. Eight patients showed a significant improvement of left ventricular performance indices. In fact, the ejection fraction (13.8%, p < 0.05), the cardiac output (24%, p < 0.001), the circumferential shortness fraction (29.9%, p < 0.05), and the fraction shortening (16.0%, p < 0.005) increased significantly, whereas the end-systolic diameter (21%, p < 0.001), the endsystolic stress (23.8%, p < 0.01) and the left ventricle ejection time (4.8%, p < 0.05) decreased significantly. Systolic and diastolic blood pressure values also underwent a significant reduction by 17% and 11% (p < 0.01 and p < 0.05 respectively). No evident correlation between the improvement of the left ventricular function and the basal renin rates was noticed. A significant improvement of parietal kinesis was observed especially in those segments which showed movement abnormalities (hypokinesia and akinesia) and in many cases this was detected by M-B Mode echocardiography. Our findings may be the result of the following factors: 1) reduction of parietal stress; 2) increased district coronary flow; 3) inhibition of tissue renin-angiotensin-aldosterone system; and 4) "scavenging" action exerted by the SH group of captopril.


Integrated PK/PD Modeling Relates Smoothened Inhibitor Biomarkers to The Heterogeneous Intratumor Disposition of Cetuximab in Pancreatic Cancer Tumor Models.
Therapeutic antibodies have shown little efficacy in the treatment of pancreatic ductal adenocarcinomas (PDAC). Tumor desmoplasia, hypovascularity, and poor perfusion result in insufficient tumor cell exposure, contributing to treatment failure. Smoothened inhibitors of hedgehog signaling (sHHi) increase PDAC tumor permeability, perfusion, and drug delivery, and provide a tool to develop a quantitative, mechanistic understanding as to how the temporal dynamics of tumor priming can impact intratumor distribution of monoclonal antibodies (mAb). A linked pharmacokinetic (PK)/pharmacodynamic (PD) model was developed to integrate the plasma and tumor PK of a sHHi priming agent with its effects upon downstream stromal biomarkers Gli1, hyaluronic acid, and interstitial fluid pressure in PDAC patient-derived xenograft (PDX) tumors. In parallel, in situ tumor concentrations of cetuximab (CTX: anti-epidermal growth factor receptor; EGFR) were quantified as a marker for tumor delivery of mAb or antibody-drug conjugates. A minimal, physiologically-based pharmacokinetic (mPBPK) model was constructed to link sHHi effects upon mechanistic effectors of tumor barrier compromise with the intratumor distribution of CTX, and CTX occupancy of EGFR in tumors. Integration of the mPBPK model of mAb deposition and intratumor distribution with the PK/PD model of tumor responses to priming not only identified physiological parameters that are critical for tumor antibody distribution, but also provides insight into dosing regimens that could achieve maximal tumor disposition of therapeutic antibodies under conditions of transient PDAC tumor permeability barrier compromise that mechanistically-diverse tumor priming strategies may achieve.


Effect of a single dose of low-level laser therapy on spontaneous and chewing pain caused by elastomeric separators.
The aim of this study was to see the effect of a single dose of low-level laser therapy on spontaneous and chewing pain after the placement of elastomeric separators. Eighty-eight patients were randomly selected for this single-blind study. Elastomeric separators were placed mesial and distal to the permanent first molars in all quadrants. Both arches were divided into experimental and control sides. The experimental sides were treated with low-level laser therapy on 3 points on the buccal mucosa for 20 seconds each, with a 940-nm gallium-aluminum-arsenic diode laser on continuous mode and power set at 200 mW. The other side received placebo laser therapy without turning on the laser. A numeric rating scale was used to assess the intensity of spontaneous and chewing pain for the next 7 days. The independent sample t test and repeated-measures analysis of variance with the post hoc Tukey test was used to analyze the results. Significant differences were found in spontaneous and chewing pain among both groups (P <0.05). A single dose of low-level laser therapy can be an efficient modality to reduce the postoperative pain associated with the placement of elastomeric separators.


Human relations: will humanism and public service survive?
Bureaucratization and specialization threaten to undermine personalized care. Will humanism and public service be sacrificed for the ethos of business?


Commercial Vinegar Tablets Do Not Display the Same Physiological Benefits for Managing Postprandial Glucose Concentrations as Liquid Vinegar.
Research evidence suggests that vinegar may effectively reduce postprandial glucose in both healthy adults and those with type 2 diabetes. There is heightened consumer interest in commercially available vinegar tablets; however, it is not known whether these products lower postprandial glycemia to the same extent as liquid vinegar. This crossover trial examined the impact of liquid vinegar versus commercial vinegar tablet ingestion at the start of a meal on the 60-minute glucose excursion postmeal in healthy adults. Twelve young men and women (22.6 ± 0.6 y; 21.2 ± 1.2 kg/m2) completed this 4-arm Latin square crossover trial. Testing was separated by one week and consisted of a test meal (64 g carbohydrate) consumed immediately following one of the four oral treatments: CON, 60 g water (control treatment); VIN, 25 g liquid vinegar (5% acidity; 1.25 g acetic acid) diluted with 35 g water; PILL, four vinegar tablets (1.50 g acetic acid) swallowed whole with 60 g water; and PILL-c, four crushed vinegar tablets dissolved in 60 g water. Capillary blood glucose was tested in the fasted state and at 30 and 60 minutes postmeal. The 60-minute glucose excursion varied significantly by treatment (iAUC: 4.9 ± 0.6, 3.4 ± 0.4, 4.9 ± 0.6, and 4.1 ± 0.5 mmol˖h/l for CON, VIN, PILL, and PILL-c, respectively; F (3, 33) = 3.037, p = 0.043; repeated measures ANOVA). Post hoc analysis revealed a 31% reduction in the glucose postmeal excursion for VIN in comparison to CON and PILL (p = 0.040 and p = 0.049, respectively). These data suggest that commercial vinegar tablets taken whole at mealtime are not as effective as liquid vinegar for reducing the postmeal glucose excursion in young, healthy adults.


[An immunofluorescent study of post-segregational gene action in rats].
Immunofluorescent analysis of the post-segregation effect in mice was carried out. Congenic strains of mice were used. Epididymal sperm of F1 hybrids was studied by means of the indirect method of labelling antibodies with fluoresceine isothiocyanate. The data obtained indicate the possibility of post--segregation effect in mammalian gametes and the possibility of exploration of immunogenetic method for alteration of Mendel's natural correlation in mammals.


Complex drivers of invasive macroalgae boom and bust in Kāne'ohe Bay, Hawai'i.
Invasive macroalgae Eucheuma sp. and Kappaphycus spp. (E/K) became a dominant benthic feature in Kāne'ohe Bay throughout the past four decades - occurring on up to 74 ha of reef area and growing up to three meters thick, which prompted intensive management action. In 2013, E/K cover began decreasing at managed and unmanaged sites. This study examined the extent and timing of the E/K decline and evaluated environmental and ecological drivers beyond management contributing to the decline. E/K continued to recede into 2017 and remains sparse in Kāne'ohe Bay today. Increasing over the sampling period, herbivore biomass was negatively correlated with E/K cover, and other significant, non-linear relationships emerged between E/K cover and coral cover, sea surface temperature, wind, and rainfall. This study uncovers several possible mechanisms explaining a boom and bust in E/K abundance, emphasizes the importance of herbivory, and highlights the resilience of coral reefs in Kāne'ohe Bay.


Species reassignment of Geotrichum bryndzae, Geotrichum phurueaensis, Geotrichum silvicola and Geotrichum vulgare based on phylogenetic analyses and mating compatibility.
The present classification of Galactomyces and its anamorph, Geotrichum, is based on various studies that used morphology, ecology, biochemistry, DNA-DNA reassociation comparisons and gene sequencing. In this study, the identities of strains of the Centraalbureau voor Schimmelcultures yeast culture collection, as well as seven strains from South Africa, were examined by analyses of the nucleotide divergence in the internal transcribed spacer regions of the nuclear rRNA gene (nrRNA) operon, the D1/D2 domains of the 26S rRNA gene and partial actin gene sequences as well as compatibility studies. The South African strains were assigned to species in the genus Galactomyces. The phylogenetic analyses and mating studies revealed that Geotrichum silvicola and Geotrichum bryndzae are synonyms of Galactomyces candidus and that Geotrichum vulgare is a synonym of Galactomyces pseudocandidus.


Long working hours directly and indirectly (via short sleep duration) induce headache even in healthy white-collar men: cross-sectional and 1-year follow-up analyses.
Headache in employees may be linked with both overwork and sleep restriction induced by long working hours. Inter-relationships among working hours, sleep duration and headache were investigated. Cross-sectional analyses for prevalent headache (n = 35,908) and 1-year follow-up analyses for incident headache (n = 19,788) were conducted in apparently healthy white-collar men aged 25-59 years. Headache (yes/no), working hours and sleep duration were based on self-administered questionnaire. After determination of relationships between working hours and sleep duration, logistic regression analysis estimated odds ratio (OR) and 95% confidence interval for prevalent and incident headache according to working hours (35-44, 45-49, 50-59 and ≥60 h/week) and sleep duration (≥7, 6-6.9, 5-5.9 and <5 h/day), and tested linear trends in OR. Additionally, interactive effects of working hours and sleep duration on OR were checked. Covariates in the analyses were age, body mass index, drinking, smoking and exercise. Prevalent and incident headache was found in 1979 (5.5%) men and 707 (3.6%) men, respectively. Working hours were inversely associated with sleep duration. OR for prevalent and incident headache rose with increasing working hours and with reducing sleep duration, regardless of influences of the covariates. Working hours and sleep duration had no interactive effects on OR for prevalent or incident headache. The results indicate that long working hours directly and indirectly (via short sleep duration) induce headache even in apparently healthy white-collar men. Headache in employees may be useful for early detection of adverse health effects by long working hours.


Improving acute medical management: Junior Doctor Emergency Prescription Cards.
Doctors commencing Foundation Year (FY) training face many stresses and challenges. FY doctors are often the first point of contact for acutely unwell and deteriorating patients. Trust guidelines are used to aid acute medical management. Accessing guidelines is often fraught with barriers. Evidence suggests aide-memoire cards can provide easier access to guidelines and management pathways. We aimed to improve prescribing accuracy and efficiency of FY doctors for acute medical conditions within Gloucestershire trust by improving access to and usability of trust guidelines. Questionnaires were distributed to FY doctors to identify acute medical conditions to include on the emergency prescription cards (EPCs). Two small double-sided cards were created containing bullet pointed trust guidelines for: hyper/hypokalaemia, status epilepticus, diabetic emergencies, arrhythmias, myocardial infarction, acute asthma, pulmonary oedema, anaphylaxis and a ward-round checklist. Feedback was used to improve EPCs prior to distribution. Pre (N=53) and post-intervention (N=46) written questionnaires were completed by FY doctors. These assessed acute clinical management including use of guidance, confidence in management, speed of prescribing and EPC "usability". To assess prescribing accuracy, prescriptions for acute medical conditions were reviewed pre (N=8) and post-intervention (N=12). The EPCs were well received (80% quite/very useful) and found "easy to use" (83%). The introduction of EPCs increased guidance use (pre-intervention 58.8%, post-intervention 71.7%), increased confidence (pre-intervention 79%, post-intervention 89%) and significantly improved prescribing speed (p=0.05). There was a significant correlation with confidence and prescribing speed (p = 0.023). The accuracy of prescribed doses improved (pre-intervention 62.5%, post-intervention 87.5% accurate) as did details regarding route / additional required information (pre-intervention 75%, post-intervention 97.7%). The EPCs support the management of unwell patients, are relevant to the workload of modern doctors practice and may improve patient care. This improvement measure could be applied to other NHS trusts and medical specialties.


Control of DNA polymerase alpha, beta and gamma activities in heat- and cold-sensitive mammalian cell-cycle mutants.
Two heat-sensitive (arrested in G1 at 39.5 degrees C) and two cold-sensitive (arrested in G1 at 33 degrees C) clonal cell-cycle mutants of the murine P-815-X2 mastocytoma line were tested for DNA polymerase alpha, beta and gamma activities. After transfer of mutant cells to the respective nonpermissive temperature, DNA polymerase alpha activities decreased more slowly than relative numbers of cells in S phase. Furthermore, numbers of DNA-synthesizing cells decreased to near-zero levels, whereas polymerase alpha activities in arrested cells were as high as 15-40% of control values. After return of arrested cells to the permissive temperature, polymerase alpha activities increased essentially in parallel with relative numbers of cells in S phase. In contrast to the changes in thymidine kinase (Schneider, E., Müller, B. and Schindler, R. (1983) Biochim. Biophys. Acta 741, 77-85), the decrease of polymerase alpha during entry of cells into proliferative quiescence thus appears to be under rather relaxed control, while after return of arrested cells to the permissive temperature the increase in polymerase alpha is tightly coupled with reentry of cells into S phase. For DNA polymerase beta and gamma activities, no obvious correlation with changes in the proliferative state of cells was detected.


Oral misoprostol for induction of labour.
Misoprostol is an orally active prostaglandin. In most countries misoprostol is not licensed for labour induction, but its use is common because it is cheap and heat stable. To assess the use of oral misoprostol for labour induction in women with a viable fetus. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (17 January 2014). Randomised trials comparing oral misoprostol versus placebo or other methods, given to women with a viable fetus for labour induction. Two review authors independently assessed trial data, using centrally-designed data sheets. Overall there were 76 trials (14,412) women) which were of mixed quality.In nine trials comparing oral misoprostol with placebo (1109 women), women using oral misoprostol were more likely to give birth vaginally within 24 hours (risk ratio (RR) 0.16, 95% confidence interval (CI) 0.05 to 0.49; one trial; 96 women), need less oxytocin (RR 0.42, 95% CI 0.37 to 0.49; seven trials; 933 women) and have a lower caesarean section rate (RR 0.72, 95% CI 0.54 to 0.95; eight trials; 1029 women).In 12 trials comparing oral misoprostol with vaginal dinoprostone (3859 women), women given oral misoprostol were less likely to need a caesarean section (RR 0.88, 95% CI 0.78 to 0.99; 11 trials; 3592 women). There was some evidence that they had slower inductions, but there were no other statistically significant differences.Nine trials (1282 women) compared oral misoprostol with intravenous oxytocin. The caesarean section rate was significantly lower in women who received oral misoprostol (RR 0.77, 95% CI 0.60 to 0.98; nine trials; 1282 women), but they had increased rates of meconium-stained liquor (RR 1.65, 95% CI 1.04 to 2.60; seven trials; 1172 women).Thirty-seven trials (6417 women) compared oral and vaginal misoprostol and found no statistically significant difference in the primary outcomes of serious neonatal morbidity/death or serious maternal morbidity or death. The results for vaginal birth not achieved in 24 hours, uterine hyperstimulation with fetal heart rate (FHR) changes, and caesarean section were highly heterogenous - for uterine hyperstimulation with FHR changes this was related to dosage with lower rates in those with lower doses of oral misoprostol. However, there were fewer babies born with a low Apgar score in the oral group (RR 0.60, 95% CI 0.44 to 0.82; 19 trials; 4009 babies) and a decrease in postpartum haemorrhage (RR 0.57, 95% CI 0.34 to 0.95; 10 trials; 1478 women). However, the oral misoprostol group had an increase in meconium-stained liquor (RR 1.22, 95% CI 1.03 to 1.44; 24 trials; 3634 women). Oral misoprostol as an induction agent is effective at achieving vaginal birth. It is more effective than placebo, as effective as vaginal misoprostol and results in fewer caesarean sections than vaginal dinoprostone or oxytocin.Where misoprostol remains unlicensed for the induction of labour, many practitioners will prefer to use a licensed product like dinoprostone. If using oral misoprostol, the evidence suggests that the dose should be 20 to 25 mcg in solution. Given that safety is the primary concern, the evidence supports the use of oral regimens over vaginal regimens. This is especially important in situations where the risk of ascending infection is high and the lack of staff means that women cannot be intensely monitored.


Banded mongooses avoid inbreeding when mating with members of the same natal group.
Inbreeding and inbreeding avoidance are key factors in the evolution of animal societies, influencing dispersal and reproductive strategies which can affect relatedness structure and helping behaviours. In cooperative breeding systems, individuals typically avoid inbreeding through reproductive restraint and/or dispersing to breed outside their natal group. However, where groups contain multiple potential mates of varying relatedness, strategies of kin recognition and mate choice may be favoured. Here, we investigate male mate choice and female control of paternity in the banded mongoose (Mungos mungo), a cooperatively breeding mammal where both sexes are often philopatric and mating between relatives is known to occur. We find evidence suggestive of inbreeding depression in banded mongooses, indicating a benefit to avoiding breeding with relatives. Successfully breeding pairs were less related than expected under random mating, which appeared to be driven by both male choice and female control of paternity. Male banded mongooses actively guard females to gain access to mating opportunities, and this guarding behaviour is preferentially directed towards less closely related females. Guard-female relatedness did not affect the guard's probability of gaining reproductive success. However, where mate-guards are unsuccessful, they lose paternity to males that are less related to the females than themselves. Together, our results suggest that both sexes of banded mongoose use kin discrimination to avoid inbreeding. Although this strategy appears to be rare among cooperative breeders, it may be more prominent in species where relatedness to potential mates is variable, and/or where opportunities for dispersal and mating outside of the group are limited.


Cell wall-deficient, L-form bacteria in the 21st century: a personal perspective.
The peptidoglycan (PG) cell wall is a defining feature of the bacteria. It emerged very early in evolution and must have contributed significantly to the success of these organisms. The wall features prominently in our thinking about bacterial cell function, and its synthesis involves the action of several dozen proteins that are normally essential for viability. Surprisingly, it turns out to be relatively simple to generate bacterial genetic variants called L-forms that completely lack PG. They grow robustly provided that lack of the cell wall is compensated for by an osmoprotective growth medium. Although their existence has been noted and studied on and off for many decades, it is only recently that modern molecular and cellular methods have been applied to L-forms. We used Bacillus subtilis as an experimental model to understand the molecular basis for the L-form switch. Key findings included the discovery that L-forms use an unusual blebbing, or tubulation and scission mechanism to proliferate. This mechanism is completely independent of the normal FtsZ-based division machinery and seems to require only an increased rate of membrane synthesis, leading to an increased surface area-to-volume ratio. Antibiotics that block cell wall precursor synthesis, such as phosphomycin, efficiently induce the L-form switch without the need for genetic change. The same antibiotics turned out to induce a similar L-form switch in a wide range of bacteria, including Escherichia coli, in which we showed that proliferation was again FtsZ-independent. Aside from further basic science, future work on L-forms is likely to focus on their possible role in chronic or recurrent infections, their use as a model in studies of the origins of life, and possibly, biotechnological applications.


Improved extraction of fluoroquinolones with recyclable ionic-liquid-based aqueous biphasic systems.
In the past few years, the improvement of advanced analytical tools allowed to confirm the presence of trace amounts of metabolized and unchanged active pharmaceutical ingredients (APIs) in wastewater treatment plants (WWTPs) as well as in freshwater surfaces. It is known that the continuous contact with APIs, even at very low concentrations (ng L-1-μg L-1), leads to serious human health problems. In this context, this work shows the feasibility of using ionic-liquid-based aqueous biphasic systems (IL-based ABS) in the extraction of quinolones present in aqueous media. In particular, ABS composed of imidazolium- and phosphonium-based ILs and aluminium-based salts (already used in water treatment plants) were evaluated in one-step extractions of six fluoroquinolones (FQs), namely ciprofloxacin, enrofloxacin, moxifloxacin, norfloxacin, ofloxacin and sarafloxacin, and extraction efficiencies up to 98% were obtained. Despite the large interest devoted to IL-based ABS as extractive systems of outstanding performance, their recyclability/reusability has seldomly been studied. An efficient extraction/cleaning process of the IL-rich phase is here proposed by FQs induced precipitation. The recycling of the IL and its further reuse without losses in the ABS extractive performance for FQs were established, as confirmed by the four consecutive removal/extraction cycles evaluated. This novel recycling strategy supports IL-based ABS as sustainable and cost-efficient extraction platforms.


Phylotyper: in silico predictor of gene subtypes.
Whole genome sequencing (WGS) is being adopted in public health for improved surveillance and outbreak analysis. In public health, subtyping has been used to infer phenotypes and distinguish bacterial strain groups. In silico tools that predict subtypes from sequences data are needed to transition historical data to WGS-based protocols. Phylotyper is a novel solution for in silico subtype prediction from gene sequences. Designed for incorporation into WGS pipelines, it is a general prediction tool that can be applied to different subtype schemes. Phylotyper uses phylogeny to model the evolution of the subtype and infer subtypes for unannotated sequences. The phylogenic framework in Phylotyper improves accuracy over approaches based solely on sequence similarity and provides useful contextual feedback. Phylotyper is a python and R package. It is available from: https://github.com/superphy/insilico-subtyping. email@example.com. Supplementary data are available at Bioinformatics online.


Critical thermal limits affected differently by developmental and adult thermal fluctuations.
Means and variances of the environmental thermal regime play an important role in determining the fitness of terrestrial ectotherms. Adaptive phenotypic responses induced by heterogeneous temperatures have been shown to be mediated by molecular pathways independent of the classic heat shock responses; however, an in-depth understanding of plasticity induced by fluctuating temperatures is still lacking. We investigated high and low temperature acclimation induced by fluctuating thermal regimes at two different mean temperatures, at two different amplitudes of fluctuation and across the developmental and adult life stages of Drosophila melanogaster For developmental acclimation, we found mildly detrimental effects of high-amplitude fluctuations for critical thermal minima, while the critical thermal maxima showed a beneficial response to higher amplitude fluctuations. For adult acclimation involving shifts between fluctuating and constant regimes, cold tolerance was shown to be dictated by developmental temperature conditions irrespective of the adult treatments, while the acquired heat tolerance was readily lost when flies developed at fluctuating temperature were shifted to a constant regime as adults. Interestingly, we also found that the effect of fluctuations at any life stage was gradually lost with prolonged adult maintenance, suggesting a more prominent effect of fluctuations during developmental compared with adult acclimation in D.melanogaster.


Azaphosphiridines: challenges and perspectives.
This mini-review describes a broad spectrum of synthetic strategies to access mono- and polycyclic azaphosphiridines; properties and theoretical calculations of free and metal-ligated are discussed. These species are characterized by a highly strained (saturated) CPN ring with three differently polarized endocyclic bonds. The latter causes a high reactivity of aza-phosphiridine complexes towards ring expansion and exchange reactions. Increasing the ring strain leads to masked FLP behaviour, i.e., small molecule activation was observed. New perspectives in homogenous catalysis are also offered, e.g., a pool of novel chiral P-heterocyclic ligands, ready to be explored, if the challenge to unligate them is solved.


The effects of cobalt and iodine supplementation of the pregnant ewe diet on immunoglobulin G, vitamin E, T3 and T4 levels in the progeny.
Sixty twin-bearing ewes were allocated to one of four dietary treatments investigating the effects of supplementary iodine or cobalt during late pregnancy on lamb serum immunoglobulin G (IgG), triiodothyronine (T3), thyroxine (T4) and vitamin E concentrations, and lamb IgG absorption efficiency. Ewes were offered grass silage ad libitum supplemented with 800 g per ewe per day of a 190 g/kg crude protein (CP) concentrate from day 126 of gestation until parturition plus one of the following supplements (n = 15 per treatment); no supplement (C); 26.6 mg iodine per day for final 3 weeks pre partum (I-3); 26.6 mg iodine/day for final week pre partum (I-1); 20 mg cobalt/day for final 3 weeks pre partum (Co-3). Lambs were blood sampled at 24 and 72 h post partum for serum IgG and vitamin E concentrations. Ten lambs from C and I-3 were blood sampled at 1 h post partum for serum IgG, vitamin E, T3 and T4 concentrations. There were no differences in serum IgG, vitamin E or T4 values (P > 0.05) at 1 h post partum between lambs born to the C and I-3 ewes. T3 levels were lower in I-3 compared with C progeny (P < 0.05). Supplemental iodine reduced colostral IgG absorption efficiency (P < 0.001) and lamb serum IgG concentrations at 24 and 72 h post partum (P < 0.001). Serum vitamin E concentration in I-3 and I-1 lambs was lower than in Co-3 lambs at 24 h post partum, while at 72 h post partum I-3, I-1 and Co-3 lambs had significantly lower concentrations than C lambs (P < 0.001). Supplementing the ewe's diet with 26.6 mg/day of iodine for the final week of pregnancy reduced lamb serum IgG concentration at 24 and 72 h post partum. The lower total and free T3 values in the progeny of I-3-treated ewes suggest interference in the synthesis and metabolism of thyroid hormones when ewes receive excessive dietary iodine for 3 weeks immediately pre partum. Based on these findings, the indications are that the toxicity level for iodine in the diet of the pregnant ewe should be lowered to 20 mg per ewe per day, equivalent to 40% of its current level. The finding that high-level cobalt supplementation during the final 3 weeks of pregnancy will have a negative effect on serum vitamin E concentration at 72 h post partum is a new and significant finding and previously has not been reported in the literature.


Primiparous and multiparous females differ in mammary gland alveolar development: implications for milk production.
Mammary gland capacity is influenced by the number of secretory cells in the gland, the activity of those cells and the size and arrangement of the alveoli that they form. Although reproductive experience has been shown to affect the total number of secretory cells in the gland, its potential effect on the structural development of lobulo-alveolar tissues has not been directly investigated. To examine whether reproductive experience affects lobulo-alveolar development, we took mammary gland biopsies at early and peak lactation from primiparous and multiparous grey seal (Halichoerus grypus) females and used histological techniques to compare cell density, alveolar density and alveolar size within secretory lobules. Primiparous females had a significantly higher cell density compared with multiparous females throughout lactation, suggesting that primiparous females have smaller, less-developed secretory cells. Primiparous females had a significantly smaller average alveolar size compared with multiparous females throughout lactation. Although alveolar density was higher in primiparous females compared with multiparous females at early lactation, there was no significant difference between the groups at peak lactation. These results suggest that the mammary gland of primiparous females may have both a lower secretory capacity and a lower storage capacity on a relative basis than those of multiparous females and demonstrate, for the first time, that reproductive experience has a significant effect on both the rate and pattern of mammary gland alveolar development and, potentially, on a female's capacity for milk production.


High-dose palliative radiotherapy for malignant pleural mesothelioma.
High-dose radiotherapy to the hemithorax for patients with malignant pleural mesothelioma is a controversial treatment. Between 2003 and 2013 our institution had a policy of giving hemithoracic radiotherapy to at least 45 Gy. This retrospective study reports survival, progression and toxicity associated with this policy. Seventy-one patients with pleural mesothelioma were irradiated with doses of 45-60 Gy. Conformal radiotherapy (3D-CRT) to the lower hemithorax was used for 17 and intensity-modulated radiotherapy (IMRT) to the whole hemithorax for 54 patients. All patients have been followed up for at least 2 years from commencement of radiotherapy. Sixty-four patients (90%) completed planned radiotherapy and seven stopped early, usually due to progressive disease. Median overall survival was 9.5 months (95% CI: 7.7-12.4) and median progression-free survival was 4.9 months (95% CI: 4.4-5.8). Eighty-seven per cent of patients progressed or died within 2 years: 25% in-field, 49% outside the RT field and 13% died without progression. Severe toxicity (grade 3-5) was observed in 53% of 3D-CRT and 78% of IMRT patients, most commonly pulmonary fibrosis 27%, radiation dermatitis 18%, dyspnoea 11%, GGT increased 11%, pneumonitis 10%, pleuritic pain 8% and fatigue 8%. There were two, possibly three, treatment-related deaths. High-dose radiotherapy to the hemithorax caused significant toxicity to most patients with no improvement in survival. Lower doses of radiotherapy to limited volumes may be useful for palliative purposes.


Dihydroceramide accumulation mediates cytotoxic autophagy of cancer cells via autolysosome destabilization.
Autophagy is considered primarily a cell survival process, although it can also lead to cell death. However, the factors that dictate the shift between these 2 opposite outcomes remain largely unknown. In this work, we used Δ9-tetrahydrocannabinol (THC, the main active component of marijuana, a compound that triggers autophagy-mediated cancer cell death) and nutrient deprivation (an autophagic stimulus that triggers cytoprotective autophagy) to investigate the precise molecular mechanisms responsible for the activation of cytotoxic autophagy in cancer cells. By using a wide array of experimental approaches we show that THC (but not nutrient deprivation) increases the dihydroceramide:ceramide ratio in the endoplasmic reticulum of glioma cells, and this alteration is directed to autophagosomes and autolysosomes to promote lysosomal membrane permeabilization, cathepsin release and the subsequent activation of apoptotic cell death. These findings pave the way to clarify the regulatory mechanisms that determine the selective activation of autophagy-mediated cancer cell death.


Analysis of the melanoma epidemic, both apparent and real: data from the 1973 through 1994 surveillance, epidemiology, and end results program registry.
The incidence of melanoma has been increasing faster than that of any other cancer in the United States. It is unclear whether the increase is related to increased surveillance or other changes in the disease. To examine changes in melanoma rates by several measures of severity of disease and to review the ways in which increased surveillance may cause lead-time bias through early detection or length bias through detection of clinically insignificant lesions as a basis for interpreting these changing rates. Population-based incidence rates for 1973 through 1994. United States Surveillance, Epidemiology, and End Results Program tumor registries. A total of 47,638 cases of melanoma among white patients aged 20 years and older. Relative incidence rates for melanoma by stage, and tumor thickness adjusted for age and sex. Localized-stage melanoma increased, but no significant change for distant-stage melanomas was seen. Among those diagnosed from 1988 through 1994, there were 22%, 26%, and 31% increases for tumor thickness less than 1.0 mm, between 1.0 and 3.0 mm, and 3.0 mm or greater, respectively. The 2-year mortality rates also increased over time. While these data show large increases in early disease (localized stage, thin tumors), they also suggest some increase in advanced disease (thick tumors, 2-year mortality). This indicates that the increasing incidence rates of melanoma are not solely caused by increased early detection and diagnosis of clinically insignificant melanomas, but may also represent a true increase in cancer rates.


Origin of the anomalous piezoelectric response in wurtzite Sc(x)Al(1-x)N alloys.
The origin of the anomalous, 400% increase of the piezoelectric coefficient in Sc(x)Al(1-x)N alloys is revealed. Quantum mechanical calculations show that the effect is intrinsic. It comes from a strong change in the response of the internal atomic coordinates to strain and pronounced softening of C33 elastic constant. The underlying mechanism is the flattening of the energy landscape due to a competition between the parent wurtzite and the so far experimentally unknown hexagonal phases of the alloy. Our observation provides a route for the design of materials with high piezoelectric response.


Genetic determinants in head and neck squamous cell carcinoma and their influence on global personalized medicine.
While sequencing studies have provided an improved understanding of the genetic landscape of head and neck squamous cell carcinomas (HNSCC), there remains a significant lack of genetic data derived from non-Caucasian cohorts. Additionally, there is wide variation in HNSCC incidence and mortality worldwide both between and within various geographic regions. These epidemiologic differences are in part accounted for by varying exposure to environmental risk factors such as tobacco, alcohol, high risk human papilloma viruses and betel quid. However, inherent genetic factors may also play an important role in this variability. As limited sequencing data is available for many populations, the involvement of unique genetic factors in HNSCC pathogenesis from epidemiologically diverse groups is unknown. Here, we review current knowledge about the epidemiologic, environmental, and genetic variation in HNSCC cohorts globally and discuss future studies necessary to further our understanding of these differences. Long-term, a more complete understanding of the genetic drivers found in diverse HNSCC cohorts may help the development of personalized medicine protocols for patients with rare or complex genetic events.


Potential risk assessment of miR-143 gene polymorphisms rs41291957 and rs353292 in colorectal cancer.
Colorectal cancer is a life-threatening and therapeutically challenging disease. Increasingly it is being deciphered that genetic and epigenetic mutations play a central role in cancer onset and progression. Excitingly, discovery of non-coding RNAs is considered to be a milestone in molecular oncology and emerging evidence is deepening our understanding about pivotal role of miRNAs in carcinogenesis. miR-143 has been experimentally verified to play an instrumental role as tumor suppressor. Recent studies suggest that single nucleotide polymorphisms rs41291957 and rs353292 in miR-143 may associate with the progression and or development of colorectal cancer. In present study 400 Pakistani subjects participated including 200 colorectal cancer patients and 200 age and gender matched healthy individuals. Blood samples and clinical information of the confirmed patients was collected from cancer diagnosis and treatment hospitals in Pakistan. The polymorphisms rs41291957 and rs353292 were genotyped in patients and controls by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and results were validated by Sanger sequencing. The association of the SNPs within the study group was analyzed by χ² test with p value < 0.05 as significant. Odds ratio was calculated with 95% confidence interval.Genetic predisposition to cancer was observed in presence of characteristic rs45291957 polymorphism. χ² test results show strongly significant association mi-RNA rs45291957 SNP with colorectal cancer p value 0.0111 (<0.05) along with the statistically significant correlation tested by odds ratio with 95% confidence interval. However, no significant correlation (p value 0.6683) could be found for the association of rs353292 with colorectal cancer in Pakistani population. The present study for the first time gave evidence of miR-143 rs41291957 involvement in colorectal cancer patients of Pakistani population. This target can be a useful molecular tool for the prognosis and treatment targets for colorectal cancer in Pakistani population. rs353292 genetic association can be explored for different cancers in Pakistan to completely rule out its role in cancer.


Agent-based modeling of the immune system: NetLogo, a promising framework.
Several components that interact with each other to evolve a complex, and, in some cases, unexpected behavior, represents one of the main and fascinating features of the mammalian immune system. Agent-based modeling and cellular automata belong to a class of discrete mathematical approaches in which entities (agents) sense local information and undertake actions over time according to predefined rules. The strength of this approach is characterized by the appearance of a global behavior that emerges from interactions among agents. This behavior is unpredictable, as it does not follow linear rules. There are a lot of works that investigates the immune system with agent-based modeling and cellular automata. They have shown the ability to see clearly and intuitively into the nature of immunological processes. NetLogo is a multiagent programming language and modeling environment for simulating complex phenomena. It is designed for both research and education and is used across a wide range of disciplines and education levels. In this paper, we summarize NetLogo applications to immunology and, particularly, how this framework can help in the development and formulation of hypotheses that might drive further experimental investigations of disease mechanisms.


Hepatitis B in renal transplant patients.
Hepatitis B virus (HBV) poses a significant challenge for both dialysis patients and kidney transplant recipients despite its decreasing rates, especially in developed countries. The best preventive method is vaccination. Patients with chronic renal disease should ideally be vaccinated prior to dialysis, otherwise, reinforced vaccination practices and close antibody titer monitoring should be applied while on dialysis. HBV infected dialysis patients who are renal transplant candidates must be thoroughly examined by HBV-DNA, and liver enzyme testing and by liver biopsy. When needed, one must consider treating patients with tenofovir or entecavir rather than lamivudine. Depending on the cirrhosis stage, dialysis patients are eligible transplant recipients for either a combined kidney-liver procedure in the case of decompensated cirrhosis or a lone kidney transplantation since even compensated cirrhosis after sustained viral responders is no longer considered an absolute contraindication. Nucleoside analogues have led to improved transplantation outcomes with both long-term patient and graft survival rates nearing those of HBsAg(-) recipients. Moreover, in the cases of immunized HBsAg(-) potential recipients with concurrent prophylaxis, we are enabled today to safely use renal grafts from both HBsAg(+) and HBsAg(-)/anti-HBc(+) donors. In so doing, we avoid unnecessary organ discarding. Universal prophylaxis with entecavir is recommended in HBV kidney recipients and should start perioperatively. One of the most important issues in HBV(+) kidney transplantation is the duration of antiviral prophylaxis. In the absence of robust data, it seems that prophylactic treatment may be discontinued in selected stable, low-risk recipients during maintenance immunosuppression and should be reintroduced when the immune status is altered. All immunosuppressive agents in kidney transplantation can be used in HBV(+) recipients. Immunosuppression is intimately associated with increased viral replication; thus it is important to minimize the total immunosuppression burden long term.


The Developmental Implications of the Use of Reproductive Technologies for Transgender People: A Comparative Cross-Section Protocol.
Today, individuals and couples with fertility issues can use advances in biomedical technologies to conceive. Transgender persons also benefit from these advances and can not only actualize their self-identified gender identities but also experience parenthood. These strategies for persons to self-actualize and to access parenthood have improved the condition of transgender persons. However, some may question the welfare of the offspring because such transfamily forms are often confusing to many. The sparse research on the psychological well-being of children of transgender people is reassuring. However, the limited empirical research justifies more studies to be conducted with an evidence-based methodology to assess whether these new methods of parenting have any adverse impact on children. The current report details the protocol we built to compare cognitive development, mental health, gender identity, quality of life, and family dynamics in children of transgender fathers and donor sperm insemination (DSI) and two control groups matched for age and gende typically developing (TD) children and children from cisgender parents and DSI. To calculate sample sizes, we hypothesize no significant difference between groups. Since 2008, married couples that include a transgender father have been able to access DSI and have started conceiving children in France. They are always invited to participate in research to assess their children's well-being. To date, the cohort includes 53 children in 37 families. We propose to carry out a cross-sectional comparative study exploring cognitive development with the Brunet-Lézine Psychomotor Development Scale or Wechsler's Intelligence Scales according to age; mental health with the Child Behaviour Checklist; gender identity with the Gender Identity Interview for Children; quality of life with the KIDSCREEN and the Adolescent Coping Questionnaire; and family dynamics with the Parental Bonding Instrument, the Inventory of Parent and Peer Attachment, the Five-Minute Speech Sample, and Corman's Family Drawing Test. To assess possible subtle differences between children's family drawings, we will use a generalization of the "lady-tasting-tea" procedure to link qualitative and quantitative approaches in psychiatric research. Twenty raters [four child and family psychoanalysts (CHILDPSY), four adult psychiatrists (ADUPSY), four biologists working in assistive reproduction technology (BIOL), four endocrinologists working with transgender individuals (ENDOC), and four students (STUD)] will be randomly shown the drawings and asked to blindly classify them using a Likert scale according to whether the child has a transgender father. After testing normality, comparisons between the three groups will be performed with appropriate statistical tests (Kruskal-Wallis, ANOVA, Chi2 or Fisher's exact test). For the "lady-tasting-tea" procedure, we will use a permutation test. The study protocol has been approved by the CERES (Comité d'Ethique de Recherche en Santé) of Paris 5 University. Registration number is 2015/31.


Dot-ELISA assessment of guinea pig antibody responses to repeated Dermacentor andersoni infestations.
Acquired resistance to ixodid tick infestation is expressed by cattle and laboratory animals. Humoral factors appear to be involved in host acquired resistance to tick bite; however, specific immune responses have yet to be fully characterized. This study examined tick resistance expressed by Hartley guinea pigs upon repeated infestation with Dermacentor andersoni, and describes longitudinal development of antigen specific immunoglobulin over approximately 180 days. Guinea pigs were infested either 4 times with D. andersoni adults, or twice with nymphs. Both infestation groups, adults and nymphs, demonstrated a significant level of resistance to re-infestation, following initial exposure. Dot enzyme-linked immunosorbent assay (Dot-ELISA) was employed to detect antibody reactive with D. andersoni salivary gland antigens (SGA). Animals infested with adults had antibody that increased at a relatively constant rate until the fourth infestation, which was differentiated by a sharp increase in titer, that was maintained for approximately 2 wk. Guinea pigs that received nymph infestations had SGA-specific antibody; however, titers were lower than those in the adult infestation group. Antibody levels continued to increase approximately 80 days beyond the final (second) infestation for this group. A direct correlation between resistance and antibody titer was not evident, since resistance was relatively stable after the second infestation in both infestation groups, and tick-specific immunoglobulin levels continually increased.


ATR: an essential regulator of genome integrity.
Genome maintenance is a constant concern for cells, and a coordinated response to DNA damage is required to maintain cellular viability and prevent disease. The ataxia-telangiectasia mutated (ATM) and ATM and RAD3-related (ATR) protein kinases act as master regulators of the DNA-damage response by signalling to control cell-cycle transitions, DNA replication, DNA repair and apoptosis. Recent studies have provided new insights into the mechanisms that control ATR activation, have helped to explain the overlapping but non-redundant activities of ATR and ATM in DNA-damage signalling, and have clarified the crucial functions of ATR in maintaining genome integrity.


BCR/ABL rearrangement with b3a3 fusion transcript in a case of childhood acute lymphoblastic leukemia.
The role of BCR/ABL isoforms and their relationship to leukemia phenotype have been of major concern. Atypical BCR/ABL mRNA transcripts lacking exon a2 have been reported in 12 cases of acute lymphoblastic leukemia (ALL) to date; among them, a b3a3 type transcript has been reported only once in the childhood ALL. Reported here is the case of a patient with Philadelphia-positive (Ph(+)) ALL expressing a b3a3 type transcript, a rare type of BCR/ABL mRNA lacking ABL exon a2 sequences. Bone marrow showed a hypercellular marrow with leukemic blasts positive for CD10, CD19, CD79a, and cytoplasmic mu, which is consistent with pre-B ALL. The G-banding and fluorescence in situ hybridization analyses indicated Ph(+). After the patient was diagnosed with ALL-L2, induction chemotherapy was performed and imatinib mesylate was thereafter given as the maintenance therapy. Sequencing analysis showed deletion of ABL a2 in the polymerase chain reaction product, which corresponded to a b3a3 fusion transcript. To our knowledge, this is the second report of an aberrant BCR/ABL product lacking ABL exon a2 in childhood ALL.


Global burden of respiratory infections due to seasonal influenza in young children: a systematic review and meta-analysis.
The global burden of disease attributable to seasonal influenza virus in children is unknown. We aimed to estimate the global incidence of and mortality from lower respiratory infections associated with influenza in children younger than 5 years. We estimated the incidence of influenza episodes, influenza-associated acute lower respiratory infections (ALRI), and influenza-associated severe ALRI in children younger than 5 years, stratified by age, with data from a systematic review of studies published between Jan 1, 1995, and Oct 31, 2010, and 16 unpublished population-based studies. We applied these incidence estimates to global population estimates for 2008 to calculate estimates for that year. We estimated possible bounds for influenza-associated ALRI mortality by combining incidence estimates with case fatality ratios from hospital-based reports and identifying studies with population-based data for influenza seasonality and monthly ALRI mortality. We identified 43 suitable studies, with data for around 8 million children. We estimated that, in 2008, 90 million (95% CI 49-162 million) new cases of influenza (data from nine studies), 20 million (13-32 million) cases of influenza-associated ALRI (13% of all cases of paediatric ALRI; data from six studies), and 1 million (1-2 million) cases of influenza-associated severe ALRI (7% of cases of all severe paediatric ALRI; data from 39 studies) occurred worldwide in children younger than 5 years. We estimated there were 28,000-111,500 deaths in children younger than 5 years attributable to influenza-associated ALRI in 2008, with 99% of these deaths occurring in developing countries. Incidence and mortality varied substantially from year to year in any one setting. Influenza is a common pathogen identified in children with ALRI and results in a substantial burden on health services worldwide. Sufficient data to precisely estimate the role of influenza in childhood mortality from ALRI are not available. WHO; Bill & Melinda Gates Foundation.


Absence of hepatitis C virus transmission in a prospective cohort of heterosexual serodiscordant couples.
To analyse hepatitis C virus (HCV) transmission in a cohort of heterosexual couples who are discordant both for HIV and for HCV. We followed an open cohort of 171 people, 152 women and 19 men, who were not initially infected by either HIV or HCV, and whose steady heterosexual partner presented antibodies to both viruses (index case). Other risk exposures were excluded. Every 6 months clinical, epidemiological, and risk behaviour information was collected, and antibodies to both viruses were determined. During 529 person years of follow up more than 40 000 vaginal or anal penetrations were recorded. 74 partners (43.3%) had vaginal and/or anal intercourse without condoms with the index case; another 15.8%, who always used condoms, declared breaking or slipping episodes during intercourse; and another 22.2% had unprotected orogenital exposures. During the follow up, over 5800 unprotected vaginal and anal contacts with the index case were estimated, as well as more than 25 000 unprotected orogenital contacts. 31 women became pregnant (two were index cases), and seroconversion to HIV occurred for one woman (1.7 per 10 000 unprotected contacts; 95% CI, 0 to 9.5), but there was no seroconversion to HCV (95% CI, 0-6.3 per 10 000 unprotected contacts). These results are consistent with a low or null transmissibility of HCV in heterosexual relations, even when the index case is HIV co-infected.


Coughing, sneezing, and aching online: Twitter and the volume of influenza-like illness in a pediatric hospital.
This study investigates the relation of the incidence of georeferenced tweets related to respiratory illness to the incidence of influenza-like illness (ILI) in the emergency department (ED) and urgent care clinics (UCCs) of a large pediatric hospital. We collected (1) tweets in English originating in our hospital's primary service area between 11/1/2014 and 5/1/2015 and containing one or more specific terms related to respiratory illness and (2) the daily number of patients presenting to our hospital's EDs and UCCs with ILI, as captured by ICD-9 codes. A Support Vector Machine classifier was applied to the set of tweets to remove those unlikely to be related to ILI. Time series of the pooled set of remaining tweets involving any term, of tweets involving individual terms, and of the ICD-9 data were constructed, and temporal cross-correlation between the social media and clinical data was computed. A statistically significant correlation (Spearman ρ = 0.23) between tweets involving the term flu and ED and UCC volume related to ILI 11 days in the future was observed. Tweets involving the terms coughing (Spearman ρ = 0.24) and headache (Spearman ρ = 0.19) individually were also significantly correlated to ILI-related clinical volume four and two days in the future, respectively. In the 2014-2015 cold and flu season, the incidence of local tweets containing the terms flu, coughing, and headache were early indicators of the incidence of ILI-related cases presenting to EDs and UCCs at our children's hospital.


Identification and characterization of enolase as a collagen-binding protein in Lactobacillus plantarum.
Collagen is a target of pathogens for adhesion, colonization, and invasion of host tissue. Probiotic bacteria can mimic the same mechanism as used by the pathogens in the colonization process, expressing cell surface proteins that specifically interact with extracellular matrix component proteins. The capability to bind collagen is expressed by several Lactobacillus isolates, including some Lactobacillus plantarum strains. In this study we report the involvement of the L. plantarum EnoA1 alfa-enolase in type I collagen (CnI) binding. By adhesion assays, we show that the mutant strain LM3-CC1, carrying a null mutation in the enoA1 gene, binds to immobilized collagen less efficiently than wild type strain. CnI overlay assay and Elisa tests, performed on the purified EnoA1, show that this protein can bind collagen both under denaturing and native conditions. By using truncated recombinant enolase proteins, we also show that the region spanning from 73rd to the 140th amino acid residues is involved in CnI binding.


Prosurvival factors derived from the embryonic brain promote adult neural stem cell survival.
Temporally distinct populations of neural stem cells (NSCs; embryonic and adult) display the cardinal stem cell properties of self-renewal and multipotentiality; however, their relative frequency and cell kinetics vary through development and into old age. We asked whether changes in NSC behavior could be accounted for by changes in environmental signals over time. We identified a prosurvival signaling cascade that enhances adult-derived NSC survival using cues released from embryonic neurons. Specifically, we demonstrate that stromal-cell-derived factor-1α (SDF-1α) released by embryonic neurons leads to upregulation of neuronal nitric oxide synthase in adult neural precursor cells. The resulting increase in nitric oxide leads to the upregulation of the stem cell factor (SCF) receptor ckit on adult NSCs (ANSCs). SCF released from embryonic neurons results in enhanced NSC survival. Using both in vitro and in vivo assays, we have demonstrated expansion of the size of the NSC pool through this pathway, indicating that ANSCs retain their ability to respond to embryonic-derived cues into adulthood.


Genetic analysis of radiation-sensitive mutations in the slime mould Dictyostelium discoideum.
The linkage of two mutations leading to increased sensitivity to ultraviolet light and 60Co gamma rays was determined in the slime mould Dictyostelium discoideum using a genetic analysis based on the parasexual cycle. Diploids were selected from a mixture of radiation-sensitive, temperature-resistant and radiation-resistant, temperature-sensitive haploids on the basis of simultaneous radiation and temperature resistance. Analysis of drug-resistant haploid segregants of the heterozygous diploids indicated that one of the radiation-sensitive mutations, radA20, was linked to linkage group I whereas the other, radB13, was linked to the recently defined linkage group VI.


A high-throughput splicing assay identifies new classes of inhibitors of human and yeast spliceosomes.
The spliceosome is the macromolecular machine responsible for pre-mRNA splicing, an essential step in eukaryotic gene expression. During splicing, myriad subunits join and leave the spliceosome as it works on the pre-mRNA substrate. Strikingly, there are very few small molecules known to interact with the spliceosome. Splicing inhibitors are needed to capture transient spliceosome conformations and probe important functional components. Such compounds may also have chemotherapeutic applications, as links between splicing and cancer are increasingly uncovered. To identify new splicing inhibitors, we developed a high-throughput assay for in vitro splicing using a reverse transcription followed by quantitative PCR readout. In a pilot screen of 3080 compounds, we identified three small molecules that inhibit splicing in HeLa extract by interfering with different stages of human spliceosome assembly. Two of the compounds similarly affect spliceosomes in yeast extracts, suggesting selective targeting of conserved components. By examining related molecules, we identified chemical features required for the activity of two of the splicing inhibitors. In addition to verifying our assay procedure and paving the way to larger screens, these studies establish new compounds as chemical probes for investigating the splicing machinery.


Design and implementation of the GLIF3 guideline execution engine.
We have developed the GLIF3 Guideline Execution Engine (GLEE) as a tool for executing guidelines encoded in the GLIF3 format. In addition to serving as an interface to the GLIF3 guideline representation model to support the specified functions, GLEE provides defined interfaces to electronic medical records (EMRs) and other clinical applications to facilitate its integration with the clinical information system at a local institution. The execution model of GLEE takes the "system suggests, user controls" approach. A tracing system is used to record an individual patient's state when a guideline is applied to that patient. GLEE can also support an event-driven execution model once it is linked to the clinical event monitor in a local environment. Evaluation has shown that GLEE can be used effectively for proper execution of guidelines encoded in the GLIF3 format. When using it to execute each guideline in the evaluation, GLEE's performance duplicated that of the reference systems implementing the same guideline but taking different approaches. The execution flexibility and generality provided by GLEE, and its integration with a local environment, need to be further evaluated in clinical settings. Integration of GLEE with a specific event-monitoring and order-entry environment is the next step of our work to demonstrate its use for clinical decision support. Potential uses of GLEE also include quality assurance, guideline development, and medical education.


A Comprehensive Peptidomic Approach to Characterize the Protein Profile of Selected Durum Wheat Genotypes: Implication for Coeliac Disease and Wheat Allergy.
The wheat varietal selection undertaken by breeders in recent decades has been tailored mainly to improve technological and productivity-related traits; however, the latter has resulted in a considerable impoverishment of the genetic diversity of wheat-based products available on the market. This pitfall has encouraged researchers to revalue the natural diversity of cultivated and non-cultivated wheat genotypes in light of their different toxic/immunogenic potential for celiac disease and wheat-allergic patients. In the present investigation, an advanced proteomic approach was designed for the global characterization of the protein profile of selected tetraploid wheat genotypes (Triticum turgidum). The approach combined proteins/peptides sequence information retrieved by specific enzymatic digestions (single and dual proteolytic enzymes) with protein digestibility information disclosed by means of in-vitro simulated human gastroduodenal digestion experiments. In both cases, the peptide pools were characterized by discovery analysis with liquid chromatography high-resolution tandem mass spectrometry, and specific amino acid sequences were identified via commercial software. The peptide list was screened for in silico toxicity/immunogenicity risk assessment, with the aid of various open-source bioinformatics tools for epitopes matching. Given the global information provided by the designed proteomic approach, the in silico risk assessment not only tackled toxicity implication for celiac disease patients, but also scouted for immunogenic sequences relevant for wheat allergic patients, achieving a comprehensive characterization of the protein profile of the selected genotypes. These latter were assessed to encrypt a variable number of toxic/immunogenic epitopes for celiac disease and wheat allergy, and as such they could represent convenient bases for breeding practices and for the development of new detoxification strategies.


Endocardial mapping of atrial fibrillation in the human right atrium using a non-contact catheter.
Endocardial mapping of atrial fibrillation in humans is limited by its low resolution and by complexities in the arrhythmia and atrial anatomy. A catheter mounted non-contact multielectrode was deployed in the right atrium of 11 patients with atrial fibrillation and used to reconstruct 3360 electrograms, superimposed onto a computer-simulated model of the endocardium, using inverse solution mathematics. This allows construction of isopotential maps of the right atrium. Patients had either sustained atrial fibrillation (n=3) for >6 months or developed atrial fibrillation during the study (n=8). Spontaneous initiation of atrial fibrillation was recorded in one patient and was demonstrated by the non-contact system to arise from two successive atrial ectopic beats from the site of a roving contact catheter. Reconstruction of electrograms recorded during atrial fibrillation was validated by comparison with contact electrograms with cross-correlation. During established atrial fibrillation, four patients predominantly had a single right atrial wave front, two had two wave fronts and five patients had three to five wave fronts for most of the time. Periods of electrical silence were seen in the right atrium in eight patients, after which, activity emerged from consistent septal sites alone, suggesting a left atrial origin. During intravenous administration of flecainide, atrial fibrillation in two patients terminated spontaneously or following pacing manoeuvres, while in the remaining patient sinus rhythm was restored via atrial tachycardia. Non-contact mapping of the right atrium has demonstrated modes of initiation and termination of atrial fibrillation, characterized different patterns of right atrial activation in atrial fibrillation and suggests that the left atrium may sustain atrial fibrillation in some patients. Simultaneous mapping of the right and left atrium is required to further elucidate the mechanisms of human atrial fibrillation.


Dexamethasone inhibits IL-1 beta gene expression in LPS-stimulated RAW 264.7 cells by blocking NF-kappa B/Rel and AP-1 activation.
In the present study, the mechanism by which dexamethasone (DEX) inhibited IL-1beta gene expression in bacterial lipopolysaccharide (LPS)-activated RAW 264.7 cells was investigated. The decrease in LPS-induced IL-1beta mRNA expression was demonstrated by quantitative reverse transcription polymerase chain reaction (RT-PCR). Since the promoter in IL-1beta gene contains binding motifs for NF-kappaB/Rel, AP-1, NF-IL6, and CREB/ATF, which appear to be important in LPS-mediated IL-1beta induction, the effects of DEX on the activation of these transcription factors were examined. Treatment of DEX to RAW 264.7 cells induced a dose-related inhibition of NF-kappaB/Rel and AP-1 in chloramphenicol acetyltransferase activity, while neither NF-IL6 nor CREB/ATF activation was affected by DEX. Treatment of RAW 264.7 cells with DEX inhibited DNA binding of NF-kappaB/Rel and AP-1 proteins to their cognate DNA sites as measured by electrophoretic mobility shift assay (EMSA). DEX treatment caused a significant reduction in nuclear c-rel, p65, and p50 protein contents, and these decreases were paralleled by the accumulation of cytoplasmic c-rel, p65, and p50. DEX treatment of RAW 264.7 cells did not inhibit the nuclear translocation of c-jun and c-fos. We found that the inhibition of IL-1beta production by DEX is not related to p38, which is important in the IL-1beta induction. These results suggest that DEX may inhibit IL-1beta gene expression by a mechanism involving the blocking of LPS-induced NF-kappaB/Rel and AP-1 activation.


Effects of various plasticizers and nanoclays on the mechanical properties of red algae film.
To manufacture red algae (RA) film, we used various plasticizers such as glycerol, sorbitol, sucrose, fructose, and polypropylene glycol (PPG), and then determined the mechanical properties of the RA films. The tensile strength (TS), elongation at break (E), and water vapor permeability (WVP) of the films containing various plasticizers ranged between 0.43 to 9.10 MPa, 10.93% to 47.17%, and 1.28 to 1.42 ng m/m2sPa, respectively. RA films containing fructose as a plasticizer had the best mechanical properties of all the films evaluated. Incorporation of nanoclay (Cloisite Na+ and 30B) improved the mechanical properties of the films. RA film with 3% Cloisite Na+ had a TS of 10.89, while RA film with 30B had a TS of 10.85 MPa; these films also had better E and WVP values than the other RA films evaluated. These results suggest that RA/nanoclay composite films are suitable for use as food packaging materials. Edible RE/nanoclay composite films prepared in the present investigation can be applied in food packaging.


Femoroacetabular impingement inducing non-union of a femoral neck fracture: a case report.
We describe a case in which femoroacetabular impingement (FAI) was identified as the cause of non-union of a femoral neck fracture and the subsequent treatment strategy. Retrospective review of a 35-year-old patient, without any risk factors for non-union, who sustained a femoral neck fracture. Pre-existing FAI was identified as the cause for the non-union of the femoral neck fracture, with successful treatment of the non-union according to established arthroscopic treatment of the hip. After treatment of the FAI, the non-union healed uneventfully within 3 months. FAI may be a less common but potential cause of delayed union or non-union in the setting of femoral neck fracture in the young.


[The role of the glutamatergic system in alcohol addiction].
New animal models have been developed which mimic several aspects of alcoholism. These models provide the basis to study the neurobiological mechanisms of "addicted behaviour". At least two different neurobiological pathways which are involved in the development and maintenance of addicted behaviour have been identified. The first pathway involves the opioidergic system and probably the mesolimbic dopaminergic system and may induce alcohol craving and relapse due to the mood enhancing, positive reinforcing effects of alcohol consumption. A second pathway involves several components of the glutamatergic system (in particular NMDA receptors) and may induce alcohol craving and relapse by negative motivational states including withdrawal and stress. In particular conditioned withdrawal and stress-induced relapse are mediated by a hypertrophic glutamatergic system. Thus it has recently been shown that the NMDA receptor modulator acamprosate inhibits conditioned abstinence behaviour in rats. Although more systematic work is needed to fully define these different neurobiological pathways involved in addicted behaviour, preclinical studies have identified low affinity non-competitive NMDA receptor antagonists as a novel potential generation of anti-relapse compounds and clinical studies have already been initiated in order to test these compounds in alcoholic patients.


Rhenium-188 based radiopharmaceuticals for treatment of liver tumours.
Rhenium-188 (188Re) is a high energy beta-emitter with a physical half life of 17 hours. Various 188Re based radiopharmaceuticals were developed to treat liver malignancies. The vast majority of studies focus on patients suffering from hepatocellular carcinoma (HCC). Most radiopharmaceuticals are based on Lipiodol as a vehicle for the rhenium-188. The radiopharmaceutical that was tested clinically in detail is the 188Re-HDD/Lipiodol, developed by the Seoul University. Clinical data derived from several phase I and II studies using 188Re-HDD/Lipiodol suggest an excellent tolerance in patients with Child-Pugh A cirrhosis. A shortcoming in some trials was the occasional low labelling efficiency of 188Re-HDD/Lipiodol. Some newer 188Re based radiopharmaceuticals claim to have consistent high labelling efficiencies, however clinical data for these compounds are scarce or lacking at this moment. Hopefully, phase I clinical data will become available for promising radiopharmaceuticals such as 188Re-SSS-Lipiodol, developed by the group of Rennes, in the upcoming years. In Dresden a very different approach is used. They labelled human serum albumin microspheres with high activities of 188Re. In a small group of patients with liver metastasis and a few HCC patients, treatment proved safe. In the present clinical field, 188Re-based radiopharmaceuticals will have to proof firmly their strength and reliability in large patient groups if they want to compete with the commercially available yttrium-90 microspheres.


Prospective evaluation of the toxicity profile of proteasome inhibitor-based therapy in renal transplant candidates and recipients.
A prospective intermediate-term evaluation of toxicities associated with bortezomib therapy for antibody-mediated rejection (AMR) and desensitization was conducted. Patients were graded for bortezomib-related toxicities: hematologic and gastrointestinal toxicities by Common Terminology Criteria for Adverse Events and peripheral neuropathy by modified Functional Assessment of Cancer Therapy questionnaire and Common Terminology Criteria for Adverse Events. Fifty-one patients treated for AMR and 19 patients treated for desensitization received 96 bortezomib cycles (1.3 mg/m(2) ×4 doses); mean (SD) follow-up was 16.3 (9.0) months. Patients treated for AMR and patients treated for desensitization were similar in age, gender, ethnicity, and baseline peripheral neuropathy. Patients treated for AMR received a mean (SD) of 4.9 (2.0) bortezomib doses in 1.3 (0.5) cycles; and patients treated for desensitization, a mean of 7.3 (1.6) doses in 1.8 (0.4) cycles. Prevalence of diabetes and anemia were higher at baseline in patients treated for AMR. In the AMR cohort, two cases of cytomegalovirus infection, two cases of BK virus infection, and one case of Epstein-Barr virus infection were observed. No cases of viral infection were observed in the desensitization cohort. Malignancies were not observed. Significant bortezomib toxicities included anemia and peripheral neuropathy, which were manageable. Anemia was more common in patients treated for AMR; and peripheral neuropathy, more common in patients treated for desensitization. Bortezomib-related toxicities in kidney transplant candidates and recipients are low in incidence and severity and vary based on treatment population.


[A developmental study of low birth weight infants in 1986].
Evolution of the newborn infant weighing 1,000-1,499 g, was studied in 1986 in the "Mariana Grajales" Gynecologic and Obstetric Teaching Provincial Hospital, Department of Neonatology, Santa Clara. Data was obtained from the clinical histories of the newborns and of their mothers. A control group was selected in order to analyze and compare the causes of low birthweight, and as causes of it were determined age and height, and as associated diseases, urinary sepsis, anemia and toxemia of pregnancy. Premature rupture of membranes was significant. Half of the infants of the group under study was severe and moderately depressed. At the somatic evolution was observed a trend to increment and a high coefficient of correlation within the three variables under study. Main nursing actions offered fit up to the attention of these neonates.


Accommodative Exercises to Lower Intraocular Pressure.
This study investigated how a conscious change in ocular accommodation affects intraocular pressure (IOP) and ocular biometrics in healthy adult volunteers of different ages. Thirty-five healthy volunteers without ocular disease or past ocular surgery, and with refractive error between -3.50 and +2.50 diopters, were stratified into 20, 40, and 60 year old (y.o.) age groups. Baseline measurements of central cornea thickness, anterior chamber depth, anterior chamber angle, cornea diameter, pupil size, and ciliary muscle thickness were made by autorefraction and optical coherence tomography (OCT), while IOP was measured by pneumotonometry. Each subject's right eye focused on a target 40 cm away. Three different tests were performed in random order: (1) 10 minutes of nonaccommodation (gazing at the target through lenses that allowed clear vision without accommodating), (2) 10 minutes of accommodation (addition of a minus 3 diopter lens), and (3) 10 minutes of alternating between accommodation and nonaccommodation (1-minute intervals). IOP was measured immediately after each test. A 20-minute rest period was provided between tests. Data from 31 subjects were included in the study. ANOVA and paired t-tests were used for statistical analyses. Following alternating accommodation, IOP decreased by 0.7 mmHg in the right eye when all age groups were combined (p = 0.029). Accommodation or nonaccommodation alone did not decrease IOP. Compared to the 20 y.o. group, the 60 y.o. group had a thicker ciliary muscle within 75 μm of the scleral spur, a thinner ciliary muscle at 125-300 μm from the scleral spur, narrower anterior chamber angles, shallower anterior chambers, and smaller pupils during accommodation and nonaccommodation (p's < 0.01). Alternating accommodation, but not constant accommodation, significantly decreased IOP. This effect was not lost with aging despite physical changes to the aging eye. A greater accommodative workload and/or longer test period may improve the effect.


Prophylactic effect of myricetin and apigenin against lipopolysaccharide-induced acute liver injury.
Liver has an important role in the initiation and progression of multiple organ failure that occurs in sepsis. Many natural active substances can be used to reduce the liver injury caused by sepsis. For this aim, the effects of myricetin and apigenin on mice model of acute liver injury was evaluated in this study. Thirty-six mice were randomly divided into six groups as; control, lipopolysaccharide (LPS) (5 mg/kg), LPS + myricetin (100 mg/kg), LPS + myricetin (200 mg/kg), LPS + apigenin (100 mg/kg), and LPS + apigenin (200 mg/kg) groups. Myricetin and apigenin were administered orally for 7 days, and LPS was administered intraperitoneally only on the 7th day of the study. 24 h after LPS application, all animals were sacrificed and serum biochemical parameters, histopathology and oxidative stress and inflammation markers of liver tissue were examined. Myricetin and apigenin pre-treatments increased serum albumin and total protein levels, liver GSH level and catalase and SOD activities and decreased serum ALT, AST, ALP, γ-GT, CRP, total and direct bilirubin levels, liver MPO activity, MDA, NOx, PGE2, TNF-α, IL-1β, and IL-6 levels, iNOS and COX-2 mRNA levels, phosphorylation of NF-κB p65, IκB, and IKK proteins but not p38, ERK, and JNK proteins in LPS-treated mice. Myricetin and apigenin administration also regained the hepatic architecture disrupted during LPS application. Myricetin and apigenin pre-treatments led to reduction of liver injury indices and oxidative stress and inflammatory events and these flavonoids has probably hepatoprotective effects in acute liver injury.


Coronary subclavian steal syndrome: an extracoronary cause of acute coronary syndrome.
Significant atherosclerotic changes of the coronary arteries are the common cause of cardiac chest pain. We report the case of an 80-year-old woman suffering from unstable angina caused by extracoronary atherosclerosis. The patient had an extensive medical history with severe coronary heart disease and cardiac bypass surgery (LIMA to LAD, two venous bypass grafts). An urgent coronary angiography was performed. The angiogram displayed the already known three-vessel disease, the bypass grafts were in a good functional condition. Subsequently, a stenosis of the proximal segment of the left subclavian artery was detected. Measurement of the pullback pressure gradient confirmed the significance of the stenosis. We suspected a "steal phenomenon" concerning the bypass graft LIMA to LAD. An ad hoc PTA with consecutive stenting (self-expandable stent) enabled a successful revascularization of the left subclavian artery without any adverse effects to the vertebral arteries. Cardiac chest pain did not occur any more. Coronary Subclavian Steal Syndrome should be considered a rare but important differential diagnosis in acute coronary syndrome after bypass surgery.


Comparison of Comorbidities in Relation to Critical Conditions among Coronavirus Disease 2019 Patients: A Network Meta-Analysis.
Severe illness and poor outcome are mainly associated with aging or certain medical comorbidities, especially chronic diseases. However, factors for unfavorable prognosis have not been well described owing to relatively small sample sizes and single-center reports. Therefore, this study aimed to compare the contribution of comorbidities in the development of critical conditions in coronavirus disease 2019 (COVID-19) patients. Pooled estimates of relative risks (RRs) and their 95% confidence intervals (CIs) were calculated by conducting a meta-analysis and network meta-analysis of 18 studies. Chronic obstructive pulmonary disease (COPD) was most strongly associated with the overall critical condition (RR = 4.22, 95% CI = 3.12 - 5.69), followed by cardiovascular disease (CVD) (RR = 3.00, 95% CI = 2.41 - 3.73), malignancy (RR = 2.91, 95% CI = 2.16 - 3.91), cerebrovascular accident (CVA) (RR = 2.86, 95% CI = 1.95 - 4.19), diabetes (RR = 2.10, 95% CI = 2.16 - 3.91), hypertension (RR = 2.02, 95% CI = 1.82 - 2.23), and chronic kidney disease (RR = 2.00, 95% CI = 1.36 - 2.94). The presence of comorbidities except for chronic liver disease and chronic kidney disease significantly increased the risk of severe infection, intensive care unit (ICU) admission, and cardiac injury in the subgroup analysis by types of critical conditions. Preexisting hypertension and diabetes additionally increased the risk of acute respiratory distress syndrome (ARDS). Among comorbidities, COPD had the highest probability of leading to severe COVID-19, ICU admission, and liver injury, while malignancy was most likely to cause ARDS and cardiac injury. In summary, preexisting COPD, CVD, CVA, hypertension, diabetes, and malignancy are more likely to worsen the progression of COVID-19, with severe infection, ICU admission requirement, and cardiac injury development.


X-ray structure determination of a vanadium-dependent haloperoxidase from Ascophyllum nodosum at 2.0 A resolution.
The homo-dimeric structure of a vanadium-dependent haloperoxidase (V-BPO) from the brown alga Ascophyllum nodosum (EC 1.1.11.X) has been solved by single isomorphous replacement anomalous scattering (SIRAS) X-ray crystallography at 2.0 A resolution (PDB accession code 1QI9), using two heavy-atom datasets of a tungstate derivative measured at two different wavelengths. The protein sequence (SwissProt entry code P81701) of V-BPO was established by combining results from protein and DNA sequencing, and electron density interpretation. The enzyme has nearly an all-helical structure, with two four-helix bundles and only three small beta-sheets. The holoenzyme contains trigonal-bipyramidal coordinated vanadium atoms at its two active centres. Structural similarity to the only other structurally characterized vanadium-dependent chloroperoxidase (V-CPO) from Curvularia inaequalis exists in the vicinity of the active site and to a lesser extent in the central four-helix bundle. Despite the low sequence and structural similarity between V-BPO and V-CPO, the vanadium binding centres are highly conserved on the N-terminal side of an alpha-helix and include the proposed catalytic histidine residue (His418(V-BPO)/His404(V-CPO)). The V-BPO structure contains, in addition, a second histidine near the active site (His411(V-BPO)), which can alter the redox potential of the catalytically active VO2-O2 species by protonation/deprotonation reactions. Specific binding sites for the organic substrates, like indoles and monochlordimedone, or for halide ions are not visible in the V-BPO structure. A reaction mechanism for the enzymatic oxidation of halides is discussed, based on the present structural, spectroscopic and biochemical knowledge of vanadium-dependent haloperoxidases, explaining the observed enzymatic differences between both enzymes.


Proteolytic modification of the amino-terminal and carboxyl-terminal regions of rat hepatic phenylalanine hydroxylase.
Activation of rat liver phenylalanine hydroxylase by limited proteolysis catalyzed by chymotrypsin was investigated with the use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and high pressure gel filtration. Both activation and proteolysis were decreased by the addition of the natural cofactor, (6R)-tetrahydrobiopterin. From chymotryptic digests of the hydroxylase carried out in the presence and absence of (6R)-tetrahydrobiopterin, several different enzyme species were isolated by high pressure gel filtration. One species (subunit Mr = 47,000) with unchanged hydroxylase activity was isolated from the chymotryptic digest in the presence of (6R)-tetrahydrobiopterin; it was derived from the native enzyme (Mr = 52,000) by cleavage of the COOH-terminal Mr = 5,000 portion of the native enzyme. In the absence of (6R)-tetrahydrobiopterin, another species (subunit Mr = 36,000) was isolated. In addition to modification at the COOH-terminal end of the molecule, this species also had lost a Mr = 11,000 fragment from the NH2-terminal end of the hydroxylase. The Mr = 11,000 fragment was shown to include the phosphorylation site of the enzyme. This Mr = 36,000 species was 30-fold more active than the native phenylalanine hydroxylase when assayed in the presence of tetrahydrobiopterin. These results suggest that the regulatory domain that inhibits hydroxylase activity in the basal state may be located at the NH2 terminus of the phenylalanine hydroxylase subunit.


Sensor technology for smart homes.
A smart home is a residence equipped with technology that observes the residents and provides proactive services. Most recently, it has been introduced as a potential solution to support independent living of people with disabilities and older adults, as well as to relieve the workload from family caregivers and health providers. One of the key supporting features of a smart home is its ability to monitor the activities of daily living and safety of residents, and in detecting changes in their daily routines. With the availability of inexpensive low-power sensors, radios, and embedded processors, current smart homes are typically equipped with a large amount of networked sensors which collaboratively process and make deductions from the acquired data on the state of the home as well as the activities and behaviors of its residents. This article reviews sensor technology used in smart homes with a focus on direct environment sensing and infrastructure mediated sensing. The article also points out the strengths and limitations of different sensor technologies, as well as discusses challenges and opportunities from clinical, technical, and ethical perspectives. It is recommended that sensor technologies for smart homes address actual needs of all stake holders including end users, their family members and caregivers, and their doctors and therapists. More evidence on the appropriateness, usefulness, and cost benefits analysis of sensor technologies for smart homes is necessary before these sensors should be widely deployed into real-world residential settings and successfully integrated into everyday life and health care services.


A bioreactor test system to mimic the biological and mechanical environment of oral soft tissues and to evaluate substitutes for connective tissue grafts.
Gingival cells of the oral connective tissue are exposed to complex mechanical forces during mastication, speech, tooth movement and orthodontic treatments. Especially during wound healing following surgical procedures, internal and external forces may occur, creating pressure upon the newly formed tissue. This clinical situation has to be considered when developing biomaterials to augment soft tissue in the oral cavity. In order to pre-evaluate a collagen sponge intended to serve as a substitute for autogenous connective tissue grafts (CTGs), a dynamic bioreactor system was developed. Pressure and shear forces can be applied in this bioreactor in addition to a constant medium perfusion to cell-material constructs. Three-dimensional volume changes and stiffness of the matrices were analyzed. In addition, cell responses such as cell vitality and extracellular matrix (ECM) production were investigated. The number of metabolic active cells constantly increased under fully dynamic culture conditions. The sponges remained elastic even after mechanical forces were applied for 14 days. Analysis of collagen type I and fibronectin revealed a statistically significant accumulation of these ECM molecules (P < 0.05-0.001) when compared to static cultures. An increased expression of tenascin-c, indicating tissue remodeling processes, was observed under dynamic conditions only. The results indicate that the tested in vitro cell culture system was able to mimic both the biological and mechanical environments of the clinical situation in a healing wound.


Stronger and durable SARS-CoV-2 immune response to mRNA vaccines in 5-11 years old children with prior COVID-19.
mRNA vaccines elicit a durable humoral response to SARS-CoV-2 in adults, whereas evidence in children is scarce. This study aimed to assess the early and long-term immune response to the mRNA vaccine in children with or without previous SARS-CoV-2 infection. In a multicentre prospective observational study, we profiled the immune response to the Pfizer BioNTech (BNT162b2) vaccine in 5-11-year-old children attending the University Pediatric Hospital of Padua and Bambino-Gesù Hospital in Rome (Italy) from December-2021 to February-2023. Blood samples were collected pre-, 1-, and 6-months after vaccination. Neutralizing antibodies (NAbs) and anti-spike-receptor-binding-domain (anti-S-RBD) IgG titers were analyzed through Plaque Reduction Neutralization Test (PRNT) and chemiluminescent immune-enzymatic assay (CLIA), respectively. Immune cell phenotypes were analyzed by flow cytometry. Sixty children (26 [43 %] female, median age = 8 years [IQR = 7-10.7]) were enrolled in the study, including 46 children with a laboratory-confirmed previous COVID-19 (SARS-CoV-2-recovered) and 14 SARS-CoV-2-naïve participants defined as the absence of antigen-specific antibodies before vaccination. SARS-CoV-2-recovered participants recorded higher anti-S-RBD IgG and Wild-type and Omicron BA.2 NAbs titers than SARS-CoV-2-naïve participants at both 1- and 6-months after vaccination. Antibody titers correlated with T (Tregs) and B (Bregs) regulatory cell frequencies in SARS-CoV-2-recovered children. Both SARS-CoV-2-recovered and SARS-CoV-2-naïve participants decreased antibody titers by approximately 100 to 250 % from 1 to 6 months. While children with immunocompromising underlying conditions developed immune responses comparable to those of healthy children, solid organ transplant recipients exhibited lower levels of NAbs and anti-S-RBD IgG titers, as well as reduced frequencies of Tregs and Bregs. mRNA vaccination triggered a higher production of specific anti-SARS-CoV-2 antibodies along with increased levels of regulatory cells in children with previous SARS-CoV-2 infection up to the following 6 months. These findings provide insights into boosting pre-existing immunity.


Passive transfer of a monoclonal antibody specific for a sialic acid-dependent epitope on the surface of Trypanosoma cruzi trypomastigotes reduces infection in mice.
Trypanosoma cruzi, the parasite that causes Chagas' disease, proliferates in the cytosol of mammalian cells. When the trypomastigote forms exit the infected cell, they become extensively sialylated because the parasite contains an enzyme called trans-sialidase. This enzyme efficiently catalyzes the transfer of bound sialic acid residues from host glycoconjugates to acceptors containing terminal beta-galactosyl residues on the parasite surface. The sialic acid acceptors are developmentally regulated mucin-like glycoproteins that are extremely abundant on the trypomastigote surface. In the present study, we determined whether passive transfer of monoclonal antibodies specific for sialic acid acceptors could reduce the acute infection induced by T. cruzi in a highly susceptible mouse strain. We found that passive transfer to naive mice of an immunoglobulin G1 monoclonal antibody directed to a sialylated epitope of these mucin-like glycoproteins significantly decreased parasitemia and the number of tissue parasites as measured by a DNA probe specific for T. cruzi. Upon challenge with trypomastigotes, mice which received this antibody also had a significant increase in survival. A statistically significant reduction in parasitemia could be accomplished with relatively small doses of immunoglobulin, and Fab fragments alone could not mediate protective immunity. The precise mechanism of parasite elimination is unknown; however, this monoclonal antibody does not lyse trypomastigotes in vitro in the presence of human complement or mouse spleen cells.