Structure in tRNA data.
260 sequences of tRNA are compared, after their classification into six categories: prokaryotic (83 sequences) and eukaryotic (83 sequences) elongators, prokaryotic (10 sequences) and eukaryotic (11 sequences), initiators, lower eukaryotic mitochondrial tRNA (53 sequences) and archaebacterial tRNA (20 sequences). Beside the presence of invariable and semi-invariable positions in tRNA, non-random base distribution in almost all positions is also evident; most of them being characteristic of each class of tRNA. Therefore, during evolution it would seem that selectional pressures do act in so called variable positions as well as invariant positions of tRNA molecules. Results are discussed in relation to possible restrictions depending on functional and/or structural constraints of tRNA.


Corrigendum: Effects of Trained Peer vs. Teacher Feedback on EFL Students' Writing Performance, Self-Efficacy, and Internalization of Motivation.
[This corrects the article DOI: 10.3389/fpsyg.2021.788474.].


Dynamic flux regulation for high-titer anthranilate production by plasmid-free, conditionally-auxotrophic strains of Pseudomonas putida.
Anthranilate, an intermediate of the shikimate pathway, is a high-value aromatic compound widely used as a precursor in the production of dyes, fragrances, plastics and pharmaceuticals. Traditional strategies adopted for microbial anthranilate production rely on the implementation of auxotrophic strains-which requires aromatic amino acids or complex additives to be supplemented in the culture medium, negatively impacting production costs. In this work, we engineered the soil bacterium Pseudomonas putida for high-titer, glucose-dependent anthranilate production by repurposing elements of the Esa quorum sensing (QS) system of Pantoea stewartii. The PesaS promoter mediated a self-regulated transcriptional response that effectively knocked-down the expression of the trpDC genes. Next, we harnessed the synthetic QS elements to engineer a growth-to-anthranilate production switch. The resulting plasmid-free P. putida strain produced the target compound at 3.8 ± 0.3 mM in shaken-flask cultures after 72 h-a titer >2-fold higher than anthranilate levels reported thus far. Our results highlight the value of dynamic flux regulation for the production of intermediate metabolites within highly-regulated routes (such as the shikimate pathway), thereby circumventing the need of expensive additives.


Molluscan neurons in culture: shedding light on synapse formation and plasticity.
From genes to behaviour, the simple model system approach has played many pivotal roles in deciphering nervous system function in both invertebrates and vertebrates. However, with the advent of sophisticated imaging and recording techniques enabling the direct investigation of single vertebrate neurons, the utility of simple invertebrate organisms as model systems has been put to question. To address this subject meaningfully and comprehensively, we first review the contributions made by invertebrates in the field of neuroscience over the years, paving the way for similar breakthroughs in higher animals. In particular, we focus on molluscan (Lymnaea, Aplysia, and Helisoma) and leech (Hirudo) models and the pivotal roles they have played in elucidating mechanisms of synapse formation and plasticity. While the ultimate goal in neuroscience is to understand the workings of the human brain in both its normal and diseased states, the sheer complexity of most vertebrate models still makes it difficult to define the underlying principles of nervous system function. Investigators have thus turned to invertebrate models, which are unique with respect to their simple nervous systems that are endowed with a finite number of large, individually identifiable neurons of known function. We start off by discussing in vivo and semi-intact preparations, regarding their amenability to simple circuit analysis. Despite the 'simplicity' of invertebrate nervous systems however, it is still difficult to study individual synaptic connections in detail. We therefore emphasize in the next section, the utility of studying identified invertebrate neurons in vitro, to directly examine the development, specificity, and plasticity of synaptic connections in a well-defined environment, at a resolution that it is still unapproachable in the intact brain. We conclude with a discussion of the future of invertebrates in neuroscience in elucidating mechanisms of neurological disease and developing neuron-silicon interfaces.


Influence of connector cross-sectional geometry on the load-bearing capacity under fatigue of implant-supported zirconia fixed partial prosthesis.
Zirconia has been reported to be suitable for multiunit monolithic restorations. However, as the connector region is the weakest part of the system, studies are needed to determine the optimal connector geometry. The purpose of this in vitro study was to evaluate the load-bearing capacity under fatigue of implant-supported fixed partial prostheses made of monolithic zirconia with different connector cross-sectional geometries. Three-unit monolithic fixed partial prostheses (from mandibular second premolar to the second molar) were fabricated in zirconia (Zenostar T) by computer-aided design and computer-aided manufacture (CAD-CAM). Different connector cross-sectional geometries, with an area of 9 mm2, were tested (n=10): round, square with rounded angles, or trapezoid with rounded angles. The prostheses were screwed over 2 implants and inserted into acrylic resin bases. The specimens were submitted to a mechanical fatigue test until failure, with load applied to the pontic using the stepwise stress methodology (initial load of 100 N for 5000 cycles, followed by an increase to 200 N for 20 000 cycles, with a subsequent increase of 50 N each step). For data analysis, fatigue failure load (FFL) and cycles for fatigue failure (CFF) were recorded. Failed structures were analyzed by fractography. The Kaplan-Meier test followed by a log-rank Mantel-Cox post hoc test was used to analyze FFL and CFF (α=.05). Weibull analysis was also performed. The round (1065 N) and trapezoid (1010 N) groups presented higher FFL values than the square one (870 N) (P≤.05). For CFF, the round connector (358 777) was better than the square (280 310) (P=.006), and the trapezoid (337 773) was statistically similar to both (P>.05). No difference among groups was observed in Weibull modulus either for CFF or FFL data. All failures were catastrophic, originating at the base of the connector and propagating toward the occlusal surface of the pontic (region under loading). The connector cross-sectional geometry significantly influenced the mechanical fatigue performance of implant-supported fixed partial prostheses made of monolithic zirconia.


Medication adherence: a key factor in effective management of rosacea.
Rosacea is a chronic condition associated with relapses. Unsuccessful treatment is predicated, in part, on suboptimal adherence with the medication regimen. Motivating long-term compliance remains a challenge. The literature on adherence with rosacea medication is scant, but data from other diseases suggest that a multifactorial approach combining nonpharmacologic and adherence-enhancing pharmacologic interventions appears to offer the greatest success. The variety of topical metronidazole formulations that are relatively well tolerated and convenient to administer has been a notable advance in rosacea management. The dermatologist, by emphasizing the importance of adherence with therapy, can do much to facilitate this most critical behavior.


MOFFeCo/B-CN composites achieve efficient degradation of antibiotics in a non-homogeneous concurrent photocatalytic-persulfate activation system.
We synthesized an MFeCoB0.4CNx% (MOF-Fe/Co nanosheets/boron-doped g-C3N4) composite catalyst for enhancing the concurrent photocatalytic-persulfate activation (CPPA) system and achieved efficient degradation of antibiotics. The role of MOF-Fe/Co is to activate persulfate, while boron-doped g-C3N4 can generate photogenerated electrons for the reduction of Co3+/Fe3+ to enhance the regeneration of the active center. The rate constant for Tetracycline degradation by the CPPA system was 4.74 and 7.54 times higher than the photocatalytic and persulfate-activated systems, respectively. This composite was shown to be practical and economically viable for antibiotic degradation. The degradation behavior was explored based on experiments, and molecular orbitals and Fukui functions were obtained by density functional theory calculations. Mechanisms were investigated using reactive oxygen species trapping studies and electron spin resonance, and the process was explained in terms of the charge population and electron density difference of MOF-Fe/Co nanosheets. The CPPA system is an ecologically benign technology for removing antibiotic-related risks to the environment and human health.


Studies on fibronectin in the skin. VII. Production in cell cultures from normal human skin.
In the present study, cell cultures of fibroblasts from normal skin have been investigated regarding the production of fibronectin. The development of multimeric insoluble fibronectin is demonstrated as small dots at the cell surface, developing into a branched meshwork of fibrous structures in parallel arrays. Soluble dimeric fibronectin is also found in the culture medium.


The effect of delayed umbilical cord clamping on the infant's beta-endorphin level, mother-infant attachment and breastfeeding.
To examine the effect of delayed umbilical cord clamping on the infant's betaendorphin level, mother-infant attachment and breastfeeding. This study had an experimental design with a control group. The study was undertaken between October and December 2017 in a maternity hospital in the east of Turkey. In total, 107 pregnant women [55 in the experimental group (delayed cord clamping) and 52 in the control group (early cord clamping)] participated in the study. The beta-endorphin level in the umbilical cord was 775.80 ± 229.35 in the experimental group and 547.91 ± 290.01 in the control group, and the difference was significant (t = 4.492, p = 0.000). Similarly, the prolactin level in the umbilical cord was 174.26 ± 47.20 in the experimental group and 119.06 ± 47.74 in the control group, and the difference was significant (t = 6.012, p = 0.000). Mother-infant attachment and breastfeeding success were higher in the experimental group. Beta-endorphin and prolactin levels in the umbilical cord, mother-infant attachment and breastfeeding success were higher in the group which underwent delayed cord clamping.


Expression of genes involved in nucleotide excision repair and sensitivity to cisplatin and melphalan in human cancer cell lines.
DNA repair has been proposed to be an important determinant of cancer cell sensitivity to alkylating agents and cisplatin (DDP). Nucleotide excision repair (NER), which represents one of the most important cellular DNA repair processes able to remove a broad spectrum of DNA lesions, is involved in the recognition and repair of the crosslinks caused by DDP and melphalan (L-PAM). In this study, the mRNA levels of the different genes involved in NER (ERCC1, XPA, XPB, XPC, XPD, XPF) were examined in a panel of eight different human cancer cell lines, together with the overall DNA repair capacity using a host cell reactivation assay of a damaged plasmid. A statistically significant correlation was observed between the relative expression of XPA/XPC (P < 0.05) and ERCC1/XPC (P < 0.05) mRNAs. No correlation was found between the DDP and L-PAM IC50S and the relative mRNA expression of the tested NER genes. When the overall cellular DNA repair capacity was studied, carcinomas seemed to have a higher repair activity than leukaemias; but this repair DNA activity correlated neither with the mRNA expression of the different NER genes nor with DDP and L-PAM IC50S. These data seem to suggest that even if the NER pathway is an important determinant for the cytotoxicity of alkylating agents, as demonstrated by the extremely high sensitivity to alkylating agents in cells lacking this repair system, other factors have to play a role in regulating the cellular sensitivity/resistance to these antitumour drugs.


Carrageenan-based multifunctional packaging films containing Zn-carbon dots/anthocyanin derived from Kohlrabi peel for monitoring quality and extending the shelf life of shrimps.
Carrageenan-based active/intelligent packaging films containing anthocyanin and ZnO-doped CD (Zn-CD) from purple Kohlrabi peels were prepared for freshness monitoring and shelf-life extension of shrimp, and the influence of additives on the films' physical, functional, and structural properties was investigated. The films showed excellent UV blocking ability (85.2% of UV-A and 99.4% of UV-B) and high antioxidant effect (∼99% for ABTS and ∼ 58.6% for DPPH radical scavenging activity) and showed strong antibacterial activity to stop the growth (100%) of L. monocytogenes and to reduce the growth of E. coli by 8.1 log CFU/mL after 12 h of incubation. In shrimp packaging experiments, the films were evident in the freshness monitoring, reduced spoilage, and increased shelf life. This study suggests that next-generation biopolymer films impregnated with biomass-derived CDs and natural colorants will provide broad directions for ensuring safety and extending shelf life to meet the accelerating demand for packaging products.


The potential for enhanced tumour localisation by poly(ethylene glycol) modification of anti-CEA antibody.
Attachment of poly(ethylene glycol) (PEG) to proteins can greatly alter their pharmacological properties, including extending the plasma half-life and reducing immunogenicity, both of which are potentially beneficial to tumour targeting. IgG, F(ab')2 and Fab' fragments of the anti-CEA antibody A5B7 were chemically modified with PEG (M(r) 5,000), labelled with 125I and their pharmacokinetics compared with the unmodified forms in the LS174T colonic xenograft in nude mice. PEG modification of the intact antibody had little effect on biodistribution, although tumour localisation was slightly reduced. In contrast, similar modification of F(ab')2 and Fab'A5B7 significantly prolonged plasma half-life and increased radioantibody accumulation in the tumour and to a lesser extent in normal tissues, but reduced tissue to blood ratios. Prior to modification, Fab' A5B7 (M(r) 50,000) cleared more rapidly from the circulation than F(ab')2 (M(r) 100,000), but after PEG attachment their biodistributions converged, while the tumour to blood ratios were reduced and resembled that of the intact antibody. The enhanced tumour accumulation, reduced normal tissue to blood ratios and potentially reduced immunogenicity of fragments after PEG attachment may therefore prove superior to either unmodified fragments or intact antibody for antibody-targeted therapy, although the increased plasma half-life may necessitate the use of a clearance mechanism.


Some features of paediatric urolithiasis in a group of Syrian children.
A group of 28 Syrian children (19 males and 9 females; age ranging from 2.5 to 12 years) were diagnosed clinically and radiologically to have upper urinary tract stones. The commonest presentations were renal colic, vomiting, haematuria, pyrexia and vague abdominal pain. Family history of renal stones was present in 21% of cases. Haematological picture and chemical analysis of blood were within the normal limits for their age and sex. Urine analysis, however, showed significantly marked increase in the 24-hour excretions of calcium and uric acid. Microscopic examination showed haematuria and pyuria in 72% of the children with urolithiasis. Chemical analysis of removed stones revealed that most of them were mixed stones of calcium oxalate and urate or/and phosphate. Pure stones of calcium oxalate or calcium phosphate were less common. Radiologically, about 95% of all stones were demonstrated by plain X-ray, while 5% only after IVP.


The theoretical impact on corrective upper limb elective services following analysis of distal radius fractures managed nonoperatively during COVID-19 pandemic.
To assess the proportion of patients with distal radius fractures (DRFs) who were managed nonoperatively during the COVID-19 pandemic in accordance with the British Orthopaedic Association BOAST COVID-19 guidelines, who would have otherwise been considered for an operative intervention. We retrospectively reviewed the radiographs and clinical notes of all patients with DRFs managed nonoperatively, following the publication of the BOAST COVID-19 guidelines on the management of urgent trauma between 26 March and 18 May 2020. Radiological parameters including radial height, radial inclination, intra-articular step-off, and volar tilt from post-reduction or post-application of cast radiographs were measured. The assumption was that if one radiological parameter exceeds the acceptable criteria, the patient would have been considered for an operative intervention in pre-COVID times. Overall, 92 patients formed the cohort of this study with a mean age of 66 years (21 to 96); 84% (n = 77) were female and 16% (n = 15) were male. In total, 54% (n = 50) of patients met at least one radiological indication for operative intervention with a mean age of 68 years (21 to 96). Of these, 42% (n = 21) were aged < 65 years and 58% (29) were aged ≥ 65 years. More than half of all DRFs managed nonoperatively during the COVID-19 pandemic had at least one radiological indication to be considered for operative management pre-COVID. We anticipate a proportion of these cases will require corrective surgery in the future, which increases the load on corrective upper limb elective services. This should be accounted for when planning an exit strategy and the restart of elective surgery services.Cite this article: Bone Joint Open 2020;1-10:612-616.


A resuscitation puzzle: acute acquired methemoglobinemia.
We treated a 32-day-old baby suffering acute acquired methemoglobinemia induced by topical application of aniline cocaine. Although acute acquired methemoglobinemia with severe cyanosis and distress is potentially fatal, this rare syndrome is easily curable if it is correctly diagnosed.


Increasing pre-excitation during exercise and isoproterenol infusion. Evidence for a catecholamine sensitive bypass tract.
A patient with atypical chest pain developed pre-excitation during exercise and isoproterenol infusion, with "ischemic" ST depression only during the pre-excited beats. Coronary angiography and myocardial lactate extraction showed no evidence of abnormal coronary vessels. Electrophysiologic study and pacing-induced tachycardia did not induce pre-excitation, whereas exercise induced progressive increase in pre-excitation. The data are consistent with an unusual form of pre-excitation, perhaps related to responsiveness of an accessory bypass tract to catecholamine stimulation. The data also show that the false positive exercise test in this syndrome is due to progressively increasing pre-excitation and therefore more abnormal repolarization.


Free-roaming dog populations: a cost-benefit model for different management options, applied to Abruzzo, Italy.
Since 1991, Italian free-roaming dogs have been under government protection and euthanasia is restricted by law. Management measures are regulated at the regional level and include: kennelling, adoptions, conversion of stray dogs into block dogs, and population control of owned dogs. "Block dogs" are free-roaming dogs that have been collected by the veterinary services, microchipped, sterilised, vaccinated, and released under the responsibility of the local municipalities. The present paper describes a cost-benefit model for different management options and applies it to two provinces in Abruzzo, central Italy. The model considers welfare, nuisance and direct costs to the municipality. Welfare is quantified based on the expert opinions of 60 local veterinarians, who were asked to assign a score for each dog category according to the five freedoms: freedom from pain, physical discomfort, disease, fear, and freedom to express normal behaviour. Nuisance was assessed only for comparisons between management options, using the number of free-roaming dogs per inhabitant as a proxy indicator. A community dog population model was constructed to predict the effect of management on the different subpopulations of dogs during a ten-year period. It is a user-friendly deterministic model in Excel, easily adaptable to different communities to assess the impact of their dog management policy on welfare, nuisance and direct monetary cost. We present results for Teramo and Pescara provinces. Today's management system is compared to alternative models, which evaluate the effect of specific interventions. These include either a 10% yearly increase in kennel capacity, an increase in adoptions from kennels, a doubling of the capture of stray dogs, or a stabilisation of the owned dog population. Results indicate that optimal management decisions are complex because welfare, nuisance and monetary costs may imply conflicting interventions. Nevertheless, they clearly indicate that management actions that would act on dog ownership patterns to reduce the number of free-roaming dogs would have the most favourable outcomes. These include reducing the reproductive capacity of the owned dog population, stronger enforcement of mandatory dog identification, reducing abandonment and increasing adoptions. This would increase welfare and free resources for implementing public campaigns. Block dogs may be an important intermediary means to reduce stray dogs, but adoption would be preferable.


Cross-Reactivity to Kynureninase Tolerizes B Cells That Express the HIV-1 Broadly Neutralizing Antibody 2F5.
2F5 is an HIV-1 broadly neutralizing Ab that also binds the autoantigens kynureninase (KYNU) and anionic lipids. Generation of 2F5-like Abs is proscribed by immune tolerance, but it is unclear which autospecificity is responsible. We sampled the BCR repertoire of 2F5 knock-in mice before and after the first and second tolerance checkpoints. Nearly all small pre-B (precheckpoint) and 35-70% of anergic peripheral B cells (postcheckpoint) expressed the 2F5 BCR and maintained KYNU, lipid, and HIV-1 gp41 reactivity. In contrast, all postcheckpoint mature follicular (MF) B cells had undergone L chain editing that purged KYNU and gp41 binding but left lipid reactivity largely intact. We conclude that specificity for KYNU is the primary driver of tolerization of 2F5-expressing B cells. The MF and anergic B cell populations favored distinct collections of editor L chains; surprisingly, however, MF and anergic B cells also frequently expressed identical BCRs. These results imply that BCR autoreactivity is the primary determinant of whether a developing B cell enters the MF or anergic compartments, with a secondary role for stochastic factors that slightly mix the two pools. Our study provides mechanistic insights into how immunological tolerance impairs humoral responses to HIV-1 and supports activation of anergic B cells as a potential method for HIV-1 vaccination.


Production of Recombinant Proteins in the Chloroplast of the Green Alga Chlamydomonas reinhardtii.
Chloroplast transformation in the green algae Chlamydomonas reinhardtii can be used for the production of valuable recombinant proteins. Here, we describe chloroplast transformation of C. reinhardtii followed by protein detection. Genes of interest integrate stably by homologous recombination into the chloroplast genome following introduction by particle bombardment. Genes are inherited and expressed in lines recovered after selection in the presence of an antibiotic. Recombinant proteins can be detected by conventional techniques like immunoblotting and purified from liquid cultures.


Electrochemical tap water softening: A zero chemical input approach.
Electrochemical water softening was proposed as a sustainable alternative for ion exchange softening, avoiding the input of salt to drinking water and the production of a concentrated brine. Here we demonstrated two novel modes of operation combining an electrochemical cell with a fluidized bed crystallizer. The first approach relied on an electrochemical cell consisting of an anode and cathode separated by a cation or anion exchange membrane. The feed water was first directed into a crystallizer where it was blended with alkaline cathode effluent. The effluent of the crystallizer, softened water, was in part recirculated to the cathode to generate alkalinity, in part to the anode compartment, where the pH was again decreased. Average removal efficiencies for calcium and magnesium of 75-86% and 7-21% respectively, could be sustainably reached, at a specific energy consumption of 7.0-10.1 kWh kg-1 CaCO3 (0.86-1.39 kWh m-3 water). This configuration allowed reagent-free water softening, albeit with an effluent with a pH between 3.0 and 3.6. In a second mode of operation, the process influent to soften was also directed to the crystallizer and recirculated over the cathode, which was separated from the anode using an anion exchange membrane. In this mode of operation, the cathode effluent was sent through the crystallizing unit, and the anode compartment was operated in closed-loop. Average calcium and magnesium removal efficiencies of 73-78% and 40-44% were obtained at specific energy consumptions of 5.8-7.5 kWh kg-1 CaCO3 (0.77-0.88 kWh m-3 water). Although the softened water had an elevated pH (∼9.4), the advantage of this configuration is concomitant removal of anions and the formation of acids/disinfectant in the anode compartment. Both methods of operation thus showed reagent-free water softening at a relatively low specific energy consumption. These novel methods of softening could be used in remote locations where access to chemicals or discharge of ion exchange brines proves to be difficult, or in case addition of chemicals for softening is unwanted. Further research is needed to further decrease the specific energy consumption during long-term operation.


Base analogue mutagenesis in yeast and its modulation by pyrimidine deoxynucleotide pool imbalances: incorporation of bromodeoxyuridylate and iododeoxyuridylate.
Cells of the yeast, Saccharomyces cerevisiae, which are auxotrophic for thymidylate (tmpl) can also incorporate analogues of thymidylate. When the base analogue, 5-bromodeoxyuridylate, is incorporated into tmpl yeast cells it is lethal and mutagenic. Both lethality and mutation induction can be drastically altered by perturbation of the pyrimidine nucleotide pools. Analysis of mutation induction, bromodeoxyuridylate incorporation into DNA, and cell viability under various conditions revealed: (1) lethality and mutagenesis can be uncoupled, (2) thymidylate enhances mutagenesis and deoxycytidylate suppresses it, (3) mutation induction is not correlated with the magnitude of bromodeoxyuridylate incorporation into DNA. Therefore, in yeast, the pyrimidine nucleotide pools have a powerful effect on bromodeoxyuridylate mutagenesis. Both bromodeoxyuridylate and iododeoxyuridylate are extensively incorporated into the DNA of tmpl yeast cells; however, iododeoxyuridylate is non-mutagenic. Replication proceeds at the same rate in the presence of the natural substrate or either analogue. When cells are supplied with thymidylate and bromodeoxyuridylate together, there is no discrimination against bromodeoxyuridylate as a DNA precursor. However, in the presence of thymidylate and iododeoxyuridylate, there is a 3 to 1 discrimination against iododeoxyuridylate as compared to thymidylate.


Biallelic variants in HECT E3 paralogs, HECTD4 and UBE3C, encoding ubiquitin ligases cause neurodevelopmental disorders that overlap with Angelman syndrome.
Pathogenic variants in genes encoding ubiquitin E3 ligases are known to cause neurodevelopmental syndromes. Additional neurodevelopmental disorders associated with the other genes encoding E3 ligases are yet to be identified. Chromosomal analysis and exome sequencing were used to identify the genetic causes in 10 patients from 7 unrelated families with syndromic neurodevelopmental, seizure, and movement disorders and neurobehavioral phenotypes. In total, 4 patients were found to have 3 different homozygous loss-of-function (LoF) variants, and 3 patients had 4 compound heterozygous missense variants in the candidate E3 ligase gene, HECTD4, that were rare, absent from controls as homozygous, and predicted to be deleterious in silico. In 3 patients from 2 families with Angelman-like syndrome, paralog-directed candidate gene approach detected 2 LoF variants in the other candidate E3 ligase gene, UBE3C, a paralog of the Angelman syndrome E3 ligase gene, UBE3A. The RNA studies in 4 patients with LoF variants in HECTD4 and UBE3C provided evidence for the LoF effect. HECTD4 and UBE3C are novel biallelic rare disease genes, expand the association of the other HECT E3 ligase group with neurodevelopmental syndromes, and could explain some of the missing heritability in patients with a suggestive clinical diagnosis of Angelman syndrome.


Glucocorticoids impair type I IFN signalling and enhance rhinovirus replication.
Inhaled corticosteroids (ICS) are recommended treatments for all degrees of asthma severity and in combination with bronchodilators are indicated for COPD patients with a history of frequent exacerbations. However, the long-term side effects of glucocorticoids (GCs) may include increased risk of respiratory infections, including viral triggered exacerbations. Rhinovirus (RV) infection is the main trigger of asthma and COPD exacerbations. Thus, we sought to explore the influence of GCs on viral replication. We demonstrate the ICS fluticasone propionate (FP) and two selective non-steroidal (GRT7) and steroidal (GRT10) glucocorticoid receptor (GR) agonists significantly suppress pro-inflammatory (IL-6 and IL-8) and antiviral (IFN-λ1) cytokine production and the expression of the interferon-stimulated genes (ISGs) OAS and viperin in RV-infected bronchial epithelial cells, with a consequent increase of viral replication. We also show that FP, GRT7 and GRT10 inhibit STAT1 Y701 and/or STAT2 Y690 phosphorylation and ISG mRNA induction following cell stimulation with recombinant IFN-β. In addition, we investigated the effects of the ICS budesonide (BD) and the long-acting β2 agonist (LABA) formoterol, alone or as an ICS/LABA combination, on RV-induced ISG expression and viral replication. Combination of BD/formoterol increases the suppression of OAS and viperin mRNA observed with both BD and formoterol alone, but an increase in viral RNA was only observed with BD treatment and not with formoterol. Overall, we provide evidence of an impairment of the innate antiviral immune response by GC therapy and the potential for GCs to enhance viral replication. These findings could have important clinical implications.


Very late thrombosis of paclitaxel-eluting stent.
Drug-eluting stents (DES) have been proven to be effective in reducing restenosis after percutaneous coronary interventions, but they are associated with a risk of late thrombotic occlusion with adverse clinical events. We report a case of an angiographically confirmed paclitaxel-eluting stent (PES) thrombosis which occurred 36 months after successful stent implantation. To the best of our knowledge, this is the most delayed case of PES thrombosis described so far.


Effect of a natural contaminant on foam fractionation of bromelain.
Foam fractionation is a simple, inexpensive method for separating and purifying proteins. Typically, a dilute bromelain solution with a pH ranging from 2.0 to 7.0 foams very well when bubbles are introduced into a foam fractionation column. It was observed, however, that the dilute enzyme solution only foamed between approximately pH 2.0 and 3.0 when the inner wall of the fractionation column was coated with a natural contaminant (okra residue). We studied the separation ratio and the protein mass recovery to explore the effect of a natural antifoaming agent on the foam fractionation of a dilute bromelain solution. The control variables used in this process were the initial bulk solution pH, which ranged from 2.0 to 7.0, and the superficial air velocity, which varied between 1.7 and 6.2 cm/s.


Influence of delta-hexachlorocyclohexane (δ-HCH) to Phytophthora ×alni resistant Alnus glutinosa genotypes - Evaluation of physiological parameters and remediation potential.
Hexachlorocyclohexanes (HCHs) are persistent organochlorine pesticides with the adverse effects on human health and the environment. The effect of delta-isomer of hexachlorocyclohexane (δ-HCH) on germination, growth parameters and physiological parameters was studied in different Alnus glutinosa (L.) Gaertn. progeny of resistant genotypes to pathogen Phytophthora ×alni. Two experiments were performed: a short-term experiment to determine the effect of δ-HCH on total germination (GT), germination energy (GE), speed of germination (SG), shoot length and biomass of seedlings, and a long-term experiment devoted to remediation aspects. In addition, changes in the hormonal system of alders were monitored in both cases. Significant differences were found between the treated and control group in most of the evaluated characteristics. Also, the content of studied phytohormones differs between groups. Furthermore, the obtained results indicate genetically determined variability in response to δ-HCH. Of the six tested, the Březové and Tuřany progeny seem to be suitable candidates for phytoremediation because of the adaptation to stress conditions or high remediation efficiency. The rest of tested progeny seems to be unsuitable due to higher mortality, lower remediation efficiency and higher levels of stress hormones resulting in significant decrease in biomass and plant height. Moreover, results indicate the role of the plant as a remediation accelerator, probably through released exudates, and a positive effect on the soil microbiome as the presence of plants increased the remediation efficiency by 20.85 - 35.89%. The obtained research findings may be helpful in better understanding the processes involved in removing these pesticides from the soil. Further research should be focused on rhizosphere microbiome, mechanism of in-plant isomerization and metabolites identification.


Growth and energetics of a trichopteran larva feeding on fresh submerged and terrestrial plants.
We studied the growth and assimilation of larvae of Anabolia nervosa, a lotic trichopteran shredder, on diets of fresh tissue of the submerged macrophyte Potamogeton perfoliatus and fresh leaves of the trees Alnus glutinosa and Fagus sylvatica. Fifth-instar larvae were fed these three diets for 23 days (20 May-12 June) in the laboratory. Instantaneous growth rate and gross growth efficiency were almost the same for larvae fed Potamogeton and Alnus, but lower for larvae fed Fagus. The larvae consumed 3 times more fresh Potamogeton than Alnus leaves, but the same amounts in terms of dry mass. The consumption of fresh Fagus was initially twice that of the two other food items but dropped rapidly during the experiment, following the marked seasonal decline of leaf nitrogen content. Allocation patterns of fat and protein differed among food sources. Larvae grown on Alnus accumulated more fat than larvae grown on Potamogeton, while the reverse was true for protein accumulation. The assimilation efficiency based on carbon units was the same (44%) for all three food items and was among the highest reported for shredding caddis larvae. The study demonstrates that fresh tissue of Potamogeton perfoliatus provides a valuable food source to the facultative shredder herbivore Anabolia nervosa.


A short screening test for visual neglect in stroke patients.
to extend a preliminary study of the internal structure of six measures comprising the 'conventional' subtests of the Behavioural Inattention Test (BIT) in order to develop a short screening test for visual neglect. discriminant function analysis of the 15 tests constituting BIT. rehabilitation centre. 59 selected from consecutive stroke patients entering the centre. absent prior history of major CNS disorder, at least 1 week post-stroke, right-handed for writing, absent significant visual impairment other than visual field deficit, no psychiatric overlay or generalized intellectual deterioration, and ability to comprehend and respond to the visuomotor tasks administered. COMPARISON GROUP: 50 non-brain-damaged subjects (hospital employees, members of University subject panel, and local community volunteers). the 15 tests constituting the BIT. two subtests, letter cancellation and star cancellation, were the most sensitive measure, identifying 74% of neglect patients with no false positives. letter and star cancellation offer an adequate yet brief screening test for determining which patients might benefit from administration of the complete neglect test battery.


Mucoadhesive formulation of Bidens pilosa L. (Asteraceae) reduces intestinal injury from 5-fluorouracil-induced mucositis in mice.
Gastrointestinal mucositis induced during cancer treatment is considered a serious dose-limiting side effect of chemotherapy and/or radiotherapy. Frequently, interruption of the cancer treatment due to this pathology leads to a reduction in cure rates, increase of treatment costs and decrease life quality of the patient. Natural products such as Bidens pilosa L. (Asteraceae), represent a potential alternative for the treatment of mucositis given its anti-inflammatory properties. In this study, B. pilosa glycolic extract was formulated (BPF) with poloxamer, a mucoadhesive copolymer, was used for treatment of 5-fluorouracil (5-FU)-induced mucositis in mice. As expected, animals only treated with 5-FU (200 mg/kg) presented marked weight loss, reduction of intestinal villi, crypts and muscular layer, which was associated with severe disruption of crypts, edema, inflammatory infiltrate and vacuolization in the intestinal tissue, as compared to the control group and healthy animals only treated with BPF. On the other hand, the treatment of intestinal mucositis-bearing mice with BPF (75, 100 or 125 mg/kg) managed to mitigate clinical and pathologic changes, noticeably at 100 mg/kg. This dose led to the restoration of intestinal proliferative activity through increasing Ki-67 levels; modulated the expression of Bax, Bcl2 and p53 apoptotic markers protecting intestinal cells from cell death. Moreover, this treatment regulated lipid peroxidation and inflammatory infiltration. No acute toxic effects were observed with this formulation. This work demonstrated that BPF was safe and effective against 5-FU-induced intestinal mucositis in mice. Additional studies are already in progress to further characterize the mechanisms involved in the protective effects of this technological formulation toward the development of a new medicine for the prevention and treatment of intestinal injury in patients undergoing chemotherapy/radiotherapy.


Translation-rotation coupling and the kinematics of non-slip boundary conditions: A rough sphere between two sliding walls.
A non-slip constraint between a particle and a wall is applied at the microscopic level of collision dynamics using the rough sphere model. We analyze the consequences of the translation-rotation coupling of a rough sphere confined between two parallel planar walls and establish that shearing the walls past each other (i) preferentially deposits energy into the rotational degree of freedom and (ii) results in a bounded oscillation of the energy of the confined particle.


Age-related treatment strategy and long-term outcome in acute myocardial infarction patients in the PCI era.
Older age, as a factor we cannot affect, is consistently one of the main negative prognostic values in patients with acute myocardial infarction. One of the most powerful factors that improves outcomes in patients with acute coronary syndromes is the revascularization preferably performed by percutaneous coronary intervention. No data is currently available for the role of age in large groups of consecutive patients with PCI as the nearly sole method of revascularization in AMI patients. The aim of this study was to analyze age-related differences in treatment strategies, results of PCI procedures and both in-hospital and long-term outcomes of consecutive patients with acute myocardial infarction. Retrospective multicenter analysis of 3814 consecutive acute myocardial infarction patients divided into two groups according to age (1800 patients ≤ 65 years and 2014 patients > 65 years). Significantly more older patients had a history of diabetes mellitus and previous myocardial infarctions. The older population had a significantly lower rate of coronary angiographies (1726; 95.9% vs. 1860; 92.4%, p < 0.0001), PCI (1541; 85.6% vs. 1505; 74.7%, p < 0.001), achievement of optimal final TIMI flow 3 (1434; 79.7% vs. 1343; 66.7%, p < 0.001) and higher rate of unsuccessful reperfusion with final TIMI flow 0-1 (46; 2.6% vs. 78; 3.9%, p = 0.022). A total of 217 patients (5.7%) died during hospitalization, significantly more often in the older population (46; 2.6% vs. 171; 8.5%, p < 0.001). The long-term mortality (data for 2847 patients from 2 centers) was higher in the older population as well (5 years survival: 86.1% vs. 59.8%). Though not significantly different and in contrast with PCI, the presence of diabetes mellitus, previous MI, final TIMI flow and LAD, as the infarct-related artery, had relatively lower impact on the older patients. Severe heart failure on admission (Killip III-IV) was associated with the worst prognosis in the whole group of patients, though its significance was higher in the youngers (HR 6.04 vs. 3.14, p = 0.051 for Killip III and 12.24 vs. 5.65, p = 0.030 for Killip IV). We clearly demonstrated age as a strong discriminator for the whole population of AMI patients. In a consecutive AMI population, the older group (>65 years) was associated with a less pronounced impact of risk factors on long-term outcome. To ascertain the coronary anatomy by coronary angiography and proceed to PCI if suitable regardless of age is crucial in all patients, though the primary success rate of PCI in the older age is lower. Age, when viewed as a risk factor, was a dominant discriminating factor in all patients.


The effect of pyrroline-5-carboxylic acid on nucleotide metabolism in erythrocytes from normal and glucose-6-phosphate dehydrogenase-deficient subjects.
Pyrroline-5-carboxylate, the intermediate in the interconversion of proline, ornithine, and glutamate, increases 5-phosphoribosyl 1-pyrophosphate (PP-ribose-P) and purine nucleotide formation in intact human erythrocytes. We proposed that: 1) pyrroline-5-carboxylate is converted to proline by pyrroline-5-carboxylate reductase with concomitant oxidation of NADPH, 2) NADP+ augments glucose-6-phosphate dehydrogenase activity, and 3) production of ribose-5-phosphate via the pentose shunt is increased. Since glucose-6-phosphate dehydrogenase plays a central role in this proposed mechanism, we examined the responsiveness of glucose-6-phosphate dehydrogenase-deficient erythrocytes to pyrroline-5-carboxylate. We compared erythrocytes from four Sardinian glucose-6-phosphate dehydrogenase-deficient subjects and four Sardinian normal controls. Without pyrroline-5-carboxylate treatment, the levels of pentose shunt activity, PP-ribose-P, and inosine monophosphate were comparable in the two populations. However, the response to pyrroline-5-carboxylate in erythrocytes from normal and glucose-6-phosphate dehydrogenase-deficient subjects was markedly different. In normal erythrocytes, pyrroline-5-carboxylate treatment increased pentose shunt activity 600%, PP-ribose-P formation 250%, and the incorporation of hypoxanthine into inosine monophosphate 260%. In contrast, pyrroline-5-carboxylate had no effect on glucose-6-phosphate dehydrogenase-deficient erythrocytes. These findings strongly support our proposed mechanism for the pyrroline-5-carboxylate effect on nucleotides. Furthermore, the markedly different capacities for nucleotide synthesis in the two populations with pyrroline-5-carboxylate treatment suggest a role for pyrroline-5-carboxylate-mediated modulation of nucleotide metabolism in normal cells.


Alpha cells, the main source of IL-1β in human pancreas.
Interleukin-1β (IL-1β) is known to trigger beta cell dysfunction in vitro and could potentially play a role during the pathogenesis of type 1 diabetes and type 2 diabetes. However, several clinical trials attempting to block IL-1β function have had minimal success. We therefore re-investigated local expression of IL-1β in human diabetic and non-diabetic pancreata. We obtained pancreatic tissue sections from the Network for Pancreatic Organ Donors with Diabetes (nPOD) including non-diabetic (n = 9), non-diabetic auto-antibody positive (AAb+, n = 5), type 1 diabetes (n = 6), and type 2 diabetes (n = 6) donors. Islets were systematically investigated for the presence of IL-1β mRNA by in situ hybridization and IL-1β protein by indirect immunofluorescence. We found that intra-islet IL-1β was produced at comparable level in both non-diabetic and diabetic donors. Interestingly, the main source for IL-1β was alpha cells but not beta cells. Our findings call into question the role of IL-1β in the diabetic pancreas as it has been proposed in previous literature. Additionally, our results regarding the localization of IL-1β should lead to further investigation into the role of IL-1β in the physiology of pancreatic alpha cells.


Erratum.
Journal of Heredity, Volume 105, Issue 6 . The issue should start on page 835. The publisher regrets this error. The online version of this issue has been corrected.


Prevalence of chronic obstructive pulmonary disease and associated factors among small-holder fish vendors along coastal areas in Tanzania.
In Tanzania little is known about how the respiratory system of small-holder fish vendors is affected by occupational exposure to biomass smoke and other associated factors. This study assessed the prevalence of lung obstruction and associated factors among small-holder fish vendors along coastal areas in Tanzania. A cross-sectional descriptive study was conducted in Bagamoyo and Kunduchi fish markets along coastal areas of Tanzania. Environmental air pollutant levels and composition were measured using a hand-held device. A standardized questionnaire was used to assess respiratory symptoms while EasyOne spirometer was used to test for lung function among small-holder fish vendors. Chronic Obstructive Pulmonary Disease (COPD) was defined as FEV1/FVC below the lower limit of normal. Data were analyzed using STATA Version 17. Descriptive statistics was performed and logistic regression analysis was used to determine factors that are associated with poor lung function presented as crude and adjusted odds ratio and their 95% confidence intervals. A total of 103 participants were included in the study who were predominantly males 82 (79.6%). The participants' mean age was 35.47 (± 8.77 SD) years. The hourly average concentration levels of PM1, PM2.5, PM10, and CO exposure during fish frying were 653.6 (± 206.3 SD) μg/m3, 748.48 (± 200.6 SD) μg/m3, 798.66 (± 181.71 SD) μg/m3 and 62.6 (± 12.3 SD) ppm respectively which are higher than the WHO recommended limits. The prevalence of COPD was found to be 32.04% (95% CI 0.23-0.42). Most of the participants reported respiratory symptoms like coughing, wheezing, sputum production and breathlessness during performing their daily activities. Findings suggest that three out of ten participants had COPD and the major environmental air pollutants (PMs and CO) concentration levels were too high, suggesting that occupational exposure to biomass smoke may be a risk factor. This calls for effective approaches to reduce exposure and prevent known acute and chronic respiratory diseases that are associated with such exposure to air pollutants. Also the study calls for follow up or cohort studies to be conducted in this area.


Novel crossed corticothalamic projections after neonatal cerebral hemispherectomy. A quantitative autoradiography study in cats.
This is a quantitative study of remodeling of the corticothalamic projections in cats with a left cerebral hemispherectomy performed neonatally or in adulthood. Kittens were lesioned at a mean postnatal age of 10 days and compared, as adults, to adult-lesioned cats of prolonged survival time. All cats received injections of [3H]leucine-proline in the right pericruciate cortex and were sacrificed 5 days later. Injection site and terminal field areas were reconstructed from autoradiography-processed tissue. For the 3 animal groups the label filled a similar extent of cortical areas 4 gamma and 3a. Computerized procedures were used to count labeled particles from multiple bilateral sites of ventral thalamic nuclei (VTN), intralaminar nuclei and midline/paramedial regions at 2 thalamic coronal planes. In control cats the only labeling in the hemithalamus contralateral to the injection was in the intralaminar nuclei and medial-most region of VTN but the counts were low. In contrast, the contralateral hemithalamus of neonatal-lesioned cats showed: a significant increase in terminal field density relative to control cats throughout the VTN, at both coronal planes, and with a distribution of labeling different to the intact side (suggesting heterotypic growth); an increase in counts of labeled particles in intralaminar nuclei which was significant for one sampling site; and an increase in counts for midline/paramedial regions which was significant for all sites at the caudal plane. Adult hemispherectomized cats were similar to intacts except for a tendency to increased VTN counts. These novel terminals were interpreted as collateral sprouting of corticofugal fibers from the intact motor cortex which crossed the midline to reinnervate the decorticate hemithalamus. Findings are discussed in the context of our reported behavioral and anatomical results.


The effect of pregnancy on metabolic responses during rest, immersion, and aerobic exercise in the water.
To examine the effects of advancing pregnancy on metabolic responses, 12 women, who were recruited early in pregnancy, were studied during 20 minutes of immersion in 30 degrees C water, followed by 20 minutes of exercise in the water (60% of predicted maximal capacity) and 20 minutes of lateral supine recovery. Each subject completed the trials during the fifteenth, twenty-fifth, and thirty-fifth weeks of pregnancy, as well as a control period 8 to 10 weeks post partum. Resting oxygen uptake increased with advancing pregnancy. Resting oxygen uptake was higher in the water than on land but was not altered by pregnancy. Exercise oxygen uptakes were similar for all trials, but the work load required to elicit the VO2 decreased during the thirty-fifth week of pregnancy. Exercise heart rates followed the same pattern as oxygen uptake. Lactate concentrations declined with advancing pregnancy after exercise. Blood glucose levels were normal for pregnancy but declined slightly during exercise. Blood triglyceride levels were elevated with exercise, with a tendency to increase with advancing pregnancy. Resting plasma cortisol concentrations increased with pregnancy but remained lower during immersion and exercise. These results suggest that pregnancy significantly alters metabolic responses to exercise in the water.


Nano zinc oxide-sodium alginate antibacterial cellulose fibres.
In the present study, antibacterial cellulose fibres were successfully fabricated by a simple and cost-effective procedure by utilizing nano zinc oxide. The possible nano zinc oxide was successfully synthesized by precipitation technique and then impregnated effectively over cellulose fibres through sodium alginate matrix. XRD analysis revealed the 'rod-like' shape alignment of zinc oxide with an interplanar d-spacing of 0.246nm corresponding to the (101) planes of the hexagonal wurtzite structure. TEM analysis confirmed the nano dimension of the synthesized zinc oxide nanoparticles. The presence of nano zinc oxide over cellulose fibres was evident from the SEM-EDS experiments. FTIR and TGA studies exhibited their effective bonding interaction. The tensile stress-strain curves data indicated the feasibility of the fabricated fibres for longer duration utility without any significant damage or breakage. The antibacterial studies against Escherichia coli revealed the excellent bacterial devastation property. Further, it was observed that when all the parameters remained constant, the variation of sodium alginate concentration showed impact in devastating the E. coli. In overall, the fabricated nano zinc oxide-sodium alginate cellulose fibres can be effectively utilized as antibacterial fibres for biomedical applications.


Teratogenic and dominant lethal studies of patulin in mice.
Teratogenicity of daily intraperitoneal (i.p.) injections of 1.5 or 2.0 mg/kg of patulin on days 6 through 17 of pregnancy and mutagenic effects of acute i.p. exposure of 3.0 mg/kg of patulin on male germ cells were evaluated. Resorption of all implanted embryos occurred at 2.0 mg/kg/day of patulin, while a significant reduction in the average body weight of 19-day-old fetuses from patulin-treated mothers, compared to control fetuses, was noticed at 1.5 mg/kg/day without any lethal effects on the implanted embryos. Patulin was embryocidal, possibly fetotoxic but was neither teratogenic nor mutagenic to mice.


Xanthichthysgreenei, a new species of triggerfish (Balistidae) from the Line Islands.
Xanthichthysgreenei sp. n. is described from six specimens, 97-154 mm standard length (SL) collected from mesophotic coral ecosystems (90-100 m) at Kiritimati (Christmas Island), Line Islands, part of the Republic of Kiribati in the Central Pacific. Of the six species of Xanthichthys, it is most similar to the Atlantic Xanthichthysringens and the Indo-West Pacific Xanthichthyslineopunctatus, sharing with these species the character of three pigmented cheek grooves. It is distinctive in its low body scale row count (33-35, other Xanthichthys species with 39 or more), small size (maximum SL 154 mm, other species over 225 mm), and color pattern of scattered dark spots sub-dorsally and no other spots or lines on body.


A facile synthesis of Au-nanoparticles decorated PbI2 single crystalline nanosheets for optoelectronic device applications.
This research communication presents a rapid and facile microwave-assisted synthesis of single crystalline nanosheets (SCNSs) of hexagonal lead iodide (PbI2) decorated with Au nanoparticles, a potential optoelectronics material. Homogeneous low dimensional AuNP decoration in PbI2 resulted in a new absorption band at ~604 nm and a shift in band gap from 3.23 to 3.00 eV. The significant enhancement of photoluminescent (PL) intensity observed in the AuNP-PbI2 SCNSs is attributed to the coupling of the localized surface plasmon resonanzce of AuNP leading to improved excitation and emission rates of PbI2-SCNSs in the region of the localized electromagnetic field. The Au-PbI2 SCNSs display a compelling increment in photoconductivity, and its fabricated photodetector showed a stable and switchable photo-response. Due to ease of synthesis and enhanced photoconductivity along with appealing PL features, Au-PbI2 SCNS has the potential to be used as a material of choice when fabricating an optoelectronic devices of high performance.


Clinical impact of carbon-ion radiotherapy on hepatocellular carcinoma with Child-Pugh B cirrhosis.
Hepatocellular carcinoma (HCC) patients with Child-Pugh (CP)-B not eligible for surgery nor other focal therapy options due to impaired liver function, have very limited treatment options. This study aims to retrospectively investigate the toxicity and efficacy of Carbon-ion radiotherapy (C-ion RT) on HCC with CP-B patients. Patients with CP-B, no extrahepatic metastasis, and treated with C-ion RT between May 2000 and March 2020 were retrospectively extracted and included in this study. Sixty-nine lesions of 58 patients were included. The median follow-up duration was 20.5 (2.7-108) months. During follow-up, recurrence was observed in 43 patients, including 2 local recurrences and 39 intrahepatic recurrences beyond the irradiation field. A grade 3 acute hepatotoxicity was observed in one patient during the observation period. No acute or late adverse event of grade ≥4 was observed. Overall survival was 80.4% and 46.0% at 1 and 2 years, respectively, and the median survival time was 22.6 months. Local control rate was 96.4% at both 1 and 2 years, and progression-free survival was 38.6% and 6.9% at 1 and 2 years, respectively, with a median of 9.7 months. The C-ion RT showed low toxicity and good local effect in patients with HCC and CP-B. Therefore, C-ion RT could be an appropriate treatment for patients with HCC with poor liver function.


Results of programmes for antenatal detection of thalassemia in reducing the incidence of the disorder.
The characteristics and the effectiveness of programmes designed to prevent beta-thalassemia major present in high frequency in several areas of the world such as Cyprus, Greece and Sardinia are reviewed. All these programmes are based on heterozygote detection, counselling and foetal diagnosis. The target population for screening have been couples at marriage, conception or early pregnancy. Awareness of the problem and involvement of the population was achieved via mass-media or personal approaches through lectures or discussions. Parent's Association were consulted and have been actively involved. Information leaflets have been made available to prospective couples at several critical areas. Education on thalassemias was introduced into the school curriculum. Counselling was based on a private interview at which the several options available were discussed with the individual carrier or the couple. Prenatal diagnosis was chosen by the large majority of couples counselled. All these programmes resulted in a decline of thalassemia major births by 50-97%. The reasons for residual cases were mostly lack of information and, less frequently, misdiagnoses or refusal of fetal diagnoses.


Ovarian papillary cystadenocarcinomas in a green iguana (Iguana iguana).
The clinical, gross morphological and histological characteristics of bilateral ovarian papillary cystadenocarcinomas in an adult female green iguana (Iguana iguana) are documented in this report. Histologically, the tumour consisted of an invasive mass of small glandular structures, irregular arboriform papillae and cystic structures arising from the ovarian surface epithelium. Metastatic implants were extensively disseminated along the serosa of the coelomic viscera, mesentery and fat bodies. Tumour morphology and metastatic behaviour indicated a close resemblance to cystadenocarcinoma in other species. Primary ovarian neoplasia is rare in reptiles; documented cases include tumours arising from surface epithelium, germ cells and sex cord stroma.


The MILLER banding procedure as a treatment alternative for dialysis access steal syndrome: a single institutional experience.
To describe a single institutional experience with minimally invasive limited ligation endoluminal-assisted revision (MILLER) for treatment of dialysis access steal syndrome (DASS). Twenty patients were retrospectively identified that underwent 30 MILLER band procedures for DASS at our institution from March 2010 to December 2014. Technical success was defined by successful creation of MILLER band with preservation of flow for hemodialysis. Clinical success was defined as complete resolution of signs and symptoms with preservation of dialysis access in a 1-month postprocedural period. Primary MILLER band patency, postintervention-assisted primary access patency, and postprocedure secondary access patency are reported. Technical success was achieved in all patients. Clinical success was achieved in 75% of patients after one banding procedure and in 95% of patients after two banding procedures. One patient experienced access thrombosis following the initial banding procedure which was subsequently treated and did not lead to loss of access. MILLER band patency was 83% at 1 month and 77% at 6 months. Postintervention-assisted primary patency was 95%, 93%, and 92% at 3 months, 6 months, and 1 year, respectively. Postintervention secondary patency was 86%, 68%, and 59% at 3 months, 6 months, and 1 year, respectively. MILLER banding offers a less-invasive alternative to surgical therapy that appears to be safe and permits preservation of dialysis access.


Non-osmotic effects of external sugars on the cell water content and transport of cations across the plasmalemma inHydrodictyon reticulatum.
Saccharides added to the cultivation medium influence the properties of the cell wall of the strictly autotrophic green algaHydrodictyon reticulatum, most probably in such a way that they interfere with the processes of growth and repair of microfibrils. Natural monosaccharides (glucose, fructose, galactose, mannose, sucrose, cellobiose, raffinose) reduce the cell water content, increase the intracellular concentration mostly of both potassium and sodium cations and reduce the ohmic resistance of the cell membrane.


Portable coagulometer for vitamin K-antagonist monitoring: the patients' point of view.
The aim of this study was to know the patients' point of view on the monitoring of vitamin K-antagonist (VKA) therapy by means of a point of care testing (POCT), ie, using a portable coagulometer by self-testing at home. At first, patients had prothrombin time (PT) international normalized ratio (INR) monitoring at a thrombosis center; afterward, they were shifted to self-testing at home. An interview was done to evaluate the patients' point of view on the two monitoring periods. A total of 92 oral anticoagulated patients were enrolled. The questionnaire contained nine questions that elicited a maximum of five closer answers that were arranged in increasing levels of satisfaction: very little, little, enough, much and completely. Percentage of time in therapeutic range (TTR) and adverse events were compared during the two periods of conventional monitoring and self-testing. The period of conventional monitoring was shorter than that of self-testing (p<0.0001), and the median TTR was satisfactory but lower than that of self-testing (p<0.0001). A total of 85% of the patients were satisfied with self-testing at home. In all, 83% and 73% (p=0.06) of patients felt comfortable about side effects while measuring the PT INR at both home and the thrombosis center, respectively. During the self-testing period, quality of life was improved in 87% of the patients. The cost of test strips was medium-high for 89% of the patients, and 75% of them stated that it was worth improving their quality of life. A switch from VKA to a direct oral anticoagulant (DOAC) was proposed to 24% of the patients, but 68% of them declined because they felt more comfortable monitoring their oral anticoagulant therapy by POCT. VKA monitoring using POCT at home may play a role in improving the patients' quality of life and may be considered as an alternative to the use of DOAC at least in certain settings of patients.


Electric shock sign after digital nerve injury; report of a case and review of literature.
Digital nerve lacerations are rationally common, especially following penetrating injuries. The majority of patients suffer from numbness as the main complaint. However, electric shock pain is a rare manifestation of partial nerve injury. A 65-year-old woman with partial digital nerve injury and an electric shock sign due to a 1 cm laceration on the volar side of the proximal phalanx three weeks earlier. The surgical exploration revealed a neuroma-like mass in the digital nerve; however, the continuity of the nerve was grossly intact. Due to the superficiality of digital nerves, penetrating nerve injuries are rationally common in this area. Laceration of the digital nerve could lead to loss of sensation, clumsiness, and even loss of function. Gradually, neuroma formation would add a local pain and electric shock sign to the mentioned symptoms. Electric shock sign is characteristic for partial digital nerve injury with unknown mechanism. It seems the symptoms would be progressive and more complicated with neuroma formation. V.


Psychosocial aspects of children and families of children treated with automated peritoneal dialysis.
The aim of this study was to analyze psychosocial aspects of chronic kidney disease (CKD) in children treated with automated peritoneal dialysis (APD). The study assessed 41 children > 2 (range 2.1-18) years of age and their parents. Data concerning the illness and sociodemographic parameters were collected. Patients completed the Paediatric Quality of Life Inventory (PedsQL) and their parents the PedsQL-proxy version, General Health Questionnaire (GHQ-12), Berlin Social Support Scales (BSSS), and Caregiver's Burden Scale (CBS). Parents rated their children's overall health-related quality of life (QoL) as well as their physical and emotional functioning lower than the patients themselves. The majority of primary caregivers had a medium level of the total burden index in the CBS and higher values in the scales need for support and perceived available support than in the received support (BSSS). In the GHQ-12, 51.2% of primary caregivers had scores >2 points, which indicated the possible occurrence of abnormal mental functioning. Financial support for patients' families is necessary. Parents who provide primary care to children on PD require, above all, emotional support and assistance in self-fulfilment. More than half of them may have impaired mental function. There is the strong need to provide continuous psychological care for caregivers. Differences in perception of the children's activity in varied areas by the patients themselves and their caregivers may contribute to further problems within families.


Septation of the univentricular heart. Transatrial approach.
The technique of septation of the univentricular heart via a transatrial approach is described in detail. Clinical details of four patients having Type A-III univentricular heart document the feasibility of working through the right-sided atrioventricular valve to place a cloth prosthesis into the ventricle to divide it into approximately equal chambers. Improved hemodynamics were observed in the postoperative period, which probably were related to retaining the integrity of the ventricular wall. This approach seems best suited to the ventricular chamber estimated to be large by echocardiography without severe pulmonary outflow tract obstruction. Techniques for enlargement of the pulmonary outflow tract are also described.


Tissue, age, sex, and disease patterns of matrisome expression in GTEx transcriptome data.
The extracellular matrix (ECM) has historically been explored through proteomic methods. Whether or not global transcriptomics can yield meaningful information on the human matrisome is unknown. Gene expression data from 17,382 samples across 52 tissues, were obtained from the Genotype-Tissue Expression (GTEx) project. Additional datasets were obtained from The Cancer Genome Atlas (TCGA) program and the Gene Expression Omnibus for comparisons. Gene expression levels generally matched proteome-derived matrisome expression patterns. Further, matrisome gene expression properly clustered tissue types, with some matrisome genes including SERPIN family members having tissue-restricted expression patterns. Deeper analyses revealed 382 gene transcripts varied by age and 315 varied by sex in at least one tissue, with expression correlating with digitally imaged histologic tissue features. A comparison of TCGA tumor, TCGA adjacent normal and GTEx normal tissues demonstrated robustness of the GTEx samples as a generalized matrix control, while also determining a common primary tumor matrisome. Additionally, GTEx tissues served as a useful non-diseased control in a separate study of idiopathic pulmonary fibrosis (IPF) matrix changes, while identifying 22 matrix genes upregulated in IPF. Altogether, these findings indicate that the transcriptome, in general, and GTEx in particular, has value in understanding the state of organ ECM.


From Campers to Counselors: a Resource for Prospective Genetic Counseling Students.
When thinking about the future of the genetic counseling field, one place to start is with prospective genetic counseling graduate school applicants. Although resources and mentorship opportunities exist for genetic counselors entering the field, the process of deciding on a career, applying to graduate programs, and being admitted can be daunting. As members of the profession, we should take responsibility for ensuring that individuals have the information and resources necessary to make an educated decision about whether genetic counseling is the correct path for them and to take the initial steps along this path. In this article, we present our Genetic Counseling Boot Camp as a model for other genetic counselors to use in developing their own local programs. This type of program can benefit prospective genetic counselors as they begin their professional journeys and can also provide value for the organizers and presenters who are already seasoned in the field.


Cell-based treatment of osteochondral defects in the rabbit knee with natural and synthetic matrices: cellular seeding determines the outcome.
Matrix-associated transplantation of cartilage constructs is an appealing method in cartilage repair. Three different matrices seeded with allogenic chondrocytes were compared in an osteochondral defect model in the rabbit. An investigation was conducted to identify the best matrix for cell-based treatment of osteochondral defects in the rabbit knee joint. Osteochondral defects (diameter 3 mm) were created in the trochlea and the femoral condyles of 33 New Zealand White rabbits, which were then treated with bioartificial cartilage constructs. The cartilage constructs were created in vitro using three different resorbable carrier materials (two fleece matrices: one of PLLA, and one composite of polydioxanon/ polyglactin, as well as one consisting of lyophilized dura) cultured with isolated allogenic chondrocytes. The defects were evaluated macroscopically, by histological and immunhistological techniques, and by scanning electron microscopy after 6 weeks, 6 months, and 12 months. The chondrocyte-seeded constructs were compared to defects treated with carrier material alone as well as to untreated control defects. There was a significant improvement in defect repair quality in the transport materials, which were cultured with chondrocytes prior to implantation (P < 0.0005). No significant differences were observed between the three carrier matrices, and no significant differences were seen between the unseeded matrices and the untreated control defects. There is no difference in the outcome between the three tested matrices in the treatment of osteochondral defects in the rabbit knee. The results of this in vitro experiment are promising and with refinement may lead to useful clinical therapies.


Reactive Oxygen Species-Responsive Polymeric Prodrug Nanoparticles for Selective and Effective Treatment of Inflammatory Diseases.
It is challenging to manage inflammatory diseases using traditional anti-inflammatory drugs due to their limited efficacy and systemic side effects, which are a result of their lack of selectivity, poor stability, and low solubility. Herein, it reports the development of a novel nanoparticle system, called ROS-CA-NPs, which is formed using polymer-cinnamaldehyde (CA) conjugates and is responsive to reactive oxygen species (ROS). ROS-CA-NPs exhibit excellent drug stability, tissue selectivity, and controlled drug release upon oxidative stress activation. Using mouse models of chronic rheumatoid arthritis and acute ulcerative colitis, this study demonstrates that the systemic administration of ROS-CA-NPs results in their accumulation at inflamed lesions and leads to greater therapeutic efficacy compared to traditional drugs. Furthermore, ROS-CA-NPs present excellent biocompatibility. The findings suggest that ROS-CA-NPs have the potential to be developed as safe and effective nanotherapeutic agents for a broad range of inflammatory diseases.


Adaptation of Ocular Opponency Neurons Mediates Attention-Induced Ocular Dominance Plasticity.
Previous research has shown that ocular dominance can be biased by prolonged attention to one eye. The ocular-opponency-neuron model of binocular rivalry has been proposed as a candidate account for this phenomenon. Yet direct neural evidence is still lacking. By manipulating the contrast of dichoptic testing gratings, here we measured the steady-state visually evoked potentials (SSVEPs) at the intermodulation frequencies to selectively track the activities of ocular-opponency-neurons before and after the "dichoptic-backward-movie" adaptation. One hour of adaptation caused a shift of perceptual and neural ocular dominance towards the unattended eye. More importantly, we found a decrease in the intermodulation SSVEP response after adaptation, which was significantly greater when high-contrast gratings were presented to the attended eye than when they were presented to the unattended eye. These results strongly support the view that the adaptation of ocular-opponency-neurons contributes to the ocular dominance plasticity induced by prolonged eye-based attention.


Capillary electrophoresis for capture and concentrating of target nucleic acids by affinity gels modified to contain single-stranded nucleic acid probes.
Selective capture and pre-concentration of target nucleic acids from relatively complicated samples may provide a method to facilitate introduction to a microfluidic-based detection system to improve detection limits. An acrylamide polymer gel modified with Acrydite that contained 20mer oligonucleotide probe was prepared and loaded into a capillary column. The results indicated that the amount of probe DNA that was captured into the acrylamide was about 40% of the starting monomer, and different quantities of probe could therefore be coupled into the gel. The gel was passivated by pre-treatment with non-complementary DNA oligonucleotide to block non-selective adsorption sites, and the gel was determined to be stable for multiple cycles of use. The probe could hybridize with target sequences that were introduced by electrokinetic injection from a sample solution. The target could be freed from the polymer gel by use of a combination of heating, chaotropic salt and voltage conditions. Target capture efficiency was up to 90% when using samples that did not saturate probe sites in the columns, and recovery of target from the gel could be as high as 95%.


Retention forces of 14-unit zirconia telescopic prostheses with six double crowns made from zirconia--an in vitro study.
The aim of this study was to evaluate the retention forces (RFs) of zirconia double-crown systems, with primary and secondary crowns made from zirconia in vitro. Ten maxillary models with six abutment teeth were prepared. Sixty inner crowns were fabricated from pre-sintered zirconia with a taper of 0°. Ten 14-unit telescopic prostheses (removable partial dentures, RDPs) were fabricated, using the same computer-aided design/computer-aided manufacturing system as that used for the inner crowns. The removal test was performed in a standardized setup using a universal testing device at a crosshead speed of 10 mm/min. Ten separation cycles were carried out for each single primary crown as well as for each 14-unit RDP in the presence of artificial saliva. The results were imported into a statistic program and analysed by a one-way ANOVA and post hoc tests. The level of significance was set at 5 %. The mean RFs of the single double-crown systems were in the range of 0.611-2.895 N, whereas the RFs for the whole RDP varied between 8.1 and 13.6 N. RF was dependent on the abutment tooth (p < 0.001) and on the model (p < 0.001). The results of this study indicate that the manufacturing of full-zirconia double-crown systems is possible as well as reproducible. The RFs are comparable to those reported from casted and electroformed double-crown systems. It has been shown that the RFs of the presented telescopic system are comparable to existing double-crown systems.


Teaching the wounded healer.
The wounded healer remains easily hidden in a profession that implicitly, and at times actively, encourages a denial of illness. The wounded healer is, at the very least, less likely to encourage healthy habits in patients, and at the very worst, can become an impaired physician. In training, however, the wounded healer can be identified and treated before entering clinical practice. It would seem, then, that medical school and residency is an optimal time to identify and treat the wounded healer.


Heterologous expression of genes encoding bacterial light-harvesting complex II in Rhodobacter capsulatus and Rhodovulum sulfidophilum.
In the present work we report the high-level expression of foreign genes encoding the light-harvesting (LHII) membrane-spanning polypeptides in photosynthetic bacteria. To do this we first constructed three deletion strains of Rhodovulum (Rhv.) sulfidophilum in which all or part of the puc operon, encoding the peripheral light-harvesting proteins, is missing. To investigate the heterologous expression of the light-harvesting polypeptides from Rb. capsulatus in Rhv. sulfidophilum and vice versa we have reintroduced functional foreign LH genes into these and equivalent strains of Rhodobacter (Rb.) capsulatus. In some cases very high levels of expression were obtained (85%) of those observed in the wild type), while in other cases much lower expression was observed; possible reasons for these differences are discussed. The heterologously expressed proteins were shown to contain normal pigment-binding sites and to be normally and functionally integrated within the host photosynthetic apparatus. The results indicate that heterologous proteins are able to assemble properly and enter into the same protein-protein interactions as their analogs originally present in the host strain.


Impact of Baseline Imaging of Non-Culprit Coronary Lesions on Adverse Events: Insight From LRP Study.
Intravascular ultrasound (IVUS) and near-infrared spectroscopy (NIRS) can identify vulnerable coronary atherosclerotic plaques. We aimed to compare the presence or absence of baseline intravascular imaging of non-culprit lesions and their subsequent adverse events. We identified patients from the Lipid Rich Plaque (LRP) study who had a non-culprit-lesion adverse event and divided them into 2 cohorts: those with lesions detected with NIRS-IVUS imaging at baseline and those with lesions not imaged at baseline. Overall, 73 patients had an adverse event (99 coronary segments) during the 24-month follow-up period. Among them, 41 patients (56.2%) had a non-culprit-lesion adverse event related to a coronary segment imaged at baseline, and 32 patients (43.8%) had a non-culprit-lesion adverse event adjudicated to a segment that was not scanned at baseline. Angiographic core laboratory analysis suggested that unscanned lesions were more often in the right coronary artery (~50%); branches of the left coronary artery, i.e., diagonal or left obtuse marginal arteries (~20%); smaller vessels; or more tortuous vessels; and less often in the left anterior descending or distal locations. There was a weak trend for acute severe events (adjudicated myocardial infarction and acute coronary syndrome) in patients with lesions not scanned at baseline (50.0% versus 36.6%, p = 0.250). In patients with follow-up non-culprit-lesion adverse events, nearly half were not imaged with NIRS-IVUS at baseline. Because events related to non-imaged lesions were at least as severe as events related to imaged lesions, future clinical trials and clinical protocols should be designed to minimize this issue. The Lipid-Rich Plaque Study (LRP), https://clinicaltrials.gov/ct2/show/NCT02033694, NCT02033694.


[Comparative evaluation of late results of the Hoffmeister-Finsterer gastrectomy and the method of valve anastomoses in peptic ulcer].
Comparative assessment of the remote results of Hoffmeister-Finsterer gastrectomy with creation of valve anastomoses has demonstrated rare occurrence of postoperative complications owing to anastomoses. The operation is simple as concerns operative techniques and can be recommended as operation of choice in the treatment of peptic ulcer.


Recent Advances in Organic Photovoltaic Materials Based on Thiazole-Containing Heterocycles.
Organic solar cells (OSCs) have achieved great progress, driven by the rapid development of wide bandgap electron donors and narrow bandgap non-fullerene acceptors (NFAs). Among a large number of electron-accepting (A) building blocks, thiazole (Tz) and its derived fused heterocycles have been widely used to construct photovoltaic materials, especially conjugated polymers. Benefiting from the electron deficiency, rigidity, high planarity, and enhanced intra/intermolecular interactions of Tz-containing heterocycles, some related photovoltaic materials exhibit proper energy levels, optimized molecular aggregation, and active layer morphology, leading to excellent photovoltaic performance. This review focuses on the progress of Tz-based photovoltaic materials in the field of OSCs. First, the Tz-based donor and acceptor photovoltaic materials are reviewed. Then, the materials based on promising Tz-containing heterocycles, mainly including thiazolo[5,4-d]thiazole (TzTz), benzo[1,2-d:4,5-d']bis(thiazole) (BBTz), and benzo[d]thiazole (BTz) are summarized and discussed. In addition, the new emerging Tz-fused structures and their application in OSCs are introduced. Finally, perspectives and outlooks for the further development of Tz-containing heterocycle-based photovoltaic materials are proposed.


Protective mechanism of the EZH2/microRNA-15a-5p/CXCL10 axis in rats with depressive-like behaviors.
Enhancer of zeste homolog 2 (EZH2), microRNA-15a-5p (miR-15a-5p), and chemokine C-X-C ligand 10 (CXCL10) have been studied in many diseases. However, the investigation of the EZH2/miR-15a-5p/CXCL10 axis in depression is not sufficient. Our study aimed to investigate the regulatory functions of the EZH2/miR-15a-5p/CXCL10 axis in rats with depressive-like behaviors. The rat model of depression-like behaviors was established by chronic unpredictable mild stress (CUMS), and EZH2, miR-15a-5p, and CXCL10 expression levels in rats with depression-like behaviors were detected. The silenced EZH2 or enhanced miR-15a-5p recombinant lentivirus was injected into the rats with depression-like behaviors to assess the changes in behavioral tests, hippocampal pathological structure, levels of inflammatory cytokines in the hippocampus, and hippocampal neuron apoptosis. The regulatory relationships among EZH2, miR-15a-5p, and CXCL10 were measured. miR-15a-5p expression was reduced, and EZH2 and CXCL10 expression levels were elevated in rats with depressive-like behaviors. Downregulation of EZH2 or elevation of miR-15a-5p improved depressive behavior, and inhibited hippocampal inflammatory response and hippocampal neuron apoptosis. EZH2 promoted histone methylation at the promoter of miR-15a-5p, and miR-15a-5p bound to CXCL10 to inhibit its expression. Our study summarizes that EZH2 promotes the hypermethylation of the miR-15a-5p promoter, thereby promoting CXCL10 expression. Upregulation of miR-15a-5p or inhibition of EZH2 can improve the symptoms in rats with depressive-like behaviors.


Direct cardiac massage without major thoracotomy: feasibility and systemic blood flow.
Open-chest cardiac massage (OC-CM) provides higher blood pressure and flow than closed-chest compression and may improve the probability of successful resuscitation from cardiac arrest. Its clinical use has been limited by its requirement for a major thoracotomy. The present pilot study tested the technical feasibility of performing effective direct cardiac massage without a major thoracic incision, by using a simple, manually-powered plunger-like device, inserted through a small thoracic incision, to cyclically compress the cardiac ventricles. The method was termed minimally-invasive direct cardiac massage (MID-CM). Systemic blood flow using MID-CM was compared to that with OC-CM, by both direct systemic hemodynamic measurements, cumulative metabolic indicators of the ratio of whole body oxygen delivery and oxygen consumption, and a metabolic index of pulmonary blood flow. In 12 large swine, baseline systemic and pulmonary hemodynamic measurements were performed. Arterial and mixed venous blood gases and metabolic indicators of systemic blood flow were measured. Ventricular fibrillation was induced and after 4 min, animals underwent either bimanual OC-CM (N = 6) or MID-CM (N = 6). At 10, 20 and 30 min, hemodynamic and metabolic measurements were repeated. Systemic Blood Pressure: Aortic systolic and diastolic blood pressures were reduced from baseline levels with both OC-CM and MID-CM. No difference in pressure was noted between OC-CM and MID-CM groups. Pulmonary Artery Pressure: Pulmonary artery systolic pressure was elevated from baseline during OC-CM and MID-CM. Pulmonary artery diastolic pressures remained constant throughout the resuscitation period in both groups. No differences in pulmonary systolic or diastolic pressure were noted between OC-CM and MID-CM groups. A trend towards higher pulmonary systolic pressures appeared with MID-CM. Thermodilution Blood Flow: Cardiac index fell from baseline levels with OC-CM and MID-CM. No difference in cardiac index was noted between OC-CM and MID-CM groups. Metabolic Indices: Mixed venous O2 saturation decreased from baseline levels during resuscitation in both experimental groups, with a further decrease at 30 min compared to 10- and 20-min levels. No difference was noted between OC-CM and MID-CM groups at any point. Arterial pH was reduced from baseline levels at 30 min in both groups compared to baseline but no difference was noted between groups.(ABSTRACT TRUNCATED AT 400 WORDS)


[Differentiation of spermatic antigens and vaginal discharge in mixed spots].
The technique of differentiation of ABO antigens in a mixed spot of sperm and vaginal discharge based on higher electrophoretic mobility of spermatic protein compared to that of vaginal secretion is proposed.


Chemotherapy with epirubicin and paclitaxel for breast cancer during pregnancy: case report and review of the literature.
Breast cancer diagnosed during pregnancy is a challenging clinical situation. Little data are currently available about chemotherapy in pregnant women with this malignancy. We report the case of a 36-year-old pregnant woman with a T2N1M0 breast cancer who received sequential chemotherapy including epirubicin (120 mg/m2 every three weeks for four cycles) and paclitaxel (175 mg/m2 every three weeks for three cycles) from the 14th to the 32nd week of gestation. The patient delivered a normal female baby by caesarean section at the 36th week. The immunohistochemical examination of the placenta showed a diffuse, strong P-glycoprotein expression. Thirty-six months after the delivery, the mother was disease-free and the infant showed normal development and growth. Sequential chemotherapy including epirubicin and paclitaxel should be taken into consideration as adjuvant treatment for pregnant women with high-risk breast cancer. The strong placental expression of P-glycoprotein may play a major role in limiting fetal exposure to anthracyclines and taxanes.


Angle-scanning SPR imaging for detection of biomolecular interactions on microarrays.
In this paper we describe the use of a commercial surface plasmon resonance (SPR) imaging instrument for monitoring the binding of biomolecules on user-defined regions of interest of a microarray. By monitoring the angle shift of the SPR-dip using a continuous angle-scanning mode instead of monitoring the change in reflectivity at a fixed angle, a linear relationship with respect to the mass density change on the surface will remain over a wide dynamic angle range of 8 degrees. Peptides (2.4 kDa) and proteins (150 kDa) were both spotted on the same sensor chip to illustrate that both, low and high molecular weight ligands with initial large differences in off-set SPR angles, can be applied within the same experiment. By using a fluorescently labeled antibody, SPR results can be confirmed by means of fluorescence microscopy after completion of a SPR experiment. SPR imaging in angle-scanning operation provides sensitive, accurate, and label-free detection of analyte binding on microarrays containing different molecular weight ligands.


Binding of antibodies to functional epitopes on the pore formed by Escherichia coli hemolysin in cells and model membranes.
Escherichia coli hemolysin (HlyA) inserts into target membranes producing a cation-selective pore. We approached the problem of determining which portions of this protein remain exposed on the side of attack by applying specific antibodies. Results obtained with resealed erythrocyte ghosts and planar phospholipid membranes were compared. The effects of one polyclonal and four monoclonal anti-hemolysin antibodies (mAbs) were studied. Using ghosts we found one mAb which strongly reduced the ion-permeability through the preinserted HlyA channels and one which clearly increased it. Experiments with planar bilayers corroborated these results by showing that the former mAb effectively promoted the closed state of the channel whereas the latter forced the HlyA channel into an open configuration. Anti-hemolysin polyclonal antibodies initially stimulated but then prevented channel opening, indicating they contained clones able to act on both these channel determinants. They were effective only when applied on the same side as the hemolysin indicating that the epitopes were exposed to that side. Finally, the antigenic epitopes of three of the mAbs were localised on the HlyA molecule by using different mutants (amber and frame shift mutants and hemolysin gene hybrids).


Accuracy of cytokeratin 18 (M30 and M65) in detecting non-alcoholic steatohepatitis and fibrosis: A systematic review and meta-analysis.
Association between elevated cytokeratin 18 (CK-18) levels and hepatocyte death has made circulating CK-18 a candidate biomarker to differentiate non-alcoholic fatty liver from non-alcoholic steatohepatitis (NASH). Yet studies produced variable diagnostic performance. We aimed to provide summary estimates with increased precision for the accuracy of CK-18 (M30, M65) in detecting NASH and fibrosis among non-alcoholic fatty liver disease (NAFLD) adults. We searched five databases to retrieve studies evaluating CK-18 against a liver biopsy in NAFLD adults. Reference screening, data extraction and quality assessment (QUADAS-2) were independently conducted by two authors. Meta-analyses were performed for five groups based on the CK-18 antigens and target conditions, using one of two methods: linear mixed-effects multiple thresholds model or bivariate logit-normal random-effects model. We included 41 studies, with data on 5,815 participants. A wide range of disease prevalence was observed. No study reported a pre-defined cut-off. Thirty of 41 studies provided sufficient data for inclusion in any of the meta-analyses. Summary AUC [95% CI] were: 0.75 [0.69-0.82] (M30) and 0.82 [0.69-0.91] (M65) for NASH; 0.73 [0.57-0.85] (M30) for fibrotic NASH; 0.68 (M30) for significant (F2-4) fibrosis; and 0.75 (M30) for advanced (F3-4) fibrosis. Thirteen studies used CK-18 as a component of a multimarker model. For M30 we found lower diagnostic accuracy to detect NASH compared to previous meta-analyses, indicating a limited ability to act as a stand-alone test, with better performance for M65. Additional external validation studies are needed to obtain credible estimates of the diagnostic accuracy of multimarker models.


New Atrophic Acne Scar Classification: Reliability of Assessments Based on Size, Shape, and Number.
Post-acne atrophic scarring is a major concern for which standardized outcome measures are needed. Traditionally, this type of scar has been classified based on shape; but survey of practicing dermatologists has shown that atrophic scar morphology has not been well enough defined to allow good agreement in clinical classification. Reliance on clinical assessment is still needed at the current time, since objective tools are not yet available in routine practice.
 Evaluate classification for atrophic acne scars by shape, size, and facial location and establish reliability in assessments.
 We conducted a non-interventional study with dermatologists performing live clinical assessments of atrophic acne scars. To objectively compare identification of lesions, individual lesions were marked on a high-resolution photo of the patient that was displayed on a computer during the clinical evaluation. The Jacob clinical classification system was used to define three primary shapes of scars 1) icepick, 2) boxcar, and 3) rolling. To determine agreement for classification by size, independent technicians assessed the investigators' markings on digital images. Identical localization of scars was denoted if the maximal distance between their centers was ≤ 60 pixels (approximately 3 mm). Raters assessed scars on the same patients twice (morning/afternoon). Aggregate models of rater assessments were created and analyzed for agreement.
 Raters counted a mean scar count per subject ranging from 15.75 to 40.25 scars. Approximately 50% of scars were identified by all raters and ~75% of scars were identified by at least 2 of 3 raters (weak agreement, Kappa pairwise agreement 0.30). Agreement between consecutive counts was moderate, with Kappa index ranging from 0.26 to 0.47 (after exclusion of one outlier investigator who had significantly higher counts than all others). Shape classifications of icepick, boxcar, and rolling differed significantly between raters and even for same raters at consecutive sessions (P<.001 and P=0.4, respectively). Analysis showed only 65% of scars were identical in both sessions. We also found that there is a threshold of detection in terms of size, with poor agreement among investigators for very small scars (<2 mm). The repeatability of identification of scars ≥ 2.0 mm was acceptable, and we found that increasing scar size was positively correlated with agreement. Reliability was improved when only scars >2 mm were included. For smaller scars (<2 mm), inter-rater reliability was poor.
 While intuitively it makes sense that describing scar morphology could guide treatment, we have shown that shape-based evaluations are subjective and do not readily yield strong agreement. Until there is a more objective way to evaluate morphology that is readily available to practicing clinicians, we propose that size should be considered a primary characteristic for scar classification systems. We further suggest classification of <2 mm, 2-4 mm, and >4 mm based on how the size would likely affect diagnostic and therapeutic choices. Finally, we recommend that scars <2 mm not be included in a clinical classification but should be evaluated by an objective method that may be refined in the future.

 J Drugs Dermatol. 2016;15(6):693-702.


Virtual histology-intravascular ultrasound assessment of lesion coverage after angiographically-guided stent implantation in patients with ST Elevation myocardial infarction undergoing primary percutaneous coronary intervention.
An occlusion or severe stenosis (angiographic culprit lesion) of the infarct-related artery is frequently located at the site of the maximum thrombus burden, whereas the origin of the plaque rupture (the true culprit) can be situated proximal or distal to it. The aim of this study was to examine stent coverage of true culprit lesions in 20 patients who underwent primary percutaneous coronary intervention and had Thrombolysis In Myocardial Infarction (TIMI) grade 3 flow restored in the infarct-related artery by angiographically guided direct stenting. Images of lesions were obtained using virtual histology-intravascular ultrasound before and after intervention (blinded to the operator). Plaque rupture sites were identified by intravascular ultrasound in 12 lesions (60%), 11 proximal and 1 distal to the minimum luminal area (MLA). Maximum necrotic core sites were found proximal to the MLA in 16 lesions, at the MLA in 3 lesions, and distal to the MLA in 1 lesion. Plaque rupture sites were fully covered by stents in 11 lesions. Virtual histology-intravascular ultrasound-derived thin-cap fibroatheroma longitudinal geographic misses were found in 10 lesions, 7 in the proximal reference segment and in 3 patients in the proximal and distal reference segments. In conclusion, in about 50% of patients who undergo primary percutaneous coronary intervention for ST-segment elevation myocardial infarction with optimal angiographic results, the stent does not fully cover the maximum necrotic core site related to the culprit lesion.


Ex vivo expansion of tumor-infiltrating lymphocytes from nasopharyngeal carcinoma patients for adoptive immunotherapy.
Establishing Epstein-Barr virus(EBV)-specific cytolytic T lymphocytes(EBV-CTLs) from peripheral blood mononuclear cells(PBMCs) for adoptive immunotherapy has been reported in EBV-associated malignancies including Hodgkin's lymphoma and nasopharyngeal carcinoma(NPC). In the current study, we performed ex vivo expansion of tumor-infiltrating lymphocytes(TILs) obtained from NPC biopsy specimens with a rapid expansion protocol using anti-CD3 monoclonal antibody(OKT3), recombinant human interleukin(IL)-2, and irradiated PBMCs from healthy donors to initiate the growth of TILs. Young TIL cultures comprised of more than 90% of CD3+ T cells, a variable percentage of CD3+CD8+ and CD3+CD4+ T cells, and less than 10% of CD3-CD16+ natural killer cells, a similar phenotype of EBV-CTL cultures from PBMCs. Interestingly, TIL cultures secreted high levels of the Th1 cytokines, interferon gamma (IFNγ) and tumor necrosis factor-alpha (TNF-α), and low levels of the Th2 cytokines, IL-4 and IL-10. Moreover, young TILs could recognize autologous EBV-transformed B lymphoblast cell lines, but not autologous EBV-negative blast cells or allogeneic EBV-negative tumor cells. Taken together, these data suggest that ex vivo expansion of TILs from NPC biopsy tissue is an appealing alternative method to establish T cell-based immunotherapy for NPC.