Solubilization of a Poorly Soluble B-Raf (Rapidly Accelerated Fibrosarcoma) Inhibitor: From Theory to Application.
The oral bioavailability of a drug candidate is influenced by its permeability, metabolism, and physicochemical properties. Among the physicochemical properties, solubility and dissolution rate often are the most critical factors affecting the oral bioavailability of a compound. The increasing challenge for the pharmaceutical industry is to achieve reasonable oral bioavailability of poorly water-soluble drug candidates. G-F is a potent and selective B-Raf (rapidly accelerated fibrosarcoma) inhibitor with poor water solubility and moderate permeability, which resulted in an absorption-limited exposure in preclinical safety studies. The intrinsic solubility of G-F is 8 μg/mL (i.e., 0.0188 nM). In this study, pH adjustment combined with cosolvency, micellization, or complexation was applied as a technique to enhance the solubility of G-F. pH 9.5 and 4 buffers were selected to combine with the solubilization agents based on G-F's acidic pKa of 7.47. The solubilization power of each solubilization agent was determined based on the experimental data. The solubility G-F can be increased up to 4000-fold in a selected combination. The advantage of combination over individual solubilization agent was demonstrated. In this study, the understanding of the solubilization power of each solubilization agent played an important role in the formulation development of this development candidate.


The effect of beam shape on physical parameters of head and neck simultaneous-integrated boost intensity-modulated radiation therapy.
To evaluate the influence of the beam shape created by X-rays with "flat beams" and without "flattening-filter-free [FFF] beams" a flattening filter, and the isocenter locations for FFF beams on the treatment of a large irradiated volume for tumours. The increase of dose rate and the decrease of out-of-field dose can be expected for FFF beams and lead to effective and safety radiotherapy. On the other hand, the bell-shaped dose profile is thought to be a factor of negating these advantages. Treatment plans for 15 patients with head and neck cancer were created using XiO (Elekta, Stockholm AB, Sweden) in fixed-gantry step-and-shoot delivery under the same dose constraints. Seven fields of FFF beams with 7 MV and flat beams with 6 MV were used with the technique of intensity-modulated radiation therapy (IMRT). We compared the dose homogeneity and conformity of targets and dose constraints for organs as the plan quality and evaluated physical parameters: monitor unit (MU) values, number of segments and their locations from the isocenter in beam's-eye-view. No significant differences were found in the plan quality. The isocenter locations do not affect the physical parameters for FFF beams. It has been confirmed that the number of segments and MU values were 40% higher with FFF beams than with flat beams (p < 0.05). This study demonstrates flat dose distribution is more suitable for IMRT with large and complex targets.


Hepatoprotective activity of scopoletin, a constituent of Solanum lyratum.
Scopoletin (7-hydroxy-6-methoxycoumarin), a coumarin, was isolated from the aerial part of Solanum lyratum Thunb. by the activity-guided fractionation employing carbon tetrachloride-intoxicated primary cultured rat hepatocytes as a screening system. Its hepatoprotective activity was first evaluated by measuring the release of glutamic pyruvic transaminase and sorbitol dehydrogenase from carbon tetrachloride-intoxicated rat hepatocytes into the culture medium. Scopoletin significantly reduced the releases of glutamic pyruvic transaminase and sorbitol dehydrogenase from the carbon tetrachloride-intoxicated primary cultured rat hepatocytes by 53% and 58%, respectively, from the toxicity in a dose-dependent manner over concentration ranges of 1 microM to 50 microM. Further studies revealed that at the concentration of 10 microM, scopoletin significantly preserved glutathione content by 50% and the activity of superoxide dismutase by 36% and also inhibited the production of malondialdehyde to the degree as seen in the control.


Gene therapy in the cornea: 2005--present.
Successful restoration of vision in human patients with gene therapy affirmed its promise to cure ocular diseases and disorders. The efficacy of gene therapy is contingent upon vector and mode of therapeutic DNA introduction into targeted cells/tissues. The cornea is an ideal tissue for gene therapy due to its ease of access and relative immune-privilege. Considerable progress has been made in the field of corneal gene therapy in last 5 years. Several new gene transfer vectors, techniques and approaches have evolved. Although corneal gene therapy is still in its early stages of development, the potential of gene-based interventions to treat corneal abnormalities has begun to surface. Identification of next generation viral and nanoparticle vectors, characterization of delivered gene levels, localization, and duration in the cornea, and significant success in controlling corneal disorders, particularly fibrosis and angiogenesis, in experimental animal disease models, with no major side effects have propelled gene therapy a step closer toward establishing gene-based therapies for corneal blindness. Recently, researchers have assessed the delivery of therapeutic genes for corneal diseases and disorders due to trauma, infections, chemical, mechanical, and surgical injury, and/or abnormal wound healing. This review provides an update on the developments in gene therapy for corneal diseases and discusses the barriers that hinder its utilization for delivering genes in the cornea.


Fluoride levels in river water from the volcanic regions of Cauca (Colombia).
Fluoride concentrations of 0.5 mg/L in drinking water are considered useful for dental caries prevention. However, fluoride concentrations higher than 1.2 mg/L in water can pose a risk of dental fluorosis due to high exposure to fluoride. The objective is to determine the fluoride concentration in water from aqueducts of different Colombian municipalities of Cauca (Popayán, Coconuco, and Puracé) to assess the fluoride dietary intake from the consumption of this water. A total of 66 water samples have been taken from Popayán, Coconuco, and Puracé. Fluoride content was determined by fluoride ion-selective electrode (ISE) potentiometry. The fluoride concentrations recorded in Coconuco and Puracé water were ≤ 0.002 mg/L. The mean fluoride content recorded in the Popayán water was 0.42 mg/L, with its highest concentration in Cauca River water (0.83 mg/L). Considering the admissible intake values, the water from Popayán confers remarkable fluoride intakes, especially in children with high percentages of contribution to the admissible daily intake (46.7% to 7- to 12-month children and 41.5% to 1- to 3-year children). The fluoride content in the water of Coconuco and Puracé does not reach an optimal value (< 0.5 mg/L) for the protective effect against dental caries, while the water of the main Cauca River basin does reach the optimal value. Likewise, the intake of fluoride from the consumption of the analyzed water does not confer any health risk. However, the implementation of monitoring systems for fluoride levels is recommended in order to safeguard the consumer's health.


Formation of Stilbene Azo-Dimer by Direct Irradiation of p-Azidostilbene†.
Triplet arylnitrenes may provide direct access to aryl azo-dimers, which have broad commercial applicability. Herein, the photolysis of p-azidostilbene (1) in argon-saturated methanol yielded stilbene azo-dimer (2) through the dimerization of triplet p-nitrenostilbene (3 1N). The formation of 3 1N was verified by electron paramagnetic resonance spectroscopy and absorption spectroscopy (λmax ~ 375 nm) in cryogenic 2-methyltetrahydrofuran matrices. At ambient temperature, laser flash photolysis of 1 in methanol formed 3 1N (λmax ~ 370 nm, 2.85 × 107 s-1 ). On shorter timescales, a transient absorption (λmax ~ 390 nm) that decayed with a similar rate constant (3.11 × 107 s-1 ) was assigned to a triplet excited state (T) of 1. Density functional theory calculations yielded three configurations for T of 1, with the unpaired electrons on the azido (TA ) or stilbene moiety (TTw , twisted and TFl , flat). The transient was assigned to TTw based on its calculated spectrum. CASPT2 calculations gave a singlet-triplet energy gap of 16.6 kcal mol-1 for 1 N; thus, intersystem crossing of 1 1N to 3 1N is unlikely at ambient temperature, supporting the formation of 3 1N from T of 1. Thus, sustainable synthetic methods for aryl azo-dimers can be developed using the visible-light irradiation of aryl azides to form triplet arylnitrenes.


Dual effects of neuroprotection and neurotoxicity by general anesthetics: role of intracellular calcium homeostasis.
Although general anesthetics have long been considered neuroprotective, there are growing concerns about neurotoxicity. Preclinical studies clearly demonstrated that commonly used general anesthetics are both neuroprotective and neurotoxic, with unclear mechanisms. Recent studies suggest that differential activation of inositol 1,4,5-trisphosphate receptors, a calcium release channel located on the membrane of endoplasmic reticulum (ER), play important role on determining the fate of neuroprotection or neurotoxicity by general anesthetics. General anesthetics at low concentrations for short duration are sublethal stress factors which induce endogenous neuroprotective mechanisms and provide neuroprotection via adequate activation of InsP3R and moderate calcium release from ER. On the other hand, general anesthetics at high concentrations for prolonged duration are lethal stress factors which induce neuronal damage by over activation of InsP3R and excessive and abnormal Ca(2+) release from ER. This review emphasizes the dual effects of both neuroprotection and neurotoxicity via differential regulation of intracellular Ca(2+) homeostasis by commonly used general anesthetics and recommends strategy to maximize neuroprotective but minimize neurotoxic effects of general anesthetics.


An Analysis of Self-Reported Barriers to Type 1 Diabetes Care in a Pediatric Population in British Columbia, Canada.
Our aim in this study was to identify patient-level barriers to attending pediatric type 1 diabetes mellitus (T1DM) clinic and to better understand the demographic and clinical characteristics of these reporting barriers. Patients were recruited from pediatric T1DM clinics throughout British Columbia. Barriers to attending clinic were identified through a survey. Demographic and clinical characteristics of patients who reported difficulty attending clinic appointments were compared with those who did not. Of the 197 study participants, 31% reported difficulty attending appointments. Commonly reported barriers were distance to clinic and missing work. Younger child age and residing in northern regions increased the odds of reporting a barrier, whereas residing on Vancouver Island decreased odds of reporting a barrier. There were no differences in glycated hemoglobin levels between the 2 groups. Approximately 1 in 3 patients identified challenges in attending T1DM appointments in British Columbia. Further research is needed to determine whether similar challenges exist in other provinces.


Brief report: What types of Internet guidance do patients want from their physicians?
To understand what patients expect from physicians regarding information seeking on the Internet. Self-administered survey. Waiting rooms of 4 community-based primary care offices. Of 494 patients invited to participate, 330 completed the survey for a response rate of 67%. Of 177 respondents who used the Internet for health information, only 15% agreed that physicians should ask them about their Internet searches. Most (62%) agreed that physicians should recommend specific web sites where patients can learn more about their health care. Primary care physicians should recognize that many patients would like guidance as they turn to the Internet for medical information. Physicians can utilize quality assessment tools and existing resources that facilitate referring patients to authoritative, commercial-free, patient-oriented medical information on the Internet.


Effect of social distancing on injury incidence during the COVID-19 pandemic: an interrupted time-series analysis.
To evaluate the effects of social distancing on the incidence and characteristics of injuries during the COVID-19 pandemic. This cross-sectional study used the National Emergency Department Information System (NEDIS) database. Injured patients who visited all 402 emergency departments (EDs) between 29 February and 29 May 2020 (after-distancing), and in the corresponding period in 2019 (before distancing) to control for seasonal influences. The study outcome was the incidence of injury. Using the interrupted time-series analysis models, we analysed weekly trends of study outcomes in both periods (before and after distancing), the step change (the effect of intervention), and the slope change over two periods (the change in the effect over time). The incidence rates of injury per 100 000 person-days were 11.2 and 8.6 in the before-distancing and after-distancing periods, respectively. In the after-distancing period, the incidence rate of injury decreased (step change -3.23 (95% CI -4.34 to -2.12) per 100 000 person-days) compared with the before-distancing period, while the slope change was 0.10 (95% CI 0.04 to 0.24). The incidence rate ratios of all injuries and intentional injuries for the after-distancing period were 0.67 (95% CI 0.60 to 0.75) and 1.28 (95% CI 1.18 to 1.40), respectively, compared with the before-distancing period. Fewer injuries occurred after the implementation of social distancing programme compared with the same period in the previous year. However, this effect gradually decreased postimplementation.


The electric fence to cell-cycle progression: Do local changes in membrane potential facilitate disassembly of the primary cilium?: Timely and localized expression of a potassium channel may set the conditions that allow retraction of the primary cilium.
Kv10.1 is a voltage-gated potassium channel relevant for tumor biology, but the underlying mechanism is still unclear. We propose that Kv10.1 plays a role coordinating primary cilium disassembly with cell cycle progression through localized changes of membrane potential at the ciliary base. Most non-dividing cells display a primary cilium, an antenna-like structure important for cell physiology. The cilium is disassembled when the cell divides, which requires an increase of Ca2+ concentration and a redistribution of phospholipids in its basal region, both of which would be facilitated by local hyperpolarization. Cells lacking Kv10.1 show impaired ciliary disassembly and delayed entrance into mitosis. Kv10.1 is predominantly expressed during G2/M, a critical period for ciliary resorption, and localizes to the ciliary base and vesicles associated with the centrosome. This could explain the influence of Kv10.1 in cell proliferation, as well as phenotypic features of patients carrying gain of function mutations in the gene.


Ciprofloxacin exposure and adverse pregnancy outcomes: A Danish nationwide cohort study.
To examine the association between maternal exposure to ciprofloxacin and the risk of miscarriage and major malformations. A nationwide register-based cohort study. Data were obtained from the Medical Birth Registry, the National Hospital Registry, the Danish National Prescription Registry and Statistics Denmark. Data were collected in the period between 1997 and 2016 and included all registered pregnancies that ended in an elective termination, miscarriage, stillbirth or a live birth. Exposure was defined as redeeming one or more prescriptions of ciprofloxacin. Miscarriage was defined as a diagnosis given before 22 weeks without any medical intervention. Major malformations were classified according to EUROCAT 1.4. We matched ciprofloxacin-exposed pregnancies to unexposed pregnancies on the propensity score in a ratio 1:4. To estimate the hazard ratio (HR) of miscarriage a Cox proportional hazard regression model was used. A log binomial model was used to estimate the relative risk ratio (RR) of major malformations. HR of miscarriage and the RR of major malformations. A total of 1 650 649 pregnancies were identified. Of these, 10 250 (2050 ciprofloxacin-exposed) and 6100 (1220 ciprofloxacin-exposed) were included in the miscarriage and major malformation analysis, respectively. The HR of miscarriage was 0.99 (95% confidence interval [CI] 0.84-1.17). For major malformation, the RR was 1.01 (95% CI 0.72-1.40). For the organ-specific major malformations and the sensitivity analyses, no significant increased risks were identified. We demonstrated no association between miscarriage and maternal ciprofloxacin exposure within the first 22 weeks of pregnancy, or between major malformations and maternal exposure during the first trimester. No association between maternal ciprofloxacin exposure and adverse pregnancy outcomes.


Subtyping of uropathogenic Escherichia coli according to the pathogenicity island encoding uropathogenic-specific protein: comparison with phylogenetic groups.
Phylogenetic analysis has been used widely to characterize extraintestinal pathogenic Escherichia coli in molecular epidemiological studies. We have recently reported a putative pathogenicity island (PAI), carrying uropathogenic-specific protein (usp) and a unique mosaic structure of small open reading frames following usp, providing four subtypes of PAIusp classified from their sequential patterns. A total of 427 E. coli isolates from uncomplicated urinary tract infections (194 cystitis, 76 pyelonephritis, and 107 prostatitis) and 50 fecal isolates were examined for phylogenetic grouping and PAIusp subtyping as well as the prevalence of virulence factors (VF) and O serogroups. Both phylogenetic group B2 and usp-positive strains were equally predominant in cystitis, pyelonephritis and prostatitis (B2, 80.9%, 86.8%, and 86.9%; usp, 79.4%, 93.4%, and 88.8%, respectively). Furthermore, each PAIusp subtype was shown to be closely associated with several VF genes as well as several common O serogroups of uropathogenic E. coli. In molecular epidemiological studies, PAIusp subtyping will provide additional informative findings of E. coli strains belonging to phylogenetic group B2.


A personal construct approach to perception of disclosure targets.
Investigating the perception of targets for self-disclosure from the perspective of the psychology of personal constructs, it was predicted that respondents from a mixed-nationality group would (a) characterize friends more than acquaintances in terms of 'superordinate' personality-descriptive dimensions, (b) 'differentiate' friends more than acquaintances by extensive application of constructs to them, and (c) 'polarize' targets for high disclosure by construing them sharply and meaningfully in terms of personally significant dimensions of meaning. All predictions received significant support for 34 college students (Mdnage 22 yr.) who responded in writing to a questionnaire. The results suggest the fruitfulness of further examining self-disclosure and friendship formation from a vantage point which emphasizes the perceptual and interpretive framework of an adult person.


MS3 eliminates ratio distortion in isobaric multiplexed quantitative proteomics.
Quantitative mass spectrometry-based proteomics is highly versatile but not easily multiplexed. Isobaric labeling strategies allow mass spectrometry-based multiplexed proteome quantification; however, ratio distortion owing to protein quantification interference is a common effect. We present a two-proteome model (mixture of human and yeast proteins) in a sixplex isobaric labeling system to fully document the interference effect, and we report that applying triple-stage mass spectrometry (MS3) almost completely eliminates interference.


Angiographic measurement of posterior fossa in the Japanese.
Measurement employing the concept of the centroid is the most accurate way to study the radiographic anatomy of the posterior fossa. The normal measurements in Japanese subjects and as reported by Ross and du Boulay correlated well. We have developed a new radiographic centroid based on the morbid anatomy of the posterior fossa. The normal measurement in the Japanese by this method is also presented.


Osteogenic effects of the bioactive small molecules and minerals in the scaffold-based bone tissue engineering.
Reconstruction of the damaged bone is a striking challenge in the medical field. The bone grafts as a current treatment is associated with inherent limitations; hence, the bone tissue engineering as an alternative therapeutic approach has been considered in the recent decades. Bone tissue engineering aims at replacing the lost tissue and restoring its function by recapitulating the natural regeneration process. Concerted participation and combination of the biocompatible materials, osteoprogenitor/ stem cells and bioactive factors closely mimic the bone microenvironment. The bioactive factors regulate the cell behavior and they induce the stem cells to osteogenic differentiation by activating specific signaling cascades. Growth factors (GFs) are the most important bioactive molecules and mediators of the natural bone repair process. Although these soluble factors have approved applications in the bone regeneration, however, there are several limitations such as the instability, high dose requirements, and serious side effects which could restrict their clinical usage. Alternatively, a new generation of bioactive molecules with the osteogenic properties are used. The non-peptide organic or inorganic molecules are physiologically stable and non-immunogenic due to their small size. Many of them are obtained from the natural resources and some are synthesized through the chemical methods. As a result, these molecules have been introduced as the cost-effective osteogenic agents in the bone tissue regeneration. In this paper, three groups of these bioactive agents including the organic small molecules, minerals and metallic nanoparticles have been investigated, considering their function in accelerating the bone regeneration. We review the recent in vitro and in vivo studies that utilized the osteogenic molecules to promote the bone formation in the scaffold-based bone tissue engineering systems.


Molecular structure and vibrational spectra of 3-and 4-amino-2-bromopyridine by density functional methods.
In this work, experimental and theoretical study on the molecular structure and the vibrational spectra of 3-amino-2-bromopyridine (Compound I) and 4-amino-2-bromopyridine (Compound II) are presented. The vibrational frequencies of these compounds were obtained theoretically by DFT/B3LYP employing the 6-311G(2df,2p) basis set for optimized geometries and were compared with Fourier Transform Infrared solid phase spectra (FTIR) in the region of 400-4000 cm(-1), FT-Raman spectra in the region of 100-4000 cm(-1) and with solution phase spectra in the region of 400-4000 cm(-1). Complete vibrational assignment, analysis and correlation of the fundamental modes for these compounds have been carried based on the potential energy distribution (PED). The vibrational harmonic frequencies were scaled using scale factors, yielding a good agreement between the experimentally recorded and the theoretically calculated values. Finally the calculated HOMO and LUMO energies are localized on the entire ring and substituents.


Should tumour necrosis factor antagonist safety information be applied from patients with rheumatoid arthritis to psoriasis? Rates of serious adverse events in the prospective rheumatoid arthritis BIOBADASER and psoriasis BIOBADADERM cohorts.
Information on the safety of tumour necrosis factor (TNF) antagonists frequently arises from their use in rheumatic diseases, their first approved indications, and is later applied to psoriasis. Whether the risk of biological therapy is similar in psoriasis and rheumatoid arthritis has been considered a priority research question. To compare the safety profile of anti-TNF drugs in patients with rheumatoid arthritis and psoriasis. We compared two prospective safety cohorts of patients with rheumatoid arthritis and psoriasis that share methods (BIOBADASER and BIOBADADERM). There were 1248 serious or mortal adverse events in 16 230 person-years of follow-up in the rheumatoid arthritis cohort (3171 patients), and 124 in the 2760 person-years of follow-up of the psoriasis cohort (946 patients). Serious and mortal adverse events were less common in patients with psoriasis than in rheumatoid arthritis (incidence rate ratio of serious adverse events in psoriasis/rheumatoid arthritis: 0·6, 95% confidence interval 0·5-0·7). This risk remained after adjustment for sex, age, treatment, disease, hypertension, diabetes, hypercholesterolaemia and simultaneous therapy with methotrexate (hazard ratio 0·54, 95% confidence interval 0·47-0·61), and after excluding patients receiving corticosteroids. Patients with rheumatoid arthritis showed a higher rate of infections, cardiac disorders, respiratory disorders and infusion-related reactions, whereas patients with psoriasis had more skin and subcutaneous tissue disorders and hepatobiliary disorders. Patients with rheumatoid arthritis clinical practice have almost double the risk of serious adverse events compared with patients with psoriasis, with a different pattern of adverse events. Safety data from rheumatoid arthritis should not be fully extrapolated to psoriasis. These differences are likely to apply to other immune-mediated inflammatory diseases.


Zinc and copper concentration do not differentiate bipolar disorder from major depressive disorder.
The aim of this study was to compare the zinc and copper concentration in the group of patients with bipolar disorder (BD) and major depressive disorder (MDD). 110 patients with the diagnosis of BD and 114 with MDD were qualified to the study. To assess the levels of microelements, the flame atomic absorption spectrometry (FAAS) was used in the case of zinc and the electrothermal atomic absorption spectrometry (ETAAS) was used in the case of copper. There were no differences between concentration of zinc and copper in remission and depressive phase between patients with BD and MDD. Additionally, there were also no statistically significant differences in comparisons including type I and II, early or late phase of BD and MDD. The lack of differences in zinc and copper concentrations between patients with bipolar disorder and major depressive disorder might indicate that those disorders have similar etiology.


Positive alliesthesia after insulin.
Volunteers experienced sucrose solution as more pleasant 36-48 min after insulin, than after saline control. These changes in affective estimates correlate negatively with blood sugar at 30 min and positively at 50 min after the insulin injection.


Relative bioavailability of three newly developed albendazole formulations: a randomized crossover study with healthy volunteers.
This study of healthy volunteers shows that the relative bioavailability of albendazole formulations that use arachis oil-polysorbate 80 or hydroxypropyl-beta-cyclodextrin as an excipient was enhanced 4.3- and 9.7-fold compared to the results seen with commercial tablets. Administration of macrogol suppositories did not result in measurable plasma concentrations of albendazole sulfoxide.


COVID-19 and vaccination against SARS-CoV-2 in patients with neuromyelitis optica spectrum disorders.
Reports on outcomes of COVID-19 in patients with neuromyelitis optica spectrum disorder (NMOSD) are scarce, as well as those related to the safety profile of the vaccines in this population. The aim of this survey is to present demographic and clinical characteristics of patients with NMOSD who developed COVID-19 and safety data of the COVID-19 vaccines in these persons. This study comprise all patients from the Hospital registry of NMOSD, of the Clinic of Neurology in Belgrade, who fulfilled the 2015 NMOSD diagnostic criteria, and who after invitation by phone call, from April 10 to May 10, 2021, accepted to participate and provide information regarding COVID-19 and vaccination against Sars-CoV-2 (n = 53). Sixteen out of 53 enrolled NMOSD patients were diagnosed with COVID-19. In three cases (18.8%), severity of COVID-19 clinical manifestations warranted hospitalization, and one of these patients, died due to COVID-19 (case fatality ratio = 6.25%), after invasive mechanical ventilation. The remaining two patients had grade II COVID -19 severity and were hospitalized because of pneumonia, not requiring supplemental oxygen. Median EDSS in patients requiring hospitalization was 4.5, and in the non-hospitalized group, it was 3.0. Nine out of 53 patients received two doses of vaccine against Sars-Cov-2 (8 Sinopharm and one Pfizer). Pain at the site of application was the only vaccine-related adverse effect. Our survey indicates overall favourable COVID-19 outcome and encouraging safety profile of the vaccines in persons with NMOSD, in our cohort. Prospective studies are warranted to confirm these data.


Local heating of radiation belt electrons to ultra-relativistic energies.
Electrically charged particles are trapped by the Earth's magnetic field, forming the Van Allen radiation belts. Observations show that electrons in this region can have energies in excess of 7 MeV. However, whether electrons at these ultra-relativistic energies are locally accelerated, arise from betatron and Fermi acceleration due to transport across the magnetic field, or if a combination of both mechanisms is required, has remained an unanswered question in radiation belt physics. Here, we present a unique way of analyzing satellite observations which demonstrates that local acceleration is capable of heating electrons up to 7 MeV. By considering the evolution of phase space density peaks in magnetic coordinate space, we observe distinct signatures of local acceleration and the subsequent outward radial diffusion of ultra-relativistic electron populations. The results have important implications for understanding the origin of ultra-relativistic electrons in Earth's radiation belts, as well as in magnetized plasmas throughout the solar system.


Assessing Cutaneous Mosaicism at the Molecular Level.
Mosaicism results from postzygotic alterations during embryogenesis leading to genetically distinct populations of cells within individuals and has been historically recognized by phenotypes with visible, often patterned manifestations. Before the advent of molecular profiling assays and high-throughput sequencing, it was challenging to study mosaicism in human disease; however, the study of mosaic disorders has recently revealed unexpected and novel pathways for disease pathogenesis. In this paper, we will review the techniques for discovery of disease-causing alleles using Proteus syndrome; phakomatosis pigmentokeratotica; linear porokeratosis; and vacuoles, E1 enzyme, X-linked, autoinflammatory somatic syndrome as models. These tools represent powerful approaches for dissecting the genetic basis for human disorders.


Neutropenia associated with large granular lymphocytes responsive to corticosteroids in vitro and in vivo.
We describe a patient with neutropenia associated with increased circulating large granular lymphocytes (LGL). Absolute neutropenia was accompanied by the absence of myeloid precursor cells in the bone marrow. No myeloid progenitor cells (CFU-C) could be detected by in vitro colony culture. The peripheral blood was also remarkable for the presence of a population of large granular lymphocytes demonstrable by conventional staining. These cells in flow microfluorometry studies expressed antigens Leu 4 (T-cell antigen receptor), Leu 7 (natural killer cell marker), Leu 2 (suppressor cell marker), and HLA-DR (activation marker); they lacked Leu 1 (a pan-T cell antigen), Leu 3 (helper cell marker) and Tac (interleukin 2 receptor). Hematopoietic colony formation in vitro improved with addition of corticosteroids to the culture medium or elimination of the LGL population with complement-mediated cytotoxicity. Anti-neutrophil antibodies were present prior to and following therapy. Clinically, administration of prednisone resulted in a normalization of the total white blood cell count and absolute polymorphonuclear cell number, an increase into the normal range of the number of CFU-C, and elimination of the LGL population. In this case of steroid-responsive LGL-associated neutropenia, laboratory studies suggested direct suppression of myelopoiesis by steroid-responsive LGL.


New functionally selective muscarinic agonists.
The muscarinic pharmacology of two novel agonists related to McN-A-343, 4-F-PyMcN and 4-F-PyMcN+, has been studied by the use of pharmacological and radioligand binding techniques. Both compounds were potent agonists at M1 receptors in rabbit vas deferens (pEC50 = 6.24 and 6.96) and rat duodenum (pEC50 = 5.47 and 6.38), but very weak partial agonists or competitive antagonists at guinea-pig cardiac M2 and ileal M3 receptors. There was no receptor reserve for 4-F-PyMcN in rabbit vas deferens, for which the potency (pEC50 = 6.24) and apparent affinity (pKA = 5.99 and 6.21) were similar. 4-F-PyMcN+ showed only limited binding selectivity between four muscarinic receptor binding assays with apparent affinity constants (pKi) of 5.8, 5.2, 5.6 and 5.7 for M1, M2, M3 and M4 muscarinic receptor subtypes. The two novel functionally M1-selective agonists may provide useful tools with which to study muscarinic receptor mechanisms. The non-quaternary compound, 4-F-PyMcN, might become a starting point for the development of drugs that selectively affect M1 receptors involved in central cholinergic function.


The Changing Face of Technologically Integrated Neurosurgery: Today's High-Tech Operating Room.
Over the last decade, surgical technology in planning, mapping, optics, robotics, devices, and minimally invasive techniques has changed the face of modern neurosurgery. We explore the current advances in clinical technology across all neurosurgical subspecialties, examine how clinical practice is being shaped by this technology, and suggest what the operating room of tomorrow may look like.


Formulation and characterization of DNA-polyethylenimine-dextran sulfate nanoparticles.
Polyethylenimine (PEI) is a promising non-viral gene delivery polymer that produces high transfection efficiency both in vitro and in vivo. The use of PEI, however, is hindered by its toxicity, reflecting its polycationic nature. In an attempt to decrease this charge-dependent cytotoxicity, a polyanionic polymer, dextran sulfate (DS), has been incorporated into self-assembling PEI-DNA complexes with zinc as stabilizing agent. Spherical particles with a mean particle size of approximately 200 nm and a polydispersity index of 0.2 were achieved using the following optimal conditions: PEI solutions at pH 8, PEI/DS mass ratios of >or=2, and 25 microM zinc sulfate. Plasmid DNA was completely condensed within the nanoparticles as confirmed by an ethidium bromide accessibility assay. This result correlates well with DNase protection studies which find partial protection of the DNA nanoparticles from degradation by the enzyme. The DNA was incorporated into the PEI-DS particles with a high efficiency (>95%) and maintained a primarily supercoiled B-form as determined by gel electrophoresis and circular dichroism. The cytotoxicity of the DNA nanoparticles appeared to decrease as the amount of DS in the formulation was increased and they produced moderate transfection activities that were only modestly inhibited by the presence of serum.


Evidence that high potency foot-and-mouth disease vaccine inhibits local virus replication and prevents the "carrier" state in sheep.
The ability of a single administration of a high, medium and low potency foot-and-mouth disease (FMD) vaccine to decrease or inhibit local virus replication and excretion in the oropharynx of sheep following aerosol challenge with homologous live virus 14 days later was examined. Unvaccinated sheep showed signs of clinical FMD, whereas all of the vaccinated sheep, regardless of antigen payload, were protected against clinical disease and development of viraemia. Virological and serological results confirmed that there had been no local virus replication in the oropharynx of sheep from the high potency vaccine group in contrast to moderate or substantial virus replication in the oropharynx of the low potency vaccinated or unvaccinated sheep respectively. The vaccines showed no evidence of promoting a local mucosal antibody response at the time of virus challenge, but were capable of stimulating a systemic gamma interferon response, the level of which was related to the antigen payload. This suggests that the systemic gamma interferon response could be a useful indicator of the ability of a FMD vaccine to elicit a sterile immunity and indicates that further work is warranted to investigate the role of systemic gamma interferon in this immunity. This is the first experiment to clearly show that high potency, high payload, FMD vaccines are capable of inhibiting local virus replication and consequently persistence and the carrier state in this target species.


The signaling pathways and targets of traditional Chinese medicine and natural medicine in triple-negative breast cancer.
Triple-negative breast cancer (TNBC) has a poorer prognosis than other subtypes due to its strong invasion and higher risk of distant metastasis. Traditional Chinese medicine (TCM) and natural medicine have the unique advantages of multitargets and small side-effects and may be used as long-term complementary and alternative therapies. The present article summarizes the classical signaling pathways and potential targets by the action of TCM and natural medicine (including extracts, active constituents and formulas) on TNBC and provides evidence for its clinical efficacy. The literature information was acquired from the scientific databases PubMed, Web of Science and CNKI from January 2010 to June 2020, and it was designed to elucidate the internal mechanism and role of TCM and natural medicine in the treatment of TNBC. The search key words included "Triple negative breast cancer" or "triple negative breast carcinoma", "TNBC" and "traditional Chinese medicine" or "Chinese herbal medicine", "medicinal plant", "natural plant", and "herb". We described the antitumor activity of TCM and natural medicine in TNBC based on different signaling pathways. Plant medicine and herbal formulas regulated the related gene and protein expression via pathways such as PI3K/AKT/mTOR, MAPK and Wnt/β-catenin, which inhibit the growth, proliferation, migration, invasion and metastasis of TNBC cells. The inhibitory effect of TCM and natural medicine on tumors was reflected in multiple levels and multiple pathways, providing reasonable evidence for new drug development. To make TCM and natural medicine widely and flexibly used in clinical practice, the efficacy, safety and mechanism of action need more in-depth experimental research.


Increased disease activity is associated with altered sleep architecture in an experimental model of systemic lupus erythematosus.
The aim of this study was to evaluate sleep patterns during the course of the disease in (NZB/NZW)F1 mice, an experimental model of systemic lupus erythematosus (SLE). Female mice were implanted with electrodes for chronic recording of sleep-wake cycles during the entire experimental phase (9, 19, and 29 weeks of age). The disease course was also assessed. At each time-point, blood samples were collected from the orbital plexus to evaluate serum antinuclear antibodies (ANA), which are important serologic parameters of disease evolution. Pain perception was also evaluated. During the dark phase, (NZB/NZW)F, mice aged 19 weeks spent more time in sleep, and, as a consequence, the total waking time was lower when compared with earlier periods. An augmented number of sleep-stage transitions and microarousals were observed at the 29th week of life in both light and dark phases. At this same time-point, the mice showed lower pain thresholds than they had at 9 weeks of life. The disease status was confirmed; the entire group of mice at 29 weeks of life showed positive ANA with high titer levels. The sleep-recording data showed that, during the progress and severe phases of the disease (19 and 29 wks of age, respectively), sleep architecture is altered. According to these results, increased sleep fragmentation, disease activity, and pain sensitivity are features observed in these mice, similar to symptoms of SLE.


Toxicity of the 13 priority pollutant metals to Vibrio fisheri in the Microtox chronic toxicity test.
The Microtox Acute Toxicity Test has been successfully used to measure the toxicity of metals and other pollutants at high concentrations (ppm) in selected environmental samples. However, metals and other toxicants are often found in much lower concentrations (ppb) in many municipal wastewaters and receiving waters. In order to assess the toxicity of these pollutants in these samples, a more sensitive toxicity assay is needed. The Microtox chronic toxicity test has been developed to measure the sublethal effect of toxicants over multiple generations of the test species, Vibrio fisheri. In this study, the toxicity of the 13 priority pollutant metals [i.e. As, Se, Cd, Cr (III and VI), Cu, Pb, Sb, Ag, Tl, Zn, Be, Hg and Ni] to V. fisheri was evaluated using the Microtox chronic toxicity test. In this test, the inhibitory concentration (IC), lowest observable effect concentration (LOEC), and no observable effect concentration (NOEC) were obtained after 22-h of incubation at 27+/-1 degrees C, by comparing the light output of the control to that of the test sample. Among the 13 priority pollutant metals, beryllium (Be) was found to be the most toxic in the test (LOEC=0.742-1.49 microg/l) while thallium (Tl) was the least toxic (LOEC=3840-15300 microg/l). The LOECs for copper (as Cu) and lead (Pb) in reagent (ASTM Type I) water were 6.78-13.6 microg/l and 626-1251 microg/l, respectively. The toxicity of copper sulfate (as Cu) in reagent water was shown and significantly reduced with the addition of natural organic matter (fulvic acid) or EDTA to the sample. The LOEC values for the 13 priority pollutant metals in this test were comparable to or lower than those reported for commonly used aquatic toxicity tests, such as the Ceriodaphnia dubia assay.


[The connection of the electronic structure and biological activity of various derivatives of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine].
Electron structure of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and some of its analogs--the substrates of monoaminooxidase--substituted by phenyl cycles was studied by semiempiric quantum-chemical CNDOR, MINDOB methods. The relationship between the obtained electron and conformation parameters (orientation of the phenyl ring in particular) and biological activity of the compounds under consideration is discussed. A comparative analysis of the distribution pattern of the electron density for the MPTP molecule calculated by the above methods showed a good agreement between the results obtained.


Identification of molecular subtypes based on chromatin regulator and tumor microenvironment infiltration characterization in papillary renal cell carcinoma.
Papillary renal cell carcinoma (pRCC) is the second most common histological type of renal cell carcinoma. The prognosis of local pRCC is better than that of ccRCC, but the situation has changed greatly after pRCC metastasis. Chromatin regulators (CRs) are indispensable in epigenetic regulation, and their abnormal expression in tumors leads to the occurrence and development of tumor. However, the role of CRs in pRCC has not been studied yet. 291 samples were obtained from TCGA-KIPR cohort. Unsupervised clustering analysis was utilized to divide the patients of pRCC into two subtypes. Lasso Cox regression analysis was performed to construct a CRs_score model for predicting OS. The unique characteristics of different molecular subtypes were determined by TME cell infiltration analysis, GO and KEGG analysis and drug sensitivity analysis. We also carried out drug sensitivity experiments in vitro to verify the effect of signature genes on drug sensitivity to sunitinib. We described the transcriptional and genetic alteration of 19 prognosis-related CRs genes in 291 cases of TCGA-KIRP cohort. We identified two distinct molecular subtypes, which have significant differences in prognosis, clinicopathological features and tumor immune microenvironment (TME). Then, four signature genes were selected by lasso regression analysis to construct a CRs_score for predicting OS, and its predictive ability for patients with pRCC was verified. A nomogram was established to improve the clinical applicability of CRs_score. We found that there was a significant difference in the proportion of immune cell infiltration between high- and low-CRs_score. In addition, CRs_score was significantly correlated with chemosensitivity. Finally, we found that SK-RC-39 cell lines were more sensitive to sunitinib after knocking down the signature gene CDCA3, PDIA4, or SUCNR1. Our comprehensive analysis of CRs gene in pRCC showed that CRs gene plays a potential role in TME, prognosis and drug resistance in pRCC. These findings may lay a foundation for further study of the regulatory role of CRs gene in pRCC, and provide a new method for evaluating prognosis and developing more effective targeted therapy.


Deciphering the host-pathogen protein interface in chikungunya virus-mediated sickness.
Successful infection with chikungunya virus (CHIKV) depends largely on the ability of this virus to manipulate cellular processes in its favour through specific interactions with several host factors. The knowledge of virus-host interactions is of particular value for understanding the interface through which therapeutic strategies could be applied. In the current study, the authors have employed a computational method to study the protein interactions between CHIKV and both its human host and its mosquito vector. In this structure-based study, 2028 human and 86 mosquito proteins were predicted to interact with those of CHIKV through 3918 and 112 unique interactions, respectively. This approach could predict 40 % of the experimentally confirmed CHIKV-host interactions along with several novel interactions, suggesting the involvement of CHIKV in intracellular cell signaling, programmed cell death, and transcriptional and translational regulation. The data corresponded to those obtained in earlier studies for HIV and dengue viruses using the same methodology. This study provides a conservative set of potential interactions that can be employed for future experimental studies with a view to understanding CHIKV biology.


Analysis of laser lipolysis effects on human tissue samples obtained from liposuction.
The use of the neodymium: yttrium-aluminum-garnet (Nd:YAG) laser as an auxiliary tool for lipoplasty has refined the traditional technique. The primary limitation of the traditional technique-large volume blood loss-is addressed by the use of the Nd:YAG laser. This report describe's the technique of laser- assisted liposuction, then reviews and compares the histologic and morphometric effects with those of the traditional method. The Nd:YAG laser was used to produce the desired injury to the adipose tissue after sufficient contact. Laser lipolysis was performed on the patient's right flank in two different areas, with the application of 1,000 and 3,000 J of accumulated energy (in), respectively. Subsequently, another cannula was introduced to suction the liquid content obtained by laser lipolysis. This tissue product was sent to the pathology laboratory for morphologic and morphometric analysis, and for comparison with material obtained from traditional lipoplasty performed on the left flank of the same patient. The histopathologic examination of adipose tissue after laser lipolysis showed cell swelling and less bleeding, as compared with the traditional method. The morphometric analysis showed that the mean diameter of the major adipocyte was 95.69 microm with laser lipolysis using 1,000 J (in), 82.63 microm using 3,000 J (in), and 84.54 microm with the traditional method The traditional method produced less reversible cellular damage (swelling) than laser lipolysis using 1,000 J (in). The area receiving 3,000 J (in) showed major irreversible damage (cytoplasmatic retraction and disruption of membranes). For this reason, the mean diameter of the adipocyte was less.


Biologically active epitopes of bovine leukemia virus glycoprotein gp51: their dependence on protein glycosylation and genetic variability.
A panel of monoclonal antibodies to the bovine leukemia virus envelope glycoprotein (BLV gp51) has previously demonstrated the association of the biological activities of the virus (infectivity, syncytia induction) with three out of eight epitopes of gp51. In BLV-infected cells, the unglycosylated homolog of the precursor to the BLV envelope glycoproteins (gPr72env) is a 47,000-MW polypeptide. Immunoprecipitation studies with monoclonal antibodies show that the neutralizing antibody-inducing sites, although present in gPr72env, are not conserved in the 47,000-MW unglycosylated homolog. Finally, it is demonstrated that the neutralizing antibody-inducing sites of gp51 are subject to antigenic variation among BLV isolates of the same or different geographical origins.


The experiences and beliefs of people with severe haemophilia and healthcare professionals on pain management, and their views of using exercise as an aspect of intervention: a qualitative study.
To explore the experiences, views and beliefs of people with severe haemophilia and healthcare professionals (HCPs) on approaches for pain management, as well as their views on exercise being used as an aspect of management. Taking a qualitative inquiry approach using focus groups and semi-structured interviews, participants included people with severe haemophilia living with chronic pain and haemophilia HCPs. Data were analysed using reflexive thematic analysis. Fourteen men with haemophilia with a median age of 47 (range 23-73) and six haemophilia HCPs agreed to participate. Of the people with haemophilia, 11 attended two focus groups and three were interviewed over telephone. Healthcare professionals were interviewed face-to-face. Two themes were conceptualised from the data: (i) haemophilia management and pain management is discordant (imbalance between good haemophilia care but poor pain management, historical medico-social influences on pain management, the need for trust); (ii) uncertain about exercise but clear on what matters (conflicting views on exercise, the need for proof of safety, personalised care). Options for effective pain management remain limited and what is used is heavily influenced by beliefs and experience. Exercise as a treatment option in pain management is conceptually acceptable for people with haemophilia. Effective pain management requires understanding of individual beliefs and fears, and a personalised approach supported by knowledgeable, trusted clinicians.Implications for rehabilitationMusculoskeletal joint pain and its relationship with bleeding in people with haemophilia continues to be a management challenge.Current pain management strategies are of limited effectiveness with little evidence of an approach that reflects the multi-modal pain experience.Whilst exercise and rehabilitation approaches are conceptually possible for people with severe haemophilia, barriers remain regarding perception of overall safety and effectiveness.People with severe haemophilia may consider exercise as part of a pain management strategy if it is individualised, and they are supported to do it by clinicians who understand them and their haemophilia.


CT, MRI, and autopsy findings in brain of a patient with MELAS.
Brain autopsy findings in a 14-year-old patient with mitochondrial myopathy, encephalopathy, lactic acidosis, and strokelike episodes were compared with those of computed tomography (CT) and magnetic resonance imaging (MRI). Pathologic examinations revealed extensive laminar necrosis bordered by gliotic tissues throughout the cerebral cortices. Moderate losses of myelin and fibrous gliosis were also observed in the subcortical and deep white matter. These lesions were demonstrated as low-density areas on CT and as high-signal areas on T2-weighted MRI. MRI revealed the lesions more distinctively and precisely than CT. Neither CT nor MRI could reveal abnormalities in the basal ganglia, including vascular proliferation and calcium deposits in the blood vessels.


Effect of exposure time on the accuracy and reliability of cone beam computed tomography in the assessment of dental implant site dimensions in dry skulls.
To investigate the accuracy and reliability of implant site measurements, recorded from low-dose cone beam computed tomography (CBCT) images. CBCT reformatted images of five skulls were obtained using 40, 20 and 7 s exposure protocols. From these protocols, edentulous ridge dimensions were recorded by two observers and compared with measurements recorded directly from the bone. The measurement errors and intra- and inter-examiner reliability were calculated for each exposure protocol and compared with each other. The mean absolute errors from the 40, 20 and 7 s protocols were 0.50, 0.46, and 0.51 mm, respectively. The intra-examiner reliability scores were 0.996, 0.995 and 0.998, respectively. The inter-examiner reliability scores were 0.993, 0.998 and 0.994, respectively. There was no significant difference in accuracy or reliability between the three protocols. In imaging of dry skulls, lowering the CBCT exposure time from 40 s to 20 or 7 s does not affect the reliability or accuracy of implant site measurements.


Numerical Study of Laminar Flow and Convective Heat Transfer Utilizing Nanofluids in Equilateral Triangular Ducts with Constant Heat Flux.
This study numerically investigates heat transfer augmentation using water-based Al₂O₃ and CuO nanofluids flowing in a triangular cross-sectional duct under constant heat flux in laminar flow conditions. The Al₂O₃/water nanofluids with different volume fractions (0.1%, 0.5%, 1%, 1.5%, and 2%) and CuO/water nanofluids with various volume fractions (0.05%, 0.16%, 0.36%, 0.5%, and 0.8%) are employed, and Reynolds numbers in the range of 700 to 1900 in a laminar flow are considered. The heat transfer rate becomes more remarkable when employing nanofluids. As compared with pure water, at a Peclet number of 7000, a 35% enhancement in the convective heat transfer coefficient, is obtained for an Al₂O₃/water nanofluid with 2% particle volume fraction; at the same Peclet number, a 41% enhancement in the convective heat transfer coefficient is achieved for a CuO/water nanofluid with 0.8% particle volume concentration. Heat transfer enhancement increases with increases in particle volume concentration and Peclet number. Moreover, the numerical results are found to be in good agreement with published experimental data.


Experimental validation of Monte Carlo dosimetry for therapeutic beta emitters with radiochromic film in a 3D-printed phantom.
Validation of dosimetry software, such as Monte Carlo (MC) radiation transport codes used for patient-specific absorbed dose estimation, is critical prior to their use in clinical decision making. However, direct experimental validation in the clinic is generally not performed for low/medium-energy beta emitters used in radiopharmaceutical therapy (RPT) due to the challenges of measuring energy deposited by short-range particles. Our objective was to design a practical phantom geometry for radiochromic film (RF)-based absorbed dose measurements of beta-emitting radionuclides and perform experiments to directly validate our in-house developed Dose Planning Method (DPM) MC code dedicated to internal dosimetry. The experimental setup was designed for measuring absorbed dose from beta emitters that have a range sufficiently penetrating to ∼200 μm in water as well as to capture any photon contributions to absorbed dose. Assayed 177 Lu and 90 Y liquid sources, 13-450 MBq estimated to deliver 0.5-10 Gy to the sensitive layer of the RF, were injected into the cavity of two 3D-printed half-cylinders that had been sealed with 12.7 μm or 25.4 μm thick Kapton Tape. A 3.8 × 6 cm strip of GafChromic EBT3 RF was sandwiched between the two taped half-cylinders. After 2-48 h exposures, films were retrieved and wipe tested for contamination. Absorbed dose to the RF was measured using a commercial triple-channel dosimetry optimization method and a calibration generated via 6 MV photon beam. Profiles were analyzed across the central 1 cm2 area of the RF for validation. Eleven experiments were completed with 177 Lu and nine with 90 Y both in saline and a bone equivalent solution. Depth dose curves were generated for 177 Lu and 90 Y stacking multiple RF strips between a single filled half-cylinder and an acrylic backing. All experiments were modeled in DPM to generate voxelized MC absorbed dose estimates. We extended our study to benchmark general purpose MC codes MCNP6 and EGSnrc against the experimental results as well. A total of 20 experiments showed that both the 3D-printed phantoms and the final absorbed dose values were reproducible. The agreement between the absorbed dose estimates from the RF measurements and DPM was on average -4.0% (range -10.9% to 3.2%) for all single film 177 Lu experiments and was on average -1.0% (range -2.7% to 0.7%) for all single film 90 Y experiments. Absorbed depth dose estimates by DPM agreed with RF on average 1.2% (range -8.0% to 15.2%) across all depths for 177 Lu and on average 4.0% (range -5.0% to 9.3%) across all depths for 90 Y. DPM absorbed dose estimates agreed with estimates from EGSnrc and MCNP across the board, within 4.7% and within 3.4% for 177 Lu and 90 Y respectively, for all geometries and across all depths. MC showed that absorbed dose to RF from betas was greater than 92% of the total (betas + other radiations) for 177 Lu, indicating measurement of dominant beta contribution with our design. The reproducible results with a RF insert in a simple phantom designed for liquid sources demonstrate that this is a reliable setup for experimentally validating dosimetry algorithms used in therapies with beta-emitting unsealed sources. Absorbed doses estimated with the DPM MC code showed close agreement with RF measurement and with results from two general purpose MC codes, thereby validating the use of this algorithms for clinical RPT dosimetry.


Discovery of Two Novel Oxidases Using a High-Throughput Activity Screen.
Discovery of novel enzymes is a challenging task, yet a crucial one, due to their increasing relevance as chemical catalysts and biotechnological tools. In our work we present a high-throughput screening approach to discovering novel activities. A screen of 96 putative oxidases with 23 substrates led to the discovery of two new enzymes. The first enzyme, N-acetyl-D-hexosamine oxidase (EC 220.127.116.11) from Ralstonia solanacearum, is a vanillyl alcohol oxidase-like flavoprotein displaying the highest activity with N-acetylglucosamine and N-acetylgalactosamine. Before our discovery of the enzyme, its activity was an orphan one - experimentally characterized but lacking the link to amino acid sequence. The second enzyme, from an uncultured marine euryarchaeota, is a long-chain alcohol oxidase (LCAO, EC 184.108.40.206) active with a range of fatty alcohols, with 1-dodecanol being the preferred substrate. The enzyme displays no sequence similarity to previously characterised LCAOs, and thus is a completely novel representative of a protein with such activity.


[Association of sarcoidosis and primary biliary cirrhosis: review of the literature apropos of a new case].
We report a case of sarcoidosis associated with primary biliary cirrhosis in a 56-year old woman, with a 5 years' follow up. Sarcoidosis was ascertained by chest X ray, gallium scintigraphy, broncho-alveolar washing and liver biopsy (intralobular epithelioid granulomas). The diagnosis of primary biliary cirrhosis was based on IgM level increase (4 g/l), AMA positivity (1/1000) and liver biopsy assessment (non suppurative chronic cholangitis). Such an association is rare (12 detailed cases in literature), brings out pathophysiological questions and points to an additional mechanism of cholestasis in patients with sarcoidosis.


Foot Osteoarthritis Frequency and Associated Factors in a Community-Based Cross-Sectional Study of White and African American Adults.
Few studies have explored foot osteoarthritis (OA) in the general population. The purpose of this study was to determine the frequency of foot OA and identify associated factors in a cross-sectional analysis of a large community-based cohort. Data were from the 2013-2015 study visit of the Johnston County OA Project. Radiographic OA of the foot was defined using the La Trobe radiographic atlas (≥2 osteophytes or joint space narrowing in at least 1 of 5 joints). Symptomatic OA of the foot was defined as foot radiographic OA with pain, aching, or stiffness in the same foot. At the foot-level, separate logistic regression models with generalized estimating equations to account for intraperson correlations were performed to examine associations of foot radiographic OA or symptomatic OA with age, body mass index (BMI), sex, race, educational attainment, and previous foot injury. Of 864 participants with available data (mean age 71 years, mean BMI 30 kg/m2 , 68% women, 33% African American, 13% <12 years of schooling), 22% had foot radiographic OA, 20% had foot symptoms, and 5% had foot symptomatic OA. Radiographic, but not symptomatic, foot OA was more common in African American than White participants. Participants with obesity, compared to normal weight, had >2 times the odds of radiographic OA and >5 times the odds of symptomatic OA in adjusted models. Foot radiographic OA and foot symptoms were common in the sample, but both conditions simultaneously (i.e., symptomatic OA) occurred infrequently. Notably, obesity was linked with foot symptomatic OA, perhaps implicating metabolic or mechanical influences.


The Association of the Neutrophil-to-Lymphocyte Ratio, Platelet-to-Lymphocyte Ratio, Lymphocyte-to-Monocyte Ratio and Systemic Inflammation Response Index with Short-Term Functional Outcome in Patients with Acute Ischemic Stroke.
The aim of this study was to explore the relationship between functional prognosis and the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR) and systemic inflammatory response index (SIRI) in patients with acute ischemic stroke (AIS) at discharge. A total of 861 patients with AIS were enrolled between January 2019 and December 2021. Blood cell counts were collected on admission. Logistic regression analysis was performed to assess the relationship between NLR, PLR, LMR, SIRI and adverse functional outcomes (modified Rankin scale score of 3-6) at discharge. We also used receiver operating characteristic (ROC) curves to estimate the overall ability of NLR, PLR, LMR and SIRI to judge short-term functional outcomes. Associations between NLR, PLR, LMR, and SIRI with length of hospital stay were analyzed by Spearman correlation test. A total of 194 patients (22.5%) had poor functional outcomes at discharge. Multivariate logistic regression analysis showed that NLR (odds ratio [OR], 1.060; 95% confidence interval [CI] 1.004-1.120, P=0.037), PLR (OR, 1.003; 95% CI 1.000-1.005, P=0.018), LMR (OR, 0.872; 95% CI 0.774-0.981, P=0.023) and SIRI (OR, 1.099; 95% CI 1.020-1.184, P=0.013) were independent factors for poor functional outcome. The odds ratios of the highest versus lowest quartiles of NLR, PLR and SIRI were 2.495 (95% CI 1.394-4.466), 1.959 (95% CI 1.138-3.373) and 1.866 (95% CI 1.106-3.146), respectively. The odds ratio of the lowest versus highest quartile of LMR was 2.300 (95% CI 1.331-3.975). The areas under the curve (AUCs) of the NLR, PLR, LMR, and SIRI to discriminate poor functional prognosis were 0.644, 0.587, 0.628, and 0.651, respectively. NLR, LMR, and SIRI were related with the length of hospital stay (P<0.05). NLR, PLR, LMR, and SIRI were associated with functional outcome at discharge in AIS patients. NLR, LMR and SIRI were related to hospitalization days in patients with AIS.


Chitosan fibers with improved biological and mechanical properties for tissue engineering applications.
The low mechanical properties of hydrogel materials such as chitosan hinder their broad utility for tissue engineering applications. Previous research efforts improved the mechanical properties of chitosan fiber through chemical and physical modifications; however, unfavorable toxicity effects on cells were reported. In this paper, we report the preparation of chitosan fibers with improved mechanical and biocompatibility properties. The structure-property relationships of extruded chitosan fibers were explored by varying acetic acid (AA) concentration, ammonia concentration, annealing temperature and degree of heparin crosslinking. Results showed that optimizing AA concentration to 2vol% improved fiber strength and stiffness by 2-fold. Extruding chitosan solution into 25wt% of ammonia solution reduced fiber diameters and improved fiber strength by 2-fold and stiffness by 3-fold, due to an increase in crystallinity as confirmed by XRD. Fiber annealing further reduced fiber diameter and improved fiber strength and stiffness as temperature increased. Chitosan fibers crosslinked with heparin had increased diameter but lower strength and stiffness properties and higher breaking strain values. When individual parameters were combined, further improvement in fiber mechanical properties was achieved. All mechanically improved fibers and heparin crosslinked fibers promoted valvular interstitial cells (VIC) attachment and growth over 10 day cultures. Our results demonstrate the ability to substantially improve the mechanical properties of chitosan fibers without adversely affecting their biological properties. The investigated treatments offer numerous advantages over previous physical/chemical modifications and thus are expected to expand the utility of chitosan fibers with tunable mechanical properties in various tissue engineering applications.


Renal handling of uric acid in normal children by means of the pyrazinamide and sulfinpyrazone tests.
Thirteen normal children have been studied with the Sulfinpyrazone and Pyrazinamide tests, in order to evaluate the mechanisms of urate tubular management. A decrease in presecretory reabsorption, and an increase in tubular secretion have been identified, with regard to what has already been reported in adults. These facts could explain the higher fractional excretion of uric acid found in childhood.


ALS patients request more information about cognitive symptoms.
Once thought to impact only voluntary motor function, ALS/Motor neuron disease (MND) is now seen as a multi-system disorder in which a minority of patients experience mild cognitive dysfunction or frontotemporal dementia. Despite clinical guidelines advocating supplying complete information to patients, educational materials on ALS often state that the mind is unaffected. We sought to establish how much patients and caregivers understand about ALS, what they have been told to expect by their physician, and if they would have appreciated more complete information. A two-part survey was administered online. An 'ALS quiz' gauged participants' knowledge of physical and psychological aspects of ALS. A second questionnaire assessed which symptoms patients had discussed with their clinician and explored patients' desire to receive information on psychological effects. A total of 247 ALS patients and 87 caregivers participated. Participants knew less about psychological symptoms than physical ones (72% correct responses versus 82%; paired t((333)) = -5.04, P < 0.001). Patients commonly reported being told by their doctor about physical symptoms such as problems walking (85%) or stiffness/cramps (74%) but not psychological issues like emotional lability (46%) or cognitive change (11%). The majority of patients (62%) and carers (71%) indicated a desire to be informed that cognitive change or dementia might occur. ALS is a multi-system disorder. However, despite a desire for more information from patients and their carers, healthcare professionals continue to primarily address only the physical consequences of the disease.


Validity of accelerometry for predicting physical activity and sedentary time in ambulatory children and young adults with cerebral palsy.
/Objectives: This study aimed to validate five published ActiGraph (AG) cut-off points for the measurements of physical activity (PA) and sedentary time (ST) in ambulatory children and young adults with cerebral palsy (CP). Additionally, four energy expenditure (EE) prediction equations based on AG counts and activPAL (AP) steps were examined in this population, using oxygen uptake (VO2) as the criterion. Four male and six female participants with CP (Gross Motor Function Classification System levels I-III, ages 9-21 years) completed seven activities while simultaneously wearing an AG, AP monitor and indirect calorimetry unit. VO2 was measured on a breath-by-breath basis using the indirect calorimetry and was converted into EE using metabolic equivalents. AG counts were classified as sedentary, light PA (LPA) or moderate-to-vigorous PA (MVPA) using five cut-off points: Puyau, Evenson, Romanzini, Clanchy and Baque. The predicted EE was computed using three AG-based equations (Freedson, Trost and Treuth) and an AP step-based equation. Based on 1920 available data points from the 10 participants, Baque (r = 0.896, κ = 0.773) and Clanchy (r = 0.935, κ = 0.721) AG cut-off points classified PA and ST most accurately. All the equations overestimated EE during sitting activities and underestimated EE during rapid walking. The Freedson, Treuth and AP equations exhibited systematic bias during rapid walking, as their differences from the criterion measure increased progressively with increasing activity intensity. The AG accurately classified PA and ST when the Baque and Clanchy cut-off points were used. However, none of the available AG or AP equations accurately predicted the EE during PA and ST in children and young adults with CP. Further development is needed to ensure that both devices can estimate EE accurately in this population.


[Aplasia cutis congenita of the scalp (5 observations)].
Aplasia cutis congenita (ACC) is an uncommon congenital malformation. It is characterized by defects of the skin that occur most frequently on the scalp along the midline, but can also be localized on the trunk, face and limbs, usually with a symmetrical distribution. When it is localized in the skull, it can extend to the dura mater, with only the thin pia mater to protect the brain. We report a retrospective study during a period of 10 years and we report 5 cases of ACCV hospitalized in the pediatric service in CHU Hédi Chaker and in maxillo-facial surgery service in CHU Habib Bourguiba, Sfax. We studied the epidemiologic, clinical, and therapeutic aspects in our patients. The average age at the admission was 5 days (2-8 days). A consanguinity was found in 2 cases. The clinical examination revealed cutaneous and osseous structures aplasia located in frontoparietal zone in 3 patients and in parieto-occipital zone in a patient. A hypoplasia of the toes was noted in 3 cases and a hypoplasia of the 3rd finger of the 2 hands in a case. Plain X-ray skull (3 cases) showed the osseous defect in all the cases. The cerebral IRM (2 cases) showed osseous and cutaneous defect in two cases and a lipome of the corpus callosum in one patient. A surgical repair using a cutaneous graft was performed for 3 patients. A patient died on the 16th day of life from a haemorrhage of the longitudinal sinus. The evolution was favourable in 4 cases with a cicatrisation of good quality but with subsequent alopecia. ACC of the scalp is a rare and often sporadic affection. Our experience confirms that fatal bleeding from the longitudinal sinus can occur during the 1st weeks of life.


Effects of Petroselinum crispum extract on pancreatic B cells and blood glucose of streptozotocin-induced diabetic rats.
This study investigated both morphologically and biochemically whether parsley (Petroselinum crispum), which is used as a folk remedy to decrease blood glucose, has any antidiabetic effect on pancreatic B cells of rats. Parsley extract was given to male diabetic rats. In the diabetic group given parsley extract, it was detected that the number of secretory granules and cells in islets and other morphologic changes were not different from the control diabetic group, while the blood glucose levels in the diabetic group given the plant extract were reduced in comparison to the diabetic group. In addition, a decrease was observed in the weight of the control diabetic group and the diabetic group given the plant extract. It is suggested that the plant therapy can provide blood glucose homeostasis and cannot regenerate B cells of the endocrine pancreas.


Biochemical properties of beta-glucosidase in leukocytes from patients and obligated heterozygotes for Gaucher disease carriers.
Gaucher's disease (GD) is a disorder caused by the deficiency of lysosomal beta-glucosidase, an enzyme that participates in the degradation of glycosphingolipids. Deficiency of this enzyme results in the storage of glucocerebrosides in lysosomes of macrophage. No studies are available in the literature comparing biochemical and kinetic behavior of this enzyme in leukocytes and fibroblasts from normal individuals, obligate heterozygotes and patients with GD. The behavior of beta-glu in terms of optimum pH, heat stability, Km and Vmax in leukocytes from patients with GD and obligated heterozygotes with different genotypes and normal individuals were characterized. Optimum pH was similar in all groups analyzed. In terms of Km and Vmax, several differences among heterozygotes and homozygotes groups and among these groups and normal enzyme were observed. Enzyme from all groups were inactivated when preincubated at 60 degrees C, but some enzymes were more stable than other. Results showed a different behavior of the enzyme in the 3 groups under analysis. Such behavior varied according to individual mutation. The catalytic gradient presented by beta-glu allowed the correlation of N370S mutation-which presented more stable biochemical properties-with the non-neurological clinical condition of the disease and the catalytically less stable mutation (D409H), with the neurological clinical condition of GD. This study contributes to a better understanding of the repercussion of the different mutations on the protein function, thus allowing to predict the severity of such complex metabolic disorder and to anticipate the most appropriate intervention for each case specifically.


The effect on the tensile bond strength of orthodontic brackets of titanium tetrafluoride (TiF4) application after acid etching.
Titanium tetrafluoride (TiF4) was applied to acid-etched enamel surfaces to study the effect on surface structure and tensile bond strength of orthodontic brackets. Three groups of 20 premolars each were compared in debonding tests. Group I received a 1% and Group II received a 4% topical TiF4 treatment after acid etching. Group III served as control, that is, no fluoride solution was applied to the enamel after acid etching. Each treatment lasted 60 seconds followed by a thorough rinsing with water for 30 seconds. Brackets were bonded on all teeth with the same procedure. All premolars were then placed in a water bath at 37 degrees C. Ten teeth from each group were debonded after 24 hours, and the remaining teeth were debonded after 6 months. Results indicated that the application of 1% and 4% TiF4 solutions on etched enamel surfaces did not alter the tensile bond strength significantly after 24 hours and 6 months. The scanning electron microscope (SEM) examination of the surface structure demonstrated the presence of extensive reaction products, which covered enamel surfaces and appeared to interfere with etch patterns. It was concluded that the application of TiF4 solutions to etched enamel surfaces before bonding will not have an adverse effect on the tensile strength of orthodontic attachments.


Computer monitoring of auditory brainstem responses.
A relatively low cost, bedside, portable, microcomputer based system for long-term monitoring of human auditory evoked responses in critical care environments is described, and test data are presented. The system enables the user to acquire responses, detect significant peaks (usually peaks I, III, and V), store information and display trends (latency vs time graphs) automatically. Prior to processing, responses are digitally filtered so that no phase distortion is introduced. A peak detection algorithm takes into consideration both the different morphologies of the IV-V wave complex and the time-varying characteristics of the ABR waveform. Evaluations using normal subjects and intensive care unit patients show the clinical potential of automated long-term auditory brainstem monitoring.


Resolution of chronic inflammatory demyelinating polyneuropathy-associated central nervous system lesions after treatment with intravenous immunoglobulin.
Chronic inflammatory demyelinating polyneuropathy (CIDP) is a condition affecting the peripheral nervous system; however, it has been associated with central nervous system (CNS) involvement. The natural history and response to treatment of these CNS lesions are unknown. We report the case of a patient with CIDP which met research criteria for definite CIDP and associated symptomatic CNS lesions. She had resolution of her CNS-based clinical and radiographic findings with intravenous immunoglobulin (IVIG) therapy. IVIG is a reasonable treatment option when symptomatic, CIDP-associated CNS lesions are present.


Synthesis and anti-acetylcholinesterase activity of scopoletin derivatives.
A series of scopoletin derivatives incorporated with the pyridinium moiety was synthesized and evaluated for their acetylcholinesterase (AChE) inhibitory activity by the colorimetric Ellman's method. A 2-fluorobenzylpyridinium derivative was the most potent among the tested compounds, with an IC50 value of 0.215±0.015μM, which was greatly improved from that of scopoletin. Docking studies revealed that the scopoletin portion of the mentioned compound was bound to the peripheral anionic site of the AChE, whereas the N-benzylpyridinium residue to the catalytic anionic site.


Auto-MHC class II-reactive T cell line obtained from MRL/+mice suffering from "lpr-GVHD". I. Characterization of surface phenotypes, specificities and functions in vitro.
When MRL/Mp-(+)/+ (MRL/+) mice are lethally irradiated and then reconstituted with bone marrow or spleen cells from MRL/Mp-lpr/lpr (MRL/lpr) mice, they develop a graft-versus-host disease (GVHD)-like syndrome, colloquially known as "lpr-GVHD". To analyze the roles of the MRL/lpr T cells in the development of "lpr-GVHD" and autoimmune diseases, several T cell lines were established from the spleen cells of MRL/+ mice suffering from "lpr-GVHD". The surface phenotypes, specificities, and functions of a representative clone (l/+T1) of the cloned T cell lines were characterized. The l/+T1 cells showed Thy-1.2+, L3T4+ and T3+, but Lyt-2- and B220- phenotypes. Proliferative response was observed by co-culturing the cells with spleen cells from MRL/+, MRL/lpr, AKR/J, and C3H/HeN mice, but not from BALB/c or C57BL/6 mice. Furthermore, the l/+ T1 cells responded to spleen cells of B10.BR and B10.A but not B10.D2 mice. The proliferative response of l/+ T1 cells to MRL/+ spleen cells was inhibited by anti-I-Ek (but not anti-I-Ak or anti-Kk) antibodies, suggesting that the specificity of l/+T1 cell culture enhanced the proliferative response only in the presence of appropriate stimulators. Treatment of stimulator cells with J11d.2 + C (but not anti-Thy-1.2 + C or 33D1 + C) abolished the stimulatory effect, indicating that B cells are effective stimulator cells for auto-MHC class II-reactive l/+T1 cells. When MRL/+ splenic B cells were co-cultured with l/+T1 cells, both B cell proliferation and IgM production were observed. In addition, IgM-class rheumatoid factor and anti-ssDNA antibody activities were found in the supernatants of MRL/+ splenic B cells co-cultured with l/+T1 cells. These results are discussed in relation to "lpr-GVHD" and autoimmunity in MRL/lpr mice.


[The effect of high concentration of sodium and potassium ion on motility of human sperm preserved in electrolyte-free solution].
To assess the effect of high concentration of sodium and potassium ion on the motilities of human sperm following the preservation in electrolyte-free solution. Semen samples were obtained from patients attending our infertility clinic. The motilities of their sperm were more than 40%. Ejaculated sperm were washed by using the electrolyte-free Percoll gradient and then preserved in electrolyte-free solution at 4 degrees C for a week. The preserved sperm was incubated and analyzed after the addition of electrolyte solutions under acid or alkaline condition. The sperm were continuously preserved for a day at room temperature and analyzed. The sperm was by the addition of 35-135 mM Na+ or 5-135 mM K+ in acid solutions. Under the alkaline conditions, sperm were reactivated without Na+ or K+. However, the addition of 135 mM Na+ or K+ significantly decreased in sperm motilities. The some sperm in the solution containing low concentration of Na+ or K+ ions maintained their motilities for a day at room temperature. However, all sperm lost their motilities in the 135 mM Na+ or K+ solutions. The high concentration of Na+ or K+ ions had detrimental effects on motilities of sperm following the preservation in electrolyte-free solution.


Characteristics of health-state utilities used in cost-effectiveness analyses: a systematic review of published studies in Asia.
Cost-utility analysis (CUA) is the preferred form of economic evaluation in many countries. As one of the key data inputs in cost-utility models, health state utility (HSU) has a crucial impact on CUA results. In the past decades, health technology assessment has been expanding rapidly in Asia, yet research examining the methodology and process used to generate cost-effectiveness evidence is scarce. The aim of this study was to examine the reporting of the characteristics of HSU data used in CUAs in Asia and how the characteristics have changed over time. A systematic literature search was performed to identify published CUA studies targeting Asian populations. Information was extracted for both the general characteristics of selected studies and the characteristics of reported HSU data. For each HSU value identified, we extracted data for four key characteristics, including 1) estimation method; 2) source of health-related quality of life (HRQoL) data; 3) source of preference data; and 4) sample size. The percentage of nonreporting was calculated and compared over two time periods (1990-2010 vs 2011-2020). A total of 789 studies were included and 4,052 HSUs were identified. Of these HSUs, 3,351 (82.7%) were from published literature and 656 (16.2%) were from unpublished empirical data. Overall, the characteristics of HSU data were not reported in more than 80% of the studies. Of HSUs whose characteristics were reported, most of them were estimated using the EQ-5D (55.7%), Asian HRQoL data (91.9%), and Asian health preferences (87.7%); 45.7% of the HSUs was estimated with a sample of 100 or more individuals. All four characteristics showed improvements after 2010. Over the past two decades, there has been a significant increase in CUA studies targeting Asian populations. However, HSU's characteristics were not reported in most of the CUA studies, making it difficult to evaluate the quality and appropriateness of the HSUs used in those cost-effectiveness studies.


Hyperplastic callus formation in osteogenesis imperfecta.
A case of a child with the hyperplastic callus formation in association with osteogenesis imperfecta is presented. The soft tissue mass arising at the site of femoral fracture was clinically and radiologically suspected to be a hyperplastic callus, although a possibility of osteosarcoma could not be completely excluded. A drill biopsy of the soft tissue mass was performed which showed osteocartilaginous callus. The soft tissue mass disappeared following biopsy. This occurrence is also reported in previously described cases.


Nuclear magnetic resonance: an overview of its spectroscopic and imaging applications in pediatric patients.
Magnetic resonance (MR) is a technique that permits the noninvasive evaluation of morphologic features and function based on the distribution of protons and other selected elements. We present a basic description of MR and illustrate both 31P-MR spectroscopy and proton imaging applications in pediatric patients. The applications of these techniques are diverse, and are presented concisely in an attempt to give pediatricians an overview of this new technology and its potential role in patient management.


Development of active jejunal glucose absorption in broiler chickens.
Growth in chickens, especially meat-type chickens (broilers), is extremely rapid, but studies on the regulatory mechanism of intestinal glucose absorption with growth are few, contradictory, and unclear. Here, we investigated the regulation of intestinal glucose absorption with growth in broiler chickens using oral glucose gavage, intestinal Evans blue transit, intestinal glucose absorption, scanning electron microscopy, and glucose absorption- and cell junction-related gene expression analyses. Peak blood glucose levels after oral glucose gavage occurred at 10 and 50 min in chickens at 1 wk (C1W) and 5 wk (C5W) of age, respectively. The area under the curve for glucose levels was greater for the C5W than the C1W (P = 0.035). The stain ratio in the small intestine in the C5W was lower than that in the C1W (P = 0.01), but there were no differences in the tissue regions stained with Evans blue and the migration distance of Evans blue from Meckel's diverticulum. In everted sac and Ussing chamber experiments, we observed reduced intestinal glucose uptake and electrogenic glucose absorption in the jejunum of the C5W. Phloridzin, an inhibitor of sodium/glucose cotransporter 1 (SGLT1), suppressed the glucose-induced short-circuit current in the C1W (P = 0.016) but not the C5W. Although the addition of NaCl solution stimulated the glucose-induced short-circuit current in the C1W, no differences between the treatments were observed (P = 0.056), which was also the case in the C5W. Additionally, tissue conductance was diminished in the C5W compared with that in the C1W. Moreover, in the C5W, the intestinal tract was more developed and the jejunal villi were enlarged. In conclusion, glucose absorption throughout the intestine could be greater in C5W than in C1W; however, reduced SGLT1 sensitivity, decreased ion permeability, and intestinal overdevelopment lead to decreased local glucose absorption in the jejunum with growth in broiler chickens. These data provide a detailed analysis of intestinal glucose absorption in growing broiler chickens, and can contribute to the development of novel feeds.


Peptides based on CcdB protein as novel inhibitors of bacterial topoisomerases.
The ccd toxin-antitoxin system of the F plasmid encodes CcdB, a protein that poisons the essential Escherichia coli DNA gyrase, unique type IIA topoisomerase able to introduce negative supercoils into DNA. Based on CcdB structure, a series of linear peptide analogues were obtained by the solid-phase methodology. One of these peptides (CcdBET2) displayed inhibition of the supercoiling activity of bacterial DNA gyrase with a concentration required for complete inhibition (IC(100)=10 microM) lower than the wild type CcdB. For Topo IV, a second type IIA bacterial topoisomerase, CcdBET2 was better inhibited the relaxation activity with an IC(100) of 5 microM (wt CcdB>10 microM). The replacement of Gly, present in the three C-terminal amino acid residues, by Glu, abolished the capacity to inhibit the gyrase but not the Topo IV activities. These findings demonstrate that the mechanism by which CcdBET2 inhibits DNA gyrase is different of the mechanism by which inhibits Topo IV. Therefore, CcdBET2 is a new type II topoisomerase inhibitor with specificity for Topo IV.


Pulmonary plasma cell granuloma.
Two cases of pulmonary plasma cell granuloma in children are reported. The main features of this rare entity are described.


Patellar tendinopathy: an overview of prevalence, risk factors, screening, diagnosis, treatment and prevention.
Patellar tendinopathy (PT), or jumper's knee, is an overuse injury that occurs in professional, as well as recreational, athletes. This condition is a noncontact injury, typically characterized by gradually increasing pain in the patellar tendon. It is prevalent in participants of several sports, but it occurs mostly in jumping sports. The diagnosis of PT is primarily clinical; however, imaging techniques can be useful as well. Risk factors differ between sexes, playing conditions, the kind of sport, playing level and personal characteristics. Screening is an essential tool to assess PT. This condition affects athletic performance and often persists for years. The use of preventative methods is imperative because of the persistence of this condition, especially in elite athletes who sometimes end their career after long and failed treatments. There are a wide variety of treatment and rehabilitation options available, the majority of which are non-operative, such as eccentric exercises, cryotherapy, platelet-rich plasma (PRP) injections, and anti-inflammatory strategies. If conservative treatment fails, surgery is the next most preferable step. Even though there are many surgical treatment methods, there is no clear evidence on what is the most effective approach to address PT. Taking this into consideration, as well as the extent of this clinical entity, novel therapeutic techniques, as well as screening and prevention methods, are expected to emerge in the near future.


Cromolyn does not modulate human allergic skin reactions in vivo.
Cromolyn pretreatment frequently reduces antigen inhalation-induced bronchospasm possibly by inhibiting mast cell degranulation and mediator release. However, the local effects of cromolyn on type I hypersensitivity skin reactions are not well understood. We studied the effect of local cromolyn on antigen-induced skin reaction, histamine release, cellular inflammatory response, and ultramicroscopic changes of mast cells in 10 ragweed-sensitized subjects. Results showed that cromolyn 2% (nonirritant dose) does not modulate ragweed-induced skin whealing response, histamine release, and ultramicroscopic changes of mast cells. Thus, unlike the situation in the tracheobronchial tree, allergic skin reactions and associated events are not inhibited by local cromolyn application.


Variation in the lymphocyte transformation assay. Slope analysis of cell dose-response curves.
Blood samples from a single sheep were cultured at a number of different cell concentrations and stimulated with three lymphocyte mitogens, phytohaemagglutinin, concanavalin A and pokeweed mitogen. The response to stimulation, measured as log thymidine uptake (log cpm) was found to be a linear function of log cell concentration (log cell dose). Samples taken on different dates, varied significantly in the gradients of the log cpm: log cell dose plots. These alterations of gradient constituted the major source of variation of cpm in cultures at optimum cell concentration. It is concluded that the optimal mitotic response of lymphocyte cultures is determined more by the relative proportions of interactive cell types in the response than by the frequency of responder units. Also this proportion is subject to wide, spontaneous variation in the normal animal.


DNA complex-releasing system by injectable self-setting apatite cement.
The therapeutic effect of plasmid DNA complexes is not satisfactory because of the short duration of their gene expression. However, an efficient DNA slow-release system has not been established owing to the low dispersion stability of the DNA/polycation (or cationic lipid) complexes. We have found that hyaluronan (HA) could deposit onto the DNA/polycation complexes, stabilize their dispersion and prepare very small particles. We have also reported that the injectable self-setting apatite cement has high biocompatibility and biodegradability. Thus, durable gene expression systems using injectable apatite cement, including DNA/polycation/HA complexes, were expected. Small DNA/polyethyleneimine (PEI)/HA complex particles were prepared by a lyophilizing and rehydration process, and the in vitro release ratio of DNA complex from the apatite cements by MLC-6 cells was examined. Apatite cement slurry with collagen including plasmid DNA (pDNA) complex [encoding granulocyte macrophage-colony stimulating factor (GM-CSF)] was injected close to the tumor subcutaneously inoculated into mice. The therapeutic effect and degradation ratio of the apatite cement were evaluated. Very fine DNA/PEI/HA complex particles kept being dispersed in the apatite cement. The DNA complexes were continuously released from the apatite cement by MLC-6 cells. Single injection of the apatite cement-including pDNA-GM-CSF complex induced complete disappearance of tumor in 60% of mice. Smooth degradation of the apatite cement was observed in the mice in which a high therapeutic effect was seen. Single injection of the apatite cement-including pDNA-GM-CSF complex showed a high therapeutic effect on solid tumor, and thus appears to be promising as a sustained gene expression device.


[A crustacean-specific neurotoxin from the venom of the black widow spider Latrodectus mactans tredecimguttatus].
A method of the isolation of a crustacea-specific neurotoxin from the venom of the Latrodectus mactans tredecimguttatus spider by means of ion exchange chromatography on Mono Q and Mono S columns and hydrophobic chromatography on Phenyl-Superose column has been developed. LD50 of the toxin has been elucidated.


Antifungal activity of the plant Dodonaea viscosa var. angustifolia on Candida albicans from HIV-infected patients.
was to determine the minimum inhibitory concentration (MIC) and the time taken by Dodonaea viscosa var. angustifolia (PLE) a South African medicinal plant, chlorhexidine gluconate (CHX) and triclosan (TRN) to kill Candida albicans. 41 strains of Candida albicans were investigated, 20 from HIV-positive patients, 20 from HIV-negative subjects and Candida albicans ATCC 90028. The MICs of an acetone extract of PLE, CHX and TRN were measured using a microtitre double dilution technique, and the time taken to kill 99.5% of the strains was determined. The MICs of PLE, CHX and TRN were 6.25-25, 0.008-0.16 and 0.0022-0.009 mg/ml, respectively. PLE killed all the test strains within 30s and CHX 40% of the isolates from HIV-positive patients and 20% of strains from HIV-negative subjects in 1 min. During the same time TRN killed 55% and 35% of isolates from HIV-positive and HIV-negative patients. Dodonaea viscosa var. angustifolia has antifungal properties and is more effective than commercially available mouthrinses.


cGAS-STING signaling in ischemic diseases.
Double stranded DNA (dsDNA) is known to act as a damage-associated molecular pattern (DAMP) that stimulates the body's innate immune response. In general, cyclicGMP-AMP(cGAMP)synthase(cGAS), a DNA sensor, detects these disease-causing DNA and activates the stimulator of interferon gene (STING), which in turn phosphorylates interferon regulatory factor 3 (IRF3), triggering the synthesis of type I interferon (IFN). During this process, the cGAS-STING pathway interacts with different modes of cell death, including autophagy, apoptosis, pyroptosis, and necroptosis. Importantly, cGAS might get stimulated by self-DNA, such as nuclear DNA (nuDNA) and mitochondrial DNA (mtDNA), which ensures a close association between the cGAS-STING signaling pathway and autoimmune responses. Following an ischemic attack, damaged or necrotic cells release large amounts of self-DNA that subsequently activate cGAS, resulting in a range of consequences related to an injury. The present study presents an overview of studies focused on cGAS-STING signaling and cell death, and summarizes the findings of this pathway with regard to ischemia or ischemia/reperfusion (I/R) in different organs of the body, including heart, brain, liver, kidney, and intestine.


Alpha2-antiplasmin is involved in the production of transforming growth factor beta1 and fibrosis.
Fibrotic disease occurs in most tissues. Transforming growth factor (TGF)-beta is the major inducer of fibrosis. The fibrinolytic system is considered to play an important role in the degradation of extracellular matrices. However, the detailed mechanism of how this system affects fibrosis remains unclear. We examined experimental fibrosis in mice with a deficiency of alpha(2)-antiplasmin (alpha2AP), which is a potent and specific plasmin inhibitor. We found that the lack of alpha2AP attenuated bleomycin-induced TGF-beta(1) synthesis and fibrosis. In addition, the production of TGF-beta(1) from the explanted fibroblasts of alpha2AP(-/-) mice decreased dramatically as compared to that in wild-type mice. Moreover, we found that alpha2AP specifically induces the production of TGF-beta(1) in fibroblasts. The lack of alpha2AP attenuated TGF-beta(1) synthesis, thereby resulting in attenuated fibrosis. This is the first report to describe the crucial role that alpha2AP plays in TGF-beta(1) synthesis during the process of fibrosis. Our results provide new insights into the role of alpha2AP in fibrosis.