Atrial natriuretic peptide down-regulates LPS/ATP-mediated IL-1β release by inhibiting NF-kB, NLRP3 inflammasome and caspase-1 activation in THP-1 cells.
Atrial natriuretic peptide (ANP) is an hormone/paracrine/autocrine factor regulating cardiovascular homeostasis by guanylyl cyclase natriuretic peptide receptor (NPR-1). ANP plays an important role also in regulating inflammatory and immune systems by altering macrophages functions and cytokines secretion. Interleukin-1β (IL-1β) is a potent pro-inflammatory cytokine involved in a wide range of biological responses, including the immunological one. Unlike other cytokines, IL-1β production is rigorously controlled. Primarily, NF-kB activation is required to produce pro-IL-1β; subsequently, NALP3 inflammasome/caspase-1 activation is required to cleave pro-IL-1β into the active secreted protein. NALP3 is a molecular platform capable of sensing a large variety of signals and a major player in innate immune defense. Due to their pleiotropism, IL-1β and NALP3 dysregulation is a common feature of a wide range of diseases. Therefore, identifying molecules regulating IL-1β/NALP3/caspase-1 expression is an important step in the development of new potential therapeutic agents. The aim of our study was to evaluate the effect of ANP on IL-1β/NALP3/caspase-1 expression in LPS/ATP-stimulated human THP1 monocytes. We provided new evidence of the direct involvement of ANP/NPR-1/cGMP axis on NF-kB/NALP3/caspase-1-mediated IL-1β release and NF-kB-mediated pro-IL-1β production. In particular, ANP inhibited both NF-kB and NALP3/caspase-1 activation leading to pro- and mature IL-1β down-regulation. Our data, pointing out a modulatory role of this endogenous peptide on IL-1β release and on NF-kB/NALP3/caspase-1 activation, indicate an important anti-inflammatory and immunomodulatory effect of ANP via these mechanisms. We suggest a possible employment of ANP for the treatment of inflammatory/immune-related diseases and IL-1β/NALP3-associated disorders, affecting millions of people worldwide.


The acoustic properties, centered on 20 MHZ, of an IEC agar-based tissue-mimicking material and its temperature, frequency and age dependence.
The purpose of this study was to characterize the ultrasonic properties of agar-based tissue-mimicking materials (TMMs) at ultrasound frequencies centered around 20 MHz. The TMM acoustic properties measured are the amplitude attenuation coefficient alpha (dB cm(-1)MHz(-1)), the speed of sound (ms(-1)) and the backscattered power spectral density (distribution of power per unit frequency normalized to the total received power) characteristics of spectral slope (dB MHz(-1)), y-axis intercept (dB) and reflected power (dB). The acoustic properties are measured over a temperature range of 22 to 37 degrees C. An intercomparison of results between two independent ultrasound measurement laboratories is also presented. A longitudinal study of the acoustic properties over a period of two years is also detailed, and the effect of water immersion on the acoustic properties of TMM is measured. In addition, the physical parameters of mass density rho (kg m(-3)) and specific heat capacity C (J kg(-1) K(-1)) are included. The measurement techniques used were based on the substitution technique using both broadband and narrowband pulses centered on 20 MHz. Both the attenuation coefficient and speed of sound (both group and phase) showed good agreement with the expected values of 0.5 dB cm(-1) MHz(-1) and 1540 ms(-1), respectively, with average values over the three-year period of 0.49 dBcm(-1)MHz1 (SD +/- 0.05) and 1540.9 ms(-1) (SD +/- 8.7). These results also showed agreement between the two independent measurement laboratories. Speed of sound and attenuation coefficient were shown to change with temperature with rates of + 2.1 m s(-1) degrees C(-1) and -0.005 dB cm(-1) MHz(-1) degrees C(-1), respectively. Attenuation changed linearly with frequency at the high frequency range of 17 to 23 MHz, and speed of sound was found to be independent of frequency in this range. The spectral slope of relative backscattered power for the material increased with frequency at typically 1.5 dB MHz(-1). This compared favorably with theoretical spectral slope values, calculated for a variety of scatterer sizes, albeit at a lower frequency range. It is also noticed that, on extrapolation back to lower frequencies, the backscatter is comparable with that measured at 7 MHz. Overall, this non-commercial agar-based TMM is shown to perform as expected at the higher frequency range of 17 to 23 MHz and is seen to retain its acoustic properties of attenuation and speed of sound over a three-year period.


Human Data Supporting Glyburide in Ischemic Stroke.
The SUR1-TRPM4 channel is a critical determinant of edema and hemorrhagic transformation after focal ischemia. Blockade of this channel by the small molecule glyburide results in improved survival and neurological outcome in multiple preclinical models of ischemic stroke. A robust, compelling body of evidence suggests that an intravenous formulation of glyburide, RP-1127, can prevent swelling and improve outcome in patients with stroke. Retrospective studies of diabetic stroke patients show improved outcomes in patients who are continued on sulfonylureas after stroke onset. An early phase II study using magnetic resonance imaging and plasma biomarkers supports the conclusion that RP-1127 may decrease swelling and hemorrhagic transformation. Finally, the ongoing phase II RP-1127 development program has demonstrated continued safety as well as feasibility of enrollment and tolerability of the intervention. Continued efforts to complete the ongoing phase II study and definitive efficacy studies are needed to bring a candidate pharmacotherapy to a population of severe stroke patients that currently have no alternative.


Heterogeneity among H-2Kk antigens. Differential expression on lymphoid cells.
Evidence is presented for heterogeneity in the expression of H-2Kk determinants specific for two monoclonal antibodies, H100-30.R3 and 11-4.1. Heterogeneity was detectable by differences in cellular distribution on thymocytes and activated T cells, but not by sequential immunoprecipitation or differential redistribution studies. This result contrasts with earlier evidence of heterogeneity, demonstrable by these latter methods, between H-2Kk determinants specific for H100-30.R3 and another antibody, H100-27.R9.


A Mobile Phone App Intervention Targeting Fruit and Vegetable Consumption: The Efficacy of Textual and Auditory Tailored Health Information Tested in a Randomized Controlled Trial.
Mobile phone apps are increasingly used to deliver health interventions, which provide the opportunity to present health information via different communication modes. However, scientific evidence regarding the effects of such health apps is scarce. In a randomized controlled trial, we tested the efficacy of a 6-month intervention delivered via a mobile phone app that communicated either textual or auditory tailored health information aimed at stimulating fruit and vegetable intake. A control condition in which no health information was given was added. Perceived own health and health literacy were included as moderators to assess for which groups the interventions could possibly lead to health behavior change. After downloading the mobile phone app, respondents were exposed monthly to either text-based or audio-based tailored health information and feedback over a period of 6 months via the mobile phone app. In addition, respondents in the control condition only completed the baseline and posttest measures. Within a community sample (online recruitment), self-reported fruit and vegetable intake at 6-month follow-up was our primary outcome measure. In total, 146 respondents (ranging from 40 to 58 per condition) completed the study (attrition rate 55%). A significant main effect of condition was found on fruit intake (P=.049, partial η(2)=0.04). A higher fruit intake was found after exposure to the auditory information, especially in recipients with a poor perceived own health (P=.003, partial η(2)=0.08). In addition, health literacy moderated the effect of condition on vegetable intake 6 months later (P<.001, partial η(2)=.11). A higher vegetable intake was found for recipients with high health literacy after exposure to the textual or auditory intervention compared to the control condition (contrasts P=.07 and P=.004, respectively). In the case of relatively low health literacy, vegetable intake was the highest in the control condition (contrasts text control: P=.03; audio control: P=.04). This study provides evidence-based insight into the effects of a mobile health app. The app seems to have the potential to change fruit and vegetable intake up to 6 months later, at least for specific groups. We found different effects for fruit and vegetable intake, respectively, suggesting that different underlying psychological mechanisms are associated with these specific behaviors. Based on our results, it seems worthwhile to investigate additional ways to increase fruit and vegetable intake in recipients with low health literacy. International Standard Randomized Controlled Trial Number (ISRCTN): 23466915; http://www.isrctn.com/ISRCTN23466915 (Archived by WebCite at http://www.webcitation.org/6hTtfSvaz).


Magnetic, fluorescent and hybrid nanoparticles: From synthesis to application in biosystems.
Multifunctional nanoparticles have emerged as an outstanding candidate for a new generation of biomedical applications, mainly due to their remarkable properties and biocompatibility. Individual reports on multi-metal, semiconducting and superparamagnetic nanoparticles (SPIONs), elucidating on each's unique intrinsic properties, have demonstrated that the biological application of such materials is highly dependent of their size, shape, surface nature and core nature. However, reviews combining nanoparticles with multiple properties, as fluorescence and paramagnetism, as well as, biocompatibility, toxicology and biodegradability are yet seldom. This review highlights the highest output advances, of the last decade, on synthetic procedures for the design of multifunctional magneto-luminescent hybrid nanosystems based on quantum dots, SPIONs and mesoporous silica nanoparticles, as well as, surface modifications and their role for biological applications.


Cocaine withdrawal produces behavioral disruptions in rats.
There is currently no laboratory or clinical evidence from animal or human studies documenting a withdrawal syndrome associated with cocaine dependence, although many users report that withdrawal disturbances are responsible for their repeated use of the drug. In the present study rats self-administered i.v. cocaine and a sweetened drinking solution. When cocaine access was terminated there was a marked suppression in operant behavior reinforced by the sweetened solution, and this withdrawal disruption was immediately reversed when cocaine was reinstated. There were no physical signs of withdrawal, and food intake increased when cocaine was withdrawn. The results suggest that sensitive behavioral tests reveal aspects of drug dependence that may account for persistent abuse.


Characterization of interfacial property of a two-layered plate using a nonlinear low-frequency Lamb wave approach.
This work investigates the feasibility of using a nonlinear low-frequency Lamb wave approach for characterizing the interfacial property of a two-layered plate. Compared with the case of the exact phase-velocity matching, the approximate phase-velocity matching in the low-frequency region can still guarantee the cumulative second-harmonic generation (SHG) of primary Lamb wave propagation, which overcomes the drawbacks arising from the inherent dispersion and multimode features of Lamb wave propagation. For a given two-layered plate, the appropriate mode pair at low frequency consisting of primary Lamb wave and double-frequency Lamb wave (DFLW), which satisfies the approximate phase-velocity matching and nonzero energy flux, is selected to ensure that the amplitude of the generated second harmonic grows within the maximum cumulative distance (MCD). Meanwhile, the numerical analyses indicate that the variation of the SHG efficiency is maximized at the MCD during the interfacial degradation. Using the nonlinear ultrasonic measurement-based experimental setup, the time-domain signal of the second harmonic generated at different propagation distances is conveniently extracted, and then the relative nonlinear acoustic parameter curve consistent with the theoretical prediction is obtained. For examining the influence of interfacial property on the SHG effect of low-frequency Lamb wave propagation, the different annealing cycles of the thin adhesive layer (acrylics) are used to simulate minor changes in the interfacial property of the given two-layered plate. It is found that the relative nonlinear acoustic parameter at the MCD decreases monotonically and sensitively with the increment of annealing cycle number, which verifies the quantitative correlation between the SHG efficiency of low-frequency Lamb wave propagation and the degree of the interfacial degradation. The consistency between the numerical analysis and the experimental measurement shows the potential of using the SHG effect of low-frequency Lamb wave propagation to characterize a minor change in the interfacial properties of a layered composite plate.


[Electron-microscopic characteristics of nodular variants of prolymphocytic lymphosarcomas].
Electron microscopic examinations of the lymph nodes tissue in prolymphocytic lymphosarcomas with nodular growth established that tumor cells forming neoplastic nodules retain the ultrastructural characteristics of their analogues (cells with cleaved melei) in unaltered or reactive germ centers of lymphatic follicles.


Silver nanoparticles induced with aqueous black carpenter ant extract selectively inhibit the growth of Pseudomonas aeruginosa.
Aqueous black carpenter ant extract (ABCAE) was used to synthesize silver nanoparticles (AgNPs). The ABCAE was rich in water-soluble compounds such as hydrophilic polypeptides that behaved as both reducing and stabilizing agents for generating AgNPs from Ag+ ion precursors. The diameter of the observed AgNPs was mostly in the range of 20-60 nm. The AgNPs were tested as an antibacterial agent for the growth inhibition of two pathogenic bacteria (Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 27661) and one common bacteria (Escherichia coli K12 ATCC 10798). Disk diffusion test showed that the AgNPs selectively inhibited the growth of P. aeruginosa but not for the other two species, suggesting the potential application of the green-chemically synthesized AgNPs as a selective antibacterial agent without harming other beneficial bacteria.


In vitro protein binding of teicoplanin to neonatal serum.
The protein-binding ability of teicoplanin (TEIC) to neonatal serum was investigated and the following results were obtained. (1) The protein binding ability of TEIC at concentrations ranging from 10 to 100 microg/ml, to neonatal serum was 80.5%-71.9% and was generally low as compared with that for adult serum. (2) The protein binding ability of TEIC (20 microg/ml) to albumin in neonatal serum was not concentration-dependent, and the correlation between the protein-binding ability and albumin concentration was low. Namely, it was suggested that the protein-binding ability of TEIC to neonatal serum was not dependent merely on the albumin concentration, and there was a possibility that the protein-binding ability also depended on qualitative changes in albumin. (3) The protein-binding ability of TEIC (20 microg/ml) to total bilirubin and free bilirubin was not concentration-dependent, and the correlation was extremely low, indicating that there is a low possibility that the protein-binding ability of TEIC induces an increase in free bilirubin.


Examination of three theories for mechanisms of cation-induced bioflocculation.
Research from different studies has been used to support three different theories pertaining to the role of cations in bioflocculation. These theories are the alginate theory. Derjaguin, Landau, Verwey, and Overbeek (DLVO) theory, and divalent cation bridging (DCB) theory. The objectives of this research were to examine the role of cations in bioflocculation to determine which theory, if any, best describes cation induced bioflocculation. Experiments were performed using laboratory scale activated sludge systems with bactopeptone as a feed. The feed was supplemented with either calcium, magnesium, or sodium at increasing concentrations. Floc properties were analyzed in each reactor during steady state periods. The addition of calcium or magnesium to the feed individually resulted in improvements in SVI, CST, SRF, cake solids and floc strength and each of these divalent cations produced similar results. The addition of sodium to the feed resulted in a deterioration in floc properties relative to a control reactor. Analysis of these results suggest that the DCB theory best explains the role of cations. The discrepancies between different studies were examined and are thought to be a result of different experimental procedures in different studies and in particular the use of short-term batch tests versus continuous flow reactor studies. In addition, the implications of DCB theory suggests that activated sludge systems should attempt to lower the ratio of monovalent to divalent cations to improve floc properties and treatment performance.


Synthesis of graphene oxide/polyethyleneimine sponge and its performance in the sustainable removal of Cu(II) from water.
Modification of graphene oxide (GO) with polyethyleneimine (PEI) has been studied to develop a GO/PEI sponge material that not only performs well in the adsorption of Cu(II) but also is easily separated from water. The results showed that GO had excellent affinity for PEI, and GO/PEI prepared at pH 9.0 using PEI with a MW of 70,000 was shown to be a good adsorbent for Cu(II). This GO/PEI was characterized with SEM, XRD, XPS, FTIR and TG analyses and was investigated for Cu(II) adsorption further. The adsorption isotherm data of Cu(II) were fitted well with the Langmuir model, from which the maximum adsorption of GO/PEI was calculated to be 150.9 mg/g at pH 5.5. This was much higher than that of GO and two commercial resins. GO/PEI showed high selectivity towards Cu(II). In GO/PEI, the contributions of amino groups on PEI and negative charges on GO were 79.2% and 20.8%, respectively. Cu(II) adsorption on GO/PEI decreased with decreasing pH, and 1 M HCl caused nearly complete desorption of the adsorbed Cu(II). Experimental results of five cycles of adsorption-desorption indicated that this material could be reused. Column studies showed that GO/PEI performed well in terms of both Cu(II) adsorption and stability in water.


Yacón (Smallanthus sonchifolius) prevented inflammation, oxidative stress, and intestinal alterations in an animal model of colorectal carcinogenesis.
Yacón (Smallanthus sonchifolius) roots store carbohydrate in the form of prebiotic fructooligosaccharides (FOS), which improve intestinal health. Yacon has the potential to prevent the intestinal barrier alterations associated with colorectal cancer (CRC). This study aimed to investigate the preventive effects of yacón flour (YF) on alterations promoted by CRC induced by 1,2-dimethylhydrazine in rats. CRC increased tumor necrosis factor alpha levels (group CY = 10.2 ± 0.72; group C = 9.6 ± 1.0; group Y = 5.8 ± 0.54; group S = 5.95 ± 0.6 pg mL-1 ) and short-chain fatty acid production, and decreased total antioxidant capacity (group CY = 4.7 ± 0.72; group C = 3.3 ± 0.3; group Y = 4.1 ± 0.47; group S = 6.7 ± 0.78 U mL-1 ). Furthermore, YF treatment reduced intraluminal pH (group CY = 6.45 ± 0.47; group C = 7.65 ± 0.44; group Y = 6.75 ± 0.46; group S = 8.13 ± 0.2), lactulose/mannitol ratio, tumor necrosis factor-alpha (TNF-α)/interleukin (IL)-10 ratio, and increased secretory immunoglobulin A (group CY = 9.48 ± 1.46; group C = 10.95 ± 3.87; group Y = 15.95 ± 7.36; group S = 9.19 ± 1.52), but did not affect IL-10, IL-12, and TNF-α levels nor the IL-12/IL-10 ratio. YF as a source of fructooligosaccharides may help to maintain the integrity of intestinal health, which is altered in induced CRC in rats. © 2020 Society of Chemical Industry.


Mechanical characterization of a fiberoptic microneedle device for controlled delivery of fluids and photothermal excitation.
Current clinical approaches for treating pancreatic cancer have been demonstrated as ineffective at improving midterm survival. A primary obstacle to local drug delivery is the desmoplastic nature of the peritumoral environment, which acts as a significant barrier to circulating macromolecules. To address this need, our group presents a sharp fiberoptic microcatheter capable of accessing the pancreas through transduodenal endoscope and penetrating a tumor to locally co-deliver photothermal and fluid-based therapies. Experiments sought to characterize the mechanical penetration capabilities and fluid mechanics of the fiberoptic microneedle design. A refined off-center fusion splicing technique was developed for joining a multimode fiber to the annular core of a light-guiding capillary, allowing light transmission with minimal optical loss. A novel and frugal technique for assessing the penetration force of the microneedle was conducted in a bovine gelatin tissue phantom with a Young's modulus stiffer than the high range for pancratic tissue or tumor. Buckling forces for different microneedle lengths were measured and compared against theoretical values obtained from Euler's Critical Load equation under fixed-pinned column conditions. Hydraulic resistance of different capillary lengths was evaluated and compared against the theoretical values from Hagen-Poiseuille's law, allowing assessment of contributions from different segments of the device. The results demonstrated that the microcatheter can robustly and repeatably penetrate a soft tissue phantom chosen to be a conservative model of pancreatic tissue for penetration properties. Experiments showed that a 1.5 N insertion force was required for phantom penetration with a 45° beveled needle at a 5 mm unsupported length, while the critical buckling load was measured to be approximately 4 N. In addition, the design was demonstrated to efficiently transport 1064 nm light and aqueous fluids with a 70-75% light coupling efficiency and 12,200 Pa.s/μl hydraulic resistance, respectively. These findings motivate the FMD's further development as a treatment platform for pancreatic cancer.


Fetal lung maturation: human amniotic fluid phosphatidate phosphohydrolase activity through normal gestation and its relation to the lecithin/sphingomyelin ratio.
In order to evaluate the relationship between the increase in amniotic fluid phosphatidate phosphohydrolase (PAPase) specific activity and the increase in the lecithin/sphingomyelin (L/S) ratio during normal human pregnancy, PAPase specific activity and the L/S ratio were measured in 171 amniotic fluid samples obtained from 164 women who were at 17 to 42 weeks' gestation. The increase in PAPase specific activity in amniotic fluid is parallel to the increase in the L/S ratio. The correlation between PAPase specific activity and the L/S ratio in amniotic fluid from all gestational ages is highly significant. The relationship of PAPase specific activity in amniotic fluid to PAPase specific activity in gastric fluid was investigated in a study of 97 newborn infants. A highly significant correlation was found between these two values. To ascertain if a relationship exists between the specific activity of PAPase in amniotic fluid and the subsequent development of hyaline membrane disease (HMD), 223 neonates who were delivered within 72 hours of amniotic fluid collection were studied. Only one infant developed HMD out of 170 with amniotic fluid PAPase specific activity equal to or greater than 50 nmoles of orthophosphate released X mg.-1 of protein X hr.-1. On the other hand, the finding of an amniotic fluid PAPase specific activity of less than 50 nmoles was of little value in predicting lung immaturity. We believe that these findings are also supportive of the view that PAPase and surfactant are released from the type II pneumocyte as a closely related structural unit, viz., the lamellar body.


Defects in the regulation of B cell apoptosis are required for the production of citrullinated peptide autoantibodies in mice.
Protein deimination, a process to modify arginine residues to citrulline by the addition of a neutral oxygen group, is associated with apoptosis. The presence of autoantibodies recognizing citrullinated peptides is highly specific to rheumatoid arthritis (RA) and is therefore a useful marker for the early diagnosis of RA. In this study, we explored whether anti-cyclic citrullinated peptide (anti-CCP) autoantibodies are produced in several experimental models of autoimmune diseases in mice. The levels of anti-CCP autoantibodies were analyzed by enzyme-linked immunosorbent assay in several lupus-prone strains of mice, in animals with type II collagen (CII)-induced arthritis, and after induction of neonatal tolerance to alloantigens. We observed the production of these autoantibodies in 2 different lupus-prone mice, MRL-lpr/lpr and (NZW x B6)F(1)-hbcl-2 transgenic mice, characterized by the presence of abnormalities in the regulation of B cell apoptosis. Other genetic defects, determining autoimmune susceptibility, present in MRL and NZW mice were additionally required for anti-CCP autoantibody production. The induction of autoantibodies in normal BALB/c mice injected at birth with semiallogeneic spleen cells from (BALB/c x B6)F(1)-hbcl-2 transgenic mice suggested that these additional autoimmune defects may be related, at least in part, to the establishment of abnormal interactions between T cells and B cells. In addition, anti-CCP autoantibodies were not produced in the course of CII-induced arthritis, an experimental model of RA in mice. Our study provides evidence for the association between defects in the regulatory cell death machinery of B lymphocytes and the production of certain autoantibody specificities.


Sympathetic neurotransmitter metabolism in Hirschsprung's disease.
Tyrosine hydroxylase activity was measured in high speed supernatants obtained from full thickness segments of aganglionic and ganglionic colon of three children with Hirschsprung's disease. Tyrosine hydroxylase activity expressed as pmole DOPA/mg protein/min was 0.93 +/- 0.16 in ganglionic and 2.67 +/- 0.21 in aganglionic colon. Tyrosine hydroxylase activity in ganglionic colon rose to 2.29 +/- 0.11 following calcium stimulation (100 muM) but could not be further increased in aganglionic colon. Addition of norepinephrine (2 X 10(-4) M) to tissue homogenates inhibited tyrosine hydroxylase activity in ganglionic colon by 57 +/- 8% but only by 14 +/- 3% in aganglionic colon, suggesting that the enzyme present in aganglionic colon is insensitive to feedback inhibition by endogenous norepinephrine. The elevation of tyrosine hydroxylase activity in aganglionic colon and its insensitivity to calcium stimulation and norepinephrine inhibition is further evidence of sympathetic overactivity in the aganglionic colon and suggests a basic enzymatic abnormality in the pathogenesis of Hirschsprung's disease.


The pyrophosphate-dependent phosphofructokinase of the protist, Trichomonas vaginalis, and the evolutionary relationships of protist phosphofructokinases.
The pyrophosphate-dependent phosphofructokinase (PPi-PFK) of the amitochondriate protist Trichomonas vaginalis has been purified. The enzyme is a homotetramer of about 50 kDa subunits and is not subject to allosteric regulation. The protein was fragmented and a number of peptides were sequenced. Based on this information a PCR product was obtained from T. vaginalis gDNA and used to isolate corresponding cDNA and gDNA clones. Southern analysis indicated the presence of five genes. One open reading frame (ORF) was completely sequenced and for two others the 5' half of the gene was determined. The sequences were highly similar. The complete ORF corresponded to a polypeptide of about 46 kDa. All the peptide sequences obtained were present in the derived sequences. The complete ORF was highly similar to that of other PFKs, primarily in its amino-terminal half. The T. vaginalis enzyme was most similar to PPi-PFK of the mitochondriate heterolobosean, Naegleria fowleri. Most of the residues shown or assumed to be involved in substrate binding in other PPi-PFKs were conserved in the T. vaginalis enzyme. Direct comparison and phylogenetic reconstruction revealed a significant divergence among PPi-PFKs and related enzymes, which can be assigned to at least four distantly related groups, three of which contain enzymes of protists. The separation of these groups is supported with a high percentage of bootstrap proportions. The short T. vaginalis PFK shares a most recent common ancestor with the enzyme from N. fowleri. This pair is clearly separated from a group comprising the long (>60-kDa) enzymes from Giardia lamblia, Entamoeba histolytica pfk2, the spirochaetes Borrelia burgdorferi and Trepomena pallidum, as well as the alpha- and beta-subunits of plant PPi-PFKs. The third group ("X") containing protist sequences includes the glycosomal ATP-PFK of Trypanosoma brucei, E. histolytica pfk1, and a second sequence from B. burgdorferi. The fourth group ("Y") comprises cyanobacterial and high-G + C, Gram-positive eubacterial sequences. The well-studied PPi-PFK of Propionibacterium freudenreichii is highly divergent and cannot be assigned to any of these groups. These four groups are well separated from typical ATP-PFKs, the phylogenetic analysis of which confirmed relationships established earlier. These findings indicate a complex history of a key step of glycolysis in protists with several early gene duplications and possible horizontal gene transfers.


Addressing nutrient shortfalls in 1- to 5-year-old Irish children using diet modeling: development of a protocol for use in country-specific population health.
Dietary habits formed in early childhood can track into later life with important impacts on health. Food-based dietary guidelines (FBDGs) may have a role in improving population health but are lacking for young children. We aimed to establish a protocol for addressing nutrient shortfalls in 1- to 5-y-old children (12-60 mo) using diet modeling in a population-based sample. Secondary analysis of 2010-2011 Irish National Pre-School Nutrition Survey data (n = 500) was conducted to identify typical food consumption patterns in 1- to 5-y-olds. Nutrient intakes were assessed against dietary reference values [European Food Safety Authority (EFSA) and Institute of Medicine (IOM)]. To address nutrient shortfalls using diet modeling, 4-d food patterns were developed to assess different milk-feeding scenarios (human milk, whole or low-fat cow milk, and fortified milks) within energy requirement ranges aligned with the WHO growth standards. FBDGs to address nutrient shortfalls were established based on 120 food patterns. Current mean dietary intakes for the majority of 1- to 5-y-olds failed to meet reference values (EFSA) for vitamin D (≤100%), vitamin E (≤88%), DHA (22:6n-3) + EPA (20:5n-3) (IOM; ≤82%), and fiber (≤63%), whereas free sugars intakes exceeded recommendations of <10% energy (E) for 48% of 1- to 3-y-olds and 75% of 4- to 5-y-olds. "Human milk + Cow milk" was the only milk-feeding scenario modeled that predicted sufficient DHA + EPA among 1- to 3-y-olds. Vitamin D shortfalls were not correctable in any milk-feeding scenario, even with supplementation (5 µg/d), apart from the "Follow-up Formula + Fortified drink" scenario in 1- to 3-y-olds (albeit free sugars intakes were estimated at 12%E compared with ≤5%E as provided by other scenarios). Iron and vitamin E shortfalls were most prevalent in scenarios for 1- to 3-y-olds at ≤25th growth percentile. Using WHO growth standards and international reference values, this study provides a protocol for addressing nutrient shortfalls among 1- to 5-y-olds, which could be applied in country-specific population health.


Enteroclysis in the diagnosis of chronic unexplained gastrointestinal bleeding.
In a six-year period (1977-83), lesions were identified by enteroclysis in 26 patients with melena or recurrent gastrointestinal bleeding undiagnosed by other modalities. These included nine Meckel's diverticula, three metastatic lesions, three primary carcinomas, one lipoma, four leiomyomas, five surgically created blind pouches, one carcinoid, and one idiopathic dilatation of the ileum. Our experience suggests that, when the standard diagnostic procedures used to investigate chronic gastrointestinal blood loss are unrevealing, enteroclysis should be performed. The method is fast, accurate, is done in one sitting, and can be productive in the diagnostically difficult patient.


Monthly variation of Dermatophagoides allergens and its influence on respiratory allergy in a high altitude environment (Quito, 2800 m a.s.l. in Andean Ecuador).
Seasonal variation of Dermatophagoides allergens and its influence in allergic respiratory airway diseases has not been investigated in Andean cities. The objective of this study was to evaluate those parameters in a city located in the Andean mountains. Der p1 and Der f1 were measured in dust samples from mattresses in 13 houses in Quito (2800 m above sea level). Samples were collected monthly from August 2004 to July 2005. Patients presenting to a local outpatient allergy clinic with asthma and rhinitis and isolated allergy to Dermatophagoides were analysed to determine if a correlation existed between seasonal Der allergen levels and the number of patients presenting with allergies. High levels of dust mites and humidity were observed throughout the year. The highest geometrical mean values of allergens were detected in April (Der p1, 10.15 μg/g) and May (Der f1, 13.03 μg/g), whilst the lowest levels were detected in August (Der p1, 4.26 μg/g), and September (Der f1, 1.4 μg/g). Of the 361 patients examined, 182 were allergic to Dermatophagoides, (45.6% asthmatics, 97.8% rhinitics, and 43.4% with both diseases). Patient presentation spiked in August, and from February to May. However, there was not a significant correlation between mite allergen concentrations and humidity or the number of patients presenting with allergies. Dust samples from mattresses in Quito revealed high concentrations of Der p1 and Der f1. We observed a trend towards increased presentation of asthmatic and rhinitic patients in the months with highest levels of allergens.


Factors associated with repeated influenza vaccine uptake among aged care staff in an Australian sample from 2017 to 2019.
Influenza vaccination is strongly recommended every year for aged care staff to protect themselves and minimise risk of transmission to residents. This study aimed to determine the factors associated with repeated annual influenza vaccine uptake among Australian aged care staff from 2017 to 2019. Demographic, medical and vaccination data collected from the staff, who participated in an observational study from nine aged care facilities under a single provider in Sydney Australia, were analysed retrospectively. Based on the pattern of repeated influenza vaccination from 2017 to 2019, three groups were identified: (1) unvaccinated all three years; (2) vaccinated occasionally(once or twice) over three years; and (3)vaccinated all threeyears. Multinomial logistic regression analysis was performed to better understand the factors associated with the pattern of repeated influenza vaccination. From a total of 138 staff, between 2017 and 2019, 28.9 % (n = 40) never had a vaccination, while 44.2 % (n = 61) had vaccination occasionally and 26.8 % (n = 37) had vaccination all three years. In the multinomial logistic regression model, those who were<40 years old (OR = 0.57, 95 % CI: 0.19-0.90, p < 0.05) and those who were current smokers (OR = 0.20; 95 % CI: 0.03-0.76, p < 0.05) were less likely to have repeated vaccination for all three years compared to the unvaccinated group. Those who were<40 years old (OR = 0.61; 95 % CI: 0.22-0.68, p < 0.05) and those who were born overseas (OR = 0.50; 95 % CI:0.27-0.69, p < 0.05) were more likely to be vaccinated occasionally compared to the unvaccinated group. The significant predictors of repeated vaccine uptake across the three-year study period among aged care staff were age, smoking status and country of birth (Other vs Australia). Targeted interventions towards the younger age group (<40 years old), smokers and those who were born overseas could improve repeated influenza vaccination uptake in the aged care workforce.


Distinct dopaminergic systems in ACTH-induced grooming.
Excessive grooming behavior induced in rats by intraventricular injection of ACTH1-24 was inhibited by selective pharmacological manipulation of two functionally distinct types of dopaminergic terminals (DAe and DAi) in the neostriatum or the nucleus accumbens. It is concluded that when ACTH1-24 produces the behavioral response it modulates the activity of both the DAe and the DAi systems.


Evaluating the effect of clofazimine against Mycobacterium tuberculosis given alone or in combination with pretomanid, bedaquiline or linezolid.
In recent years, clofazimine (CFZ) has been regaining prominence for the treatment of tuberculosis. However, it shows limited efficacy as a single drug and optimal combination partners have not been identified. Therefore, the objective of our analysis was to evaluate the efficacy of CFZ-containing two-drug regimens with pretomanid (PMD), bedaquiline (BDQ) or linezolid (LZD) by: (i) determining their pharmacodynamic (PD) mode of interaction against Mycobacterium tuberculosis (Mtb) strain H37Rv in log- phase and acid-phase metabolic states, and against Mtb strain 18b in a non-replicating persister (NRP) metabolic state; (ii) predicting bacterial cell kill of the drugs alone and in combination; and (iii) evaluating the relationship between the interaction mode and the extent of bacterial cell kill. The results of our Greco universal response surface analysis showed that CFZ was at least additive with a clear trend towards synergy when combined with PMD, BDQ and LZD against Mtb in all explored metabolic states under in vitro checkerboard assay conditions. The results further showed that all two-drug combination regimens exerted greater bacterial kill than any of the drugs alone. CFZ alone showed the least antimicrobial efficacy amongst the evaluated drugs, and there was a lack of correlation between the mode of interaction and the extent of bacterial kill. However, we may underestimate the effect of CFZ in this screening approach owing to limited in vitro study duration and neglect of target site accumulation. Clofazimine; Pretomanid; Bedaquiline; Linezolid; Combination chemotherapy; Mycobacterium tuberculosis.


Accuracy of Orbscan optical pachymetry in corneas with haze.
To compare corneal thickness measurements with the Orbscan instrument (Orbtek, Inc.) and an ultrasonic pachymeter (Alcon Surgical) in corneas with haze. Department of Ophthalmology and Otolaryngology, University of Bari, Bari, Italy. Six eyes of 6 patients having phototherapeutic keratectomy for treatment of grade 2+ haze were assessed preoperatively and postoperatively by Orbscan optical pachymetry and ultrasonic pachymetry. Contralateral transparent untreated corneas of the same patients were used as controls. Two Orbscan and 5 ultrasound pachymetry measurements were performed at each visit and the mean values recorded. The accuracy and repeatability of the measurements were evaluated. Preoperatively, the median corneal thickness in the 6 eyes with haze was 512.7 microm by ultrasonic pachymetry and 375.0 microm by optical pachymetry. The mean difference of 143.1 microm was statistically significant (Wilcoxon signed rank test, P= .0277). Postoperatively, the median pachymetry was 439.0 microm and 433.0 microm, respectively; the mean difference was 10.7 microm (P= .0277). In the control group, the median ultrasonic pachymetry was 530.9 microm and the median optical pachymetry, 529.0 microm; the mean difference was 4.7 microm (P= .1730). Precision was good in all groups; the standard deviation was less than 6.0 microm. The Orbscan system showed a decreased accuracy in measuring corneal thickness when clinically significant haze was present. The repeatability of the measures was good, suggesting that the cause of error lies in the optical acquisition process.


Implementing Higher Phototherapy Thresholds for Jaundice in Healthy Infants 35 Plus Weeks.
To determine the impact of higher bilirubin thresholds on testing and treatment of healthy infants during the neonatal period. This quality improvement study included infants born at ≥35 weeks gestation and admitted to the well-baby nursery between July 2018 and December 2020. We assessed the transition from infants treated according to the 2004 AAP guidelines (pregroup) with those following the Northern California Neonatal Consortium guidelines (postgroup). We examined the proportion of infants receiving phototherapy and total serum bilirubin (TSB) assessments as outcome measures. We examined critical hyperbilirubinemia (TSB above 25 mg/dL or TSB within 2 mg/dL of threshold for exchange transfusion), exchange transfusion, and readmission for jaundice as balancing measures. We compared the differences in outcomes over time using Statistical Process Control p charts. Balancing measures between the pre and postgroups were compared using χ square tests and t-tests. In our population of 6173 babies, there was a significant shift in the proportion receiving phototherapy from 6.4% to 4%. There were no significant changes in incidences of bilirubin >25 mg/dL (0 of 1472 vs 7 of 4709, P = .37), bilirubin within 2 mg/dL of exchange transfusion thresholds (4 of 1472 vs 5 of 4709, P = .15), exchange transfusion (0 of 1472 vs 1 of 4709, P = .70) or readmission for phototherapy (2.9% versus 2.4%, P = .30), between the 2 groups. Higher thresholds for phototherapy treatment of neonatal hyperbilirubinemia can decrease the need for phototherapy without increasing critical hyperbilirubinemia or readmission rate.


Widespread lipoplex-mediated gene transfer to vascular endothelial cells and hemangioblasts in the vertebrate embryo.
We report here a method that allows fast, efficient, and low-cost screening for gene function in the vascular system of the vertebrate embryo. Through intracardiac delivery of nucleic acids optimally compacted by a specific cationic lipid, we are able to induce in vivo endothelial cell-specific gain-of-function during development of the vascular network in the chick embryo. When the nucleic acids are delivered during the period of intraembryonic hematopoiesis, aortic hemangioblasts, the forerunners of the hematopoietic stem cells known to derive from the aortic endothelium, are also labeled. Similarly, we show that siRNA could be used to induce loss-of-function in vascular endothelial cells. This gene transfer technique was also applied to the mouse embryo with a high efficiency. The present method allows large-scale analysis and may represent a new and versatile tool for functional genomics.


[Accelerations of the fetal cardiac rhythm and fetal movements. Significance of their association recorded on the cardiotocogram by the Tococinon].
Recording all fetal movements on the cardiotocogram using tococinon showing forty minutes observations adds a new dimension to the old and new methods of monitoring the fetus. A group of 53 pregnancies without any risk was compared to a group of 37 pregnancies of high risk in which were also found sub-groups as designated by the aetiology and the severity of the fetal risk. Furthermore we have carried out a longterm study of 5 cases over six days. We propose a choice of the best parameters to be used according to their reliability, accuracy, sensitivity and predictability. The number of accelerations that coincided with the movements that were recorded was not very reliable or sensitive but it was accurate enough and predictive. The number of movements that accompanied the accelerations when added to the overall activity did become precise and predictive. Adding the two together make it more accurate and sensitive. New methods of monitoring are suggested, and these appear to be more simple and more efficient.


Clinical Characteristics and Risk Factors of Left Ventricular Thrombus after Acute Myocardial Infarction: A Matched Case-control Study.
Left ventricular thrombus (LVT) is reported to be a common complication in acute myocardial infarction (AMI) patients. And it has the potential to cause systemic embolism. This retrospective study was to present the current situation of LVT in clinical practice, as well as to evaluate the clinical characteristics and the risk factors of LVT after AMI. LVT cases (n = 96) were identified from 13,732 AMI (non-ST elevation myocardial infarction was excluded) patients in Fuwai Hospital's electronic medical records system from January 2003 to January 2013. The controls (n = 192) were gender- and age-matched AMI patients without LVT during this period. A conditional logistic regression (fitted by the Cox model) was performed to identify the independent risk factors. The incidence of LVT after AMI was 0.7%. Univariate analysis indicated that the anterior myocardial infarction (especially extensive anterior myocardial infarction), lower left ventricular ejection fraction (LVEF), LVEF ≤40%, severe regional wall motion abnormalities (RWMA), pericardial effusion, and left ventricular aneurysm were all related to LVT after AMI. The independent risk factors obtained from the conditional logistic regression analysis were lower LVEF (odds ratio (OR) = 0.891, 95% confidence interval (CI ): 0.828-0.960), extensive anterior myocardial infarction (OR = 6.403, 95% CI: 1.769-23.169), severe RWMA (OR = 7.348, 95% CI: 1.323-40.819), and left ventricular aneurysm (OR = 6.955, 95% CI: 1.673-28.921). This study indicated that lower LVEF, extensive anterior myocardial infarction, severe RWMA, and left ventricular aneurysm were independent risk factors of LVT after AMI. It also suggested that further efforts are needed for the LVT diagnosis after AMI in clinical practice.


Corrigendum: Changes in primary somatosensory cortex following allogeneic hand transplantation or autogenic hand replantation.
[This corrects the article DOI: 10.3389/fnimg.2022.919694.].


An mRNA structure that controls gene expression by binding S-adenosylmethionine.
Riboswitches are metabolite-binding RNA structures that serve as genetic control elements for certain messenger RNAs. These RNA switches have been identified in all three kingdoms of life and are typically responsible for the control of genes whose protein products are involved in the biosynthesis, transport or utilization of the target metabolite. Herein, we report that a highly conserved RNA domain found in bacteria serves as a riboswitch that responds to the coenzyme S-adenosylmethionine (SAM) with remarkably high affinity and specificity. SAM riboswitches undergo structural reorganization upon introduction of SAM, and these allosteric changes regulate the expression of 26 genes in Bacillus subtilis. This and related findings indicate that direct interaction between small metabolites and allosteric mRNAs is an important and widespread form of genetic regulation in bacteria.


Downregulation of miR-4443 and miR-5195-3p in ovarian cancer tissue contributes to metastasis and tumorigenesis.
Ovarian cancer (OC) is one of the most fatal malignancies in women. High mortality rate may be due to problems with diagnosis in the early stages. The use of new biomarkers for faster diagnosis and selection of more efficient therapies is one of the main concerns in this area. miRNAs are non-coding and conserved molecules that are involved in regulating gene expression throughout different cell processes. Few studies have been conducted on the effects of miR-4443 and miR-5195-3p in cancer. Therefore, to determine the role of these miRNAs in OC, this study was directed to investigate the expression rate in OC tissue samples and its relationship with clinical factors. Expression levels of miR-4443 and miR-5195 were evaluated in 45 ovarian tumor and 45 ovarian non-tumor tissue samples paraffin embedded using qPCR. Expression was investigated by miRNA-specific primers and then statistical analysis was performed to determine the significance. In the next step, the relationship between clinopathologic factors and miRNA expression was investigated. The results showed that miR-4443 decreased in OC in metastatic and serous OC samples (0.154-fold, P < 0.0001). As well as, significant reduction in miR-5195-3p was observed in cancer samples (0.373-fold, P < 0.0001) and its reduction was associated with metastasis. As a result, the two studied miRNAs may contribute to suppressing tumor, so that decrease in their expression is associated with increased cell proliferation and invasion. Further investigation can help to suggest these miRNAs as diagnostic biomarkers or therapeutic targets in OC.


Release of osmolytes from perfused rat liver on perivascular nerve stimulation: alpha-adrenergic control of osmolyte efflux from parenchymal and nonparenchymal liver cells.
The effects of perivascular nerve stimulation and phenylephrine on osmolyte release were studied in the intact perfused rat liver and isolated liver parenchymal cells (PC) and nonparenchymal cells. In the perfused liver, electrical stimulation of perivascular nerves (20 Hz/2 ms/20 V) led to a phentolamine-sensitive increase of cell hydration by 6.5% +/- 1.2% (n = 3) and a transient phentolamine-sensitive stimulation of taurine and inositol, but not betaine, release. These nerve effects were mimicked by phenylephrine, but not prostaglandin F2alpha, and were not affected by sodium nitroprusside (SNP) or ibuprofen. Nerve stimulation-induced taurine, but not inositol, release was inhibited by 4, 4'-di-isothiocyanatostilbene-2,2'-disulphonic acid (DIDS) (50 micromol/L). Single-cell fluorescence studies with isolated liver PC, Kupffer cells (KC), sinusoidal endothelial cells (SEC), and hepatic stellate cells (HSC) revealed that phenylephrine induced an increase in cytosolic free Ca2+ only in PC and HSC, but not in KC and SEC, whereas extracellular uridine triphosphate (UTP) produced Ca2+ transients/oscillations in all liver cell types studied. Phenylephrine had no effect on osmolyte release from isolated KC and SEC, but increased taurine (but not inositol) release from PC and inositol (but not taurine) efflux from HSC. The data suggest that: 1) liver cell hydration and-consecutively-osmolyte content are modulated by hepatic nerves via an alpha-adrenergic mechanism, which does not involve eicosanoids or hemodynamic changes; 2) that PC and HSC are the primary targets for nerve-dependent alpha-adrenergic activation, whereas 3) KC and SEC probably do not express alpha-adrenoceptors coupled to Ca2+ mobilization or osmolyte efflux.


Responses of microalgae under different physiological phases to struvite as a buffering nutrient source for biomass and lipid production.
Microalgae cultivation for biodiesel production is promising, but the high demand for nutrients, such as nitrogen and phosphorus, remains a limiting factor. This study investigated effects of struvite, a low-cost nutrient source, on microalgae production under different physiological phases. Changes in element concentrations were determined to characterize the controllable nutrient release properties of struvite. Results showed that nutrient elements could be effectively supplemented by struvite. However, responses of microalgae under different growth stages to struvite varied obviously, achieving the highest biomass (0.53 g/L) and the lowest (0.32 g/L). Moreover, the microalgal lipid production was obviously increased by adding struvite during the growth phase, providing the first evidence that struvite could serve as an alternative buffering nutrient source to culture microalgae. The integration of microalgae cultivation with struvite as a buffering nutrient source provides a novel strategy for high ammonia nitrogen wastewater treatment with microalgae for biodiesel production.


Association between lung cancer risk and single nucleotide polymorphisms in the first intron and codon 178 of the DNA repair gene, O6-alkylguanine-DNA alkyltransferase.
The association between lung cancer risk and 2 polymorphisms, rs12268840 and rs2308327 (codon K178R), in the DNA repair protein, O(6)-alkylguanine-DNA alkyltransferase, which are associated with interindividual differences in activity, have been investigated in 3 hospital-based case-control studies. Genotyping was carried out on 617 subjects of whom 255 had lung cancer. In 2 of the 3 series, there was a significant inverse association between the 178R allele and case status (p < 0.05). In a meta-analysis, the odds ratio (95% CI) associated with the 178R allele relative to the 178K allele was 0.64 (0.45-0.92, p = 0.01) and 0.51 (0.24-1.11, p = 0.09) in fixed effects and random effects models, respectively. In a pooled analysis, after adjustment for sex, age, pack years and series, the OR (95% CI) for a heterozygote was 0.67 (0.45-1.01) and for a 178R homozygote was 0.10 (0.01-0.94); the trend for a decreased risk with the number of R alleles was significant (p = 0.008). This trend was particularly pronounced in heavy smokers (trend test p = 0.003), but not significant in light smokers (p = 0.73). There was no evidence of an association between rs12268840 and lung cancer risk. These results suggest that the R allele may protect against lung cancer, specifically in heavy smokers, an effect that may result from this polymorphism affecting the function of the MGMT protein and/or levels in MGMT activity.


Persistent Diarrhea and Eosinophilic Esophagitis Resulting From Chronic Cannabidiol Usage for Refractory Epilepsy.
Cannabidiol is used in the care of treatment-resistant epilepsy. It has been associated with varying side effects, ranging from somnolence to diarrhea and weight loss. We present a patient on chronic cannabidiol therapy who had persistent diarrhea, abdominal pain, weight loss, and esophageal eosinophilia that improved with cannabidiol dose adjustment.


Type I collagen preparations inhibit DNA synthesis in glial cells of the peripheral nervous system.
The mechanisms underlying cessation of glial proliferation in the developing peripheral nervous system are obscure. One possibility, as yet little explored, is that mitotic inhibitory signals play a part in regulating glial cell numbers. In this study we demonstrate that type I collagen preparations from several different sources can inhibit the rate of DNA synthesis in purified populations of enteric glia and both short-term and long-term secondary Schwann cells in dissociated cell cultures. When these cells are grown on gelled or dried type I collagen substrata, they proliferate at substantially lower rates than on polylysine substrata. In contrast, type III or V collagen preparations do not inhibit glial DNA synthesis and laminin, fibronectin, type IV collagen, and secreted matrix from bovine corneal endothelial cells all stimulate thymidine incorporation. The inhibitory effect is not observed with heat denatured type I collagen preparations, but is seen equally in serum-containing medium, in medium containing fibronectin-free serum, or in serum-free medium, suggesting that the interaction of collagen with the cells requires structurally intact collagen molecules and does not occur via intermediary linkage to fibronectin. The inhibition on collagen is accompanied by a shape change from a more flattened morphology to a narrow spindle form. The labeling index of a rat Schwannoma cell line, 33B, is not inhibited on type I collagen substrata. These results demonstrate that type I collagen preparations inhibit the DNA synthesis levels of early postnatal peripheral glial cells in vitro. It remains to be determined whether this effect occurs via direct collagen-cell membrane interactions or whether it depends on accessory molecules, perhaps present in the collagen preparations themselves, since these are not purified to absolute homogeneity.


Safety and tolerability study of sotrovimab (VIR-7831) prophylaxis against COVID-19 infection in immunocompromised individuals with impaired SARS-CoV-2 humoral immunity.
Multiple vaccines have been approved since August 2021 to prevent infection with SARS-CoV-2; however, 20-40% of immunocompromised people fail to develop SARS-CoV-2 spike antibodies after COVID-19 vaccination and remain at high risk of infection and more severe illness than non-immunocompromised hosts. Sotrovimab (VIR-7831) is a monoclonal neutralizing antibody that binds a conserved epitope on the SARS-CoV-2 spike protein. It is neither renally excreted nor metabolized by P450 enzymes and therefore unlikely to interact with concomitant medications (e.g., immunosuppressive medications). In this open-label feasibility study protocol, we will define the optimal dose and dosing interval of sotrovimab as pre-exposure prophylaxis for immunocompromised individuals as well as its safety and tolerability in this population specifically. We will enroll 93 eligible immunocompromised adults with a negative or low-positive (< 50 U/mL) SARS-CoV-2 spike antibody. In phase 1, the first 10 patients will participate in a lead-in pharmacokinetics (PK) cohort study to determine the optimal dosing interval. Phase 2 will expand this population to 50 participants to examine rates of infusion-related reactions (IRR) with a 30-min 500 mg sotrovimab IV infusion. Phase 3 will be an expansion cohort for further assessment of the safety and tolerability of sotrovimab. In phase 4, the first 10 patients receiving 2000 mg IV of sotrovimab on the second sotrovimab infusion day will comprise a lead-in safety cohort that will inform the duration of observation following administration of the drug. The patients will be followed for safety and COVID-19 events for 36 weeks after the second dose. In a previous phase III randomized, placebo-controlled pivotal trial, there were no significant differences in the prevalence of adverse events in patients receiving sotrovimab vs. placebo. Thus, we propose an open-label feasibility study protocol of sotrovimab as pre-exposure prophylaxis for immunocompromised individuals to evaluate its PK in immunocompromised individuals with impaired SARS-CoV-2 humoral immunity and define optimal dosing intervals. We also aim to determine COVID-19 infections over the study period and self-reported quality of life measures throughout the study. ClinicalTrials.gov Identifier: NCT05210101.


Discontinuous microduplications at chromosome 10q24.31 identified in a Chinese family with split hand and foot malformation.
Split hand/foot malformation (SHFM) is a congenital disorder characterized by a cleft of the hands and/or feet due to dificiency of central rays. Genomic rearrangement at 10q24 has been found to cause nonsyndromic SHFM (SHFM3). Four patients and fourteen unaffected individuals from a four-generation Chinese pedigree with typical SHFM3 phenotypes were recruited for this study. After informed consent was obtained, genome-wide copy number analysis was performed on all patients and two normal family members using the Affymetrix Cytogenetics Whole-Genome 2.7M Array. The results were then confirmed by real-time quantitative polymerase chain reaction in all available individuals of this pedigree. Candidate genes were further screened for mutation through sequence analyses. Copy number analysis showed a microduplication at chromosome 10q24.31-q24.32 co-segregating with the SHFM phenotype. Compared to other known genomic duplications for SHFM3, the duplication described here contains two discontinuous DNA fragments. The minimal centromeric duplicated segment of 259 kb involves LBX1, POLL and a disrupted BTRC. The minimal telomeric duplication of 114 kb encompasses DPCD and one part of FBXW4. No coding and splice-site mutations of candidate genes in the region were found. Genomic duplications at chromosome 10q24.3, which were identified in the current study, provide further evidence for limb-specific cis-regulatory sequences in this region, highlighting the importance of chromosome 10q24.31-q24.32 in limb development and SHFM pathogenesis.


Influence of early rearing on lymphocyte proliferation responses in juvenile rhesus monkeys.
Lymphocyte proliferation responses and natural killer cell activity were evaluated in 35 juvenile rhesus monkeys derived from five different rearing conditions. Nursery-reared monkeys had proliferation responses which were significantly higher than those of mother-reared subjects. Reexamination of the nursery-reared monkeys 1.5 years later indicated that an abnormally high response to concanavalin A was still evident at 2.5 years of age, but both PHA and PWM responses had shown an age-appropriate decrease into the normal range for this species. Proliferation responses in monkeys that had been weaned early from their mothers at 6 months of age were also higher than values for control monkeys that remained with their mothers, but below those of the nursery-reared monkeys. In contrast, monkeys that had received multiple separations from the mother between 3 and 7 months of age showed lymphocyte proliferation responses that were below normal. These results indicate that early rearing conditions can have a lasting effect on certain immune responses in the developing primate.


Evidence for a role for protein kinase C in the modulation of bombesin-activated cellular signalling in human breast cancer cells.
The phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) is a potent activator of protein kinase C (PKC) and is known to affect a variety of biochemical processes in human breast cancer cells. In the present study we have employed MCF-7 cells to investigate the effects of TPA on inositol lipid signalling, the putative pathway leading to PKC activation and intracellular Ca2+ mobilization. Phosphoinositide hydrolysis in MCF-7 cells was stimulated by bombesin (BN) as evidenced by increases in both inositol phosphate production and cytidine diphosphate diacylglycerol (CDP-DG) accumulation. Pretreatment of MCF-7 cells with TPA caused attenuation of both these BN-induced responses. This inhibitory action of TPA on inositol phosphate production was mimicked by diacylglycerol analogues and was reversed by staurosporine, H-7 and tamoxifen, all known inhibitors of PKC. Furthermore, putative down-regulation of PKC by prolonged TPA pretreatment also reversed the inhibitory action of TPA and enhanced BN-induced phosphoinositide hydrolysis. TPA also inhibited BN-induced increases in cytosolic Ca2+ concentration ([Ca2+]i) and caused a dose-dependent inhibition of epidermal growth factor (EGF) binding in MCF-7 cells. However, EGF receptor occupancy was unaffected by BN. These data support an inhibitory role for PKC in the regulation of phosphoinositide hydrolysis and [Ca2+]i in breast cancer cells and provide a potential mechanism for feedback regulation of this signalling pathway in these cells.


Effectiveness of pacemaker tele-monitoring on quality of life, functional capacity, event detection and workload: The PONIENTE trial.
The purpose of the present study was to assess the effectiveness of the remote monitoring (RM) of older adults with pacemakers on health-related quality of life, functional capacity, feasibility, reliability and safety. The PONIENTE study is a controlled, non-randomized, non-blinded clinical trial, with data collection carried out during the pre-implant stage and after 12 months. Between October of 2012 and November of 2013, 82 patients were assigned to either a remote monitoring group (n = 30) or a conventional hospital monitoring (HM) group (n = 52). The EuroQol-5D (EQ-5D) and the Duke Activity Status Index were used to measure health-related quality of life and functional capacity, respectively. Baseline characteristics and number of hospital visits were also analyzed. The baseline characteristics of the two study groups were similar for both the EQ-5D (RM 0.74, HM 0.67; P = 0.404) and the Duke Activity Status Index (RM 21.42, HM 19.95; P = 0.272). At the 12-month follow up, the EQ-5D utility score was improved for both groups (RM 0.91, HM 0.81; P = 0.154), unlike the EQ-5D Visual Analog Scale (P = 0.043). The Duke Activity Status Index score was similar to the baseline score. The number of in-hospital visits was 27% lower (3 vs 4; P < 0.001) in the remote group as compared with the hospital group. The PONIENTE trial suggests that the remote monitoring of pacemakers in older adults is an equivalent option to hospital monitoring, in terms of health-related quality of life and functional capacity. Furthermore, it allows for the early detection of clinical and pacemaker-related adverse events, and significantly reduces the number of in-hospital visits. Geriatr Gerontol Int 2016; 16: 1188-1195.


Final 4-year results of the RAINBOW real-world study: intravitreal aflibercept dosing regimens in France in treatment-naïve patients with neovascular age-related macular degeneration.
The purpose of this study is to evaluate real-world treatment outcomes in patients with neovascular age-related macular degeneration (nAMD) treated with intravitreal aflibercept (IVT-AFL) in routine clinical practice in France. RAINBOW (NCT02279537) was an ambispective, observational, 4-year study assessing IVT-AFL effectiveness, treatment patterns, and safety in patients with nAMD in France. Treatment-naïve patients prescribed IVT-AFL and treated according to local practice (pro re nata or treat-and-extend) were eligible. Three treatment cohorts were retrospectively identified based on their treatment pattern within the first 12 months: regular (3 initial monthly IVT-AFL injections received within 45-90 days after the first injection in month 0 and followed by injections every 2 months), irregular with the initial monthly injections, and irregular without the initial monthly injections. The primary endpoint was mean change in best-corrected visual acuity (BCVA) from baseline to month 12. The 48-month results are described here. Overall, the study included 516 patients (each with one study eye), and 30.2% of patients completed 48 months of IVT-AFL treatment. Mean change in BCVA from baseline (56.5 letters) to month 48 for patients with an assessment at both time points was + 1.1 (regular cohort, n = 47), + 0.1 (irregular cohort with initial monthly injections, n = 115), and - 1.3 letters (irregular cohort without initial monthly injections, n = 26), representing a decrease from the gains achieved at month 12. Mean number of IVT-AFL injections received by month 48 in the treatment cohorts was 14.9, 13.7, and 11.9, respectively. The safety profile of IVT-AFL was consistent with previous studies. In RAINBOW, the 48-month results demonstrate a lack of long-term effectiveness of IVT-AFL treatment of nAMD due to progressive undertreatment in routine clinical practice in France. These real-world findings highlight the importance of 3 initial monthly IVT-AFL injections followed by continuous proactive treatment beyond the first year to achieve optimal functional outcomes. ClinicalTrials.gov Identifier: NCT02279537.


Effect of adrenaline and phorbol myristate acetate or bacterial lipopolysaccharide on stimulation of pathways of macrophage glucose, glutamine and O2 metabolism. Evidence for cyclic AMP-dependent protein kinase mediated inhibition of glucose-6-phosphate dehydrogenase and activation of NADP+-dependent 'malic' enzyme.
Adrenaline has recently been shown to stimulate both glucose metabolism and H2O2 release by macrophages but the activity of the key pentose phosphate pathway enzyme, glucose-6-phosphate dehydrogenase (which generates the NADPH crucial for the reduction of molecular oxygen), was reduced under these conditions [Costa Rosa, Safi, Cury and Curi (1992) Biochem. Pharmacol. 44, 2235-2241]. We report here that adrenaline activates another NADPH-producing enzyme, NADP(+)-dependent 'malic' enzyme, while also inhibiting glucose-6-phosphate dehydrogenase, via cyclic AMP-dependent protein kinase (PKA) activation. Regulation of glucose-6-phosphate dehydrogenase activity by PKA has not been reported elsewhere. The sparing of some glucose from pentose phosphate pathway consumption may be important in the provision of glycerol 3-phosphate which in the macrophage may be required for new phospholipid synthesis. Glutamine oxidation was also stimulated by adrenaline thus providing increased substrate (malate) for NADP(+)-dependent 'malic' enzyme and therefore shifting some of the burden of NADPH production from glucose to glutamine metabolism. We also report a novel synergistic effect of adrenaline and some bacterial products and/or gamma-interferon in stimulating secretory and metabolic pathways in macrophages which may be a part of a larger network of signals that lead to enhanced macrophage activity.


The conformation of myosin head domains in rigor muscle determined by X-ray interference.
In the absence of adenosine triphosphate, the head domains of myosin cross-bridges in muscle bind to actin filaments in a rigor conformation that is expected to mimic that following the working stroke during active contraction. We used x-ray interference between the two head arrays in opposite halves of each myosin filament to determine the rigor head conformation in single fibers from frog skeletal muscle. During isometric contraction (force T(0)), the interference effect splits the M3 x-ray reflection from the axial repeat of the heads into two peaks with relative intensity (higher angle/lower angle peak) 0.76. In demembranated fibers in rigor at low force (<0.05 T(0)), the relative intensity was 4.0, showing that the center of mass of the heads had moved 4.5 nm closer to the midpoint of the myosin filament. When rigor fibers were stretched, increasing the force to 0.55 T(0), the heads' center of mass moved back by 1.1-1.6 nm. These motions can be explained by tilting of the light chain domain of the head so that the mean angle between the Cys(707)-Lys(843) vector and the filament axis increases by approximately 36 degrees between isometric contraction and low-force rigor, and decreases by 7-10 degrees when the rigor fiber is stretched to 0.55 T(0).


The use of PVP as a polymeric carrier to improve the plasma half-life of drugs.
To achieve an optimum drug delivery such as targeting or controlled release utilizing bioconjugation with polymeric modifier, the conjugate between drugs and polymeric modifiers must be designed to show desirable pharmacokinetic characteristics in vivo. In this study, we assessed the biopharmaceutical properties of various nonionic water-soluble polymers as polymeric drug carriers. Polyvinylpyrrolidone (PVP) showed the longest mean resident time (MRT) after i.v. injection of all nonionic polymers with the same molecular size. In fact, tumor necrosis factor-alpha (TNF-alpha) bioconjugated with PVP (PVP-TNF-alpha) circulated longer than TNF-alpha bioconjugated with polyethylene glycol (PEG-TNF-alpha) with the same molecular size. Each nonionic polymeric modifier showed a different tissue distribution. Dextran was accumulated in the spleen and liver. Polydimethylacrylamide (PDAAm) tended to distribute in the kidney. However, PVP showed the minimum volume of tissue distribution. These results suggested that PVP is the most suitable polymeric modifier for prolonging the circulation lifetime of a drug and localizing the conjugated drug in blood.


[Possible role of ammonium sulfate in inducing invertase biosynthesis in plants].
The induction of invertase from beetroot plants and wheat shoots was studied using the "ageing tissues" technique. It was found that ammonium sulfate at concentration 0.001 M enhances invertase induction. This effect of ammonium sulfate was not revealed when the preparations were pretreated by actidione, puromycin and chloramphenicol. A possibility of activation of invertase induction by ammonium sulfate is discussed.


Effect of large pore size of multifunctional mesoporous microsphere on removal of heavy metal ions.
Pore size of mesoporous materials is crucial for their surface grafting. This article develops a novel multifunctional microsphere with a large pore size mesoporous silica shell (ca. 10.3 nm) and a magnetic core (Fe₃O₄), which is fabricated using cetyltrimethylammonium bromide (CTAB) as pore-forming agents, tetraethyl orthosilicate (TEOS) as silicon source through a sol-gel process. Compared with small pore size mesoporous silica magnetic microspheres (ca. 2-4 nm), the large pore size one can graft 447 mg/g amino groups in order to adsorb more heavy metal ions (Pb(2+): 880.6 mg/g, Cu(2+): 628.3mg/g, Cd(2+): 492.4 mg/g). The metal-loaded multifunctional microspheres could be easily removed from aqueous solution by magnetic separation and regenerated easily by acid treatment. The results suggest that the large pore size multifunctional microspheres are potentially useful materials for high effectively adsorbing and removing different heavy metal ions in aqueous solution.


From the Hawaiian classroom to the wards: A model of effective pedagogy for clinicians, trainees, and patients.
Clinical teachers receive little training in the actual practice of teaching, often lacking any pedagogical framework by which to approach learners. The Center for Research in Education, Diversity, and Excellence (CREDE) offers an evidence-based pedagogical framework from our experience in primary and secondary education that can transform the clinician's teaching practice. The CREDE standards of effective pedagogy provide concrete guidance for a clinician's interactions with trainees, peers, and patients. This framework can support clinical teachers to be more effective when working with the diverse set of trainees, peers, and patients that our fields must engage.


Effect of statins on proteolytic activity in the wall of abdominal aortic aneurysms.
The aim of this study was to examine the effect of statin treatment on the activity of proteases in the wall of abdominal aortic aneurysms (AAAs). The activities of matrix metalloproteinases (MMPs) 9 and 3, cathepsins B, H, K, L and S, and the cystatin C level were measured in extracts of AAA wall taken from 82 patients undergoing AAA repair; 21 patients were receiving statin treatment before surgery. All values were standardized against soluble protein (SP) concentration in the extract, and reported as median (interquartile range) or mean(s.e.m.). The two groups had similar demographics. Reduced activity of MMP-9 (43 (34-56) versus 80 (62-110) pg per mg SP; P < 0.001), cathepsin H (183 (117-366) versus 321 (172-644) nmol 4-methylcoumarin-7-amide released per mg SP; P = 0.016) and cathepsin L (102 (51-372) versus 287 (112-816) micromol 7-amino-4-trifluoromethylcoumarin released per mg SP; P = 0.020) was found in the statin-treated aortas compared with AAAs from patients not taking a statin. The statin-treated group had lower MMP-3 activity, but this did not reach statistical significance (P = 0.053). Cystatin C levels were higher in statin-treated aortas than in controls (41.3(3.1) versus 28.9(2.1) ng per mg SP; P = 0.003). Statins decreased the activity of proteases that have been implicated in aneurysm disease.


General lymphangiomatosis in a child.
The lymphographic findings in a 4 year old girl with general lymphangiomatosis are presented. The number of iliac and lumbar lymphatics had increased markedly and the lymph nodes were enlarged with very loose structure and with contrast medium distributed in scattered droplets and filiform channels.


The major histocompatibility complex (CyLA) of the cynomolgus monkey. I. Serologic definition of 21 specificities.
Thirty-seven lymphocytotoxic antisera, 27 of which were raised by immunization with skin grafts and blood from partially matched donors, were tested against cells obtained from 218 unrelated animals and 205 offspring from a colony of cynomolgus monkey (Macaca fascicularis). Evidence was obtained for the presence of at least 21 specificities defined by cluster analysis and segregation within families. Allelic relationships between 16 specificities was suggested by segregation patterns, the absence of triplets and statistical analysis of association in the unrelated population sample. The data support a two-locus model, with tentative assignment of seven specificities to the A locus and six to the B locus. That these lymphocyte alloantigens constitute the major histocompatibility complex (MHC) of the cynomolgus monkey is suggested by analogy with other known MHCs and by the increased survival times of skin grafts between paternally matched half sibs compared to haplodistinct full sibs.


Two-Parameter Mobility Assessments Discriminate Diverse Regulatory Factor Behaviors in Chromatin.
Enzymatic probes of chromatin structure reveal accessible versus inaccessible chromatin states, while super-resolution microscopy reveals a continuum of chromatin compaction states. Characterizing histone H2B movements by single-molecule tracking (SMT), we resolved chromatin domains ranging from low to high mobility and displaying different subnuclear localizations patterns. Heterochromatin constituents correlated with the lowest mobility chromatin, whereas transcription factors varied widely with regard to their respective mobility with low- or high-mobility chromatin. Pioneer transcription factors, which bind nucleosomes, can access the low-mobility chromatin domains, whereas weak or non-nucleosome binding factors are excluded from the domains and enriched in higher mobility domains. Nonspecific DNA and nucleosome binding accounted for most of the low mobility of strong nucleosome interactor FOXA1. Our analysis shows how the parameters of the mobility of chromatin-bound factors, but not their diffusion behaviors or SMT-residence times within chromatin, distinguish functional characteristics of different chromatin-interacting proteins.


Specific nasal symptoms and symptom-provoking factors may predict increased risk of developing COPD.
In a 1992 questionnaire study, we found that certain nasal symptoms and symptom-provoking factors were associated with prevalence of self-reported chronic bronchitis/emphysema (CBE). In this follow-up study, we examined whether any nasal features could predict an increased incidence of self-reported physician's diagnosis of CBE/chronic obstructive pulmonary disease (COPD). In 2000, a survey was performed similar to the one in 1992. Of a paired follow-up group of 4933 participants aged 28-67 years, 4280 (86.8%) returned the questionnaire. Odds ratios (ORs) for cumulative incidence (between 1992 and 2000) of self-reported physician-diagnosed CBE/COPD and asthma, respectively, were calculated by logistic regression with adjustment for age, gender and smoking habits. Reports of thick, yellow nasal discharge and nasal blockage in 1992 predicted incidence of CBE/COPD: OR 2.3 (1.2-4.2) and 1.8 (1.1-2.8) respectively. Moreover, nasal symptoms provoked by exposure to damp/cold air and tobacco smoke predicted CBE/COPD: OR 3.4 (1.9-6.0) and 2.5 (1.4-4.2). Nasal itching and nasal symptoms provoked by exposure to grass pollen and furred animals predicted incidence of asthma. These results suggest that certain nasal symptoms and nasal symptom-provoking exposures, different from those commonly associated with asthma, may predict increased risk of developing CBE/COPD. This supports the possibility of nasal co-morbidity in COPD.


Dopamine receptors on isolated membranes of rat lymphocytes.
Using [3H]-dopamine (DA) as the ligand of choice, we have shown that rat lymphocytes possess DA binding sites. The binding of [3H]-DA was specific, saturable, and of high affinity. The specific binding in membranes obtained from Con A-activated lymphoblasts was greater than the binding in respective unactivated lymphoid cells, and displaced by lower concentrations of unlabeled ligand. [3H]-DA was displaced specifically by DA, norepinephrine, and apormorphine. The binding sites were insensitive to neuroleptic drugs such as sulpiride, bromocryptine, and haloperidol. On the basis of these findings, we have suggested that lymphocytes possess a receptor site for catecholamines and the receptor for DA might be a D-3 receptor type.


Extracellular vesicles: Pharmacological modulators of the peripheral and central signals governing obesity.
Obesity and its metabolic resultant dysfunctions such as insulin resistance, hyperglycemia, dyslipidemia and hypertension, grouped as the "metabolic syndrome", are chronic inflammatory disorders that represent one of the most severe epidemic health problems. The imbalance between energy intake and expenditure, leading to an excess of body fat and an increase of cardiovascular and diabetes risks, is regulated by the interaction between central nervous system (CNS) and peripheral signals in order to regulate behavior and finally, the metabolism of peripheral organs. At present, pharmacological treatment of obesity comprises actions in both CNS and peripheral organs. In the last decades, the extracellular vesicles have emerged as participants in many pathophysiological regulation processes. Whether used as biomarkers, targets or even tools, extracellular vesicles provided some promising effects in the treatment of a large variety of diseases. Extracellular vesicles are released by cells from the plasma membrane (microvesicles) or from multivesicular bodies (exosomes) and contain lipids, proteins and nucleic acids, such as DNA, protein coding, and non-coding RNAs. Owing to their composition, extracellular vesicles can (i) activate receptors at the target cell and then, the subsequent intracellular pathway associated to the specific receptor; (ii) transfer molecules to the target cells and thereby change their phenotype and (iii) be used as shuttle of drugs and, thus, to carry specific molecules towards specific cells. Herein, we review the impact of extracellular vesicles in modulating the central and peripheral signals governing obesity.


Chest-wall collapse as a complication of atelectasis.
A case of chest-wall collapse following atelectasis of the lung is reported. The presumed mechanism is abnormal compliance of the chest wall.


Colorimetric detection of L-histidine based on the target-triggered self-cleavage of swing-structured DNA duplex-induced aggregation of gold nanoparticles.
A rapid, highly sensitive and selective colorimetric assay is presented for visually detecting L-histidine. It is based on L-histidine-triggered self-cleavage of DNA duplex-induced gold nanoparticle (AuNP) aggregation. The citrate-capped AuNPs easily aggregate in a high concentration of salt environment. However, in the presence of L-histidine aptamers (DNA1 and DNA2), the partial strands of DNA1 and DNA2 hybridize to form a DNA duplex with a swing structure. The swing-like DNA duplexes are adsorbed on the surface of AuNPs to improve the stability of AuNPs, and the AuNPs also are better dispersed in high-salt media. When L-histidine is added to the solutions, it catalyzes the self-cleavage of DNA1 to form many single-stranded DNA (ssDNA) fragments. These ssDNA segments are adsorbed on the AuNPs and weaken the stability of AuNPs. Hence, the AuNPs aggregate in high-salt environment, and this results in a red-to-blue color change. Under the optimized conditions, L-histidine can be determined with a limit of detection of 3.6 nM. In addition, the sensor was successfully applied to the determination of L-histidine in spiked serum samples. Graphical abstract Schematic of a rapid and homogeneous colorimetric L-histidine assay. It combines L-histidine-triggered self-cleavage of the swing-like DNA duplexes and self-cleavage of DNA-induced AuNP aggregation.


Zellweger Spectrum Disorder: Ophthalmic Findings from a New Natural History Study Cohort and Scoping Literature Review.
Individuals with Zellweger spectrum disorder (ZSD) manifest a spectrum of clinical phenotypes but almost all have retinal degeneration leading to blindness. The onset, extent, and progression of retinal findings have not been well described. It is crucial to understand the natural history of vision loss in ZSD to define reliable endpoints for future interventional trials. Herein, we describe ophthalmic findings in the largest number of ZSD patients to date. Retrospective review of longitudinal data from medical charts and review of cross-sectional data from the literature. Sixty-six patients with ZSD in the retrospective cohort and 119 patients reported in the literature, divided into 4 disease phenotypes based on genotype or clinical severity. We reviewed ophthalmology records collected from the retrospective cohort (Clinicaltrials.gov NCT01668186) and performed a scoping review of the literature for ophthalmic findings in patients with ZSD. We extracted available ophthalmic data and analyzed by age and disease severity. Visual acuity (VA), posterior and anterior segment descriptions, nystagmus, refraction, electroretinography findings, visual evoked potentials, and OCT results and images. Visual acuity was worse at younger ages in those with severe disease compared with older patients with intermediate to mild disease for all 78 participants analyzed, with a median VA of 0.93 logarithm of the minimum angle of resolution (Snellen 20/320). Longitudinal VA data revealed slow loss over time and legal blindness onset at an average age of 7.8 years. Funduscopy showed retinal pigmentation, macular abnormalities, small or pale optic discs, and attenuated vessels with higher prevalence in milder severity groups and did not change with age. Electroretinography waveforms were diminished in 91% of patients, 46% of which were extinguished and did not change with age. OCT in milder patients revealed schitic changes in 18 of 23 individuals (age range 1.8 to 30 years), with evolution or stable macular edema. In ZSD, VA slowly deteriorates and is associated with disease severity, serial electroretinography is not useful for documenting vision loss progression, and intraretinal schitic changes may be common. Multiple systematic measures are required to assess retinal dystrophy accurately in ZSD, including functional vision measures. Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.


[Characterization of a closed-set logatome test : Documentation of audiometric data: discrimination function and reproducibility].
Logatomes, nonsensical combinations of consonants and vowels, are suitable for a precise capture and analysis of individual phonemes as fundamental modules of speech in audiometric diagnostics. The aim of this prospective study was to capture the audiometric characteristics of a closed-set logatome test. The slope of the discrimination function at the speech reception threshold (SRT) and the reproducibility were analyzed. A set of 102 intensity varied and randomized logatomes were presented in the form of consonant-vowel-consonant to 25 hearing unimpaired adults. The measurements were performed in a free field setting and were each repeated after a 2-week interval. The subjects were requested to repeat the heard logatome in a closed response test of 10 items per sound item on a touchscreen. The slope of the mean discrimination function at the SRT was on average 4%/dB; however, the mean discrimination function slope was steeper for the initial consonant than for the final one. The differences of the test and retest results at the SRT showed a standard deviation of 13% for consonants. These differences were normally distributed. There were no significant differences between test and retest. The slope of the discrimination function at the SRT appeared to be shallow but was comparable to established word tests. Finally, there was no evidence of a learning effect in the retest, which emphasizes the low redundancy of the speech material and makes it an attractive complementary option to routine audiometric diagnostics.


Treatment of Patients with Distant Metastases from Phyllodes Tumor of the Breast.
Here, the treatment methods and results of patients with phyllodes tumor of the breast (PT) with distant metastases at a single institution are presented. A retrospective analysis was performed on a group of 295 patients with PT treated from 1952 to 2010. Distant metastases developed in 37 (12.5 %) patients; 3/160 (1.9 %) patients had benign PT, 6/36 (16.7 %) were considered borderline, and 28/99 (28.3 %) had malignant PT. Most frequently, the metastases were located in the lungs; 28 (75.7 %), bone 7 (18.9 %), brain 4 (10.8 %), and liver 2 (5.4 %). Metastases occurred on overage 21 months (2-57) after surgery. Patients with lung metastases were generally treated with monochemotherapy or polychemotherapy. In one patient Testosterone and in two patients resection of metastases combined with Doxorubicin were used. Patients with bones or brain metastases were treated with palliative radiotherapy only or combined with Doxorubicin. The mean survival (MS) from diagnosis of distant metastases (DM) was 7 months (2-17). The longest mean survival in patients with bones metastases was 11.8 months, the worst survival was for patients with brain metastases--2.8 months. Hormone therapy appeared to have low efficacy (MS: 2 months) as well as monochemotherapy (MS: 3-5 months). Improved MS was obtained using Doxorubicin (7 months) and Doxorubicin with Cisplatin, Cyclophosphamide, or Ifosfamide (9 months). The prognosis of patients with DM from PT is poor. The role of surgery and irradiation of such patients is very limited. There appears to be no role for the use of hormone therapy. This study showed that polychemotherapy with Doxorubicin and Ifosfamide suggest that it might be more effective than once thought.


A chromatin remodeling checkpoint of diet-induced macrophage activation in adipose tissue.
The interplay between the environment and the immune cells is linked to metabolic homeostasis under physiologic and pathophysiologic conditions. Diabetes mellitus type 2 (T2D) is considered an immune-related inflammatory disorder, in which the adipose tissue macrophages (ATMs) are key players orchestrating metabolic chronic meta-inflammation and contributing to the pathogenesis of metabolic disease. However, the molecular regulators that integrate the environmental signals to control ATM activation and adipose inflammation during obesity and T2D remain unclear. Epigenetic mechanisms constitute important parameters in metabolic homeostasis, obesity and T2D via the integration of the environmental factors to the transcriptional regulation of gene programs. In a very recent study published in Diabetes by Kong et al., BAF60a has been identified as a key chromatin remodeling checkpoint factor that associates obesity-associated stress signals with meta-inflammation and systemic homeostasis. Furthermore, this work uncovers Atf3 as an important downstream effector in BAF60a-mediated chromatin remodeling and transcriptional reprogramming of macrophage activation in adipose tissue. The findings of this research may contribute to the development of new therapeutic approaches for obesity-induced metabolic inflammation and associated metabolic disorders.


Transmission of Mycobacterium tuberculosis in a family proved by genotyping.
Molecular genetic methods have been applied in various epidemiologic studies including investigations of disease acquisition by contact. This report describes the use of various molecular genetic methods in tracing possible household transmission of tuberculosis by contact. Four Mycobacterium tuberculosis strains, each from four members of a family, were first isolated and identified in the clinical laboratory of the Chest Hospital and were submitted to the National Reference Laboratory of Mycobacteriology for further confirmation and genotyping. In this study, IS6110 restriction fragment length polymorphism (RFLP), spacer oligonucleotide typing (spoligotyping) and mycobacterial interspersed repetitive units-variable number tandem repeats (MIRU-VNTR), and rpoB gene sequencing were used for genotyping. All four strains were found to have identical spoligotypes, MIRU-VNTR patterns, and similar IS6110 RFLP profiles. The results of the drug susceptibility test and of rpoB sequencing showed that all four strains were rifampicin resistant. Household transmission through close contact was thus proved by genotyping. We conclude that all four family members were infected with the same lineage of M. tuberculosis.


Effects of electroacupuncture on cold allodynia in a rat model of neuropathic pain: mediation by spinal adrenergic and serotonergic receptors.
The present study was performed to examine the effects of electroacupuncture (EA) on cold allodynia and its mechanisms related to the spinal adrenergic and serotonergic systems in a rat model of neuropathic pain. For the neuropathic surgery, the right superior caudal trunk was resected at the level between S1 and S2 spinal nerves innervating the tail. Two weeks after the nerve injury, EA stimulation (2 or 100 Hz) was delivered to Zusanli (ST36) for 30 min. The behavioral signs of cold allodynia were evaluated by the tail immersion test [i.e., immersing the tail in cold water (4 degrees C) and measuring the latency to an abrupt tail movement] before and after the stimulation. And then, we examined the effects of intrathecal injection of prazosin (alpha1-adrenoceptor antagonist, 30 microg), yohimbine (alpha2-adrenoceptor antagonist, 30 microg), NAN-190 (5-HT1A antagonist, 15 microg), ketanserin (5-HT2A antagonist, 30 microg), and MDL-72222 (5-HT3 antagonist, 12 microg) on the action of EA stimulation. Although both 2 Hz and 100 Hz EA significantly relieved the cold allodynia signs, 2 Hz EA induced more robust effects than 100 Hz EA. In addition, intrathecal injection of yohimbine, NAN-190, and MDL-72222, but not prazosin and ketanserin, significantly blocked the relieving effects of 2 Hz EA on cold allodynia. These results suggest that low-frequency (2 Hz) EA is more suitable for the treatment of cold allodynia than high-frequency (100 Hz) EA, and spinal alpha2-adrenergic, 5-HT1A and 5-HT3, but not alpha1-adrenergic and 5-HT2A, receptors play important roles in mediating the relieving effects of 2 Hz EA on cold allodynia in neuropathic rats.


Colony assay with human tumor xenografts, murine tumors and human bone marrow. Potential for anticancer drug development.
The colony formation of human tumor xenografts from nude mice, of murine tumors, and of human bone marrow (CFU-C) has been investigated in vitro using a modification of the double-layer agar assay described by Hamburger and Salmon. Systematic modification of growth conditions and careful selection of viable tumor tissue enhanced the growth rate (at least 30 colonies per dish) of human tumor xenografts to 86% (98/114). The median plating efficiency was 0.07% which is comparable to the results observed by others using fresh human tumors. The growth of human bone marrow was stimulated with a placenta-conditioned medium, which allowed growth of granulocytic stem cell colonies (CFU-C). The median plating efficiency of the bone marrow was 0.08%. The murine tumors P388, L1210, B16 melanoma, Lewis lung carcinoma and colon carcinoma 38 grew very well in vitro. Excluding the Lewis lung carcinoma, the plating efficiency of these tumors was markedly higher than that of the human tumor xenografts and human bone marrow. The colony assay may have potential as a secondary screening system for identifying new active structures and also for indicating which tumor types are most responsive to a new antitumor agent. We test new structures in 20 well-selected human tumor xenografts and in the P388 mouse leukemia in dose-response relationships. The two most responsive xenograft tumors are subsequently studied in vivo in nude mice in order to determine if a new compound presents antitumor activity in an in vivo organism at a dose around the LD10 level. If a remission or at least no change is observed in the subcutaneously growing tumor, the new compound undergoes large disease-oriented testing usually in 60 xenografts. The in vivo studies are necessary in determining whether a compound has a more specific effect on tumor cells than on the dose-limiting normal tissue. The comparison of in vitro/in vivo activity allows an assessment of the relevant in vitro dose based on in vivo pharmacological behavior of a drug. It seems justifiable to apply the conclusions of this approach to the clinical setting because mouse toxicity data, e.g. the LD10, correspond well to the maximal tolerable doses in man. Moreover, for compounds whose dose-limiting toxicity is bone marrow suppression, the comparison of drug dosages effective in vitro on human bone marrow and tumor xenografts may prove helpful. The proposed testing strategy has been applied to TGU and Tiazofurin. At the relevant dosages TGU exhibited very limited activity in 67 human tumor xenografts studied.(ABSTRACT TRUNCATED AT 400 WORDS)