Total joint replacement in HIV positive patients.
Recent HIV therapies have improved life expectancy in HIV positive patients. For the purpose of the following retrospective investigation, we analyzed the results of total joint replacement in HIV positive patients. This study exemplifies orthopaedic treatment options and perioperative problems in HIV positive patients. Our population included a high proportion of hemophilic patients. Between 1988 and 2000, we performed 55 endoprosthetic procedures (20 total hip replacements (THR), 33 total knee replacements (TKR), two shoulder replacements) in 41 patients suffering form HIV. Thirty patients are afflicted with hemophilia, seven patients were intravenous drug addicts. The mean follow-up was 81 months (2-14) years. Patients were seen annually; either the Harris Hip Score or the Knee Society Rating System was applied. The following septic complications were observed: a mycotic abscess of both hips 5/10 months after bilateral THR, two early infections following coxitis in patients with intravenous drug abuse, and one further case of septic loosening after 15 months in one patient after THR. Furthermore, one aseptic loosening of a THR after 14 months in a hemophilic patient was seen. After TKR, two early infections in patients with intravenous drug addiction were seen. The total complication rate was 12.7%. A coherency between the infection rate and the CD4+ count was not seen. An analysis of the results shows that the complications occurred in patients living under difficult social circumstances. Whereas total joint replacement in hemophilic patients with or without HIV seems to be a fairly safe procedure concerning the postoperative infection rate, intravenous drug abuse increases the risk. Functional outcome does not differ from an HIV negative population both in the TKR and THR groups.


Expression of TEM-induced damage to postmeiotic stages of spermatogenesis of the mouse during early embryogenesis. II. Cytological investigations.
After treatment of postmeiotic stages of spermatogenesis of the mouse with TEM (0.2 or 0.4 mg/kg), dose- and stage-of-spermatogenesis-dependent frequencies of cytogenetic aberrations can be observed in early embryos. The frequencies of first-cleavage metaphases exhibiting structural aberrations (all of the chromosome type), the frequencies of 2 cell embryos and of 4--8 cell embryos containing nuclei accompanied by micronuclei or nuclei connected by bridges show a close correlation to frequencies of preimplantation loss of embryos recorded in a dominant lethal test. The frequencies of morulae/blastulae exhibiting blastomeres with micronuclei show a close correlation to the frequencies of total (pre- and post-implantation) loss of embryos. This indicates delayed expression of TEM-induced chromosomal damage which could persist undetected in very early stages of embryogenesis.


The plasma membrane Ca2+ ATPase of animal cells: structure, function and regulation.
Most important processes in cell life are regulated by calcium (Ca2+). A number of mechanisms have thus been developed to maintain the concentration of free Ca2+ inside cells at the level (100-200nM) necessary for the optimal operation of the targets of its regulatory function. The systems that move Ca2+ back and forth across membranes are important actors in its control. The plasma membrane calcium ATPase (PMCA pump) which ejects Ca2+ from all eukaryotic cell types will be the topic of this contribution. The pump uses a molecule of ATP to transport one molecule of Ca2+ from the cytosol to the external environment. It is a P-type ATPase encoded by four genes (ATP2B1-4), the transcripts of which undergo different types of alternative splicing. Many pump variants thus exist. Their multiplicity is best explained by the specific Ca2+ demands in different cell types. In keeping with these demands, the isoforms are differently expressed in tissues and cell types and have differential Ca2+ extruding properties. At very low Ca2+ concentrations the PMCAs are nearly inactive. They must be activated by calmodulin, by acid phospholipids, by protein kinases, and by other means, e.g., a dimerization process. Other proteins interact with the PMCAs (i.e., MAGUK and NHERF at the PDZ domain and calcineurin A in the main intracellular domain) to sort them to specific regions of the cell membrane or to regulate their function. In some cases the interaction is isoform, or even splice variant specific. PMCAs knock out (KO) mice have been generated and have contributed information on the importance of PMCAs to cells and organisms. So far, only one human genetic disease, hearing loss, has been traced back to a PMCA defect.


[Car driving and psychiatry].
Among the specialties involved in the order of 31 August 2010, psychiatry is in Chapter IV alongside addictive behavior and drug use may impair the ability of the driver. As well as for personal vehicles for professional vehicles the incompatibility of health with driving exists when clinical factors can interfere with the skills required of the driver. There would simply absolute incompatibility for psychoses in active phase. In the other phases of psychosis is at the discretion of specialist as for illiteracy or social maladjustment. The role of the authorized psychiatrist is therefore always subjective. This article also makes room for attention-deficit disorder with hyperactivity (ADHD), not listed, but the subject of numerous articles in the English literature.


[Sleep and circadian rhythms in ageing].
With age, the consolidation of nocturnal sleep decreases, daytime napping increases, and sleep occurs earlier. Sleep regulation is dependent on the interaction between a circadian pacemaker (biological clock) and the sleep homeostat (sleep pressure increasing with duration of time awake). We have shown that in the healthy elderly, the amplitude of circadian rhythms (e. g. melatonin secretion) declines, as does slow wave sleep, parallel with an increase in afternoon sleepiness and a tendency to fall asleep in the early evening when younger subjects do not. Light is the major zeitgeber to stabilise the biological clock: older subjects require sufficient light exposure during daytime and in the evening, and should take no or only brief naps during the day to improve sleep.


Sequence of centromere separation: orderly separation of multicentric chromosomes in mouse L cells.
Mouse L cells have many dicentric chromosomes and one with eight centromeres. All eight centromeres behave similarly until midmetaphase when most centromeres split into two units each in apparently quick succession but out-of-phase. This premature separation leaves one or perhaps two closely located centromeres intact, which separate at late metaphase-anaphase, drawing the two chromatids to opposite poles. Such dominance of one centromere over all others, though unexplained, ensures the lack of any mitotic abnormality such as bridges or fragments. These observations show that all the centromeres are retained as functional primary constrictions except for a change in functional regulation when more than one centromere are located on a chromosome.


Activity coefficients of salts in highly concentrated protein solutions. I. Alkali chlorides in isoionic bovine serum albumin solutions.
In order to understand the thermodynamic state of simple salts in living cells, the mean activity coefficients of LiCl, NaCl, KCl, RbCl, CsCl were determined in concentrated isoionic bovine serum albumin (BSA) solutions by use of the EMF method with ion exchange membrane electrodes. The protein concentration range extended up to 22 wt%, whereas the salt concentration was kept constant at 0.1 mole per kilogram water. These solutions may be regarded as crude but appropriate model systems for the cytoplasm of cells as far as type and magnitude of the macromolecular component influence on the chemical potential of the salts is concerned. The mean stoichiometric activity coefficients of the alkali chlorides in the isoionic BSA solutions decreased linearly with the protein molality; this decrease, however, did not exceed ca. 10% compared with the pure 0.1 molal salt solutions. Only very small differences in the behavior of the different alkali chlorides were observed. The results may be interpreted by the superposition of the effects of specific Cl- ion binding to BSA and BSA bound "non-solvent" water with probably electrostatic long range interactions of the BSA(Cl-)nu polyions with the salt ions in solution. The resulting mean activity coefficients, corrected for ion binding and non-solvent water, showed a very slight linear dependence on the protein concentration. The departure from the value in the pure 0.1 molal salt solutions did not exceed +/- 2%.


Need for Systematic Magnetic Resonance Imaging Interpretation and Reporting after Partial Prostate Gland Ablation.
Interpretation guidelines for prostate multiparametric magnetic resonance imaging findings in the post-ablation setting are not available yet; this poses a significant challenge for accurate risk stratification in this clinical setting. Here we propose the development and implementation of a post-ablation Prostate Imaging-Reporting and Data System that would improve patient surveillance and management after prostate ablation for localized prostate cancer.


Optimization of medium for enhancement of ε-poly-L-lysine production by Streptomyces sp. M-Z18 with glycerol as carbon source.
The goal of this study was to increase ε-Poly-L-Lysine (ε-PL) production by Streptomyces sp. M-Z18 through optimization of the culture medium. Glycerol, (NH4)2SO4 and K2HPO4 were the key factors in the fermentation according to the results obtained from the Plackett-Burman experimental design. The optimal concentration of the three factors was further investigated according to the central composite design and determined to be 59.48, 4.43 and -0.49 g/L, respectively. Under optimal conditions, a ε-PL concentration of 2.27±0.15 g/L was obtained. Compared with glucose, fermentation of glycerol afforded 2.8-fold and 6.3-fold the ε-PL titer and yield, respectively. Therefore, glycerol may be an efficient carbon source for ε-PL production by Streptomyces sp. M-Z18.


Innovative programming in school mental health. Search for a methodology.
Entering a system it often regarded as the most difficult aspect of consultation, especially when entry is initiated by the consultant. Issues involved in seeking entry into a school system are considered from the perspective of a university-based consultant interested in developing, delivering, and evaluating innovative approaches to school mental health problems. Experience with a number of school systems leads to several conclusions about the entry process. Foremost is the existence of a difficult, time-consuming period of negotiations involving various school personnel; because of their varying perspectives and different interests, problems emerge and must be resolved successfully in the course of negotiations if a consultation project is to develop. Often this is not possible, and it is therefore suggested that the consultant seeking a school setting approach several systems concurrently.


Optimal soil Eh, pH for simultaneous decrease of bioavailable Cd, As in co-contaminated paddy soil under water management strategies.
The co-contamination with cadmium (Cd) and arsenic (As) in the paddy soil is the most seriously combined pollution of toxic elements in China, and it is rather difficult to decrease bioavailable Cd and As levels in soil because of the opposite ionic forms of bioavailable Cd (cation) and As (anion). This study explored the optimal conditions of Eh and pH in different soils for simultaneous decrease of Cd and As bioavailabilities in the soil-rice system through soil culture and rice pot experiments under water management strategies. The results showed that near neutral soil pH (7.0) were eventually observed under long-term flooding conditions. Under unflooded conditions, soil pH is the dominant factor influencing bioavailabilities of Cd and As, while under flooded conditions, Eh becomes the most important factor. Pot experiments showed that flooding significantly reduced the Cd concentration in rice grains from 54.5% to 95.5%, but concomitantly increased rice As concentration substantially (214%-302%). By evaluating the trade-off value between the bioavailabilities of Cd and As in the soil, the minimal trade-off value was obtained when the soil Eh was -130 mV and the pH was 6.8.


The global prevalence ptxP3 lineage of Bordetella pertussis was rare in young children with the co-purified aPV vaccination: a 5 years retrospective study.
The global prevalent ptxP3 strains varies from about 10% to about 50% of circulating B. pertussis population in different areas of China. To investigate the difference of vaccination status between different genotypes in the circulating B. pertussis after 10 years of acellular pertussis vaccine (aPV) used in China. The nasopharyngeal swabs and isolates of B. pertussis from these patients were used to perform genotyping of antigen genes. We use antibiotic susceptibility test against erythromycin and sequencing methods for site 2047 of 23S rRNA to determine the resistance status. The ptxP1 allele with erythromycin resistant (ER) B. pertussis infection (total of 449 subjects) consisted of 84.70 to 96.70% from 2012 to 2016 in this study. Vaccinated with co-purified aPV was found in 133(133/403,33.0%), 1(1/9,11.1%) and 2(2/21,9.5%) in ptxP1/fhaB3-ER, ptxP1/fhaB2-ES and ptxP3/fhaB2-ES B. pertussis infected children each, which showed a significant difference (χ2 = 6.87, P = 0.032). The ptxP3-ES B. pertussis was rare while the ptxP1-ER B. pertussis was steadily increased in Xi'an, China from 2012 to 2016, where co-purified aPV was prevalent used. This pose a hypothesis that the co-purified aPV might protect against ptxP3 strains more efficient, which generated a rare chance for ptxP3 strains to be under the antibiotic pressure and further developed to be erythromycin resistance. A further cohort study and the mechanisms of the additional antigen proteins of co-purified aPV protected against B. pertussis should be consideration.


Implicit learning of convective organization explains precipitation stochasticity.
Accurate prediction of precipitation intensity is crucial for both human and natural systems, especially in a warming climate more prone to extreme precipitation. Yet, climate models fail to accurately predict precipitation intensity, particularly extremes. One missing piece of information in traditional climate model parameterizations is subgrid-scale cloud structure and organization, which affects precipitation intensity and stochasticity at coarse resolution. Here, using global storm-resolving simulations and machine learning, we show that, by implicitly learning subgrid organization, we can accurately predict precipitation variability and stochasticity with a low-dimensional set of latent variables. Using a neural network to parameterize coarse-grained precipitation, we find that the overall behavior of precipitation is reasonably predictable using large-scale quantities only; however, the neural network cannot predict the variability of precipitation (R2 ∼ 0.45) and underestimates precipitation extremes. The performance is significantly improved when the network is informed by our organization metric, correctly predicting precipitation extremes and spatial variability (R2 ∼ 0.9). The organization metric is implicitly learned by training the algorithm on a high-resolution precipitable water field, encoding the degree of subgrid organization. The organization metric shows large hysteresis, emphasizing the role of memory created by subgrid-scale structures. We demonstrate that this organization metric can be predicted as a simple memory process from information available at the previous time steps. These findings stress the role of organization and memory in accurate prediction of precipitation intensity and extremes and the necessity of parameterizing subgrid-scale convective organization in climate models to better project future changes of water cycle and extremes.


The influence of seed apparency, nutrient content and chemical defenses on dietary preference in Dipodomys ordii.
Physical, nutritional and defensive qualities of seeds differ in the extent to which they influence granivore preference. In a study aimed to quantifying those differences, Ord's kangaroo rats (Dipodomys ordii) were found to prefer the seeds of just three of twenty-nine species: Cryptantha crassisepala, Oryzopsis hymenoides and Salsola kali. Oryzopsis hymenoides was most preferred during the early plant growth season (April-July); preference for S. kali peaked during late (August-November) and dormant (December-March) seasons; and greatest preference for C. crassisepala occurred during dormant and early seasons. Regression of forage ratios, averaged across seasons, against seed length, mass, abundance, patchiness, percent nitrogen, energy content, and chemical defenses showed seed length to be the most important predictor of seed preference. Seed length combined with nitrogen (protein) content and levels of two defensive compounds, saponins and non-protein amino acids, to account for 68% of the variation in seed preference. The importance of seed length rather than biomass indicated that there are limits to the ability of D. ordii to detect small seeds and that small size facilitated escape of dispersed seeds. Seasonality in preference suggested, however, that seed escape was encountered by predispersal harvesting of newly maturing seeds still on plants. Maximization of protein intake contradicted previously published observations, but presumably reflected low nitrogen availability. In addition to small size, the presence of saponins or non-protein amino acids in seeds was sufficient to negate the positive influence of higher protein content.


Morphological parameters of fourth lumbar spinous process palpation: a three-dimensional reconstruction of computed tomography.
The localization of lumbar fourth spinous process (L4-SP) is an important anatomical landmark, and identifying its accurate position is essential for the diagnosis and treatment of waist diseases. Five hundred participants were scanned with positive and lateral computed tomography (CT), which aimed to clarify anatomic characteristics of L4-SP. Anatomical parameters of the surface localization of L4-SP were measured and recorded through a three-dimensional (3D) reconstruction. Five hundred participants were classified into three types according to the position of BC with the iliac spine. There are just 266 that the line between the highest point of the iliac spine on both sides located on L4-SP (type I, 53.20%), 16 above L4-SP (type II, 3.20%), and 218 below L4-SP (type III, 43.60%). BC in type I (15.92 ± 1.30 mm) is longer than type III (15.56 ± 1.32 mm). While the angle combined with AB and BC is different in the three groups, the angle in type I (173.00 ± 4.83°) is larger than that in type II (164.69 ± 5.50°) and type III (159.45 ± 8.39°). Other measurements were not found any significant differences between above. The traditional palpation for L4-SP is not absolutely exact. The accuracy rate is only 53.20%, and the errors may cause serious consequences.


Structure of the respiratory MBS complex reveals iron-sulfur cluster catalyzed sulfane sulfur reduction in ancient life.
Modern day aerobic respiration in mitochondria involving complex I converts redox energy into chemical energy and likely evolved from a simple anaerobic system now represented by hydrogen gas-evolving hydrogenase (MBH) where protons are the terminal electron acceptor. Here we present the cryo-EM structure of an early ancestor in the evolution of complex I, the elemental sulfur (S0)-reducing reductase MBS. Three highly conserved protein loops linking cytoplasmic and membrane domains enable scalable energy conversion in all three complexes. MBS contains two proton pumps compared to one in MBH and likely conserves twice the energy. The structure also reveals evolutionary adaptations of MBH that enabled S0 reduction by MBS catalyzed by a site-differentiated iron-sulfur cluster without participation of protons or amino acid residues. This is the simplest mechanism proposed for reduction of inorganic or organic disulfides. It is of fundamental significance in the iron and sulfur-rich volcanic environments of early earth and possibly the origin of life. MBS provides a new perspective on the evolution of modern-day respiratory complexes and of catalysis by biological iron-sulfur clusters.


pH-sensitive O-carboxymethyl chitosan/sodium alginate nanohydrogel for enhanced oral delivery of insulin.
Oral drug delivery is considered the most preferred mode of treatment because of its high patient compliance and minimal invasiveness. However, the oral delivery of protein drug has been a difficult problem which restricts its application due to the unstable and inefficient penetration of protein in the gastrointestinal tract. In this study, a novel OCMC/SA nanohydrogel was prepared by using of O-carboxymethyl chitosan (OCMC) and sodium alginate (SA) to solve the problem. The OCMC/SA had a typical nanostructure, which was helpful to increase the specific surface area and enhanced the bioavailability of the drugs. OCMC/SA had a high drug loading capacity and realized passive drug targeting function by responding to the different pH value of the microenvironment. It could have a certain protective effect on drugs in strong acid circumstances, while its structure got loosed and effectively released drugs in intestinal circumstances. OCMC/SA could release the drug for >12 h, and the released insulin could maintain high activity. OCMC/SA nanohydrogel showed promising results in type 1 diabetic rats, and its pharmacological bioavailability was 6.57 %. In conclusion, this study constructed a novel OCMC/SA nanohydrogel, which had a lot of exciting characteristics and provided a new strategy for oral drug delivery.


Glycodelin regulates the numbers and function of peripheral natural killer cells.
Natural killer (NK) cells comprise of ∼70% of the immune cell population in the maternal decidua and ∼15% of the mononuclear cells in the peripheral blood. The decidual NK cells capable of producing high levels of cytokines are functionally distinct from the peripheral NK cells that exhibit high cytotoxicity. The numbers of peripheral NK cells and their cytotoxicity potential have been correlated with pregnancy outcome. In the same context, glycodelin, an immunomodulatory protein, has been recognized to be essential for the establishment and maintenance of pregnancy, and its' reduced levels are associated with recurrent spontaneous abortions. We investigated the effect of glycodelin on the peripheral NK cells. Our results reveal that glycodelin suppresses the cytotoxicity of peripheral NK cells via downregulating perforin, granzyme B and IFNγ. Glycodelin also induces caspase-dependent death in only activated peripheral NK cells, the effect suggested to be mediated by glycodelin upon engaging with the CD7 cell surface receptor. Thus, during pregnancy, glycodelin modulates the function and the number of cytotoxic NK cells that pose a deleterious effect on the fetus, a semi-allograft. This study provides insights into the mechanism of the regulatory effect of glycodelin on NK cells and could possibly be exploited for the management of miscarriages.


Binding of fluorescent-labeled phosphatidylcholine to rat liver nonspecific lipid transfer protein.
The fluorescent-labeled phospholipid, 1-palmitoyl-2-[12-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]dodecan oyl] phosphatidylcholine (P-C12-NBD-PC), is highly self-quenched when contained as a high mole percent in phospholipid vesicles. The addition of nonspecific lipid transfer protein (nsLTP), which is identical to sterol carrier protein2, to P-C12-NBD-PC self-quenching vesicles results in a large increase in the fluorescence yield. Evidence is presented that demonstrates that this fluorescence increase results from the binding of P-C12-NBD-PC to one or more low dielectric sites on the nsLTP molecule and that the P-C12-NBD-PC.nsLTP complex can exist free in aqueous solution. The binding is inhibited in the presence of mersalyl, a thiol reagent, and can be reversed with excess cysteine. Transfer of the nsLTP-bound P-C12-NBD-PC to phospholipid vesicles is also demonstrated. The half-times for loading and unloading the P-C12-NBD-PC.nsLTP complex are both 3 orders of magnitude faster than the spontaneous P-C12-NBD-PC transfer between vesicles. These data demonstrate that nsLTP has the necessary properties to bind and carry lipids between membranes.


Can exercise replace airway clearance techniques in cystic fibrosis? A survey of patients and healthcare professionals.
Airway clearance techniques (ACTs) are recommended in cystic fibrosis (CF) to prevent accumulation of secretions and lung infection. "Can exercise replace chest physiotherapy for people with CF?" is one of the CF community's top 10 research questions. We conducted an online survey of the CF community to gather data on current ACT use, recommendations, reported adherence levels and exercise strategies used. There were 488 respondents: 194 (40%) people with CF (pwCF), 141 (29%) family and 153 (31%) healthcare professionals (HCPs) (mostly physiotherapists). Only 10/285 (4%) of pwCF do no exercise at present and 163/303 (54%) already incorporate exercise into ACTs. ACTs were omitted by 128/267 (48%) of pwCF when they exercised. Nearly all (110/129, 93%) of HCPs currently recommend exercise to support ACTs. A trial replacing some or all ACTs with exercise, was supported by 80/110 (73%) of HCPs, with an additional 9/110 (8%) willing to consider in selected patients.


Ovarian activation delays in peripubertal ewe lambs infected with Haemonchus contortus can be avoided by supplementing protein in their diets.
The ewe lamb nutritional and physiological state interfere with the ovarian environment and fertility. The lack or excess of circulating nutrients reaching the ovary can change its gene expression. A protein deficiency in the blood caused by an Haemonchus contortus abomasal infection is detrimental to the organism's development during puberty. The peripubertal period is a time of intensive growth that requires a high level of nutrients. An essential feature controlling pubertal arousal and female reproductive potential is ovarian follicle growth activation. Protein supplementation improves the sheep's immune response to helminthic infections. We aimed to determine if supplementing protein in infected ewe lambs' diet would impact the ovarian environment leading to earlier ovarian follicle activation than in infected not supplemented animals. We fed 18 Santa Ines ewe lambs (Ovis aries) - bred by the same ram - with either 12% protein (Control groups) or 19% protein (Supplemented groups) in their diets. After 35 days of the diet, they were each artificially infected or not with 10,000 Haemonchus contortus L3 larvae. Following 77 days of the diet and 42 days of infection, we surgically collected their left ovaries and examined their genes expression through RNA sequencing. We found that protein supplementation in infected animals led to an up-regulation of genes (FDR p-values < 0.05) and biological processes (p-value cut-off = 0.01) linked to meiotic activation in pre-ovulatory follicles and primordial follicle activation, among others. The supplemented not infected animals also up-regulated genes and processes linked to meiosis and others, such as circadian behaviour. The not supplemented animals had these same processes down-regulated while up-regulated processes related to tissue morphogenesis, inflammation and immune response. Diet's protein supplementation of peripubertal infected animals allowed them to express genes related to a more mature ovarian follicle stage than their half-sisters that were not supplemented. These results could be modelling potential effects of the interaction between environmental factors, nutrition and infection on reproductive health. When ovarian activation is achieved in a timely fashion, the ewe may generate more lambs during its reproductive life, increasing sheep breeders' productivity.


Structural Basis of the Activation of Heterotrimeric Gs-Protein by Isoproterenol-Bound β1-Adrenergic Receptor.
Cardiac disease remains the leading cause of morbidity and mortality worldwide. The β1-adrenergic receptor (β1-AR) is a major regulator of cardiac functions and is downregulated in the majority of heart failure cases. A key physiological process is the activation of heterotrimeric G-protein Gs by β1-ARs, leading to increased heart rate and contractility. Here, we use cryo-electron microscopy and functional studies to investigate the molecular mechanism by which β1-AR activates Gs. We find that the tilting of α5-helix breaks a hydrogen bond between the sidechain of His373 in the C-terminal α5-helix and the backbone carbonyl of Arg38 in the N-terminal αN-helix of Gαs. Together with the disruption of another interacting network involving Gln59 in the α1-helix, Ala352 in the β6-α5 loop, and Thr355 in the α5-helix, these conformational changes might lead to the deformation of the GDP-binding pocket. Our data provide molecular insights into the activation of G-proteins by G-protein-coupled receptors.


Optogenetic TDP-43 nucleation induces persistent insoluble species and progressive motor dysfunction in vivo.
TDP-43 is a predominantly nuclear DNA/RNA binding protein that is often mislocalized into insoluble cytoplasmic inclusions in post-mortem patient tissue in a variety of neurodegenerative disorders including Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal dementia (FTD). The underlying causes of TDP-43 proteinopathies remain unclear, but recent studies indicate the formation of these protein assemblies is driven by aberrant phase transitions of RNA deficient TDP-43. Technical limitations have prevented our ability to understand how TDP-43 proteinopathy relates to disease pathogenesis. Current animal models of TDP-43 proteinopathy often rely on overexpression of wild-type TDP-43 to non-physiological levels that may initiate neurotoxicity through nuclear gain of function mechanisms, or by the expression of disease-causing mutations found in only a fraction of ALS patients. New technologies allowing for light-responsive control of subcellular protein crowding provide a promising approach to drive intracellular protein aggregation, as we have previously demonstrated in vitro. Here we present a model for the optogenetic induction of TDP-43 proteinopathy in Drosophila that recapitulates key features of patient pathology, including detergent insoluble cytoplamsic inclusions and progressive motor dysfunction.


Characterizing fluorescent dissolved organic matter in a membrane bioreactor via excitation-emission matrix combined with parallel factor analysis.
In this study, we successfully tracked the dynamic changes in different constitutes of bound extracellular polymeric substances (bEPS), soluble microbial products (SMP), and permeate during the operation of bench scale membrane bioreactors (MBRs) via fluorescence excitation-emission matrix (EEM) combined with parallel factor analysis (PARAFAC). Three fluorescent groups were identified, including two protein-like (tryptophan-like C1 and tyrosine-like C2) and one microbial humic-like components (C3). In bEPS, protein-like components were consistently more dominant than C3 during the MBR operation, while their relative abundance in SMP depended on aeration intensities. C1 of bEPS exhibited a linear correlation (R(2)=0.738; p<0.01) with bEPS amounts in sludge, and C2 was closely related to the stability of sludge. The protein-like components were more greatly responsible for membrane fouling. Our study suggests that EEM-PARAFAC can be a promising monitoring tool to provide further insight into process evaluation and membrane fouling during MBR operation.


Dose optimisation in paediatric radiography - Using regression models to investigate the relative impact of acquisition factors on image quality and radiation dose.
To investigate the optimum pelvis X-ray acquisition factors for a 10-year-old child. Secondly, to evaluate the impact of each acquisition factor on image quality (IQ) and radiation dose. Images were acquired using a pelvis phantom and a range of acquisition parameters; e.g. tube potential, additional filtration and source-to-image distance (SID). Automatic exposure control (AEC) was used with two orientations (head towards/away from two outer chambers) and three different chamber selections. Visual IQ was evaluated using relative and absolute-VGA methods. Radiation doses were measured by placing a dosimeter on the anterior surface of the phantom. Regression analysis was used to determine optimum parameters. The optimised technique (178.8 µGy), with diagnostic IQ, was with 89kVp, 130 cm SID and with 1 mm Al + 0.1 mm Cu filtration. This technique was with the head towards the two outer AEC chambers. Regression analysis showed that SID had the lowest impact on IQ (β = 0.002 95% CI -0.001 to 0.005) and dose (β = -0.96 95% CI -0.40 to -1.53). The impact of filtration on dose (β = -76.24 95% CI -86.76 to -85.72) was higher than tube potential (β = -13.44 95% CI -14.34 to -12.53). The following impact ratios were higher on IQ than radiation dose: filtration/kVp; 11.28 times, filtration/SID; 7.01 times and kVp/SID; 0.62 times. Optimised parameters were identified as 89 kVp, 130 cm SID and with 1 mm Al + 0.1 mm Cu additional filtration. Regression analysis demonstrated that filtration and tube potential had the greatest effect on radiation dose and IQ, respectively.


Acteoside and martynoside exhibit estrogenic/antiestrogenic properties.
Acteoside and martynoside are plant phenylpropanoid glycosides exhibiting anticancer, cytotoxic and antimetastatic activities. We investigated their potential to activate estrogen receptor isoforms ERalpha and ERbeta in HeLa cells transfected with an estrogen response element (ERE)-driven luciferase (Luc) reporter gene and an ERalpha or ERbeta expression vector. Their estrogenic/antiestrogenic effects were also assessed in breast cancer cells (MCF7), endometrial cancer cells (Ishikawa) and osteoblasts (KS483), by measuring IGFBP3 levels, cell viability and number of mineralized nodules, respectively, seeking for a natural selective estrogen receptor modulator (SERM). Acteoside and martynoside antagonized both ERalpha and ERbeta (p<0.001), whereas they reversed the effect of E(2) mainly via ERalpha (p<0.001). Martynoside was a potent antiestrogen in MCF-7 cells, increasing, like ICI182780, IGFBP3 levels via the ER-pathway. In osteoblasts, martynoside induced nodule mineralization, which was abolished by ICI182780, implicating an ER-mediated mechanism. Furthermore, its antiproliferative effect on endometrial cells suggests that martynoside may be an important natural SERM. Acteoside was an antiestrogen in breast cancer cells and osteoblasts, without any effect on endometrial cells. Our study suggests that the nature is rich in selective ERalpha and ERbeta ligands, the discovery of which may lead to the development of novel neutraceutical agents.


Effectiveness of alteplase infusion for the management of prosthetic mitral valve thrombosis in paediatric age group and proposed algorithm.
The incidence of prosthetic valve implantation is increasing in the paediatric population. Prosthetic valve thrombosis leading to obstruction could potentially be a life-threatening complication. There is a debate regarding optimal management of this complication, and there is limited use of thrombolytic therapy in childhood in the setting of valve thrombosis. We aim to share our experience of successfully using fibrinolytic therapy in terms of alteplase for paediatric prosthetic mitral valve thrombosis and to propose a management algorithm. This retrospective analysis of the database was conducted at our hospital including patients who underwent thrombolysis (alteplase) for prosthetic mitral valve thrombosis from June, 2011 to June, 2021. A total of 10 patients with 20 attempts of alteplase infusion were found in our record. Alteplase was successful in 19 attempts to relieve valve thrombosis. The safe and effective dose of alteplase was between 0.1 and 0.3 mg/kg/hour. There were no associated major bleeding complications and alteplase was administered either by central or peripheral line. Thrombolysis by alteplase infusion was found to be successful in relief of prosthetic mitral valve thrombosis in paediatric population without major bleeding complications.


Niobium-based superconducting nano-device fabrication using all-metal suspended masks.
We report a novel method for the fabrication of superconducting nano-devices based on niobium. The well-known difficulties of lithographic patterning of high-quality niobium are overcome by replacing the usual organic resist mask by a metallic one. The quality of the fabrication procedure is demonstrated by the realization and characterization of long and narrow superconducting lines and niobium-gold-niobium proximity SQUIDs.


Validation of an automated real-time PCR protocol for detection and quantitation of HIV and HCV genomes in semen.
The ability to detect and quantify HIV and HCV genomes is important for checking spermatozoid preparation protocols also known as "sperm washing". But no commercial assay is available. A method was developed for detecting HIV and HCV in semen fractions using the COBAS Ampliprep and COBAS Taqman instruments. It will detect fewer than 200 copies of HIV RNA per ml of semen plasma and fewer than 200 copies/3 x 10(6) semen cells. The sensitivity for HCV is similar at more than 200 IU/ml and below 200 IU/3 x 10(6) semen cells. No inhibitor of PCR amplification was detected. This automated protocol permits a convenient, standardized testing for HIV and HCV in semen. The performance is the same as that of the previous generation of automated assays but the cost and operating time are both reduced.


Role of chromatin, environmental changes and single cell heterogeneity in non-coding transcription and gene regulation.
The number and variety of factors underlying control of gene expression have been frequently underestimated. Non-coding RNAs generated through pervasive transcription have recently been implicated in shaping the transcriptional landscape in different organisms from bacteria to higher eukaryotes, adding a previously unexpected layer of complexity to the process of gene regulation. In this review, we highlight non-coding transcription-dependent regulatory mechanisms linked to chromatin organization and environmental changes, and particular emphasis is given to single-cell approaches, which have been crucial in dissecting cell-to-cell variability. These studies have revealed that non-coding transcription can underlie the extensive heterogeneity in patterns of gene expression within a cell population.


Liver Transplantation for Colorectal Liver Metastases: Is It Appropriate?
Colorectal cancer with liver metastases is a condition with significant morbidity and mortality that affects many people around the world. Many treatments exist to target liver metastases, including surgical resection, chemotherapy, nonsurgical liver-directed therapies, and liver transplantation. The field of transplant oncology is emerging as a promising alternative to palliative chemotherapy alone in appropriately selected patients. Though few clinical trials have been completed to evaluate safety of liver transplant for colorectal cancer metastases, there are several ongoing trials to hopefully make transplant a viable option for more patients with limited options.


Zonisamide regulates basal ganglia transmission via astroglial kynurenine pathway.
To clarify the anti-parkinsonian mechanisms of action of zonisamide (ZNS), we determined the effects of ZNS on tripartite synaptic transmission associated with kynurenine (KYN) pathway (KP) in cultured astrocytes, and transmission in both direct and indirect pathways of basal ganglia using microdialysis. Interactions between cytokines [interferon-γ (IFNγ) and tumor-necrosis factor-α (TNFα)] and ZNS on astroglial releases of KP metabolites, KYN, kynurenic-acid (KYNA), xanthurenic-acid (XTRA), cinnabarinic-acid (CNBA) and quinolinic-acid (QUNA), were determined by extreme liquid-chromatography with mass-spectrometry. Interaction among metabotropic glutamate-receptor (mGluR), KP metabolites and ZNS on striato-nigral, striato-pallidal GABAergic and subthalamo-nigral glutamatergic transmission was examined by microdialysis with extreme liquid-chromatography fluorescence resonance-energy transfer detection. Acute and chronic ZNS administration increased astroglial release of KYN, KYNA, XTRA and CNBA, but not QUNA. Chronic IFNγ administration increased the release of KYN, KYNA, CNBA and QUNA, but had minimal inhibitory effect on XTRA release. Chronic TNFα administration increased CNBA and QUNA, but not KYN, KYNA or XTRA. ZNS inhibited IFNγ-induced elevation of KYN, KYNA and QUNA, but enhanced IFNγ-induced that of CNBA. TNFα-induced rises in CNBA and QUNA were inhibited by ZNS. ZNS inhibited striato-nigral GABAergic, striato-pallidal GABAergic and subthalamo-nigral glutamatergic transmission via activation of groups II and III mGluRs. ZNS enhanced astroglial release of endogenous agonists of group II mGluR, XTRA and group III mGluR, CNBA. Activated endogenous mGluR agonists inhibited transmission in direct and indirect pathways of basal ganglia. These mechanisms contribute to effectiveness and well tolerability of ZNS as an adjunct treatment for Parkinson's disease during l-DOPA monotherapy. This article is part of the Special Issue entitled 'The Synaptic Basis of Neurodegenerative Disorders'.


Bone loss after initiation of androgen deprivation therapy in patients with prostate cancer.
Although androgen deprivation therapy (ADT) for prostate cancer is associated with bone loss, little is known about when this bone loss occurs. We postulated that men on ADT would experience the greatest bone loss acutely after initiation of ADT. We conducted a 12-month prospective study at an academic medical center. We studied 152 men with prostate cancer (30 with acute ADT, < 6 months; 50 with chronic ADT, > or = 6 months; and 72 with no ADT) and 43 healthy age-matched controls. We assessed bone mineral density (BMD) of the hip, wrist, total body, and spine; body composition; and markers of bone turnover. After 12 months, men receiving acute ADT had a significant reduction in BMD of 2.5 +/- 0.6% at the total hip, 2.4 +/- 1.0% at the trochanter, 2.6 +/- 0.5% at the total radius, 3.3 +/- 0.5% at the total body, and 4.0 +/- 1.5% at the posteroanterior spine (all P < 0.05). Men with chronic ADT had a 2.0 +/- 0.6% reduction in BMD at the total radius (P < 0.05). Healthy controls and men with prostate cancer not receiving ADT had no significant reduction in BMD. Both use and duration of ADT were associated with change in bone mass at the hip (P < 0.05). Men receiving acute ADT had a 10.4 +/- 1.7% increase in total body fat and a 3.5 +/- 0.5% reduction in total body lean mass at 12 months, whereas body composition did not change in men with prostate cancer on chronic ADT or in healthy controls (P < 0.05). Markers of bone formation and resorption were elevated in men receiving acute ADT after 6 and 12 months compared with the other men with prostate cancer and controls (P < 0.05). Men in the highest tertile of bone turnover markers at 6 months had the greatest loss of bone density at 12 months. Men with prostate cancer who are initiating ADT have a 5- to 10-fold increased loss of bone density at multiple skeletal sites compared with either healthy controls or men with prostate cancer who are not on ADT, placing them at increased risk of fracture. Bone loss is maximal in the first year after initiation of ADT, suggesting initiation of early preventive therapy.


Facial and vestibulocochlear nerve avulsion at the fundus of the internal auditory canal in a child without a temporal bone fracture.
To describe a case of facial, vestibular, and cochlear nerve avulsion secondary to blunt trauma without an associated temporal bone fracture. Clinical capsule report. University hospital. A 3.5-year-old girl presented with immediate facial nerve paralysis and complete deafness after being struck by an automobile. High-resolution computed tomography demonstrated a depressed occipital bone fracture with no visible fracture of the temporal bone. Magnetic resonance imaging sequence raised the question of VIIth nerve bundle discontinuity at the distal end of the internal auditory canal. The patient underwent a posterior fossa craniotomy via a translabyrinthine approach 9 months after the initial injury, and facial and auditory nerve avulsion at the fundus was confirmed at the time of surgery. The proximal segment of the facial nerve had formed a traumatic neuroma, which was resected, and primarily anastomosed to the rerouted distal segment. Facial nerve function. Patient has regained facial function to Grade III/VI House-Brackmann with no asymmetry at rest. An unusual pattern of injury is described. We suggest that in patients presenting with facial nerve paralysis secondary to blunt trauma, without an associated temporal bone fracture, high-resolution magnetic resonance imaging is recommended to evaluate internal auditory canal discontinuity of the VIIth and VIIIth nerve complexes. A potential mechanism of avulsion is explained.


Enablers and barriers to evidence implementation in complementary medicine: A systematic review.
Despite the push for complementary medicine (CM) practitioners to engage in evidence implementation, and arguments in support of evidence-based practice (EBP), uptake of EBP amongst most CM professions remains low. This review aimed to synthesise the evidence examining the barriers and enablers to evidence implementation in CM. Any primary study examining enablers and barriers to evidence implementation in CM were eligible for inclusion. Eight databases and search engines were searched for eligible studies. Reference lists of included studies were screened, and authors of included studies were contacted to identify current or unpublished studies that met the inclusion criteria. Thirty-nine published and unpublished studies were included in this review. The seven published qualitative studies and 25 published quantitative studies were rated as moderate to high quality. Fifty-two distinct barriers and 62 discrete enablers were identified. Reported barriers were predominantly structural (e.g. limited availability of time and clinical evidence) and cognitive (e.g. skills deficits), with relatively fewer studies reporting cultural (e.g. lack of industry support) or attitudinal barriers (e.g. lack of interest in, or relevance to CM). Enablers of evidence implementation largely focussed on improving access to bibliographic databases and evidence reviews, supporting skills acquisition, and cultivating leadership and interprofessional/interagency collaboration. The findings of this review highlight the diverse barriers and enablers to evidence implementation in CM that span multiple dimensions. The interplay between these various factors highlights the complexity of evidence implementation, and the need for a targeted multistakeholder, multidimensional solution to optimise evidence-based practice in CM.


Quantification of Escherichia coli by kinetic 5'-nuclease polymerase chain reaction (real-time PCR) oriented to sfmD gene.
A kinetic 5'-nuclease polymerase chain reaction (real-time PCR) for the quantification of Escherichia coli was developed. Specific primers and a fluorogenic probe oriented to sfmD gene, encoding a putative outer membrane export usher protein, were designed. The PCR system was highly specific and sensitive for E. coli, as determined with 37 non-E. coli strains (exclusivity, 100%) and 24 E. coli strains (inclusivity, 100%). When used in real-time PCR, linear calibration lines were obtained in the range from 10(2) to 10(8) CFU ml(-1) for three E. coli strains. Salmonella Enteritidis (10(6) CFU ml(-1)) or Citrobacter freundii (10(6) CFU ml(1)) had no effect on quantification of E. coli by the method. The developed real-time PCR is suitable for rapid quantification of E. coli. In connection to an appropriate sample preparation technique, the method is suitable for food safety and technological hygiene applications.


Direct analysis of calcium in liquid infant formula via laser-induced breakdown spectroscopy (LIBS).
The present work illustrates the potential of laser-induced breakdown spectroscopy (LIBS) for the direct analysis of liquid food products. The aim of the experiment was to predict calcium content in ready-to-feed infant formula. The analysis was performed by a LIBS system coupled to a liquid sample chamber with a rotatory wheel that presents the liquid to the laser beam as a thin film. Multivariate analysis with partial least squares regression (PLSR) was performed to correlate LIBS spectral data to reference calcium contents. The obtained PLSR model exhibited a good fit and linearity, as indicated by the coefficients of determination for calibration (Rc2) and cross-validation (Rcv2), with values of 0.96 and 0.89, respectively. The robustness of the calibration model was assessed by external validation showing a root-mean-square error of prediction of 6.45 mg 100 mL-1. These results demonstrated the potential of LIBS for real-time analysis of liquid food products.


Spontaneous glomerular basement membrane changes in the golden Syrian hamster (Mesocricetus auratus): a light and electron microscope study.
Changes in apparently healthy hamsters, consistent with proteinuria, are reported, but no IgG deposits or amyloid in the glomeruli were detected. Further investigation is required into the significance and the aetiology of these, as yet, obscure alterations.


Amino acid substitution in the Cυ3 domain causes either elevation or reduction of IgY uptake into egg yolks of quail.
Egg yolks of avian species contain large quantities of immunoglobulin (Ig) Ys transferred from maternal blood circulation. However, it is unclear how maternal IgYs are incorporated into the egg yolks of maturing oocytes. The aim of this study was to identify the amino acid residues required for efficient IgY transport into the egg yolks of quail by utilizing recombinant quail IgY-Fc (qIgY-Fc). Five amino acid residues (361-365 at the Cυ3 domain) located on the Cυ3/Cυ4 interface were individually substituted for alanine residues. The mutants were then intravenously injected into laying quail, and their uptakes into egg yolks were measured by ELISA. Substitution of L362, Y363 and I364 for alanine markedly reduced qIgY-Fc uptake into the egg yolks to almost undetectable levels. With respect to the Y363 residue, neither substitution for phenylalanine nor substitution of tryptophan reduced qIgY-Fc uptake, suggesting the necessity of an aromatic side-chains at the Y363 residue. Interestingly, substitution of G365 for alanine or for other polar- or non-polar amino acids elevated qIgY-Fc uptake by 2.5-fold compared to that of the wild-type qIgY-Fc. Analyses of the blood concentrations of the two alanine mutants with a low uptake (Y363A) and a high uptake (G365A) showed that their modified uptakes were not explained by changes in blood clearance. Removal of the N-glycosylated carbohydrate chain at the Cυ3 domain by substituting an N408 residue for alanine also resulted in lowered qIgY-Fc uptake. These results emphasize the existence of a selective IgY transport system recognizing the Cυ3 domain of IgY, which raises the possibility that an IgY with high transport ability might be engineered by genetic manipulation.


Novel compound heterozygous mutations in sacsin-related ataxia.
High prevalence of a form of autosomal recessive spastic ataxia with early onset was originally described among French Canadians in the Charlevoix-Saguenay region, in northeastern Quebec. Since the responsible gene (SACS) was identified, mutations in the SACS gene have been described in Tunisia, Italy, Turkey, and Japan. The mutation sites found outside Quebec are different from the ones in Quebec. All patients outside Quebec, except one Italian patient, have been reported to have homozygous mutations. The authors report here identical twin sisters with novel compound heterozygous mutations (c.[2951_2952delAG]+[3922delT]) in the SACS gene.


The frequency of parafunctional occlusal habits compared with the incidence of mandibular displacement.
Acomparison was made in this study of the prevalence of mandibular displacement in varying degrees of complexity with the incidence of parafunctional occlusal habits and some sequelae. The prevalence of occlusal habits in all patients was high but became greater with the increased complexity of the mandibular displacement, especially an asymmetrical slide from RCP to IP. The mulifactorial etiology of parafunctional occlusal habits and their sequelae tends to make the relationships more complex than a simple "cause and effect" and therefore must be considered.


Participatory learning and action to address type 2 diabetes in rural Bangladesh: a qualitative process evaluation.
Diabetes is 7th largest cause of death worldwide, and prevalence is increasing rapidly in low-and middle-income countries. There is an urgent need to develop and test interventions to prevent and control diabetes and develop the theory about how such interventions can be effective. We conducted a participatory learning and action (PLA) intervention with community groups in rural Bangladesh which was evaluated through a cluster randomised controlled trial. There was a large reduction in the combined prevalence of type 2 diabetes and intermediate hyperglycaemia in the PLA group compared with the control group. We present findings from qualitative process evaluation research to explore how this intervention was effective. We conducted group interviews and focus group discussions using photovoice with purposively sampled group attenders and non-attenders, and intervention implementers. Data were collected before the trial analysis. We used inductive content analysis to generate theory from the data. The intervention increased the health literacy of individuals and communities - developing their knowledge, capacity and self-confidence to enact healthy behaviours. Community, household and individual capacity increased through social support and social networks, which then created an enabling community context, further strengthening agency and enabling community action. This increased opportunities for healthy behaviour. Community actions addressed lack of awareness about diabetes, gendered barriers to physical activity and lack of access to blood glucose testing. The interaction between the individual, household, and community contexts amplified change, and yet there was limited engagement with macro level, or 'state', barriers to healthy behaviour. The participatory approach enabled groups to analyse how context affected their ability to have healthy behaviours and participants engaged with issues as a community in the ways that they felt comfortable. We suggest measuring health literacy and social networks in future interventions and recommend specific capacity strengthening to develop public accountability mechanisms and health systems strengthening to complement community-based interventions. Registered at ISRCTN on 30th March 2016 (Retrospectively Registered) Registration number: ISRCTN41083256 .


Hb La Coruña [beta38(C4)Thr-->Ile]: a new hemoglobin variant leading to familial polycythemia.
Hb La Coruña [beta38(C4)Thr --> Ile] is a new hemoglobin (Hb) variant that has an increased oxygen affinity. Clinically, this Hb leads to erythrocytosis. Hb La Coruña is an electrophoretically silent variant that can be detected by reversed phase high performance liquid chromatography (HPLC) and characterized by DNA sequencing. The patient was a 22-year-old Spanish male whose family lived in La Coruña, in the northwest of Spain. His mother was also a carrier.


The effects of intermittent fasting regimens in middle-age and older adults: Current state of evidence.
Aging is associated with a host of biological changes that contribute to a progressive decline in cognitive and physical function, ultimately leading to a loss of independence and an increased risk of mortality. The unprecedented growth of the aging population has thus created an urgent need for interventions that can preserve older adults' capacity to live independently and to function well. To date, there is no conclusive evidence supporting the efficacy of an intervention to prevent or reverse physical disability in older persons at risk of functional decline. A growing body of evidence indicates that prolonged fasting periods and different types of intermittent fasting regimens can have positive effects on anthropometric and metabolic health parameters in middle-aged adults similar to that of calorie restriction. For this reason, there is increasing scientific interest in further exploring the biological and metabolic effects of intermittent fasting approaches, as well as the feasibility and safety of popular types of intermittent fasting regimens in older adults. Thus, the purpose of the present review is to describe the state of evidence of different types of intermittent fasting regimes, specifically time-restricted eating and 5:2 intermittent fasting, in the growing population of middle-aged and older adults. A small, but growing body of evidence indicates both time restricted eating (TRE) and 5:2 intermittent fasting approaches can produce modest weight loss; however, only the 5:2 approach produced clinically meaningful weight losses. Reductions in blood pressure were observed for both TRE and 5:2 fasting approaches, but the effects were not consistent across studies. The majority of studies to date, in middle-age and older adults, however, have been of short duration in small study samples. Future clinical trials with larger populations and longer intervention durations are required to better understand the risks versus benefits of different types of intermittent fasting regimens in middle-age and older individuals.


Estrous stage determination in rats by means of scanning electron microscopic images of uterine surface epithelium.
With the results presented in this paper we devised an alternative method to precisely date the rat endometrium in relation to the estrous cycle. This is done by the exclusive use of scanning electron microscopy. Owing to its short estrous cycle (4 to 6 days, depending on age), the rat is ideally suited for the examination of cyclic changes occurring in the uterine epithelium. The cycle stage of rats predated by vaginal smear cytology was verified through the measurement of hormones relevant to the estrous cycle, i.e. estradiol-17-beta (E2), progesterone (P), luteinizing hormone (LH), and follicle stimulating hormone (FSH). Based on scanning electron images surface changes could thus be correlated to the cyclic variations of blood levels of sex hormones. The appearance of pseudoglands, the most prominent aspect during the cycle, is correlated with decreasing estrogen and rising progesterone levels. Pseudoglands are formed by apoptosis and necrosis of epithelial cells, and are most numerous during estrus. They had in previous studies been classified as genuine uterine glands.


Acoustic waves in bounded anisotropic media: theorems, estimations, and computations.
This paper discusses some basic achievements in theoretical studies on acoustic wave propagation along boundaries in anisotropic solids. In particular, the following issues are reviewed: existence theorems for subsonic surface and interface waves, leaky surface acoustic waves (SAW) and their relation to "supersonic" SAWs and fast exceptional bulk waves, the resonance reflection of bulk waves in the vicinity of leaky wave branches. General conclusions are illustrated by numerical examples.


Comparing telemedicine and in-person gastrointestinal cancer genetic appointment outcomes during the COVID-19 pandemic.
The study purpose is to compare outcomes associated with completion of genetic testing between telemedicine and in-person gastrointestinal cancer risk assessment appointments during the COVID-19 pandemic. Data was collected on patients with scheduled appointments between July 2020 and June 2021 in a gastrointestinal cancer risk evaluation program (GI-CREP) that utilized both telemedicine and in-person visits throughout the COVID-19 pandemic, and a survey was administered. A total of 293 patients had a GI-CREP appointment scheduled and completion rates of in-person versus telemedicine appointments were similar. Individuals diagnosed with cancer and those with Medicaid insurance had lower rates of appointment completion. Although telehealth was the preferred visit modality, there were no differences in recommending genetic testing nor in the consent rate for genetic testing between in-person and telemedicine visits. However, of patients who consented for genetic testing, more than three times more patients seen via telemedicine did not complete genetic testing compared to those seen in-person (18.3% versus 5.2%, p = 0.008). Furthermore, telemedicine visits had a longer turnaround time for genetic test reporting (32 days versus 13 days, p < 0.001). Compared to in-person GI-CREP appointments, telemedicine was associated with lower rates of genetic testing completion, and longer turnaround time for results.


Short-term outcomes in robotic vs laparoscopic ileal pouch-anal anastomosis surgery: a propensity score match study.
Laparoscopic ileal pouch-anal anastomosis (IPAA) surgery offers improved short-term outcomes over open surgery but can be technically challenging. Robotic surgery has been increasingly used for IPAA surgery, but there is limited evidence supporting its use. This study aims to compare the short-term outcomes of laparoscopic and robotic IPAA procedures. All consecutive patients receiving laparoscopic and robotic IPAA surgery at 3 centres, from 3 countries, between 2008 and 2019 were identified from prospectively collated databases. Robotic surgery patients were propensity score matched with laparoscopic patients for gender, previous abdominal surgery, ASA grade (I, II vs III, IV) and procedure performed (proctocolectomy vs completion proctectomy). Their short-term outcomes were examined. A total of 89 patients were identified (73 laparoscopic, 16 robotic). The 16 patients that received robotic surgery were matched with 15 laparoscopic patients. Baseline characteristics were similar between the two groups. There were no statistically significant differences in any of the investigated short-term outcomes. Length of stay trend was higher for laparoscopic surgery (9 vs 7 days, p = 0.072) CONCLUSION: Robotic IPAA surgery is safe and feasible and offers similar short-term outcomes to laparoscopic surgery. Length of stay may be lower for robotic IPAA surgery, but further larger scale studies are required in order to demonstrate this.


Retroaortic left renal vein with renal vein hypertension causing hematuria.
We describe a case of retroaortic left renal vein with hematuria. In this case, pullback pressure from the retroaortic left renal vein to the inferior vena cava revealed left renal vein hypertension according to criteria of the "nutcracker phenomenon." We stress that left renal vein anomaly including retroaortic left renal vein can cause clinical symptoms such as hematuria.


Transjugular intrahepatic portosystemic shunt improves oxygenation in hepatopulmonary syndrome.
Hepatopulmonary syndrome is a complication of chronic liver disease in which arterial hypoxemia results from abnormalities in pulmonary blood flow. Severe hypoxemia can lead to clinical deterioration and death. Although the etiology is unknown, portal hypertension seems to be an important factor in the development of hepatopulmonary syndrome. No effective pharmacological therapy has been identified, but liver transplantation may be curative. Arterial hypoxemia may complicate transplant surgery, however, and resolution of the syndrome after liver transplantation is performed may be delayed. In addition, it seems that complete reversal of oxygenation abnormalities after liver transplantation is performed is unpredictable. We described a patient with hepatopulmonary syndrome who noted improvement in symptoms of dyspnea after the placement of a transjugular intrahepatic portosystemic shunt. Arterial oxygenation and calculated shunt fraction improved significantly during the follow-up period, and liver transplantation was subsequently performed without difficulty. Portal decompression using transjugular intrahepatic portosystemic shunt may represent a palliative therapy for hepatopulmonary syndrome in patients awaiting liver transplantation.


Colonoscopy in mice.
Current investigational models of murine colitis and colon cancer necessitate sacrifice of animals in order to obtain colonic tissue. The purpose of this study was to develop a safe method of murine colonoscopy that would allow serial evaluation and mucosal biopsies of the same animal. Nine mice (two C3H, two C57/BL6, and five IL-10 deficient) were studied a total of four times each over 4 weeks. Three mice [APC (Min +/-)] were examined three times each. Mice were gavaged with 1 cc of a polyethylene glycol solution on the day prior to colonoscopy. Solid chow was withheld and the mice were maintained on Pedialyte. Mice were anesthetized with ketamine and xylazine. A flexible pediatric cystoscope (2.1-mm diameter) with a single biopsy channel was introduced per anum, and the colon was gently insufflated with air to a mean pressure of less than 5 mmHg. Saline irrigation was used when necessary. A single biopsy was obtained from the rectosigmoid colon during each examination. A total of 46 examinations were carried out. One mouse died after being anesthesized for the fourth examination, and two mice [one IL-10 knockout and one APC (Min+/-)] died one day after the 3rd examination. No other complications were noted. The average length of insertion was 3 cm. Transillumination allowed for localization of the endoscope tip. Biopsies, although quite small, were sufficient for pathologic evaluation and diagnosis. Murine colonoscopy is a safe and feasible technique. It permits consecutive visual and histopathological examinations, and it allows the investigator to monitor the response of the murine colon to experimental interventions.


Site-specific acylation of a bacterial virulence regulator attenuates infection.
Microbiota generates millimolar concentrations of short-chain fatty acids (SCFAs) that can modulate host metabolism, immunity and susceptibility to infection. Butyrate in particular can function as a carbon source and anti-inflammatory metabolite, but the mechanism by which it inhibits pathogen virulence has been elusive. Using chemical proteomics, we found that several virulence factors encoded by Salmonella pathogenicity island-1 (SPI-1) are acylated by SCFAs. Notably, a transcriptional regulator of SPI-1, HilA, was acylated on several key lysine residues. Subsequent incorporation of stable butyryl-lysine analogs using CRISPR-Cas9 gene editing and unnatural amino acid mutagenesis revealed that site-specific modification of HilA impacts its genomic occupancy, expression of SPI-1 genes and attenuates Salmonella enterica serovar Typhimurium invasion of epithelial cells, as well as dissemination in vivo. Moreover, a multiple-site HilA lysine acylation mutant strain of S. Typhimurium was resistant to butyrate inhibition ex vivo and microbiota attenuation in vivo. Our results suggest that prominent microbiota-derived metabolites may directly acylate virulence factors to inhibit microbial pathogenesis in vivo.


Anti-β2-glycoprotein I autoantibodies influence thrombin generation parameters via various mechanisms.
Antiphospholipid syndrome (APS) is a systemic autoimmune disease characterised by recurrent thrombotic events, pregnancy loss and thrombocytopenia and the presence of antiphospholipid antibodies (APL). The exact pathomechanism of APS is still unknown, thus we investigated the effect of anti-β2-glycoprotein I (anti-β2GPI) on thrombin generation in different plasma samples. For the separation of anti-β2GPI IgG, overall 12 APS patients were selected. The criteria were the existence of lupus anticoagulant, and the presence of anti-CL and anti-β2GPI, the latter exceeding at least 25 times the upper reference limit. We purified anti-β2GPI IgG antibodies from APS patients by affinity chromatography and added the antibodies to normal pooled, and heterozygous forms of inherited thrombophilia plasma samples (prothrombin G20210A, factor V Leiden). To further specify the mechanism of the effect, we also used factor deficient plasmas in the thrombin generation assay. In normal pooled plasma, the anti-β2GPI significantly prolonged Lag Time according to the lupus anticoagulant effect, in contrast, it also elevated Peak Thrombin significantly, which suggests a procoagulant effect. The antibody was also able to exert this multi-faceted effect both in FVLeiden heterozygous plasma and prothrombin G20210A heterozygous polymorphism, however, the prolonging effect was more remarkable in the latter. By using factor deficient plasmas, it was found that FVII is required for the prolongation, while intrinsic factors are needed for the elevation of the Peak Thrombin. The anti-β2GPI autoantibodies exert their effect in both normal and thrombophilic plasmas via various mechanisms.


Rabies antibody prevalence and virus tissue tropism in wild carnivores in Virginia.
Carnivores trapped in a rabies control program in Virginia were examined for rabies virus and serum neutralizing antibody. Local antibody prevalence ranged from 0% to 29% in gray foxes (Urocyon cinereoargenteus). Rabies virus was pantropic in naturally infected gray foxes and a bobcat (Lynx rufus).


Analysis of 107 breast lesions with automated 3D ultrasound and comparison with mammography and manual ultrasound.
Our aim was to investigate the diagnostic potential of an automated ultrasound (US) breast scanner prototype and compare it with manual US and mammography. Ninety-seven patients with a total of 107 breast lesions had mammograms, manual US and an automated breast US scan. Multiplanar reconstructions in coronal, axial and the sagittal view were reconstructed from the automated dataset and visualized. After biopsy, all lesions were confirmed histologically. The data were evaluated according to the BIRADS (Breast Imaging Reporting and Data System) classification. The sensitivity and specificity were analyzed. The BIRADS criterion "margin" was significantly related to the overall BIRADS classification, independently of the US method being used. The sensitivity of mammography was significantly lower than of each US method (Fisher's exact test with p<0.05). There were no significant differences between the US methods. The reconstructed third (axial) image plane of the whole breast, which corresponds to a craniocaudal mammogram, can give additional information about both, site and differential diagnosis of a lesion. Although image quality was sufficient, automated US is not good enough to replace manual US at this time.


Association of physical activity with metabolic profile from adolescence to adulthood.
Physical activity benefits cardiometabolic health, but little is known about its detailed links with serum lipoproteins, amino acids, and glucose metabolism at young age. We therefore studied the association of physical activity with a comprehensive metabolic profile measured repeatedly in adolescence. The cohort is derived from the longitudinal Special Turku Coronary Risk Factor Intervention Project. At ages 13, 15, 17, and 19 years, data on physical activity were collected by a questionnaire, and circulating metabolic measures were quantified by nuclear magnetic resonance metabolomics from repeatedly assessed serum samples (age 13: n = 503, 15: n = 472, 17: n = 466, and 19: n = 361). Leisure-time physical activity (LTPA;MET h/wk) was directly associated with concentrations of polyunsaturated fatty acids, and inversely with the ratio of monounsaturated fatty acids to total fatty acids (-0.006SD; [-0.008, -0.003]; p < 0.0001). LTPA was inversely associated with very-low-density lipoprotein (VLDL) particle concentration (-0.003SD; [-0.005, -0.001]; p = 0.002) and VLDL particle size (-0.005SD; [-0.007, -0.003]; p < 0.0001). LTPA showed direct association with the particle concentration and size of high-density lipoprotein (HDL), and HDL cholesterol concentration (0.004SD; [0.002, 0.006]; p < 0.0001). Inverse associations of LTPA with triglyceride and total lipid concentrations in large to small sized VLDL subclasses were found. Weaker associations were seen for other metabolic measures including inverse associations with concentrations of lactate, isoleucine, glycoprotein acetylation, and a direct association with creatinine concentration. The results remained after adjusting for body mass index and proportions of energy intakes from macronutrients. Physical activity during adolescence is beneficially associated with the metabolic profile including novel markers. The results support recommendations on physical activity during adolescence to promote health and possibly reduce future disease risks.


Role of Oxygen Starvation in Right Ventricular Decompensation and Failure in Pulmonary Arterial Hypertension.
Right ventricular (RV) function and eventually failure determine outcome in patients with pulmonary arterial hypertension (PAH). Initially, RV responds to an increased load caused by PAH with adaptive hypertrophy; however, eventually RV failure ensues. Unfortunately, it is unclear what causes the transition from compensated RV hypertrophy to decompensated RV failure. Moreover, at present, there are no therapies for RV failure; those for left ventricular (LV) failure are ineffective, and no therapies specifically targeting RV are available. Thus there is a clear need for understanding the biology of RV failure and differences in physiology and pathophysiology between RV and LV that can ultimately lead to development of such therapies. In this paper, we discuss RV adaptation and maladaptation in PAH, with a particular focus of oxygen delivery and hypoxia as the principal drivers of RV hypertrophy and failure, and attempt to pinpoint potential sites for therapy.


Genetic Basis of Common Human Disease: Insight into the Role of Missense SNPs from Genome-Wide Association Studies.
Recent genome-wide association studies (GWAS) have led to the reliable identification of single nucleotide polymorphisms (SNPs) at a number of loci associated with increased risk of specific common human diseases. Each such locus implicates multiple possible candidate SNPs for involvement in disease mechanism. A variety of mechanisms may link the presence of an SNP to altered in vivo gene product function and hence contribute to disease risk. Here, we report an analysis of the role of one of these mechanisms, missense SNPs (msSNPs) in proteins in seven complex trait diseases. Linkage disequilibrium information was used to identify possible candidate msSNPs associated with increased disease risk at each of 356 loci for the seven diseases. Two computational methods were used to estimate which of these SNPs has a significant impact on in vivo protein function. 69% of the loci have at least one candidate msSNP and 33% have at least one predicted high-impact msSNP. In some cases, these SNPs are in well-established disease-related proteins, such as MST1 (macrophage stimulating 1) for Crohn's disease. In others, they are in proteins identified by GWAS as likely candidates for disease relevance, but previously without known mechanism, such as ADAMTS13 (ADAM metallopeptidase with thrombospondin type 1 motif, 13) for coronary artery disease. In still other cases, the missense SNPs are in proteins not previously suggested as disease candidates, such as TUBB1 (tubulin, beta 1, class VI) for hypertension. Together, these data support a substantial role for this class of SNPs in susceptibility to common human disease.


Sensitive electrochemical detection of DNA damage based on in situ double strand growth via hybridization chain reaction.
Detection of DNA damage caused by ∙OH or radiation has led to rapidly growing interest in the fields of drug development, biochemistry, clinic diagnostics, and environmental evaluation. Electrochemical methods have been applied for DNA damage detection because of their fast and sensitive response. However, most of the electrochemical methods for DNA damage detection commonly require immobilization of the strands on the electrode surface. In the present work, sensitive electrochemical monitoring of DNA damage was realized successfully on the basis of in situ DNA chain growth by use of a hybridization chain reaction (HCR) technique. By use of [Ru(NH3)6]3+ as the signal probe and the Fenton reaction as the ∙OH generator, ultrasensitive detection of DNA damage induced by ∙OH was realized successfully through differential pulse voltammetry with a linear relationship of ∙OH concentration from 15 to 750 pM and a detection limit of 12 pM. Furthermore, environmental DNA-damaging UV light was tested as the lesion source to demonstrate the practicability and reliability of the proposed HCR-based amplified signal method for DNA damage detection. By integrating the HCR technique with an electrochemical method, we provide a promising alternative approach to extend the applications of electrochemical methods in bioanalytical detection of DNA damage. Graphical abstract Sensitive electrochemical monitoring of DNA damage has been realized successfully on the basis of in situ DNA chain growth by use of hybridization chain reaction (HCR). Detection of DNA damage caused by ∙OH was realized successfully through differential pulse voltammetry with a linear relationship of ∙OH concentration from 15 to 750 pM and a detection limit of 12 pM.


Proteomic characterization of vanA-containing Enterococcus recovered from Seagulls at the Berlengas Natural Reserve, W Portugal.
Enterococci have emerged as the third most common cause of nosocomial infections, requiring bactericidal antimicrobial therapy. Although vancomycin resistance is a major problem in clinics and has emerged in an important extend in farm animals, few studies have examined it in wild animals. To determine the prevalence of vanA-containing Enterococcus strains among faecal samples of Seagulls (Larus cachinnans) of Berlengas Natural Reserve of Portugal, we developed a proteomic approach integrated with genomic data. The purpose was to detect the maximum number of proteins that vary in different enterococci species which are thought to be connected in some, as yet unknown, way to antibiotic resistance. From the 57 seagull samples, 54 faecal samples showed the presence of Enterococcus isolates (94.7%). For the enterococci, E. faecium was the most prevalent species in seagulls (50%), followed by E. faecalis and E. durans (10.4%), and E. hirae (6.3%). VanA-containing enterococcal strains were detected in 10.5% of the 57 seagull faecal samples studied. Four of the vanA-containing enterococci were identified as E. faecium and two as E. durans. The tet(M) gene was found in all five tetracycline-resistant vanA strains. The erm(B) gene was demonstrated in all six erythromycin-resistant vanA strains. The hyl virulence gene was detected in all four vanA-containing E. faecium isolates in this study, and two of them harboured the purK1 allele. In addition these strains also showed ampicillin and ciprofoxacin resistance. The whole-cell proteomic profile of vanA-containing Enterococcus strains was applied to evaluate the discriminatory power of this technique for their identification. The major differences among species-specific profiles were found in the positions corresponding to 97-45 kDa. Sixty individualized protein spots for each vanA isolate was identified and suitable for peptide mass fingerprinting measures by spectrometry measuring (MALDI/TOF MS) and their identification through bioinformatic databases query. The proteins were classified in different groups according to their biological function: protein biosynthesis, ATP synthesis, glycolysis, conjugation and antibiotic resistance. Taking into account the origin of these strains and its relation to infectious processes in humans and animals, it is important to explore the proteome of new strains which might serve as protein biomarkers for biological activity. The comprehensive description of proteins isolated from vancomycin-resistant Enterococcus faecium and E. durans may provide new targets for development of antimicrobial agents. This knowledge may help to identify new biomarkers of antibiotic resistance and virulence factors.


Evaluation of anti-malarial drugs' use in Fitche Hospital, North Shoa, Oromia Region, Ethiopia.
Retrospective evaluation of anti-malarial drugs' use in Fitche Hospital, North Shoa, Oromia Region, Ethiopia. Retrospective cross-sectional study design was conducted using selected patients cards of 1-year (January 2012-January 2013 G.C) with anti-malarial agents from January 18 to 30, 2013. The sample size was calculated by using Joint Commission on the Accreditation of Health care Organization criteria and sampling was done by using a systematic random sampling technique. One hundred and twenty-five patient cards with anti-malarial drugs were reviewed of which 32.8%, 21.6%, 15.2% belongs to age range of 20-29, 10-19, and 30-39, respectively. Chloroquine prescription accounts for 50.4% from total anti-malarial drugs. 71.2% and 78.4% of patients received antibiotics and analgesics, respectively, with anti-malarial drugs. 77.6% of drugs were prescribed by generic name while the brand name was 22.39%. The study done in Fitche Hospital revealed that the use of anti-malarial agent was not in complete agreement with the current guideline of Ethiopia despite good practice.


Isolation and in vitro translation of the messenger RNA coding for pancreatic amylase.
RNA prepared from dog pancreas polysomes or microsomes directs the synthesis of pancreas-specific proteins in heterologous cell-free translation systems. A translation product, approximately 1500 daltons larger than authentic amylase, corresponding to pancreatic amylase was identified by immunoprecipitation with anti-amylase gamma-globulin and tryptic peptide analysis. We suggest that this larger form of amylase is an amylase precursor. Using amylase immunoprecipitation of reticulocyte translation reactions as an assay, we have shown that greater than 99% of the mRNA for amylase is associated with polysomes bound to the endoplasmic reticulum. Electrophoresis of pancreatic mRNA preparations in formamide-containing polyacrylamide gels and subsequent translation of the fractions have shown that amylase mRNA is of a discrete size with a mobility equivalent to that of 18 S ribosomal RNA, and therefore significantly larger than required to code solely for the amino acid sequence of the amylase precursor.


Tuning the reduction power of supported gold nanoparticle photocatalysts for selective reductions by manipulating the wavelength of visible light irradiation.
Gold nanoparticles supported on CeO(2) were found to be efficient photocatalysts for three selective reductions of organic compounds at ambient temperatures, under irradiation of visible light; their reduction ability can be tuned by manipulating the irradiation wavelength.


Analysis of the influencing factors of energy-related carbon emissions in Kazakhstan at different stages.
This study analyzes Kazakhstan's influencing factors of energy-related carbon emissions in different stages, and the study period (1992-2014) was divided into four stages by using the logarithmic mean Divisia index (LMDI) method. In the low efficiency and high output stage, Kazakhstan had the most energy-related carbon emissions. The total energy-related carbon emissions might be positive or negative in the high efficiency and high output stage and the low efficiency and low output stage, and this was mainly determined by the energy intensity effect or the economic output effect. Different influencing factors had different effects in the different stages from 1992 to 2014. The economic output effect was the first contributor for promoting energy-related carbon emissions, and the energy intensity factor was the first contributor for suppressing energy-related carbon emissions from 1992 to 2014. Finally, policy recommendations in terms of the main influencing factors are put forward, including the low-carbon economic development mode transformation, technological innovation, and renewable energy development.


Medulloblastoma--prognostic factors and outcome of treatment: review of the Mayo Clinic experience.
From March 1965 through December 1984, 58 patients (35 male and 23 female patients; median age, 17 years) with posterior fossa (PF) medulloblastoma underwent surgical treatment and postoperative radiation therapy at our institution. Radiation fields were the craniospinal axis in 39 patients, PF plus spinal axis in 12, PF in 6, and whole brain in 1. Median radiation doses were 43 Gy (22 to 60 Gy) to the PF and 34 Gy (6.2 to 50 Gy) to the spinal axis. Overall 5- and 10-year survivals were 50% and 33%, respectively; 5- and 10-year relapse-free survivals were 46% and 32%. Treatment failed in 34 patients (59%): in 18 who had irradiation to the craniospinal axis (13 had received 50 Gy or less to the PF) and in 16 who had a radiation field of less than the craniospinal axis. A statistically significant (P less than 0.05) improvement in 10-year survival was associated with the following prognostic variables: PF dose of 50 Gy or more, whole-brain irradiation, and spinal axis irradiation. In comparison with subtotal resection, total resection was correlated with better 10-year relapse-free survival but not overall survival. All five patients with initial treatment failure only in the spine had received a radiation dose of 30 Gy or less to the spinal axis. The 2-year survival after relapse was 46% with salvage chemotherapy or irradiation in 23 patients and 0% in the 11 patients who received no further treatment (P less than 0.01).


Are there hunger and satiety factors in the blood of domestic fowls?
The presence of blood-borne hunger and satiety factors in domestic fowls was investigated in two experiments. In the first, birds that had previously been fed ad libitum or fasted for 24 h ate less after their blood had been transfused with that of fed or fasted donors than they did after two control treatments. Blood from fed donors suppressed intake more than did blood from fasted donors, but not significantly so. In the second experiment, previously fed birds ate less in the first hour after intravenous injection of reconstituted plasma from 24-h fasted donors, than after control injections of saline. The same treatments had little or no effect with subjects that had previously been fasted for 24 h. These results consistently indicate an absence of any hunger factor, but are less conclusive with regard to the presence of a satiety factor, for which there is recent evidence in fowls from central administration of plasma. It is suggested that hunger and satiety may be associated with scarcity and abundance of the same humoral factor(s), and that monitoring of blood composition may provide the necessary feedback for longer-term adjustments in food intake, modifying short-term control based on emptying and filling of part(s) of the alimentary tract.


Effect of pasture vs. concentrate diet on CLA isomer distribution in different tissue lipids of beef cattle.
This study examined the effects of feeding pasture vs. concentrate on the distribution of CLA isomers in the lipids of longissimus and semitendinosus muscle, liver and heart muscle, and subcutaneous fat in beef bulls. Sixty-four German Holstein and German Simmental bulls were randomly allocated to either an indoor concentrate system or periods of pasture feeding followed by a finishing period on a concentrate containing linseed to enhance their beef content of n-3 PUFA and CLA. The concentrations of CLA isomers in the different tissues were determined by GC and silver ion HPLC. The diet affected the distribution of individual CLA isomers in the lipids of the different tissues. The concentration (mg/100 g fresh tissue) of the most prominent isomer, cis-9,trans-11 18:2, was increased up to 1.5 times in liver and heart tissue of bulls fed on pasture as compared with concentrate. However, no diet effect was observed for cis-9,trans-11 18:2 in the lipids of longissimus muscle and subcutaneous fat. In all tissues, the second-most abundant CLA isomer in concentrate-fed bulls was trans-7,cis-9 18:2. In contrast, trans-11,cis-13 18:2 was the second-most abundant CLA isomer in all investigated tissue lipids of pasture-fed bulls. The concentration of the trans-11,cis-13 18:2 isomer was up to 15 times higher in tissues of pasture-fed bulls as compared with concentrate-fed animals. Furthermore, diet affected the concentrations of the CLA trans,trans 18:2 isomers. Pasture feeding significantly increased the concentrations of some trans,trans 18:2 isomers as compared with concentrate, predominantly trans-12,trans-14 18:2 and trans-11,trans-13 18:2. Overall, pasture feeding resulted in significantly increased concentrations of the sum of CLA isomers in the lipids of longissimus muscle, subcutaneous fat, heart and liver muscle of German Holstein and German Simmental bulls, but not in semitendinosus muscle.


Incorporating 4D into Bioprinting: Real-Time Magnetically Directed Collagen Fiber Alignment for Generating Complex Multilayered Tissues.
In vitro multilayered tissues with mimetic architectures resembling native tissues are valuable tools for application in medical research. In this study, an advanced bioprinting strategy is presented for aligning collagen fibers contained in functional bioinks. Streptavidin-coated iron nanoparticles are embedded in printable bioinks with varying concentrations of low gelling temperature agarose and type I collagen. By applying a straightforward magnetic-based mechanism in hydrogels during bioprinting, it is possible to align collagen fibers in less concentrated hydrogel blends with a maximum agarose concentration of 0.5 w/v%. Conversely, more elevated concentrations of agarose in printable blends show random collagen fiber distribution. Interestingly, hydrogel blends with unidirectionally aligned collagen fibers show significantly higher compression moduli compared to hydrogel blends including random fibers. Considering its application in the field of cartilage tissue engineering, bioprinted constructs with alternating layers of aligned and random fibers are fabricated. After 21 days of culture, cell-loaded constructs with alternating layers of aligned and random fibers express markedly more collagen II in comparison to solely randomly oriented fiber constructs. These encouraging results translate the importance of the structure and architecture of bioinks used in bioprinting in light of their use for tissue engineering and personalized medical applications.


Activated murine neutrophils induce unscheduled DNA synthesis in B lymphocytes.
Activated neutrophils induce DNA damage in neighboring cells by secreting reactive oxygen compounds into the extracellular milieu. Repair of this damage is required to prevent mutagenesis and neoplastic transformation. Conditions were established to detect the activation of excision-repair pathways (unscheduled DNA synthesis) by measuring stimulated thymidine uptake in target B lymphocytes exposed to activated neutrophils. Murine neutrophils were cocultured in serum-free medium with splenic B cells or with murine plasmacytoma cells for 2 h. Unscheduled DNA synthesis in the B cells was detected at neutrophil:target cell ratios of 1:1 to 4:1 when the neutrophils were activated with phorbol myristate acetate. Reagent H2O2 alone (> or = 6 microM) also induced UDS whereas HOCl (up to 4 mM) did not. No repair synthesis was observed within the neutrophils themselves. Control experiments indicated that the induction of UDS by neutrophils and H2O2 was not due to formation of a stable genotoxic compound from HU. On the contrary, scavenging of free H2O2 by HU probably lowered the levels of UDS that could be detected by these agents. Induction of unscheduled DNA synthesis by neutrophils and H2O2 occurred under conditions of less cytostasis than was found with other DNA-damaging agents such as 4-nitroquinoline 1-oxide or gamma-irradiation. This may reflect a heightened responsiveness of the cells to repair of damage from physiological oxidants. The results demonstrate that DNA damage induced by reactive oxygen intermediates can be repaired by nucleotide excision-repair pathways.


Microbial production of levansucrase for synthesis of fructooligosaccharides and levan.
A newly isolated thermophilic bacterial strain from Tunisian thermal source was identified as Bacillus sp. and was selected for its ability to produce extracellular levansucrase. Following the optimization of carbon source, nitrogen source, temperature and initial pH of the growth medium in submerged liquid cultures. In fact, sucrose was found to be a good inducer of levansucrase enzymes. The optimal temperature and pH of the levansucrase were 50°C and 6.5, respectively and its activity increased four folds in the presence of 50mM Fe(2+). This enzyme exhibited a remarkable stability and retained 100% of its original activity at 50°C for more than 1h at pH 6.5. The half-life of the enzyme was 1h at 90°C. Crude enzyme of Bacillus sp. rich in levansucrase was established for the synthesis of fructooligosaccharides and levan. Bacillus sp. could therefore be considered as a satisfactory and promising producer of thermostable levansucrases. Contrary to other levansucrases, the one presented in the current study was able to produce high levels of levan with high molecular weight at 50°C and having an important effect as a hypoglycemic agent which was demonstrated in our previous publications (Dahech et al., 2011 [25]) and as a hypo-cholesterolemic agent which will be investigated in further research.


The effect of 3-Methyl-4-Nitrophenol on the early ovarian follicle development in mice by disrupting the clock genes expression.
3-Methyl-4-Nitrophenol (PNMC) is the main degradation product of organophosphate insecticide fenitrothion and a major component of diesel exhaust particles, which is now becoming a widely spread environmental endocrine disruptor. Previous reports showed PNMC exposure can affect the female reproductive system and ovarian function; however, the mechanism remains unclear. The main purpose of this study is to clarify the mechanism underlying the adverse effects of neonatal PNMC treatment on ovarian functions. The neonatal female mice were exposed to 10 mg/kg PNMC and the ovaries were collected on the 7th day after birth. The changes of follicular composition in mice ovaries were analyzed by histological staining, which showed that the proportion of primordial follicles in the ovaries treated by PNMC decreased, while the proportion of secondary follicles increased. The ovarian function was also investigated by detecting the expressions of steroidogenic enzymes (Star, Cyp11a1, Hsd3b1, Cyp17a1, Cyp19a1), gonadotropin receptors (Fshr and Lhr), androgen receptor (Ar), and estrogen receptors (Esr1 and Esr2) by immunohistochemistry or/and real-time quantitative PCR. The expression of Hsd3b1, Cyp17a1 and Esr2 were increased significantly in the PNMC exposed ovaries. Moreover, the expression patterns of clock genes (Bmal1, Clock, Per1, Per2, Cry1, Cry2 and Nr1d1) were disrupted in the ovaries after PNMC exposure. Furthermore, either the expression of DNA Methyltransferase Dnmt3b, or the methylation ratio of CpG islands in the upstream of Cry1 promoter regions were significantly decreased in PNMC exposed ovaries. Altogether, these results indicate that PNMC exposure affects follicle development and ovarian function by interfering with the epigenetic modification and disrupting the expression of clock genes.


Principles of Wound Management and Wound Healing in Exotic Pets.
The care of wounds in exotic animal species can be a challenging endeavor. Special considerations must be made in regard to the animal's temperament and behavior, unique anatomy and small size, and tendency toward secondary stress-related health problems. It is important to assess the entire patient with adequate systemic evaluation and consideration of proper nutrition and husbandry, which could ultimately affect wound healing. This article summarizes the general phases of wound healing, factors that affect healing, and principles of wound management. Emphasis is placed on novel methods of treating wounds and species differences in wound management and healing.


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Because of their potential health risk the amount of the Fusariumtoxins deoxynivalenol (DON) and zearalenone (ZEA) is an important quality parameter for cereals.With regard to the fact that in the european union limiting values for these Fusarium-toxins are discussed, the analysis of cerealfood from the market makes sense in order to get a realistic image of the load of consumers. Cereals mainly get in human food chain in form of bread and other bakery products. Publications about evaluation of the influence of the breadmaking process (removal or destruction of mycotoxins by fermentation and heating) are rare and sometimes give contrary results. [3, 4]Our food control laboratory started a monitoring program in order to get more informations about the influence of this bread-making process. Therefore bread and the flours the bread made of are taken from bakeries in district of Tübingen (Germany). First results are presented.