Progressive disseminated histoplasmosis in Latin America and the Caribbean in people receiving highly active antiretroviral therapy for HIV infection: A systematic review.
Histoplasmosis is the most clinically significant mycosis in Latin America; still it has been neglected in people with human immunodeficiency virus (HIV). There is limited information about its contribution to morbidity and mortality in this population. We conducted a systematic review of scientific literature to provide an estimation of the frequency and mortality of histoplasmosis among people with HIV receiving highly active antiretroviral therapy (HAART) in Latin America, and factors associated with mortality. We searched articles in PubMed, Scopus, WHO Global health library, and Scielo using different combination of terms including "histoplasmosis" and HAART. We identified 949 articles, removed 662 duplicated; screened 287 abstracts; reviewed full text of 53 articles; and selected 15 articles that provided information on the number of patients studied, included patients receiving ART, and reported any measure of frequency estimate for qualitative synthesis. Studies were conducted in Argentina (n = 4), Brazil (n = 6), Colombia (n = 2), French Guyana and the Bahamas (=2), and Guatemala (n = 1). Heterogeneity of studies characteristics precluded any aggregated estimates. Histoplamosis was frequent in these cohort studies and mortality was high despite the use of HAART. Low CD4 counts, delayed HAART initiation and poor adherence were related to increased incidence, poor prognosis and increased mortality, respectively. Histoplasmosis may be an important contributor to mortality in people with HIV in Latin America. Diagnostic delays represent an important limitation for improving care of patients suspected to have histoplasmosis. Reducing histoplasmosis diagnostic delays and therapy initiation is needed to further decrease mortality.


[Abnormal movements in a case of extrapontine myelinolysis. Review of the literature].
We present a case of extra-pontine myelinolysis caused by acute hypernatraemia in which a complex picture of late onset extrapyramdial features, choreodystonia and parkinsonism developed. Repeated physical examinations, neurophysiological and neuroimaging studies using magnetic resonance all indicated an extra-pontine site of the lesions, which symmetrically affected the striate and to a lesser extent both thalami. We review the relevant literature available and analyze the cases described as having abnormal movements associated with a myelinolytic syndrome. Extra-pontine myelinolysis is a cause of acquired dystonia, chorea and parkinsonism, generally of late onset and with varying response to treatment. The association of hypernatraemia, hyperglycaemia and liver transplant seem to predispose to the development of extra-pontine lesions.


Current reversal in polar flock at order-disorder interface.
We studied a system of polar self-propelled particles (SPPs) on a thin rectangular channel designed into three regions of order-disorder-order. The division of the three regions is made on the basis of the noise SPPs experience in the respective regions. The noise in the two wide regions is chosen lower than the critical noise of order-disorder transition and noise in the middle region or interface is higher than the critical noise. This makes the geometry of the system analogous to the Josephson junction (JJ) in solid-state physics. Keeping all other parameters fixed, we study the properties of the moving SPPs in the bulk as well as along the interface for different widths of the junction. On increasing interface width, the system shows an order-to-disorder transition from coherent moving SPPs in the whole system to the interrupted current for large interface width. Surprisingly, inside the interface, we observed the current reversal for intermediate widths of the interface. Such current reversal is due to the strong randomness present inside the interface, which makes the wall of the interface reflecting. Hence, our study gives new interesting collective properties of SPPs at the interface which can be useful to design switching devices using active agents.


Structural mechanism of replication stalling on a bulky amino-polycyclic aromatic hydrocarbon DNA adduct by a y family DNA polymerase.
Polycyclic aromatic hydrocarbons and their nitro derivatives are culprits of the detrimental health effects of environmental pollution. These hydrophobic compounds metabolize to reactive species and attach to DNA producing bulky lesions, such as N-[deoxyguanosine-8-yl]-1-aminopyrene (APG), in genomic DNA. The bulky adducts block DNA replication by high-fidelity polymerases and compromise replication fidelities and efficiencies by specialized lesion bypass polymerases. Here we present three crystal structures of the DNA polymerase Dpo4, a model translesion DNA polymerase of the Y family, in complex with APG-lesion-containing DNA in pre-insertion and extension stages. APG is captured in two conformations in the pre-insertion complex; one is highly exposed to the solvent, whereas the other is harbored in a shallow cleft between the finger and unique Y family little finger domain. In contrast, APG is in a single conformation at the extension stage, in which the pyrene ring is sandwiched between the little finger domain and a base from the turning back single-stranded template strand. Strikingly, a nucleotide intercalates the DNA helix to form a quaternary complex with Dpo4, DNA, and an incoming nucleotide, which stabilizes the distorted DNA structure at the extension stage. The unique APG DNA conformations in Dpo4 inhibit DNA translocation through the polymerase active site for APG bypass. We also modeled an insertion complex that illustrates a solvent-exposed pyrene ring contributing to an unstable insertion state. The structural work combined with our lesion replication assays provides a novel structural mechanism on bypass of DNA adducts containing polycyclic aromatic hydrocarbon moieties.


Evidence for adaptive evolution of the G6pd gene in the Drosophila melanogaster and Drosophila simulans lineages.
Proponents of the neutral theory argue that evolution at the molecular level largely reflects a process of random genetic drift of neutral mutations. Under this theory, levels of interspecific divergence and intraspecific polymorphism are expected to be correlated across classes of nucleotide or amino acid sequences with different degrees of functional constraint, such as synonymous and replacement sites. Nucleotide sites with reduced polymorphism should show comparably reduced levels of interspecific divergence. To examine this hypothesis, we have sequenced 32 and 12 copies of the glucose-6-phosphate dehydrogenase (G6pd) gene in Drosophila melanogaster and Drosophila simulans, respectively. Both species exhibit similar levels of nucleotide polymorphism at synonymous sites. D. melanogaster shows two amino acid polymorphisms, one associated with the cosmopolitan allozyme polymorphism and a second with an allozyme polymorphism endemic to European and North African populations. In contrast, D. simulans shows no replacement polymorphism. While synonymous divergence between species is 10%, which is typical of other genes, there are 21 replacement differences. This level of amino acid sequence divergence, when contrasted with levels of amino acid polymorphism, silent polymorphism, and divergence, is in 10-fold excess over that expected under the neutral model of molecular evolution. We propose that this excess divergence reflects episodes of natural selection on G6pd resulting in fixation of advantageous amino acid mutations in these two recently separated lineages.


HULIS Enhancement of Hydroxyl Radical Formation from Fe(II): Kinetics of Fulvic Acid-Fe(II) Complexes in the Presence of Lung Antioxidants.
Oxidative stress mediated by reactive oxygen species (ROS) is a hypothesized mechanism for particulate-matter related health effects. Fe(II) is a key player in ROS formation in surrogate lung fluid (SLF) containing antioxidants. Humic-like substances (HULIS) in particulate matter such as biomass burning aerosol chelate Fe(II), but the effect on ROS formation in the presence of lung antioxidants is not known. We use Suwanee River Fulvic Acid (SRFA) as a surrogate for HULIS and investigate its effect on OH formation from Fe(II). For the first time, a chemical kinetics model was developed to explain behavior of Fe(II) and SRFA in SLF. Model and experimental results are used to find best-fit rate coefficients for key reactions. Modeling results indicate SRFA enhances Fe-mediated reduction of O2 to O2- and destruction of H2O2 to OH to 5.1 ± 1.5 and (4.3 ± 1.4) × 103 M-1 s-1 respectively. Best-fit rates for Citrate-Fe(II) mediated O2 to O2- and H2O2 to OH were 3.0 ± 0.7 and (4.2 ± 1.7) × 103 M-1 s-1 respectively. The kinetics model agrees with both the experimental results and thermodynamic model calculations of chemical speciation for 0 and 5 μg/mL SRFA, but both models are less successful at predicting further enhancements to OH formation at higher SRFA Concentrations.


Evaluation of Burkholderia cepacia Complex Bacteria Pathogenicity Using Caenorhabditis elegans.
This protocol describes two biological assays to evaluate pathogenicity of Burkholderia cepacia complex (Bcc) strains against the nematode Caenorhabditis elegans. Specifically, these two assays allow one to identify if the under-investigated Bcc strains are able to kill the nematodes by intestinal colonization (slow killing assay, SKA) or by toxins production (fast killing assay, FKA). The principal differences between the two assays rely on the different killing kinetics for worms.


Bloody nipple discharge in a 7-month-old boy.
Isolated bloody nipple discharge is rare in infancy and is usually idiopathic. Discharge commonly resolves spontaneously, and ultrasonography is a useful diagnostic technique to detect the cause of discharge. The authors report a 7-month-old boy who presented with unilateral spontaneous bloody nipple discharge for 1 month without signs of infection or mass.


Isometric strength, sprint power, and aerobic power in individuals with a spinal cord injury.
This study investigated in rather specific wheelchair tests the relationships among estimates of isometric upper-body strength (Fiso), sprint power (P30), aerobic power (VO2peak), and maximal power output (POaer) in a group of 44 men (age 34 +/- 12 yr) with longstanding spinal cord injuries ranging from C4/C5 to L5. Fiso was defined as the maximum force that could be exerted on the blocked rims of a stationary wheelchair ergometer. The estimation of P30 involved the measurement of the mean power during a 30-s all-out sprint test on the same wheelchair ergometer. VO2peak and POaer were determined as the peak oxygen uptake and highest sustained power output during a discontinuous progressive maximal exercise test on a motorized treadmill, while subjects used their own daily use wheelchair. Fiso ranged from 1.5 N.kg-1 (mean of both arms) in the group with quadriplegia to 3.4 N.kg-1 in the group with lowest-lesions, and P30 ranged from 0.5 to 1.5 W.kg-1 among the subjects. VO2peak ranged from 13.6 ml.kg-1.min-1 in the group with quadriplegia to 31.3 ml.kg-1.min-1 in the group with lowest-lesions, and POaer ranged from 0.4 to 1.1 W.kg-1. Strong positive relationships (r = 0.81-0.92) were demonstrated among all variables. Regression equations among variables were calculated: P30 = 0.51 Fiso - 0.18 (R2 = 0.75); POaer = 0.34 Fiso - 0.02 (R2 = 0.66); POaer = 0.67 P30 + 0.11 (R2 = 0.81); VO2peak = 6.52 Fiso + 4.15 (R2 = 0.76); VO2peak = 12.03 P30 + 7.43 (R2 = 0.77); VO2peak = 16.81 POaer + 6.44 (R2 = 0.84).(ABSTRACT TRUNCATED AT 250 WORDS)


[Interaction of human embryo cells derived from different tissues with lymphoblastoid cell lines].
Cells from skin and muscle of two month human embryo and from nasopharynx, tonsil, lung and kidney of four month human embryo were cocultivated with lymphoblastoid cells--B95-8 and Raji. It is found that all the epithelial cells, such as the focal growth of epithelioid cells from the tonsil and nasopharynx as well as kidney cells from the human embryo can not adhere to any kind of lymphoblastoid cells; but fibroblasts from muscle and skin of two month embryo or from nasopharynx, tonsil and lung of four month embryo have the ability of sticking to lymphoblastoid cells. However, only the fibroblastoid cells from the nasopharynx of human embryo have the ability to stick and to fuse with lymphoblastoid cells--B95-8 in cocultivation and to activate the EBV production. The mechanism of this phenomenon and its biological functions will be studied in detail.


The Diverse Roles of Phagocytes During Bacterial and Fungal Infections and Sterile Inflammation: Lessons From Zebrafish.
The immediate and natural reaction to both infectious challenges and sterile insults (wounds, tissue trauma or crystal deposition) is an acute inflammatory response. This inflammatory response is mediated by activation of the innate immune system largely comprising professional phagocytes (neutrophils and macrophages). Zebrafish (danio rerio) larvae possess many advantages as a model organism, including their genetic tractability and highly conserved innate immune system. Exploiting these attributes and the live imaging potential of optically transparent zebrafish larvae has greatly contributed to our understanding of how neutrophils and macrophages orchestrate the initiation and resolution phases of inflammatory responses. Numerous bacterial and fungal infection models have been successfully established using zebrafish as an animal model and studies investigating neutrophil and macrophage behavior to sterile insults have also provided unique insights. In this review we highlight how examining the larval zebrafish response to specific bacterial and fungal pathogens has uncovered cellular and molecular mechanisms behind a variety of phagocyte responses, from those that protect the host to those that are detrimental. We also describe how modeling sterile inflammation in larval zebrafish has provided an opportunity to dissect signaling pathways that control the recruitment, and fate, of phagocytes at inflammatory sites. Finally, we briefly discuss some current limitations, and opportunities to improve, the zebrafish model system for studying phagocyte biology.


Self assembled composites of luminescent Ru(II) metallopolymers and the Dawson polyoxometalate α-[Mo18O54(SO4)2]4-.
The interaction of two luminescent metallopolymers; [Ru(bpy)(2)(PVP)(10)](2+) and [Ru(bpy)(2)(CAIP)co-poly(7)](+), where bpy is 2,2'-bipyridyl, PVP is polyvinylpyridine, and (CAIP)co-poly(7) is poly(styrene(6)-co-p-(aminomethyl)styrene) amide linked to 2-(4-carboxyphenyl)imidazo[4,5-f] [1,10]phenanthroline, with the Dawson polyoxomolybdate α-[Mo(18)O(54)(SO(4))(2)](4-) is described. Both metallopolymers undergo electrostatic association with the polyoxometalate. From both electronic and luminescence spectroscopy the thermodynamic products were determined to be {[Ru(bpy)(2)(PVP)(10)](4.5)[Mo(18)O(54)(SO(4))(2)]}(5+) and {[Ru(bpy)(2)(CAIP)co-poly(7)](5)[Mo(18)O(54)(SO(4))(2)]}(+), i.e. in both instances, the number of ruthenium centres in the cluster exceeds the number required for charge neutralization of the molybdate centre. Association quenches the luminescence of the metallopolymer although, consistent with the excess of Ru(ii) present in the associated composites, emission is not completely extinguished even when a large excess of [Mo(18)O(54)(SO(4))(2)](4-) is present. The observed emission lifetime was not affected by [Mo(18)O(54)(SO(4))(2)](4-) therefore quenching was deemed static. The luminescent intensity data was found to fit best to a (sphere of action) Perrin model from which the radii of the quenching were calculated as 4.6 Å and 5.8 Å for [Ru(bpy)(2)(PVP)(10)](2+) and [Ru(bpy)(2)(CAIP co-poly)(7)](+) respectively. Both UV/Vis and resonance Raman data indicate the presence of a new optical transition centered around 490 nm for the composite, {[Ru(bpy)(2)(PVP)(10)](4.5)[Mo(18)O(54)(SO(4))(2)]}(5+) but not for {[Ru(bpy)(2)(CAIP)co-poly(7)](5)[Mo(18)O(54)(SO(4))(2)]}(+). This indicates strong electronic interaction between the metal centres in the former composite, which despite good thermodynamic analogy, is not observed for {[Ru(bpy)(2)(CAIP)co-poly(7)](5)[Mo(18)O(54)(SO(4))(2)]}(+). These results are consistent with photoelectrochemical studies of layer by layer assemblies of these films which indicate that the ruthenium centre sensitizes polyoxometalate photo-oxidation of benzyl alcohol in {[Ru(bpy)(2)(PVP)(10)](4.5)[Mo(18)O(54)(SO(4))(2)]}(5+) but not in {[Ru(bpy)(2)(CAIP)co-poly(7)](5)[Mo(18)O(54)(SO(4))(2)]}(+).


Pattern of recurrence of medulloblastoma after low-dose craniospinal radiotherapy.
We retrospectively evaluated relapse of medulloblastoma after low- or high-dose craniospinal radiotherapy, and after conventional or hyperfractionated posterior fossa irradiation. Ninety-two pediatric patients were treated postoperatively since 1970 at the University of California, San Francisco. Until 1989, we employed conventional fractionation with low (< or = 30 Gy) or high-dose craniospinal fields and low-dose (< or = 56 Gy) posterior fossa boosts. Recently, hyperfractionation delivered low- or high-dose to the craniospinal axis and high-dose to the posterior fossa. Most patients treated after 1979 received chemotherapy. Median follow-up was 70 months. Five-year disease-free survival was 36% (22% for poor-risk vs. 59% for good-risk patients). Five-year overall survival was 52% (43% for poor vs. 68% for good-risk). Neither the dose to the posterior fossa nor the craniospinal axis was statistically related to recurrence. Failure in the posterior fossa occurred despite boosts greater than 56 Gy. Females, over the age of 6 years, had significantly better relapse-free survival than males of the same age. Six of the 54 patients who relapsed were long-term survivors. Low-dose craniospinal radiotherapy, where the majority of patients received chemotherapy, was not associated with increased failure. High-dose posterior fossa hyperfractionation did not improve control. Long-term survival was noted in a number of patients after relapse. We recommend 60 Gy or greater with conventional fractions to the primary area, and continued study of low-dose craniospinal irradiation with adjuvant chemotherapy.


Considerations and guidance in designing equity-relevant clinical trials.
Health research has documented disparities in health and health outcomes within and between populations. When these disparities are unfair and avoidable they may be referred to as health inequities. Few trials attend to factors related to health inequities, and there is limited understanding about how to build consideration of health inequities into trials. Due consideration of health inequities is important to inform the design, conduct and reporting of trials so that research can build evidence to more effectively address health inequities and importantly, ensure that inequities are not aggravated. In this paper, we discuss approaches to integrating health equity-considerations in randomized trials by using the PROGRESS Plus framework (Place of residence, Race/ethnicity/culture/language, Occupation, Gender, Religion, Education, Socio-economic status, Social capital and "Plus" that includes other context specific factors) and cover: (i) formulation of research questions, (ii) two specific scenarios relevant to trials about health equity and (iii) describe how the PROGRESS Plus characteristics may influence trial design, conduct and analyses. This guidance is intended to support trialists designing equity-relevant trials and lead to better design, conduct, analyses and reporting, by addressing two main issues: how to avoid aggravating inequity among research participants and how to produce information that is useful to decision-makers who are concerned with health inequities.


Express-saccades of the monkey: effect of daily training on probability of occurrence and reaction time.
Two monkeys learned to make saccadic eye movements from a central fixation point to a peripheral target, when there was a temporal gap between fixation point offset and target onset. Under these conditions the animals made saccades after extremely short reaction times (less than 100 ms), so called express-saccades. With ongoing training the rate of occurrence increased (10 to 1005) and the reaction time of the express-saccades decreased (95 to 75 ms). The training effects were mediated by the amount of previously executed express-saccades and they were also spatially selective for express-saccades to that target position that had been used during training. The training effects on the express-saccades can be saturated after less than 7 days of daily training and are reversible after another 7 days of no training. The results indicate the existence of a fast-operating visuo-to-oculomotor pathway which can be quickly and reversibly modified by daily exercise.


Real-Life Clinical Experience With Cariprazine: A Systematic Review of Case Studies.
The hierarchy of evidence coming from evidence-based medicine favors meta-analyses and randomized controlled trials over observational studies and clinical cases. Nonetheless, in the field of psychiatry, where conditions are much more complex, additional evidence coming from real-world clinical practice is necessary to complement data from these gold standards. Thus, in this systematic review, the aim is to summarize the evidence coming from clinical case reports regarding cariprazine, a third-generation antipsychotic drug that has been approved for the treatment of schizophrenia and bipolar I disorder with manic, depressive or mixed features in adults. A systematic review was performed using Embase and Pubmed databases searching for English-language cases published in peer-reviewed journals between 2000 January and 2021 September with the following search terms: (cariprazin* OR "rgh-188" OR rgh188 OR vraylar OR reagila) AND ("case report*" OR "case report"/de OR "case stud*" OR "case study"/de OR "case seri*"). After the removal of duplicates, 49 articles were retrieved via the search, from which 22 were suitable for this review. These 22 articles encompassed 38 cases from which 71% described patients with schizophrenia, 16% patients with psychotic disorders, 5% patients with mood disorder and 8% described patients with other disorders such as Wernicke-Korsakoff syndrome, borderline personality disorder and obsessive-compulsive disorder with paranoid schizophrenia. The median age of patients was 31, and half of them were female. The majority of patients (76%) started cariprazine with 1.5 mg/day, and the most common maintenance dose was 4.5 mg/day (34%) and 3.0 mg/day (29%). Cariprazine was found to be safe and effective in a wide range of psychiatric conditions with different symptom profiles from acute psychotic symptoms through addiction to negative and cognitive symptoms. The results are in-line with the established evidence from clinical trials, however, they also show how cariprazine can be successfully utilized for treating certain symptoms irrespective of the indication.


Natural killer cells in pancreatic cancer stroma.
Pancreatic cancer remains one of medicine's largest areas of unmet need. With five-year survival rates of < 8%, little improvement has been made in the last 50 years. Typically presenting with advance stage disease, treatment options are limited. To date, surgery remains the only potentially curative option, however, with such late disease presentation, the majority of patients are unresectable. Thus, new therapeutic options and a greater understanding of the complex stromal interactions within the tumour microenvironment are sorely needed to revise the dismal outlook for pancreatic cancer patients. Natural killer (NK) cells are crucial effector units in cancer immunosurveillance. Often used as a prognostic biomarker in a range of malignancies, NK cells have received much attention as an attractive target for immunotherapies, both as cell therapy and as a pharmaceutical target. Despite this interest, the role of NK cells in pancreatic cancer remains poorly defined. Nevertheless, increasing evidence of the importance of NK cells in this dismal prognosis disease is beginning to come to light. Here, we review the role of NK cells in pancreatic cancer, examine the complex interactions of these crucial effector units within pancreatic cancer stroma and shed light on the increasingly attractive use of NK cells as therapy.


Multiparametric magnetic resonance imaging features of a canine glioblastoma model.
To assess glioblastoma multiforme (GBM) formation with similar imaging characteristics to human GBM using multiparametric magnetic resonance imaging (MRI) in an orthotopic xenograft canine GBM model. The canine GBM cell line J3T1 was subcutaneously injected into 6-week-old female BALB/c nude mice to obtain tumour fragments. Tumour fragments were implanted into adult male mongrel dog brains through surgery. Multiparametric MRI was performed with conventional MRI, diffusion-weighted imaging, and dynamic susceptibility contrast-enhanced perfusion-weighted imaging at one week and two weeks after surgery in a total of 15 surgical success cases. The presence of tumour cells, the necrotic area fraction, and the microvessel density (MVD) of the tumour on the histologic specimen were assessed. Tumour volume, diffusion, and perfusion parameters were compared at each time point using Wilcoxon signed-rank tests, and the differences between tumour and normal parenchyma were compared using unpaired t-tests. Spearman correlation analysis was performed between the imaging and histologic parameters. All animals showed a peripheral enhancing lesion on MRI and confirmed the presence of a tumour through histologic analysis (92.3%). The normalized perfusion values did not show significant decreases through at least 2 weeks after the surgery (P > 0.05). There was greater cerebral blood volume and flow in the GBM than in the normal-appearing white matter (1.46 ± 0.25 vs. 1.13 ± 0.16 and 1.30 ± 0.22 vs. 1.02 ± 0.14; P < 0.001 and P < 0.001, respectively). The MVD in the histologic specimens was correlated with the cerebral blood volume in the GBM tissue (r = 0.850, P = 0.004). Our results suggest that the canine GBM model showed perfusion imaging characteristics similar to those of humans, and it might have potential as a model to assess novel technical developments for GBM treatment.


Assessing the usefulness of submaximal exercise heart rates for monitoring cardiorespiratory fitness changes in elite youth soccer players.
This study aimed to assess the value of monitoring fitness in elite youth soccer players (U15 to U19 age groups) by analyzing the concomitant changes in heart rate at submaximal intensity (HR12km/h) and the velocity at a lactate concentration of 4 mmol/l (v4mmol/l). Players were tested by means of an incremental treadmill test on two occasions during the summer pre-season in two consecutive seasons. Based on data from a total of 170 test comparisons from the U15 (n = 48 test comparisons), U16 (n = 40 test comparisons), U17 (n = 46 test comparisons), and U19 (n = 36 test comparisons) age groups, the agreement between substantial changes in HR12km/h and v4mmol/l was analyzed using the threshold combination of HR12km/h = 4.5% and v4mmol/l = 6.0%. Results revealed 2% full mismatches, 36% partial agreements, and 62% full agreements for the whole sample in terms of fitness change interpretation between both variables. The respective values for the U15 to U19 age groups ranged between 0% and 5% full mismatches, 28-44% partial agreements, and 56-68% full agreements with no meaningful differences between age groups. In conclusion, our findings confirm the practical value of using HR12km/h to monitor fitness changes in elite youth soccer players when lactate sampling during incremental tests is not possible.


Modulating PtPt metal-metal interactions through conformationally switchable molecular tweezer/guest complexation.
Herein a new organoplatinum(ii)-based molecular tweezer has been designed and synthesized, which displays pH-responsive mechanical transformation from the "U"- to "W"-shaped conformation. Pt(ii)Pt(ii) metal-metal interactions have been successfully incorporated into the resulting molecular tweezer/guest recognition motifs and self-assembled polymers, which can be modulated in response to pH variation.


Antimicrobial Resistance Among Escherichia coli Isolated from Veal Calf Operations in Pennsylvania.
Antimicrobial resistance (AR) is a pressing public health concern, and agricultural operations such as dairy and beef cattle production have been implicated as potential sources of resistant bacteria or genetic elements. This study aimed to determine the prevalence of antimicrobial-resistant Escherichia coli from calf pens in 6 auction houses (56 manure composite samples) and 12 veal calf operations (240 fecal samples in 2 visits: after the calves arrived at the farm and shortly before the animals were sent to slaughter) in the Commonwealth of Pennsylvania. A total of 1567 generic E. coli were isolated and screened for resistance phenotypes. Resistant E. coli were isolated from all auction houses and farms sampled. Based on nonparametric Kruskal-Wallis tests, incremental prevalence of E. coli resistant to ampicillin, azithromycin, cefoxitin, ceftiofur, ceftriaxone, chloramphenicol, streptomycin, sulfisoxazole, trimethoprim-sulfamethoxazole, and tetracycline in the samples from auction houses and the first and second farm visits was observed (χ2 6.98-15.91, p < 0.05). Multidrug-resistant E. coli (resistant to more than three antimicrobial classes) were identified in 76.8%, 90.8%, and 100% of samples collected from the auction houses, first farm visits, and second farm visits, respectively. The presence of blaCTX-M-E. coli in 11 of the 12 farms presents the possibility of veal production environments being a reservoir for resistant genetic materials that may pose a risk to human health if they are transferred to human pathogens. Additional research on the impact of various management strategies in veal calf rearing is needed for a complete scenario of AR in these production environments.


Acquired prosopagnosia as a face-specific disorder: ruling out the general visual similarity account.
Prosopagnosia is classically defined as a disorder of visual recognition specific to faces, following brain damage. However, according to a long-standing alternative view, these patients would rather be generally impaired in recognizing objects belonging to visually homogenous categories, including faces. We tested this alternative hypothesis stringently with a well-documented brain-damaged prosopagnosic patient (PS) in three delayed forced-choice recognition experiments in which visual similarity between a target and its distractor was manipulated parametrically: novel 3D geometric shapes, morphed pictures of common objects, and morphed photographs of a highly homogenous familiar category (cars). In all experiments, PS showed normal performance and speed, and there was no evidence of a steeper increase of error rates and RTs with increasing levels of visual similarity, compared to controls. These data rule out an account of acquired prosopagnosia in terms of a more general impairment in recognizing objects from visually homogenous categories. An additional experiment with morphed faces confirmed that PS was specifically impaired at individual face recognition. However, in stark contrast to the alternative view of prosopagnosia, PS was relatively more impaired at the easiest levels of discrimination, i.e. when individual faces differ clearly in global shape rather than when faces were highly similar and had to be discriminated based on fine-grained details. Overall, these observations as well as a review of previous evidence, lead us to conclude that this alternative view of prosopagnosia does not hold. Rather, it seems that brain damage in adulthood may lead to selective recognition impairment for faces, perhaps the only category of visual stimuli for which holistic/configural perception is not only potentially at play, but is strictly necessary to individualize members of the category efficiently.


Long-acting reversible contraceptives: New evidence to support clinical practice.
General practice plays a key role in providing access to contraceptive options to reduce unintended pregnancies. The use of long-acting reversible contraceptives (LARCs) is increasing in Australia. The aim of this article is to provide an update on new evidence and guidance relating to clinical practice for LARCs. A new Pharmaceutical Benefits Scheme-listed hormonal intrauterine device (IUD), as well as updated guidelines for LARC provision and use, are reviewed. The 19.5 mg levonorgestrel IUD provides a smaller, lower-dose hormonal IUD. This article compares IUDs now available in Australia to support informed patient choice. Updated guidance for IUD users in perimenopause, as well as those using menstrual cups, is outlined. The contraceptive hormonal implant has updated product information, which includes new insertion site and arm positioning for procedures.


Surgical treatment of the boutonniere deformity in rheumatoid arthritis.
The rheumatoid boutonniere deformity does not limit hand function significantly until it becomes severe. For this reason the surgical procedures used should not risk or sacrifice existing function. For the mild cases one can improve the digital balance by a simple extensor tenotomy combined with dynamic splinting. In more advanced stages with passive correction possible, the deformity can be improved by reconstruction of the extensor mechanism. With fixed deformities one can choose between proximal interphalangeal joint fusion and arthroplasty. The decision is influenced by several factors, including the digit involved and the status of adjacent joints. After a careful evaluation of the patient's capabilities and needs, it is possible to improve these deformities by the methods discussed.


Far-reaching consequences of high level air pollution on the developing immune system.
The town of Dimitrovgrad in Bulgaria has a highly industrialized region with a developing chemical industry, cement and asbestos-cement production, as well as energy production. For over four decades, the main ecological problem was air pollution, but after 1989 a great economic crisis led to an abrupt decline in the level of air pollutants. The aim of the present study was to investigate the present state of humoral immunity in teenage children from Dimitrovgrad who lived under conditions of massive air pollution during their intrauterine development and first few years of childhood. Immunoglobulins IgA, IgM, and IgG were measured in 106 clinically healthy children (average age 12.68 +/- 0.56 y) from 3 schools in Dimitrovgrad and in 41 control subjects (average age 12.35 +/- 0.22 y) from the town of Nova Mahala, an area lacking massive sources of air pollution. Immunoglobulin concentration was determined by turbidimetry using an Optima (KONE) chemical analyzer. The average IgA, IgM, and IgG levels in children from Dimitrovgrad did not differ significantly from those of the control group.


Growth factor-functionalized silk membranes support wound healing in vitro.
Chronic wounds represent a serious problem in daily medical routine requiring improved wound care. Silk of the domesticated silkworm (Bombyx mori) has been used to form a variety of biomaterials for medical applications. We genetically engineered B. mori to produce silk functionalized with growth factors to promote wound healing in vitro. In this study FGF-, EGF-, KGF-, PDGF- or VEGF-functionalized silk membranes were compared to native B. mori silk membranes without growth factors for their ability to support wound healing in vitro. All silk membranes were cytocompatible and supported macrophage secretion of neutrophil recruiting factor CXCL1 and monocyte chemoattractant protein 1 (MCP-1). VEGF-functionalized silk significantly outperformed other growth factor-functionalized silk membranes, but not native silk in angiogenesis assays. In addition, EGF- and VEGF-functionalized silk membranes slightly enhanced macrophage adhesion compared to silk without growth factors. In wound healing assays in vitro (reduction of wound lesion), dermal equivalents showed a higher wound healing capacity when covered with EGF-, FGF- or VEGF-functionalized silk membranes compared to native, KGF- or PDGF-functionalized silk membranes. Keratinocyte migration and growth is overstimulated by KGF- and VEGF-functionalized silk membranes. In conclusion, growth factor-functionalized silk membranes prepared from genetically engineered silk worm glands are promising wound dressings for future wound healing therapies.


Nevic mitoses: a review of 1041 cases.
The presence of dermal mitotic figures is helpful in identifying malignant melanocytic lesions but occasionally occur in benign nevi. We aim to determine a "benchmark" mitotic frequency for a wide variety of nevi. We prospectively collected 1041 cases of benign melanocytic nevi and reviewed them for the presence of mitotic figures. Specimens were collected from female (62%) and male (38%) participants, ages ranged from 1 to 90 years. Nevus types included compound melanocytic nevi (CMN), intradermal melanocytic nevi, junctional melanocytic nevi, lentiginous CMN (LCMN), lentiginous junctional melanocytic nevi, blue nevi, deep penetrating nevi, and pigmented spindle cell nevi. Nevi with congenital, mildly dysplastic, and Spitzoid features were included. A total of 82 of 1041 (7.9%) nevi contained one or more mitotic figures. Most (76.1%) mitoses were found in the papillary dermis. Single mitotic figures were more common (80.4%) than 2 (15.9%) or 3 (3.7%) within a single specimen. Of all cases containing mitotic figures, the most common nevus type was CMN. The nevus type most frequently demonstrating mitotic figures was CMN. The most common body site among cases with mitotic figures was trunk (53.7%), whereas the body site with the largest proportion of nevi demonstrating mitotic figures was special site (10.9%). The percentage of nevi containing mitotic figures was nearly the same among female (7.9%) and male (7.8%) participants. Results of this large review confirm that mitotic figures, even multiple ones, do not preclude benignity in a variety of melanocytic nevi.


Overall survival and risk of second malignancies with cancer chemotherapy and G-CSF support.
The use of supportive granulocyte colony-stimulating factor (G-CSF) to reduce the risk of neutropenic complications in high-risk cancer patients is consistently recommended by several clinical practice guidelines. However, in a previous meta-analysis, G-CSF prophylaxis was associated with an increased risk of secondary malignancies while reducing long-term mortality. We present here an updated systematic review and meta-analysis. A systematic literature search was carried out to identify randomized controlled trials of cancer patients receiving conventional-dose chemotherapy, assigned to primary G-CSF support or a control group without initial G-CSF, with at least 2 years of follow-up. Studies were categorized into one of the four groups, based on the chemotherapy regimen and study design. An updated meta-analysis was carried out; relative risk (RR) and 95% confidence intervals (CIs) for all-cause mortality and secondary malignancies were calculated. Of 2604 articles screened, 14 eligible studies were identified and combined with studies identified in the previous systematic literature searches. The updated meta-analysis included a total of 68 studies presenting 71 separate comparisons. Survival was significantly improved in patients receiving primary G-CSF support, compared with patients without primary G-CSF support (mortality RR=0.92; 95% CI 0.90-0.95; ARD=-3.3%; 95% CI -4.2--2.4; P < 0.0001). The largest improvement in survival was observed with dose-dense chemotherapy regimens with G-CSF support, compared with controls receiving no G-CSF support (mortality RR=0.86; 95% CI 0.80-0.92; P < 0.0001). Patients who received primary G-CSF support experienced a significantly higher risk of secondary malignancies, compared with controls (RR=1.85; 95% CI 1.19-2.88; ARD=0.47; 95% CI 0.21-0.73; P < 0.01). Our findings demonstrate that overall survival is improved in patients receiving intensified chemotherapy with primary G-CSF support, compared with those receiving standard chemotherapy. Primary G-CSF support was also associated with a higher risk of developing secondary malignancies, including secondary acute myeloid leukemia and myelodysplastic syndrome.


The presence of two ovulatory follicles at timed artificial insemination influences the ovulatory response to GnRH in high-producing dairy cows.
This study sought to examine the impact of the presence of two co-dominant (ovulatory) follicles at the time of artificial insemination (AI) on the ovulatory response to GnRH given in a fixed-time AI protocol. The study population comprised 622 lactating dairy cows: 306 (49.2%) with a single follicle, 198 (31.8%) with two bilateral follicles (one follicle per ovary) and 118 (19%) with two unilateral follicles (same ovary). Based on odds ratios, cows with two bilateral or unilateral follicles were less likely (by factors of 0.09 and 0.11, respectively) to undergo ovulation failure compared with cows with one follicle (P = 0.01 and P = 0.02, respectively); the likelihood of ovulation failure decreased 0.75 times with every one-mm increase in follicle diameter for cows with a single follicle, whereas individual follicle diameter was not related to ovulation failure in cows with two bilateral follicles (P = 0.001). The likelihood of double ovulation decreased 0.7 times with every one-mm diameter difference between the larger and smaller follicle for all cows with two follicles (P = 0.001), whereas cows with two unilateral follicles showed a higher (P < 0.05) double ovulation rate than cows with two bilateral follicles. In 116 (58.6%) of the cows with two bilateral follicles, only the larger follicle ovulated in 59.5% cows, whereas only the smaller one ovulated in the remaining 40.5% cows. In these cows, a one-mm size difference between the larger and the smaller follicle gave rise to a 1.12-fold increase in the ovulation failure rate for the larger follicles (P = 0.0001). Cows with two bilateral follicles were more likely (by a factor of 1.5) to conceive than cows with one follicle (P = 0.001). Significant right-left differences were not found in cows with two bilateral follicles, whereas the right ovary was more active than the left in the remaining cows. Our results indicate that cows with two co-dominant follicles at AI show different ovulation patterns to those with one dominant follicle. A higher rate of ovulation failure was observed among cows with one follicle than cows with two follicles, whereas the conception rate was higher for cows with two bilateral follicles than for the remaining cows. In cows with two follicles, double ovulations along with ovulation of the smaller follicle were related to the least size difference between the larger and smaller follicle.


A comparative study of intrathecal clonidine and dexmedetomidine on characteristics of bupivacaine spinal block for lower limb surgeries.
Intrathecal clonidine or dexmedetomidine has improved the quality of spinal anesthesia, this clinical study was undertaken to assess the behavior of intrathecal clonidine as an adjuvant to bupivacaine in augmenting sensory block in patients undergoing lower limb surgeries. We studied 150 patients, between 18 and 60 years of age, patients were randomly divided into three groups of 50 each Group B: 0.5% bupivacaine 15 mg + 0.5 ml normal saline Group C: 0.5% bupivacaine 15 mg + 50 μg clonidine Group D: 0.5% bupivacaine 15 mg + 5 μg dexmedetomidine. Onset and duration of sensory block and motor block, the highest level of sensory blockade, duration of analgesia, and side-effects were assessed. The onset of motor block was faster in Group C and Group D as compared to Group B, The time to reach Bromage scale 3 was fastest in Group C followed by Group D P < 0.001. The duration of sensory, motor blockade and duration of analgesia was longer in Groups C and D as compared to Group B, longest in Group D followed by C and B. The time to regression time to S1 dermatome was. It was longest in Group D followed by Group C and then Group B. Intergroup comparison B to C, B to D and C to D was significant. Duration of analgesia was significantly prolonged in Group C and Group D (P < 0.001) with a mean duration of 309.6 ± 50.99 min in Group C and 336.8 ± 55.38 min in Group D as compared to 204.8 ± 16.81 min in Group B. Intergroup comparison B to C, B to D and Group C to D was significant. Duration was longest in Group D followed by Group C and then Group B. Supplementation of bupivacaine spinal block with a low dose of intrathecal dexmedetomidine (5 μg) or clonidine (50 μg) produces a significantly shorter onset of motor and sensory block and a significantly longer sensory and motor block than bupivacaine alone.


Predicting the impact and duration of persistent and mobile organic compounds in groundwater systems using a contaminant mass discharge approach.
This study investigated methods for predicting the duration and impact on groundwater quality from persistent and mobile organic compounds (PMOCs) at a drinking water well field affected by multiple contaminant sources. The fungicide metabolite N,N-dimethylsulfamide (DMS), which frequently occurs above the Danish groundwater quality criterion (0.1 μg/L), was used as an example. By combining contaminant mass discharge (CMD) estimations, modeling, and groundwater dating, a number of important discoveries were made. The current center of contaminant mass was located near the source area. The CMD at the well field was predicted to peak in 2040, and an effect from the investigated sources on groundwater quality could be expected until the end of the 21st century. A discrepancy in the current CMD at the well field and the estimated arrival time from the studied source area suggested an additional pesticide source, which has not yet been thoroughly investigated. The presence of the unknown source was supported by model simulations, producing an improved mass balance after inclusion of a contaminant source closer to the well field. The approach applied here was capable of predicting the duration and impact of DMS contamination at a well field at catchment scale. It furthermore shows potential for identification and quantification of the contribution from individual sources, and is also applicable for other PMOCs. Predicting the duration of the release and impact of contaminant sources on abstraction wells is highly valuable for water resources management and authorities responsible for contaminant risk assessment, remediation, and long-term planning at water utilities.


Ab initio investigation of the role of the d-states on the adsorption and activation properties of CO2 on 3d, 4d, and 5d transition-metal clusters.
We report a theoretical investigation of the adsorption and activation properties of CO2 on eight-atom 3d, 4d, and 5d transition-metal (TM) clusters based on density functional theory calculations. From our results and analyses, in the lowest energy configurations, CO2 binds via a chemisorption mechanism on Fe, Co, Ni, Ru, Rh, Pd, Os, Ir, and Pt (adsorption energy from -0.49 eV on Pt up to -1.40 eV on Os), where CO2 breaks its linearity and adopts an angular configuration due to the charge transfer from the clusters toward the C atom in the adsorbed CO2. In contrast, it binds via physisorption on Cu, Ag, and Au and maintains its linearity due to a negligible charge transfer toward CO2 and has a small adsorption energy (from -0.17 eV on Cu up to -0.18 eV on Ag). There is an energetic preference for twofold bridge TM sites, which favors binding of C with two TM atoms, which enhances the charge transfer ten times than on the top TM sites (onefold). We identified that the strength of the CO2-TM8 interaction increases when the energy values of the highest occupied molecular orbital (HOMO) of the TM8 are closer to the energy values of the lowest unoccupied molecular orbital of CO2, which contributes to maximize the charge transfer toward the molecule. Beyond the energy position of the HOMO states, the delocalization of 5d orbitals plays an important role in the adsorption strength in TM, especially for the iron group, e.g., the adsorption energies are -1.08 eV (Fe, 3d), -1.19 eV (Ru, 4d), and -1.40 eV (Os, 5d).


Correlation Between Calcification Characteristics of Carotid Atherosclerotic Plaque and Plaque Vulnerability.
To investigate the relationship between calcification characteristics of carotid atherosclerotic plaque and lipid rich necrotic core (LRNC) and intraplaque hemorrhage (IPH). Patients with severe carotid stenosis undergoing carotid endarterectomy (CEA) were selected. Ultrasound and CT angiography (CTA) were performed to evaluate the calcification characteristics of the plaque before the surgery. A total of 142 patients were included and 142 pathological specimens of postoperative plaque were obtained accordingly. There were 78 plaques (54.9%) with LRNC and 41 (28.9%) with IPH. The plaque with LRNC had higher calcification rate (93.6%) compared with the plaque with IPH (87.8%). LRNC was often found in multiple calcification (P = 0.003) and mixed type calcification (P = 0.001). Multiple calcification was more likely to combine with IPH (P = 0.008), while simple basal calcification was not likely to combine IPH (P = 0.002). Smaller granular calcification was more likely to be associated with IPH (P < 0.05). In multivariate regression analysis of IPH and calcification characteristics, simple basal calcification was still a protective factor for IPH (OR, 0.25; 95% CI, 0.09-0.66; P = 0.005), while multiple calcification was closely related to the occurrence of IPH (OR, 3.58; 95% CI, 1.49-8.61; P = 0.004). Calcification characteristics of carotid atherosclerotic plaques are closely related to the vulnerability of plaques, especially multiple calcification and mixed type calcification.


Measles virus spreads in rat hippocampal neurons by cell-to-cell contact and in a polarized fashion.
Measles virus (MV) can infect the central nervous system and, in rare cases, causes subacute sclerosing panencephalitis, characterized by a progressive degeneration of neurons. The route of MV transmission in neurons was investigated in cultured rat hippocampal slices by using MV expressing green fluorescent protein. MV infected hippocampal neurons and spread unidirectionally, in a retrograde manner, from CA1 to CA3 pyramidal cells and from there to the dentate gyrus. Spreading of infection depended on cell-to-cell contact and occurred without any detectable release of infectious particles. The role of the viral proteins in the retrograde MV transmission was determined by investigating their sorting in infected pyramidal cells. MV glycoproteins, the fusion protein (F) and hemagglutinin (H), the matrix protein (M), and the phosphoprotein (P), which is part of the viral ribonucleoprotein complex, were all sorted to the dendrites. While M, P, and H proteins remained more intracellular, the F protein localized to prominent, spine-type domains at the surface of infected cells. The detected localization of MV proteins suggests that local microfusion events may be mediated by the F protein at sites of synaptic contacts and is consistent with a mechanism of retrograde transmission of MV infection.


An educational intervention using the health belief model for improvement of oral health behavior in grade-schoolers: a randomized controlled trial.
Oral health problems especially dental caries are common in school children, and education programs may help to prevent these conditions. The aim of current study was to examine the effects of an educational program based on a Health Belief Model (HBM) to improve oral health behaviors of elementary school children. A total of 112 children ages 6-12 years old accompanied by one of their parents were randomly assigned to intervention/test and control groups. In the intervention group, five consecutive weekly educational sessions based on the HBM were provided, while the control group received only routine education delivered by the dental clinic. The Decayed, Missing, and Filled Teeth (DMFT) score, papillary bleeding index, and responses to the HBM questionnaire were assessed in the intervention and control groups at baseline and three-month follow-up after the intervention was completed. Within-group and between-group differences were examined using the Student's t-test and analysis of covariance. All HBM domains were improved at follow-up in the intervention group compared to the control group (p < 0.001). The largest change was in perceived susceptibility, whereas the smallest changes were in perceived severity and perceived benefits. The papillary bleeding index demonstrated a significant change from baseline to follow-up in the intervention group (reduction of 0.7, 95% CI = - 0.9 to - 0.5). All components of the DMFT score except missing teeth also improved in the intervention group compared to controls. However, no significant difference was found in total DMFT score between intervention and control groups. An education program based on HBM may be more effective than current methods used to educate children and their parents on optimal oral health behaviors. Administration of interventions of this type along with other school-based programs to prevent dental caries may be helpful in grade-school children.


Tilt table testing, methodology and practical insights for the clinic.
Tilt table testing (TTT) has been used for decades to study short-term blood pressure (BP) and heart rate regulation during orthostatic challenges. TTT provokes vasovagal reflex in many syncope patients as a background of widespread use. Despite the availability of evidence-based practice syncope guidelines, proper application and interpretation of TTT in the day-to-day care of syncope patients remain challenging. In this review, we offer practical information on what is needed to perform TTT, how results should be interpreted including the Vasovagal Syncope International Study classification, why syncope induction on TTT is necessary in patients with unexplained syncope and on indications for TTT in syncope patient care. The minimum requirements to perform TTT are a tilt table with an appropriate tilt-down time, a continuous beat-to-beat BP monitor with at least three electrocardiogram leads and trained staff. We emphasize that TTT remains a valuable asset that adds to history building but cannot replace it, and highlight the importance of recognition when TTT is abnormal even without syncope. Acknowledgement by the patient/eyewitness of the reproducibility of the induced attack is mandatory in concluding a diagnosis. TTT may be indicated when the initial syncope evaluation does not yield a certain, highly likely, or possible diagnosis, but raises clinical suspicion of (1) reflex syncope, (2) orthostatic hypotension (OH), (3) postural orthostatic tachycardia syndrome or (4) psychogenic pseudosyncope. A therapeutic indication for TTT in the patient with a certain, highly likely or possible diagnosis of reflex syncope, may be to educate patients on prodromes. In patients with reflex syncope with OH TTT can be therapeutic to recognize hypotensive symptoms causing near-syncope to perform physical countermanoeuvres for syncope prevention (biofeedback). Detection of hypotensive susceptibility requiring therapy is of special value.


Effective isolation of GALT cells: Insights into the intestine immune response of rainbow trout (Oncorhynchus mykiss) to different bacterin vaccine preparations.
The teleost gut is a multifunction complex structure that plays a pivotal immunological role in homeostasis and the maintenance of health, in addition to digestion of food and/or nutrient absorption. In vitro examination of the intestine leucocyte repertoire has the potential to aid our understanding of gut immune competence and allows a rapid screen of host-microorganism interactions in different immunological contexts. To explore this possibility, in the present study we investigated the response of isolated gut leucocytes to 4 bacterins of Aeromonas salmonicida, prepared from different strains, combinations and strains grown in different environments, in comparison to a Yersinia ruckeri bacterin for which a commercial/effective oral booster vaccine has been developed. To aid this study we also optimized further our method of GALT cell isolation from rainbow trout, so as to avoid mechanical clearance of the intestine contents. This drastically increased the cell yield from ~12 × 106 to ~210 × 106/fish with no change in the percent cell viability over time or presence of transcripts typical of the key leucocyte types needed for the study of immune modulation (i.e. T- and B-cells, dendritic cells and macrophages). A wide array of immune transcripts were modulated by the bacterins, demonstrating the diversity of GALT cell responses to bacterial stimulation. Indeed, the GALT leucocyte responses were sensitive enough to distinguish the different bacterial species, strains and membrane proteins, as seen by distinct kinetics of immune gene expression. However, the response of the GALT cells was often relatively slow and of a low magnitude compared to those of PBL. These results enhance our knowledge of the gut biocapacity and help validate the use of this model for screening of oral vaccine candidates.


Rapid and Accurate C-V Measurements.
We report a new technique for the rapid measurement of full capacitance-voltage (C-V) characteristic curves. The displacement current from a 100 MHz applied sine-wave, which swings from accumulation to strong inversion, is digitized directly using an oscilloscope from the metal-oxide-semiconductor (MOS) capacitor under test. A C-V curve can be constructed directly from this data but is severely distorted due to non-ideal behavior of real measurement systems. The key advance of this work is to extract the system response function using the same measurement set-up and a known MOS capacitor. The system response correction to the measured C-V curve of the unknown MOS capacitor can then be done by simple deconvolution. No de-skewing and/or leakage current correction is necessary, making it a very simple and quick measurement. Excellent agreement between the new fast C-V method and C-V measured conventionally by an LCR meter is achieved. The total time required for measurement and analysis is approximately 2 seconds, which is limited by our equipment.


Randomized clinical evaluation of the effect of chlorhexidine on postoperative sensitivity of posterior composite resin restorations.
Evaluation of postoperative sensitivity in posterior teeth restored with etchand- rinse adhesive and composite resin after application of 2% chlorhexidine solution. Thirty participants with similar dental caries on both sides of the jaws were selected. After cavity preparation, each tooth was randomized to one of the following restoration methods. Teeth in the control group were restored using etch-and-rinse resin adhesive (Single Bond, 3M ESPE) and universal microhybrid composite resin (Filtek Z250, 3M ESPE). Restoration of teeth in the experimental group was similar to the control group except that 2% chlorhexidine solution (Concepsis, Ultradent) was applied on the etched dentin for 60 seconds. Subjects were recalled for evaluation of postoperative sensitivity at 1 day, 1 week, 1 month, and 6 months. Data were analyzed by Fisher exact test (α = .05). The experimental group showed significantly less postoperative sensitivity (8 teeth) compared with the control group (20 teeth) at the 1-day recall (P < .05; Fisher exact test). Conventional restorations were comparable to teeth treated with chlorhexidine at other recall times (P > .05). Chlorhexidine significantly decreased immediate postoperative sensitivity of posterior teeth restored by composite resin.


Characteristics of drug users who do or do not have care of their children.
To compare the relative frequency of eight indicators of problem drug use and potentially adverse social circumstances in drug-using parents and non-parents and to explore whether a profile based on these characteristics differs according to whether or not dependent children live with their drug-using parent. The study utilizes a 5 year national UK treatment monitoring system data set. 61,425 users with, and 105,473 without dependent children accessing drug treatment services in England and Wales between January 1996 and December 2000. Information about parenthood and children's residence was routinely collected. Drug use and social circumstance indicators were daily heroin use, daily alcohol use, regular stimulant use, sharing of injecting equipment, living with another user, living alone, unstable accommodation, and criminal justice referral. There were clear differences between drug-using parents according to where children live. Parents with children at home and non-parents showed fewer of the indicators than parents with children in care or elsewhere. Sixty-five percent of parents with none of the indicators lived with their children, compared with only 28% of those with three indicators and 9% of those with six or more indicators. Parents with children in care or living elsewhere showed the highest prevalence for each individual indicator. Drug-using parents demonstrate a range of potentially unfavourable drug use behaviours and social circumstances, but those whose children live with them use drugs less frequently and live in more favourable conditions than those whose children live elsewhere. Protective factors may operate in family situations, while severe drug use and adverse social circumstances may result in a breakdown of family structures.


Allogenic Pure Platelet-Rich Plasma Therapy for Rotator Cuff Disease: A Bench and Bed Study.
Although platelet-rich plasma (PRP) is a popular option for rotator cuff disease, the underlying mechanism of PRP and its clinical indications are unclear. Further, some kinds of PRP might be detrimental to patients. Allogenic PRP prepared through a standardized process and fully characterized could eliminate variations in PRP as well as uncertainties regarding its use in each patient, which could provide clues about its mechanism of action and indications for its use. To assess the effects of pure PRP on tenocytes with or without inflammation in an in vitro study and to evaluate the safety and efficacy of a fully characterized pure PRP injection in patients with rotator cuff disease in a clinical study. Controlled laboratory study and cohort study; Level of evidence, 3. For the in vitro study, tenocytes were enzymatically isolated and cultured from patients with rotator cuff tear and treated with or without interleukin 1β (IL-1β) and PRP. Gene expression and protein synthesis of pro- and anti-inflammatory cytokines, enzymes and their inhibitors, matrix synthesis, and cell viability were evaluated. For the clinical study, a total of 17 patients with rotator cuff disease received ultrasonography-guided subacromial PRP injection and were followed for 6 months. Pain, range of motion, muscle strength, shoulder function, and overall satisfaction in patients were compared with the results in a propensity score-matched control group who received corticosteroid (triamcinolone acetonide 40 mg). PRP induced inflammation in the absence of inflammation and ameliorated inflammation in IL-1β-induced tendinopathic conditions by regulation of cytokines such as IL-1β, cyclooxygenase 2, microsomal prostaglandin E synthase 1, vasoactive intestinal peptide, and downstream matrix metalloproteinases. No general or local adverse events were noted with regard to allogenic PRP injection. Whereas steroid injection showed earlier improvement in some kinds of pain and functional scores, PRP generally showed comparable effects with steroid injection in all clinical outcomes at 6 months. This study showed that allogenic pure PRP had pleiotropic effects on tenocytes depending on inflammation and that it did not cause adverse events but rather decreased pain and improved shoulder function to a degree comparable with steroid injection in patients with rotator cuff disease. Allogenic PRP could be a treatment option for rotator cuff disease.


Airway problems in pregnancy.
To provide a current review of the literature regarding airway problems in pregnancy and management. Obstetrical anesthesia is considered to be a high-risk practice that exposes the anesthesiologist to increased medicolegal liability. Anesthetic management of a parturient is a challenge because it involves simultaneous care of both mother and baby. Failure to appropriately manage a difficult or failed intubation increases the risk of hypoxemic cardiopulmonary arrest and/or pulmonary aspiration, resulting in a high probability of maternal morbidity and mortality. Anesthesia is the seventh leading cause of maternal mortality in the United States. Anatomic and physiologic changes during pregnancy place the parturient at increased risk for airway management problems. It is essential to perform a thorough preanesthetic evaluation and identify the factors predictive of difficult intubation. Airway devices such as the laryngeal mask airway, ProSeal, intubating laryngeal mask airway, Combitube, and laryngeal tube are described and have been used during failed intubation in pregnant patients. Teamwork between an anesthesiologist and an obstetrician is absolutely essential for the safety of both the mother and baby. Most of us tend to agree that airway emergencies have a way of occurring at the worst possible times. It is essential that all anesthesia care practitioners must have a preconceived and well thought-out algorithm and emergency airway equipment to deal with airway emergencies during difficult or failed intubation of a parturient.


Oncogene cooperation in lymphocyte transformation: malignant conversion of E mu-myc transgenic pre-B cells in vitro is enhanced by v-H-ras or v-raf but not v-abl.
Although transgenic mice bearing a c-myc gene controlled by the immunoglobulin heavy-chain enhancer (E mu) eventually develop B-lymphoid tumors, B-lineage cells from preneoplastic bone marrow express the transgene but do not grow autonomously or produce tumors in mice. To determine whether other oncogenes can cooperate with myc to transform B-lineage cells, we compared the in vitro growth and tumorigenicity of normal and E mu-myc bone marrow cells infected with retroviruses bearing the v-H-ras, v-raf, or v-abl oncogene. The v-H-ras and v-raf viruses both generated a rapid polyclonal expansion of E mu-myc pre-B bone marrow cells in liquid culture and 10- to 100-fold more pre-B lymphoid colonies than normal in soft agar. The infected transgenic cells were autonomous, cloned efficiently in agar, and grew as tumors in nude mice. While many pre-B cells from normal marrow could also be induced to proliferate by the v-raf virus, these cells required a stromal feeder layer, did not clone in agar, and were not malignant. Most normal cells stimulated to grow by v-H-ras also cloned poorly in agar, and only rare cells were tumorigenic. With the v-abl virus, no more cells were transformed from E mu-myc than normal marrow and the proportion of tumorigenic pre-B clones was not elevated. These results suggest that both v-H-ras and v-raf, but apparently not v-abl, collaborate with constitutive myc expression to promote autonomous proliferation and tumorigenicity of pre-B lymphoid cells.


[Determination of vitamin B12 concentration by fluorescence quenching with acridine orange-rhodamine 6G energy transfer system].
An energy transfer technique between acridine orange (AO) and rhodamine 6G (R6G) was studied, and the optimum experimental conditions of energy transfer were defined. It was found that the effective energy transfer could occur between AO and R6G in the dodecylbenzene sodium sulfonate solution with Na2 HPO4-citric acid buffer solution at pH 5.0. The fluorescence intensity of AO-R6G system was diminished by vitamin B12 in an alkalescence medium. Based on the AO-R6G energy transfer system anovel fluorescence quenching method for the determination of vitamin B12 has been developed. Under optimal conditious, the linear range of calibration curves for the determination of vitamin B12 was 0-3.0 x 10(-5) mol x L(-1). The detection limits were 4.8 x 10(-7) mol x L(-1) for Vitamin B12. Among six times of determination, the relative standard deviation was 0.51%-0.64%, and the recovery was 98.40% -103.62%. The method features good recurrence, rapidity of reaction, good stability, and few interfering substances. It can be satisfactorily used in the determination of the injection content of vitamin B12.


Tuberculosis morbidity--United States, 1997.
During 1997, a total of 19,855 cases of tuberculosis (TB) (7.4 cases per 100,000 population) were reported to CDC from the 50 states and the District of Columbia, representing a 7% decrease from 1996 and a 26% decrease from 1992, when the number of cases peaked during the resurgence of TB in the United States. This report summarizes national TB surveillance data for 1997 and compares it with similar data for previous years. The findings indicate that, although the overall number of TB cases continued to decrease, trends in the number of reported cases and TB case rates differed by geographic area and population characteristics.


Enhanced Adsorption of p-Arsanilic Acid from Water by Amine-Modified UiO-67 as Examined Using Extended X-ray Absorption Fine Structure, X-ray Photoelectron Spectroscopy, and Density Functional Theory Calculations.
p-Arsanilic acid ( p-ASA) is an emerging organoarsenic pollutant comprising both inorganic and organic moieties. For the efficient removal of p-ASA, adsorbents with high adsorption affinity are urgently needed. Herein, amine-modified UiO-67 (UiO-67-NH2) metal-organic frameworks (MOFs) were synthesized, and their adsorption affinities toward p-ASA were 2 times higher than that of the pristine UiO-67. Extended X-ray absorption fine structure (EXAFS), X-ray photoelectron spectroscopy (XPS), and density functional theory (DFT) calculation results revealed adsorption through a combination of As-O-Zr coordination, hydrogen bonding, and π-π stacking, among which As-O-Zr coordination was the dominant force. Amine groups played a significant role in enhancing the adsorption affinity through strengthening the As-O-Zr coordination and π-π stacking, as well as forming new adsorption sites via hydrogen bonding. UiO-67-NH2s could remove p-ASA at low concentrations (<5 mg L-1) in simulated natural and wastewaters to an arsenic level lower than that of the drinking water standard of World Health Organization (WHO) and the surface water standard of China, respectively. This work provided an emerging and promising method to increase the adsorption affinity of MOFs toward pollutants containing both organic and inorganic moieties, via modifying functional groups based on the pollutant structure to achieve synergistic adsorption effect.


Dynamic Cooperation of Hydrogen Binding and π Stacking in ssDNA Adsorption on Graphene Oxide.
Functional nanoscale structures consisting of a DNA molecule coupled to graphene or graphene oxide (GO) have great potential for applications in biosensors, biomedicine, nanotechnology, and materials science. Extensive studies using the most sophisticated experimental techniques and theoretical methods have still not clarified the dynamic process of single-stranded DNA (ssDNA) adsorbed on GO surfaces. Based on a molecular dynamics simulation, this work shows that an ssDNA segment could be stably adsorbed on a GO surface through hydrogen bonding and π-π stacking interactions, with preferential binding to the oxidized rather than to the unoxidized region of the GO surface. The adsorption process shows a dynamic cooperation adsorption behavior; the ssDNA segment first captures the oxidized groups of the GO surface by hydrogen bonding interaction, and then the configuration relaxes to maximize the π-π stacking interactions between the aromatic rings of the nucleobases and those of the GO surface. We attributed this behavior to the faster forming hydrogen bonding interaction compared to π-π stacking; the π-π stacking interaction needs more relaxation time to regulate the configuration of the ssDNA segment to fit the aromatic rings on the GO surface.


[Male patient with anterior left hemiblock simulating anteroseptal infarct].
One case with left anterior hemiblock is described with ECG and image, simulating anteroseptal infarction. The precordial recording one intercostal space below facilitates the correct interpretation of the electrocardiogram. A short survey on the mechanism of septum activation is carried out and an explanation is given to the ECG changes, described, manifested with the appearance of Q peaks in the precordial V1-V3 leads.


A pharmacokinetic study to correlate the hypoglycemic effect of phlorizin in rats: Identification of metabolites as inhibitors of sodium/glucose cotransporters.
Phlorizin (PRZ) is a natural product that belongs to a class of dihydrochalcones. The unique pharmacological property of PRZ is to block glucose absorption or reabsorption through specific and competitive inhibitors of the sodium/glucose cotransporters (SGLTs) in the intestine (SGLT1) and kidney (SGLT2). This results in glycosuria by inhibiting renal reabsorption of glucose and can be used as an adjuvant treatment for type 2 diabetes. The pharmacokinetic profile, metabolites of the PRZ, and efficacy of metabolites towards SGLTs are unknown. Therefore, the present study on the characterization of hitherto unknown in vivo metabolites of PRZ and pharmacokinetic profiling using liquid chromatography-electrospray ionization tandem mass spectrometry (LC/ESI/MS/MS) and accurate mass measurements is undertaken. Plasma, urine, and feces samples were collected after oral administration of PRZ to Sprague-Dawley rats to identify in vivo metabolites. Furthermore, in silico efficacy of the identified metabolites was evaluated by docking study. PRZ at an intraperitoneal dose of 400 mg/kg showed maximum concentration in the blood to 439.32 ± 8.84 ng/mL at 1 h, while phloretin showed 14.38 ± 0.33 ng/mL at 6 h. The pharmacokinetic profile of PRZ showed that the maximum concentration lies between 1 and 2 h after dosing. Decreased blood glucose levels and maximum excretion of glucose in the urine were observed when the PRZ and metabolites were observed in plasma. The identification and characterization of PRZ metabolites by LC/ESI/MS/MS further revealed that the phase I metabolites of PRZ are hydroxy (mono-, di-, and tri-) and reduction. Phase II metabolites are O-methylated, O-acetylated, O-sulfated, and glucuronide metabolites of PRZ. Further docking study revealed that the metabolites diglucuronide metabolite of mono-hydroxylated PRZ and mono-glucuronidation of PRZ could be considered novel inhibitors of SGLT1 and SGLT2, respectively, which show better binding affinities than their parent compound PRZ and the known inhibitors.


Field immobilization for treatment of an unknown illness in a wild chimpanzee (Pan troglodytes schweinfurthii) at Gombe National Park, Tanzania: findings, challenges, and lessons learned.
Infectious diseases are widely presumed to be one of the greatest threats to ape conservation in the wild. Human diseases are of particular concern, and the costs and benefits of human presence in protected areas with apes are regularly debated. While numerous syndromes with fatal outcomes have recently been described, precise identification of pathogens remains difficult. These diagnostic difficulties are compounded by the fact that direct veterinary intervention on wild apes is quite rare. Here we present the unique case of a wild chimpanzee at Gombe National Park that was observed with a severe illness and was subsequently examined and treated in the field. Multiple specimens were collected and tested with the aim of identifying the pathogen responsible for the illness. Our findings represent the first extensive screening of a living wild chimpanzee, yet despite our efforts, the cause and source of illness remain unknown. Nevertheless, our findings represent valuable baseline data for the ape conservation community and for comparison with other recent findings. In addition, we present the case here to demonstrate the planning required and multiple types of expertise necessary to maximize the amount of data obtained from such a rare intervention, and to provide lessons learned for future studies.


Coingestion of protein and carbohydrate in the early recovery phase, compared with carbohydrate only, improves endurance performance despite similar glycogen degradation and AMPK phosphorylation.
The present study compared the effects of postexercise carbohydrate plus protein (CHO+PROT) and carbohydrate (CHO)-only supplementation on muscle glycogen metabolism, anabolic cell signaling, and subsequent exercise performance. Nine endurance-trained males cycled twice to exhaustion (muscle glycogen decreased from ~495 to ~125 mmol/kg dry wt) and received either CHO only (1.2 g·kg-1·h-1) or CHO+PROT (0.8/0.4 g·kg-1·h-1) during the first 90 min of recovery. Glycogen content was similar before the performance test after 5 h of recovery. Glycogen synthase (GS) fractional activity increased after exhaustive exercise and remained activated 5 h after, despite substantial glycogen synthesis (176.1 ± 19.1 and 204.6 ± 27.0 mmol/kg dry wt in CHO and CHO+PROT, respectively; P = 0.15). Phosphorylation of GS at site 3 and site 2+2a remained low during recovery. After the 5-h recovery, cycling time to exhaustion was improved by CHO+PROT supplementation compared with CHO supplementation (54.6 ± 11.0 vs. 46.1 ± 9.8 min; P = 0.009). After the performance test, muscle glycogen was equally reduced in CHO+PROT and CHO. Akt Ser473 and p70s6k Thr389 phosphorylation was elevated after 5 h of recovery. There were no differences in Akt Ser473, p70s6k Thr389, or TSC2 Thr1462 phosphorylation between treatments. Nitrogen balance was positive in CHO+PROT (19.6 ± 7.6 mg nitrogen/kg; P = 0.04) and higher than CHO (-10.7 ± 6.3 mg nitrogen/kg; P = 0.009). CHO+PROT supplementation during exercise recovery improved subsequent endurance performance relative to consuming CHO only. This improved performance after CHO+PROT supplementation could not be accounted for by differences in glycogen metabolism or anabolic cell signaling, but may have been related to differences in nitrogen balance.NEW & NOTEWORTHY Endurance athletes competing consecutive days need optimal dietary intake during the recovery period. We report that coingestion of protein and carbohydrate soon after exhaustive exercise, compared with carbohydrate only, resulted in better performance the following day. The better performance after coingestion of protein and carbohydrate was not associated with a higher rate of glycogen synthesis or activation of anabolic signaling compared with carbohydrate only. Importantly, nitrogen balance was positive after coingestion of protein and carbohydrate, which was not the case after intake of carbohydrate only, suggesting that protein synthesis contributes to the better performance the following day.


Improved classification of indeterminate biliary strictures by probe-based confocal laser endomicroscopy using the Paris Criteria following biliary stenting.
Probe-based confocal laser endomicroscopy (pCLE) using the Miami Criteria has improved classification of indeterminate biliary strictures. However, previous biliary stenting may result in their misclassification as malignant strictures. Inflammatory criteria were added to form the Paris Classification to prevent this misclassification and reduce false positives. The aim of this study was to assess if the Paris Classification was more accurate than the Miami Classification in classifying indeterminate biliary strictures after biliary stenting. This was a retrospective observational study involving 21 patients with indeterminate biliary strictures from whom 27 pCLE video sequences were obtained (20 benign and seven malignant). Patients with and without prior biliary stenting underwent pCLE. Two investigators classified the strictures as malignant or benign using the Miami and Paris Classifications. Diagnostic accuracy, sensitivity (Se), and specificity (Sp) of endoscopic retrograde-guided pCLE were compared with final histopathology. In those without biliary stenting, the Miami Criteria resulted in Se 88%, Sp 75%, positive predictive value (PPV) 64%, negative predictive value (NPV) 92%, and accuracy 79%, while the Paris Classification resulted in Se 63%, Sp 88%, PPV 71%, NPV 82%, and accuracy 79%. In those with prior biliary stenting, the Miami Criteria resulted in Se 88%, Sp 36%, PPV 23%, NPV 93%, and accuracy 45%, while the Paris Classification resulted in Se 63%, Sp 73%, PPV 31%, NPV 91%, and accuracy 71%. The kappa statistic was 0.56. The Paris Classification improved specificity and accuracy of biliary stricture classification in those who had been previously stented and decreased the rate of misclassification of benign strictures as malignant.


cDNA nucleotide sequence and primary structure of mouse uterine peptidylarginine deiminase. Detection of a 3'-untranslated nucleotide sequence common to the mRNA of transiently expressed genes and rapid turnover of this enzyme's mRNA in the estrous cycle.
Peptidylarginine deiminase is a protein-modulating enzyme which converts the arginine residues in proteins to citrulline residues. This study describes the complete primary structure of mouse peptidylarginine deiminase, which was deduced from nucleotide sequence analysis of cDNA clones plus proteochemical analysis of the purified enzyme. The composite cDNA sequence contained a 5' untranslated region of 7 bases, an open reading frame of 2019 bases that encoded 673 amino acids, a 3' untranslated region of 2662 bases, and part of a poly(A) tail. The N-terminal and C-terminal sequences of the enzyme matched the sequences deduced from nucleotide analysis. Furthermore, we determined that the N-terminal sequence was N alpha-acetyl-Met-Gln-, a sequence which has never previously been reported among N alpha-acetyl-Met proteins. The Arg 352 of the enzyme was converted to a citrulline residue and the potential Asn-linked glycosylation site (Asn542-Glu543-Ser544) had no carbohydrate moiety. Thus, mouse peptidylarginine deiminase consists of 673 amino acids with a molecular mass of 76,260. Mouse peptidylarginine deiminase mRNA has two AU-rich structures in the 3' untranslated region which exhibit a high degree of similarity to those in lymphokine, cytokine and proto-oncogene mRNA species. Since the rat enzyme (previously reported) does not possess these characteristic structures, we compared the levels of enzyme activity and mRNA in the mouse and rat uterus at four defined phases of the estrous cycle. The degradation of peptidylarginine deiminase and its mRNA proceeded significantly faster in the mouse than in the rat. We speculate that the unusual structure of the mouse enzyme and its mRNA be involved in this species-specific rapid degradation.


Intrinsic GABA neurons inhibit proenkephalin gene expression in slice cultures of rat neostriatum.
In the neostriatum, the proenkephalin gene is expressed in medium spiny GABA neurons, which project to the globus pallidus. The expression is activated by glutamatergic projections from the neocortex via NMDA receptors. In these experiments we have used slice cultures of rat neostriatum to study the role of GABA in proenkephalin gene expression. Our results show that GABA is released from neostriatal neurons and negatively regulates the proenkephalin gene expression induced by NMDA receptor stimulation. The GABAA receptors involved seem to be colocalized with NMDA receptors on the projection neurons, which express the proenkephalin gene. In further experiments, we have found that the proenkephalin gene expression is not only activated by neocortical projection neurons but also by intrinsic striatal neurons as well as by projections from the thalamus. All these glutamatergic afferents enhance the proenkephalin gene expression via NMDA receptors. Their efficacy is regulated by endogenous GABA.


[Extracorporeal blood irradiation using telecesium apparatus. II. Morphological studies in patients with chronic lymphadenosis].
The second communication of our experiences with extracorporeal blood irradiation comprises a systematical investigation of the conditions in blood picture and, in one case, of the morphological findings within lymph node punctures, during long-term treatment of two selected patients with a rather different course of the disease. One of the cases showed a typical pattern of chronic lymphadenosis which was to be influenced fairly well; the other case revealed an aggressive tendency with only a feeble response to extracorporeal blood irradiation. By means of toluidine blue staining, until now being used for the verification of nucleoli and their different shapes only in experimentations on animals, a good correlation of the nucleole analysis in the peripheral blood picture and in the lymph node biopsies with the clinical progress of the disease has been observed. Thus, this simple cytological method combined with customary clinical examinations may serve as a criterion for the judgement on the course of the disease and on the therapeutic success.


Prone Positioning in Mechanically Ventilated COVID-19 Patients: Timing of Initiation and Outcomes.
The COVID-19 pandemic led to a broad implementation of proning to enhance oxygenation in both self-ventilating and mechanically ventilated critically ill patients with acute severe hypoxic respiratory failure. However, there is little data on the impact of the timing of the initiation of prone positioning in COVID-19 patients receiving mechanical ventilation. In this study, we analyzed our proning practices in mechanically ventilated COVID-19 patients. There were 931 total proning episodes in 144 patients, with a median duration of 16 h (IQR 15-17 h) per proning cycle. 563 proning cycles were initiated within 7 days of intubation (early), 235 within 7-14 days (intermediate), and 133 after 14 days (late). The mean change in oxygenation defined as the delta PaO2/FiO2 ratio (ΔPF) after the prone episode was 16.6 ± 34.4 mmHg (p < 0.001). For early, intermediate, and late cycles, mean ΔPF ratios were 18.5 ± 36.7 mmHg, 13.2 ± 30.4 mmHg, and 14.8 ± 30.5 mmHg, with no significant difference in response between early, intermediate, and late proning (p = 0.2), respectively. Our findings indicate a favorable oxygenation response to proning episodes at all time points, even after >14 days of intubation. However, the findings cannot be translated directly into a survival advantage, and more research is needed in this area.


DAZL regulates proliferation of human primordial germ cells by direct binding to precursor miRNAs and enhances DICER processing activity.
Understanding the molecular and cellular mechanisms of human primordial germ cells (hPGCs) is essential in studying infertility and germ cell tumorigenesis. Many RNA-binding proteins (RBPs) and non-coding RNAs are specifically expressed and functional during hPGC developments. However, the roles and regulatory mechanisms of these RBPs and non-coding RNAs, such as microRNAs (miRNAs), in hPGCs remain elusive. In this study, we reported a new regulatory function of DAZL, a germ cell-specific RBP, in miRNA biogenesis and cell proliferation. First, DAZL co-localized with miRNA let-7a in human PGCs and up-regulated the levels of >100 mature miRNAs, including eight out of nine let-7 family, miR21, miR22, miR125, miR10 and miR199. Purified DAZL directly bound to the loops of precursor miRNAs with sequence specificity of GUU. The binding of DAZL to the precursor miRNA increased the maturation of miRNA by enhancing the cleavage activity of DICER. Furthermore, cell proliferation assay and cell cycle analysis confirmed that DAZL inhibited the proliferation of in vitro PGCs by promoting the maturation of these miRNAs. Evidently, the mature miRNAs up-regulated by DAZL silenced cell proliferation regulators including TRIM71. Moreover, DAZL inhibited germline tumor cell proliferation and teratoma formation. These results demonstrate that DAZL regulates hPGC proliferation by enhancing miRNA processing.


Genome-wide identification of haploinsufficiency in fission yeast.
Abnormal phenotypes resulting from haploinsufficiency (HI) are due to the loss of one allele. Recent studies in budding yeast have shown that HI originates from insufficient protein levels or from a stoichiometric imbalance between subunits of protein complexes. In humans, however, HI often involves transcription factors. Therefore, the species differences in HI and the molecular mechanisms of species-specific HI remain under investigation. In this study, HI in fission yeast was systematically surveyed. HI in fission yeast affected genes related to signaling and to basic cellular processes, as observed in budding yeast. These results suggest that there are species differences in HI and that the HI that occurs in fission yeast is intermediate to and HI in budding yeast and humans.


[New European Information and Communication Platform Striving for Improved Care for Colorectal Cancer Patients - www.crcprevention.eu].
The article introduces a new information and communication platform that focuses on the promotion of colorectal cancer prevention and on the improvement of care for colorectal cancer patients. In addition to the platforms objectives, the article also presents the conclusions of the first year of the "European Colorectal Cancer Days" international conference held annually in Brno since 2012. Further information and news are available at www.crcprevention.eu.


Taxol and taurine protect the renal tissue of rats after unilateral ureteral obstruction: a stereological survey.
Blockage of the urinary tract induces changes in renal structure including tubular dilatation or atrophy, tubular cell death, inflammatory processes, and progressive interstitial fibrosis with the loss of renal parenchyma. The present study was conducted to survey the protective effects of Taxol and taurine on the renal structure after unilateral ureteral obstruction (UUO). UUO was induced in three groups of rats (n=6) who then received distilled water, Taxol (0.3 mg/kg/d), or taurine (7.5 mg/kg/d). Stereological methods were used to gather quantitative as well as comparative data. Less than -8% of the volume of the glomeruli, proximal convoluted tubules (PCT), distal convoluted tubules (DCT), Henle's loop, and collecting ducts were preserved after UUO. After treatment of the UUO rats with Taxol, between -32% and 88% of the parameters mentioned above remained intact, and after treatment of the UUO rats with taurine, between -16% and 46% of the parameters remained intact (p<0.01). Compared with the untreated UUO animals, the volume of necrotic and fibrotic tissues decreased -53% and -63% in the UUO rats treated with Taxol and taurine, respectively (p<0.01). Less than -3% of the lengths of the renal tubules (PCT, DCT, Henle's loop, and collecting) were preserved in the UUO rats. After treatment with Taxol and taurine, -61% to 70% and -43% to 53% of the length of the renal tubules were preserved, respectively (p<0.01). Taurine and Taxol are effective in preventing some structural renal damage in a direct ureteral obstruction model. Taxol was more effective in renal protection.


Labeling of monoclonal antibodies with samarium-153 for combined radioimmunoscintigraphy and radioimmunotherapy.
The labeling of a monoclonal antibody K-1-21 with 153Sm has been investigated using the bifunctional chelate cyclic diethylenetriaminepentaacetic acid (DTPA) anhydride. Labeling efficiencies greater than 60% were obtained using high specific activity [153Sm]chloride and a cDTPAa:MAb conjugation ratio of 20:1. The resultant labeled antibody had a s.a. greater than 150 MBq.mg-1 and a % retained immunoreactivity greater than 90%. Imaging and biodistribution studies in a rat model demonstrated that specific uptake of 153Sm-K-1-21 into s.c. implants of the target antigen could be clearly detected in scintigrams at 6 days p.i. The specific uptake (1.90 +/- 0.45% ID/g, 19.95 +/- 2.20 Implant:Blood ratio) compared favorably to 131I- and 111In-labeled K-1-21 (2.52 +/- 0.20 and 3.33 +/- 0.20% ID/g, 7.69 +/- 0.45 and 10.10 +/- 0.60 I:B, respectively). Labeling of MAbs with 153Sm for combined scintigraphy/therapy is feasible at clinically appropriate specific activities using cDTPAa, with the resultant conjugates retaining immunoreactivity and in vivo antigen localization.


Probiotic Enterococcus faecalis surface-delivering key domain of EtMIC3 proteins: immunoprotective efficacies against Eimeria tenella infection in chickens.
Enterococcus faecalis MDXEF-1 strain was used as host bacteria to deliver surface-displayed key domains BC1 and C4D of the Eimeria tenella microneme 3 (EtMIC3) protein, BC1 fusion to dendritic cell (DC) targeting peptides (DCpep) (DC-BC1) and DC-C4D, respectively. The expression of target proteins in MDXEF-1/BC1-CWA, MDXEF-1/DC-BC1-CWA, MDXEF-1/C4D-CWA, and MDXEF-1/DC-C4D-CWA was verified. The systemic immune responses induced by oral immunization with recombinant E. faecalis were recorded. The immunized chickens were challenged with 4.0 × 104 E. tenella-sporulated oocysts. The immune protections were evaluated by calculating average weight gain and oocyst reduction rate, observing gross and histopathological changes in the cecal tissues, and quantifying levels of inflammatory cytokines in the cecal tissues. The results showed that proteins BC1, C4D, DC-BC1, and DC-C4D were expressed on the surface of E. faecalis, respectively. Oral immunizations with recombinant E. faecalis, especially MDXEF-1/DC-BC1-CWA and MDXEF-1/DC-C4D-CWA, stimulated higher levels of antigen-specific humoral and intestinal mucosal immune responses, higher levels of Th1, Th2, and Th17-type cytokines, and higher proliferation of peripheral blood lymphocytes than phosphate-buffered saline (PBS) and vector control groups (P < 0.01). The groups immunized with four recombinant E. faecalis showed better protective effects against homologous infection than the vector control and infection control groups, and the MDXEF-1/DC-BC1-CWA group displayed the best effects. These results demonstrated that probiotic E. faecalis delivering DCpep fused with the key domain of the EtMIC3 protein could be a potential approach for the prevention of Eimeria infection. IMPORTANCE Avian coccidiosis caused by Eimeria brings huge economic losses to the poultry industry. Although live vaccines and anti-coccidial drugs were used for a long time, Eimeria infection in chicken farms all over the world commonly occurred. The exploration of novel, effective vaccines has become a research hotspot. Eimeria parasites have complex life cycles, and effective antigens are particularly critical to developing anti-coccidial vaccines. Microneme proteins (MICs), secreted from microneme organelles located at the parasite apex, are considered immunodominant antigens. Eimeria tenella microneme 3 (EtMIC3) contains four conserved repeats (MARc1, MARc2, MARc3, and MARc4) and three divergent repeats (MARa, MARb, and MARd), which play a vital role during the Eimeria invasion. Enterococcus faecalis is a native probiotic in animal intestines and can regulate intestinal flora. In this study, BC1 and C4D domains of EtMIC3, BC1 or C4D fusing to dendritic cells targeting peptides, were surface-displyed by E. faecalis, respectively. Oral immunizations were performed to investigate immune protective effects against Eimeria infection.


The Ageing Brain: Investigating the Role of Age in Changes to the Human Cerebral Microvasculature With an in silico Model.
Ageing causes extensive structural changes to the human cerebral microvasculature, which have a significant effect on capillary bed perfusion and oxygen transport. Current models of brain capillary networks in the literature focus on healthy adult brains and do not capture the effects of ageing, which is critical when studying neurodegenerative diseases. This study builds upon a statistically accurate model of the human cerebral microvasculature based on ex-vivo morphological data. This model is adapted for "healthy" ageing using in-vivo measurements from mice at three distinct age groups-young, middle-aged, and old. From this new model, blood and molecular exchange parameters are calculated such as permeability and surface-area-to-volume ratio, and compared across the three age groups. The ability to alter the model vessel-by-vessel is used to create a continuous gradient of ageing. It was found that surface-area-to-volume ratio reduced in old age by 6% and permeability by 24% from middle-age to old age, and variability within the networks also increased with age. The ageing gradient indicated a threshold in the ageing process around 75 years old, after which small changes have an amplified effect on blood flow properties. This gradient enables comparison of studies measuring cerebral properties at discrete points in time. The response of middle aged and old aged capillary beds to micro-emboli showed a lower robustness of the old age capillary bed to vessel occlusion. As the brain ages, there is thus increased vulnerability of the microvasculature-with a "tipping point" beyond which further remodeling of the microvasculature has exaggerated effects on the brain. When developing in-silico models of the brain, age is a very important consideration to accurately assess risk factors for cognitive decline and isolate early biomarkers of microvascular health.