Prevalence, extent, and independent predictors of silent myocardial infarction.
The phenomenon of silent myocardial infarction is poorly understood. We aimed to evaluate the prevalence, extent, and independent predictors of silent myocardial infarction in 2 large independent cohorts of consecutive patients without a history of myocardial infarction referred for rest/stress myocardial perfusion single photon emission computed tomography. There were 1621 patients enrolled in the derivation cohort and 338 patients in the validation cohort. Silent myocardial infarction was diagnosed in patients with a myocardial scar ≥5% of the left ventricle. In the derivation cohort, the prevalence of silent myocardial infarction was 23.3% (n = 377). The median infarct size was 10% (interquartile range [IQR] 5%-15%) of the left ventricle. The prevalence of silent myocardial infarction was 28.5% in diabetics and 21.5% in nondiabetics (P = .004). Diabetes mellitus was an independent predictor for the presence of silent myocardial infarction (odds ratio 1.5; 95% confidence interval, 1.1-1.9; P = .004). These findings were confirmed in the independent validation cohort. In the validation cohort, the prevalence of silent myocardial infarction was 26.3% (n = 89), while the prevalence was higher in diabetics (35.8%) than in nondiabetics (24%; P = .049). The median infarct size was 11.8% (IQR 5.9%-17.6%) of the left ventricle. Again, in logistic regression analysis, diabetes mellitus was a significant predictor of the presence of silent myocardial infarction. Silent myocardial infarctions are more common than previously thought. One of 4 patients with suspected coronary artery disease had experienced a silent myocardial infarction; the extent in average is 10% of the left ventricle, and it is more common in diabetics.


Correlation between gestational age, weight, or ponderosity and serum thyroxine concentration at birth.
Thyroxine levels were measured in cord blood sera from 2,1800 healthy infants born of "low-risk" mothers in Los Angeles. Variations of T4 with gestational age, birth weight, ponderosity (size related to weight), sex, and race were studied, as well as the variation of W and P with GA. Over the range of GA 30 to 47 weeks, serum T4 concentrations, W, and P each increase with GA, leveling off to approximately constant values from 42 to 47 weeks; the increases are 12, 66, and 20%, respectively. For a fixed GA, T4 increases with increasing W but is not correlated with P; W, GA, and their product are useful measurements for predicting T4, but P is not. Formulas for predicting cord T4 are given for possible use in improving computer-assisted screening programs for congenital hypothyroidism; mean T4 values (+/- I SD) are tabulated against W and GA.


Functional specificity of G alpha q and G alpha 11 in the cholinergic and glutamatergic modulation of potassium currents and excitability in hippocampal neurons.
In hippocampal and other cortical neurons, action potentials are followed by a slow afterhyperpolarization (sAHP) generated by the activation of small-conductance Ca(2+)-activated K(+) channels and controlling spike frequency adaptation. The corresponding current, the apamin-insensitive sI(AHP), is a well known target of modulation by different neurotransmitters, including acetylcholine (via M(3) receptors) and glutamate (via metabotropic glutamate receptor 5, mGluR(5)), in CA1 pyramidal neurons. The actions of muscarinic and mGluR agonists on sI(AHP) involve the activation of pertussis toxin-insensitive G-proteins. However, the pharmacological tools available so far did not permit the identification of the specific G-protein subtypes transducing the effects of M(3) and mGluR(5) on sI(AHP). In the present study, we used mice deficient in the Galpha(q) and Galpha(11) genes to investigate the specific role of these G-protein alpha subunits in the cholinergic and glutamatergic modulation of sI(AHP) in CA1 neurons. In mice lacking Galpha(q), the effects of muscarinic and glutamatergic agonists on sI(AHP) were nearly abolished, whereas beta-adrenergic agonists acting via Galpha(s) were still fully effective. Modulation of sI(AHP) by any of these agonists was instead unchanged in mice lacking Galpha(11). The additional depolarizing effects of muscarinic and glutamatergic agonists on CA1 neurons were preserved in mice lacking Galpha(q) or Galpha(11). Thus, Galpha(q), but not Galpha(11), mediates specifically the action of cholinergic and glutamatergic agonists on sI(AHP), without affecting the modulation of other currents. These results provide to our knowledge one of the first examples of the functional specificity of Galpha(q) and Galpha(11) in central neurons.


Methodology for measuring health-state preferences--I: Measurement strategies.
Values play a critical part in decision making at both the individual and policy levels. Numerous methodologies for determining the preferences of individuals and groups have been proposed, but agreement has not been reached regarding their scientific adequacy and feasibility. This is the first of a four-part series of papers that analyzes and critiques the state-of-the-art in measuring preferences, particularly the measurement of health-state preferences. In this first paper we discuss the selection of relevant attributes to comprise the health-state descriptions, and the relative merits of three measurement strategies: holistic, explicitly decomposed, and statistically inferred decomposed. The functional measurement approach, a statistically inferred decomposed strategy, is recommended because it simultaneously validates the process by which judges combine attributes, the scale values they assign to health states, and the interval property of the scale.


Myocardial perfusion imaging after coronary revascularization: a clinical appraisal.
Revascularization procedures, including percutaneous coronary intervention (PCI) and coronary artery bypass grafting (CABG), are performed in many patients with coronary artery disease. Despite the effectiveness of these procedures, different follow-up strategies need to be considered for the management of patients after revascularization. Stress myocardial perfusion single-photon emission computed tomography (MPS) is a suitable imaging method for the evaluation of patients who have undergone PCI or CABG, and it has been used in the follow-up of such patients. Radionuclide imaging is included in the follow-up strategies after PCI and CABG in patients with symptoms, but guidelines warn against routine testing of all asymptomatic patients after revascularization. After PCI, in the absence of symptoms, radionuclide imaging is recommended and indicated as appropriate after incomplete or suboptimal revascularization and in specific asymptomatic patient subsets. On the other hand, the value of MPS late after CABG in risk stratification has been demonstrated even in the absence of symptoms. Thus, given the adverse outcome associated with silent ischaemia, it can be speculated that all patients regardless of clinical status should undergo stress testing late after revascularization. Larger prospective studies are needed to assess whether stress MPS will have an impact on the outcome in asymptomatic patients after revascularization.


Author Correction: Immunologic findings precede rapid lupus flare after transient steroid therapy.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Application of real-time PCR to significantly reduce the time to obtain recombinant MVA virus.
Obtaining a pure recombinant Modified Vaccinia Ankara (MVA) virus is a multistage, time-consuming procedure. We describe a novel single-tube real-time PCR which enables determination of the amount of wild type and recombinant viruses and their ratio in plaques. Use of the real-time PCR significantly reduce the time and efforts needed to obtain purified recombinant MVA. The new approach has been applied to generate recombinant MVAs encoding different SARS-COV-2 antigens.


Degradation of β-N-methylamino-l-alanine (BMAA) by UV/peracetic acid system: Influencing factors, degradation mechanism and DBP formation.
β-N-methylamino-l-alanine (BMAA) is a cyanobacterial neurotoxin associated with human neurodegenerative diseases, and its removal in drinking water is receiving increasing attention. In this study, the degradation of BMAA in UV/peracetic acid (UV/PAA) system was investigated. BMAA degradation followed the pseudo-first-order kinetic model. The synergistic effect of UV and PAA exhibited a great potential for BMAA degradation, which was attributed to the generation of a large number of reactive radicals, of which R-C• was the most dominant contributor. We also explored the effects of different factors on BMAA degradation. The results showed that there was a positive correlation between BMAA degradation and PAA dosage, and the optimal effect was achieved at pH 7. Notably, the existence of water matrices such as bicarbonate (HCO3-), chloride ion (Cl-), humic acid (HA) and algal intracellular organic matter (IOM) all inhibited the degradation of BMAA. Based on the identified intermediates, this study suggested that reactive radicals degraded BMAA mainly by attacking the carbon-nitrogen bonds on BMAA. Besides, comparing the effect of Cl- on disinfection byproduct (DBP) formation in UV/PAA-post-PAA oxidation and UV/chlorine-post-chlorination systems, it was found that the former was more sensitive to the presence of Cl-.


Determination of bullatacin in rat plasma by liquid chromatography-mass spectrometry.
A liquid chromatography-mass spectrometry method has been developed and validated for the quantification of bullatacin, a bistetrahydrofuran annonaceous acetogenin, in rat plasma. Squamostatin-A was selected as the internal standard. Analytes were extracted from rat plasma by liquid/liquid extraction using ethyl acetate with high efficiency. The chromatographical separation was performed on an Agilent Zorbax SB-C₁₈ column (150 mm × 2.1 mm, 5 μm). The mobile phase consisted of methanol and deionized water (95:5, v/v) containing 0.01% (v/v) formic acid. The chromatographic run time was 7 min per injection and flow rate was 0.2 mL/min. The retention time was 3.22 and 5.23 min for internal standard and bullatacin, respectively. The elutes were detected under positive electrospray ionization and the target analytes quantified by selected ion monitoring mode (645.9 m/z for bullatacin and 661.9 m/z for squamostatin-A). The method was sensitive with the limit of quantitation at 0.5 ng/mL in 100 μL of rat plasma. Good linearity (r²=0.9998) was obtained covering the concentration of 0.5-2000 ng/mL. The intra- and inter-day assay precision ranged from 3.2 to 8.7% and 2.7 to 9.2%, respectively. In addition, the stability, extraction recovery and matrix effect involved in the method were also validated. This method was applied to measure the plasma bullatacin concentrations after a single tail vein intravenous administration of bullatacin in rats.


Characterization of intact microorganisms by MALDI mass spectrometry.
The application of MALDI mass spectrometry to desorb protein biomarkers from intact viruses, bacteria, fungus, and spores is the focus of this review. Instrumentation, sample collection, sample preparation, and algorithms for data analysis are summarized. Optimally these analyses should be carried out in less than five minutes. Successful applications are discussed from biotechnology, cell biology, and the pharmaceutical industry.


Stress-Buffering Role of Social Support during COVID-19.
In order to reduce the high infection rate of COVID-19, individuals began to engage in self-isolation amid a time of uncertainty and worry. Given that social support can be protective against the negative effects of distress on mental and physical health, the lack of support may negatively impact individuals during their self-isolation. Thus, the current study examined the role of self-isolation on feelings of stress, the perception and reception of social support, and mental health problems during the COVID-19 pandemic. A sample of 405 college students were asked to report on the amount of self-isolation in which they were engaging, worry about COVID-19, psychological health, and received and perceived social support. Results indicated that when the length of time in self-isolation was taken into account, perceived social support buffered the connection between worry about COVID-19 and psychological health. These results indicate that social support, worry about COVID-19, and self-isolation may influence individuals' psychological health during times of stress.


Predisposing factors for severe mitral regurgitation in idiopathic mitral valve prolapse.
To elucidate predisposing factors for severe mitral regurgitation (MR) in idiopathic mitral valve prolapse (MVP), 124 MVP patients were classified into the following categories: 55 with isolated clicks (click group), 35 with a late-systolic murmur (late-SM group), and 34 with a holosystolic murmur (holo-SM group). Their clinical and echocardiographic findings were compared with those of 26 patients with spontaneous chordal rupture (rupture group). In 22 patients in the click group, 24 in the late-SM group, and 22 in the holo-SM group, follow-up studies were performed for a mean of 4.5 years (range 1 to 13.5). The mean age was youngest in the click group and oldest in the rupture group. The click and late-SM groups showed a female predominance, but the holo-SM and rupture groups showed a male predominance. There was no difference in the incidence of systemic hypertension among the 4 groups. Most patients in the click and late-SM groups had anterior leaflet prolapse. In the holo-SM and rupture groups, however, the incidence of posterior leaflet involvement was significantly increased. The incidence of thickened mitral valve increased in order of the click (8%), late-SM (21%), holo-SM (38%), and rupture (50%) groups. Six patients in the holo-SM group developed chordal rupture with severe MR during the follow-up period. In the click and late-SM groups, however, there were no complications and no development into a holo-SM. Thus, aging, male sex, posterior leaflet prolapse, thickened mitral valve, and holo-SM were found to be important predisposing factors for severe MR in idiopathic MVP.


Why is the fusiform face area recruited for novel categories of expertise? A neurocomputational investigation.
What is the role of the Fusiform Face Area (FFA)? Is it specific to face processing, or is it a visual expertise area? The expertise hypothesis is appealing due to a number of studies showing that the FFA is activated by pictures of objects within the subject's domain of expertise (e.g., cars for car experts, birds for birders, etc.), and that activation of the FFA increases as new expertise is acquired in the lab. However, it is incumbent upon the proponents of the expertise hypothesis to explain how it is that an area that is initially specialized for faces becomes recruited for new classes of stimuli. We dub this the "visual expertise mystery." One suggested answer to this mystery is that the FFA is used simply because it is a fine discrimination area, but this account has historically lacked a mechanism describing exactly how the FFA would be recruited for novel domains of expertise. In this study, we show that a neurocomputational model trained to perform subordinate-level discrimination within a visually homogeneous class develops transformations that magnify differences between similar objects, in marked contrast to networks trained to simply categorize the objects. This magnification generalizes to novel classes, leading to faster learning of new discriminations. We suggest this is why the FFA is recruited for new expertise. The model predicts that individual FFA neurons will have highly variable responses to stimuli within expertise domains.


The appropriateness of oral fluoroquinolone-prescribing in the long-term care setting.
To evaluate the appropriateness of ciprofloxacin-prescribing in the long-term care setting. Retrospective chart review. A large academically oriented long-term care facility. Institutionalized elderly patients with a mean age of 88 years. One hundred orders were randomly selected for review from all ciprofloxacin orders initiated over a 3-year period. Criteria for appropriateness of ciprofloxacin-prescribing were developed based on a comprehensive review of the medical literature. Evaluation of appropriateness of prescribing was based on the indication for therapy and the availability of more effective and/or less expensive alternative antibiotic regimens. Only information available to the physician at the time of the order was used to judge appropriateness. Abstracted medical records were evaluated independently by a geriatrician and an infectious diseases specialist. With respect to site of infection, the urinary tract accounted for 43% of all ciprofloxacin orders; the lower respiratory tract, 28%; and skin and soft-tissue infections, 17%. Only 25% of orders were judged appropriate. Twenty-three percent of orders were judged less than appropriate based on indication, and 49% due to the availability of a more effective and/or less expensive alternative antibiotic choice. There was insufficient information in the medical record to judge 3% of the orders. These results indicate less than optimal prescribing of oral fluoroquinolones in the long-term care setting, with potential consequences including the development of resistant bacterial strains and increased health care costs.


Effect of freeze-dried poly-L-lactic acid discs mixed with bone morphogenetic protein on the healing of rat skull defects.
Porous poly-L-lactic-acid discs were prepared by a freeze-drying method (FDPLLA). During preparation of the discs, semipurified bone morphogenetic protein (BMP) was mixed with the poly-L-lactic acid (500 micrograms BMP per disc). The discs were implanted in the skull defects of rats and retrieved after 2 or 4 weeks. With the FDPLLA/BMP discs, new bone formation was observed at 2 weeks after implantation. On the other hand, plain FDPLLA discs failed to produce new bone formation even at 4 weeks after implantation. Calcium content of the retrieved FDPLLA/BMP discs was statistically higher than that of plain FDPLLA discs. These results suggest that FDPLLA may be an effective delivery system for BMP and that the FDPLLA/BMP disc may potentially have usefulness as a bone graft substitute.


Dark CO2-fixation and diurnal malic acid fluctuations in the submerged-aquatic Isoetes storkii.
In the leaves (but not corms) of the submerged aquatic Isoetes storkii malic acid concentration fluctuated from 22 μeg g FW-1 in the evening to 171 μeg g FW-1 in the morning. Associated with this was a change in titratable acidity of 152 μeg g FW-1 between morning and evening. 14C carbon was fixed in both the light and the dark, though the amount of carbon fixed in the light was more than that fixed in the dark. Autoradiographs show 88% of 14CO2 fixed in the dark is recovered after 1 h, in malic acid and the remainder in one other unidentified product, whereas these two products contain less than 15% of the 14C fixed after 1 h exposure to 14CO2 in the light. It is suggested that CAM metabolism in this aquatic species may be related to the low availability of CO2 for photosynthesis during the day in its aquatic environment and that this metabolic pathway may prove common in the genus Isoetes.


Anion-Directed Regulation of Structures and Luminescence of Heterometallic Clusters.
Anions have been used to regulate the structures and luminescence of heterometallic clusters. Introducing ClO4 - into orange-emissive, butterfly-like [(C)(Au-PPhpy2 )6 Ag4 ](BF4 )6 (1, PPhpy2 =bis(2-pyridyl)phenylphosphine) leads to the formation of red-emissive [(C)(Au-PPhpy2 )6 Ag5 (ClO4 )3 ](ClO4 )4 (2) with a novel trigonal bipyramidal structure; employing PhCO2 - gives yellow-emissive, hexagram-like [(C)(Au-PPhpy2 )6 Ag6 (PhCO2 )3 ](BF4 )5 (3). Notably, 1 exhibits weak luminescence in CH2 Cl2 /CH3 OH=1 : 1 (v : v) with a quantum yield (QY) of 0.05, whereas it was dramatically increased to 0.49 and 0.83 for 2 and 3, respectively. Theoretical calculation confirms that the involvement of anions in the electronic structures is responsible for the shifts of emission. The high QYs of 2 and 3 are attributed to the protection provided by ligands and anions. This work demonstrates that anions may serve as an extra designable factor beyond just counterions for functional metal clusters.


Development of a prognostic model for breast cancer survival in an open challenge environment.
The accuracy with which cancer phenotypes can be predicted by selecting and combining molecular features is compromised by the large number of potential features available. In an effort to design a robust prognostic model to predict breast cancer survival, we hypothesized that signatures consisting of genes that are coexpressed in multiple cancer types should correspond to molecular events that are prognostic in all cancers, including breast cancer. We previously identified several such signatures--called attractor metagenes--in an analysis of multiple tumor types. We then tested our attractor metagene hypothesis as participants in the Sage Bionetworks-DREAM Breast Cancer Prognosis Challenge. Using a rich training data set that included gene expression and clinical features for breast cancer patients, we developed a prognostic model that was independently validated in a newly generated patient data set. We describe our model, which was based on three attractor metagenes associated with mitotic chromosomal instability, mesenchymal transition, or lymphocyte-based immune recruitment.


Polymeric amylase nanoparticles as a new semi-synthetic enzyme system for hydrolysis of starch.
α-Amylase (EC 184.108.40.206; α-D-1,4,glucan glucanohydrolase) catalyzes the hydrolysis of α-D-(1,4)-glucosidic linkages in starch, glycogen, and various malto-oligosaccharides, by releasing α-anomeric products. In this study, a novel method has been developed to prepare nanoprotein particles that carry α-amylase as a monomer by using a photosensitive microemulsion polymerization process. The nanostructured α-amylase with photosensitive features have been characterized by fluorescence spectroscopy, transmission electron microscopy (TEM) and Zeta Sizer. The fluorescence intensity of amylase nanoparticles was determined to be 658 a.u. at 610 nm and the average particle size of nanoamylase was found to be about 71.8 nm. Both free α-amylase and nanoparticles were used in the hydrolysis of starch under varying reaction conditions such as pH and temperature that affect enzyme activity and the results were compared to each other. Km values were 0.26 and 0.87 mM and Vmax values were 0.36 IU mg(-1) and 22.32 IU mg(-1) for nanoenzyme and free enzyme, respectively. Then, thermal stability, storage stability and reusability were investigated and according to the results, activity was preserved 60% at 60 °C; 20% at 70-80 °C temperature values and 80% after 105 days storage. Finally after 10 cycles, the activity was preserved 90% and this novel enzymatic polymeric amylase nanoparticle has showed considerable potential as reusable catalyst.


"One-pot" preparation and adsorption performance of chitosan-based La3+/Y3+ dual-ion-imprinted thermosensitive hydrogel.
Temperature-sensitive materials are increasingly of deep interest to researchers. Ion imprinting technology is widely used in the field of metal recovery. In order to solve the problem of rare earth metal recovery, we designed a temperature-sensitive dual-imprinted hydrogel adsorption product (CDIH) with chitosan as the matrix, N-isopropylacrylamide as a thermally responsive monomer, and La3+ and Y3+ as the co-templates. The reversible thermal sensitivity and ion-imprinted structure were determined by differential scanning calorimetry, Fourier transform infrared spectrometer, Raman spectra, Thermogravimetric analysis, X-ray photoelectron spectroscopy, Scanning electron microscopy and X-ray energy spectroscopy various characterizations and analyses. The simultaneous adsorption amount of CDIH for La3+ and Y3+ was 87.04 mg/g and 90.70 mg/g, respectively. The quasi-secondary kinetic model and Freundlich isotherms model well described the adsorption mechanism of CDIH. It's worthy to mention that CDIH could be well regenerated through washing with deionized water at 20 °C, with a desorption rate of 95.29 % for La3+ and 96.03 % for Y3+. And after 10 cycles of reuse, 70 % of the adsorption amount could be maintained, revealing excellent reusability. Furthermore, CDIH expressed better adsorption selectivity to La3+ and Y3+ than its non-imprinted counterparts in a solution containing six metal ions.


Relationship between behavioral maturation measured by the "Baum" test and EEG frequency. A pilot study on monozygotic and dizygotic twins.
In a pilot study, the possible influence of the inherited electroencephalogram (EEG) pattern on aspects of psychologic maturation was investigated. The twin sample consisted of 208 pairs, 110 monozygotic (MZ) and 98 dizygotic (DZ), mostly children and adolescents. The study showed a greater similarity between MZ compared with DZ twins in an immaturity score constructed from the Baum test (Koch 1976) and in a teacher's maturity scoring of essays. For both parameters, a correlation between occipital alpha-frequency and the test criterium could be demonstrated. This correlation was in the direction expected, and it was significant statistically. This result suggests an influence of genetically determined variation of brain maturation on interindividual variability of psychologic variation. Due to shortcomings of the methods used in the study, more specific conclusions cannot be drawn; the problem needs further elucidation using modern methods.


Therapeutic uses of botulinum toxin: from facial palsy to autonomic disorders.
The therapeutic uses of botulinum toxin have been expanding due to deeper knowledge of its molecular behaviour and different mechanisms of action. To present suggested doses of Botox and Dysport for controlling the muscle hyperkinetic activity in facial palsy in the perioral area and to review other uses. An extensive updated literature review on the success and limits of the botulinum neurotoxin (BoNT) therapeutic treatments. BoNT can be considered to be the preferred single method for many disorders; it has substituted for some conventional surgical methods and it can be associated with other therapies to increase overall treatment performance. Depending on the disorder, the lack of permanent effect causes no major inconvenience.


Dissection and culturing of chick ciliary ganglion neurons: a system well suited to synaptic study.
This chapter describes the function and development of the ciliary ganglion, the potential of ciliary ganglion neurons as a cell biological tool, and their dissection, dissociation, and culturing. Ciliary ganglion neurons grow unusually rapidly on a laminin-based substratum and develop large, thin calyx terminals in culture in less than 12 h. The two neuronal classes present in the cultures can be identified by size alone. The limited number of ganglia per animal renders this ganglion a poor choice for biochemical studies based on the extraction of cultured cells. However, they are ideally suited for studies based on single-cell observation, particularly investigation of presynaptic mechanisms using fluorescence microscopy.


Nonbiological factors affecting survival in younger patients with acute myeloid leukemia.
Progress has been made in determining the biological variants of acute myelogenous leukemia (AML) and their prognostic implications. However, to the authors' knowledge, little is known regarding the impact of nonbiological factors (NBFs) on the survival of patients with AML. The impact of NBFs (marital status, insurance status, county-level income, and education) on survival was assessed along with biological factors (disease subtype, sex, age, and race/ethnicity) using a cohort of patients aged 19 to 64 years who were diagnosed with AML between 2007 and 2011 and reported to the Surveillance, Epidemiology, and End Results program registry (SEER 18). There were 5541 patients included. The median overall survival for the entire study population was 16 months. On multivariate analysis, an increased risk of death was independently linked to being a Medicaid beneficiary, uninsured, single, divorced, and residing in a county within the lower 3 quintiles of median household income. NBFs affected the risk of early (<2 months) and late mortality and their impact was confirmed among patients known to have received chemotherapy. Insurance status, marital status, and county-level income were found to independently affect the survival of younger patients with AML and should be integrated into outcome comparisons. Interventions are needed to mitigate the impact of social factors on survival among patients with AML.


Loricrin gene mutation in a Japanese patient of Vohwinkel's syndrome.
Vohwinkel's syndrome (VS) is a rare, dominantly inherited keratoderma with pseudoainhum. Recently, a mutation in loricrin gene has been reported in two VS families of British extraction. In the present study, we examined the loricrin gene mutation in a Japanese VS patient. The patient was a 20-year-old woman. She had palmoplantar keratoderma, constricting bands encircling all the fingers, fifth toes, wrist, and neck. She also had generalized mild ichthyosis and suffered from acoustic impairment. Her parents and a brother showed no skin abnormality. Histopathology of the patient revealed hyperkeratosis with parakeratosis, together with hypergranulosis. The clinical and histopathological findings were consistent with an ichthyotic (or Camisa) variant of VS. The sequence analysis of the loricrin gene revealed that the patient had a heterozygous mutation identical to that described in previous reports, i.e. a G insertion producing a frameshift at codon 231 with an abnormal C-terminus. These results clearly demonstrate that a common loricrin gene mutation underlies VS in different ethnic groups.


Combined treatment with fenretinide and indomethacin induces AIF-mediated, non-classical cell death in human acute T-cell leukemia Jurkat cells.
Currently used cytotoxic drugs in cancer therapy have a similar mechanism of action and low specificity. Applied simultaneously, they show an additive effect with strong side effects. Clinical trials with the use of different agents in cancer therapy show that the use of these compounds alone is not very effective in fighting cancer. An alternative solution could be to apply a combination of these agents, because their combination has a synergistic effect on some cancer cells. Therefore, in our investigations we examined the effects of a synthetic retinoid-fenretinide when combined with a non-steroidal anti-inflammatory drug-indomethacin on the process of apoptosis in the acute human T-cell leukemia cell line Jurkat. We demonstrate that treatment with the combination of the tested compounds induces the death of cells, that is peculiar and combines features of apoptosis as well as non-apoptotic cell death. In detail we observed, cell membrane permeabilization, phosphatydylserine exposure, no oligonucleosomal DNA fragmentation, no caspase-3 activation, but apoptosis inducing factor (AIF) nuclear translocation. Taken together these results indicate, that Jurkat cells after treatment with a combination of fenretinide and indomethacin undergo AIF-mediated programmed cell death.


Could the ketogenic diet induce a shift in thyroid function and support a metabolic advantage in healthy participants? A pilot randomized-controlled-crossover trial.
The ketogenic diet (KD) has been shown to result in body mass loss in people with disease as well as healthy people, yet the effect of the KD on thyroid function and metabolism are unknown. We aimed to determine the effects of a KD, compared with an isocaloric high-carbohydrate low-fat (HCLF) diet, on resting metabolic rate and thyroid function in healthy individuals. Eleven healthy, normal-weight participants (mean(SD) age: 30(9) years) completed this randomized crossover-controlled study. For a minimum of three weeks on each, participants followed two isocaloric diets: a HCLF diet (55%carbohydrate, 20%fat, 25%protein) and a KD (15%carbohydrate, 60%fat, 25% protein), with a one-week washout period in-between. Importantly, while on the KD, the participants were required to remain in a state of nutritional ketosis for three consecutive weeks. Crossover analyses and linear mixed models were used to assess effect of diet on body mass, thyroid function and resting metabolic rate. Both dietary interventions resulted in significant body mass loss (p<0.05) however three weeks of sustained ketosis (KD) resulted in a greater loss of body mass (mean (95%CI): -2.9 (-3.5, -2.4) kg) than did three weeks on the HCLF diet (-0.4 (-1.0, 0.1) kg, p < 0.0001). Compared to pre-diet levels, the change in plasma T3 concentration was significantly different between the two diets (p = 0.003), such that plasma T3 concentration was significantly lower following the KD diet (4.1 (3.8, 4.4) pmol/L, p<0.0001) but not different following the HCLF diet (4.8 (4.5, 5.2) pmol/L, p = 0.171. There was a significant increase in T4 concentration from pre-diet levels following the KD diet (19.3 (17.8, 20.9) pmol/L, p < 0.0001), but not following the HCLF diet (17.3 (15.7, 18.8) pmol.L, p = 0.28). The magnitude of change in plasma T4 concentration was not different between the two diets (p = 0.4). There was no effect of diet on plasma thyroid stimulating hormone concentration (p = 0.27). There was a significantly greater T3:T4 ratio following the HCLF diet (0.41 (0.27, 0.55), p < 0.0001) compared to pre-diet levels but not following the KD diet (0.25 (0.12, 0.39), p = 0.80). Although the diets were isocaloric and physical activity and resting metabolic rate remained constant, the participants lost more mass after the KD than after the HCLF diet. The observed significant changes in triiodothyronine concentration suggest that unknown metabolic changes occur in nutritional ketosis, changes that warrant further investigation. Pan African Clinical Trial Registry: PACTR201707002406306 URL: https://pactr.samrc.ac.za/.


Blood-brain barrier permeability to dipeptides and their constituent amino acids.
The penetration of two [14C]-labelled dipeptides, glycyl-L-phenylalanine and glycyl-L-leucine, and of their constituent amino acids into the brain of the rat was measured employing an intracarotid injection technique. The brain-uptakes of the dipeptides were about equal to that of sucrose suggesting a negligible extraction from the blood during the 15-s period of exposure to the peptides. Brain uptakes for L-phenylalanine and L-leucine were large and in agreement with earlier work on these amino acids; self-inhibition by unlabelled amino acids was marked as also inhibition by the typical L-transport system substrate, 2-aminobicyclo (2, 2, 1) heptane-2 carboxylic acid (BCH), whilst the substrate for the A-system, N-methyl-L-aminoisobutyric acid (MeAIB) was without effect. Uptake of L-phenylalanine and L-leucine was not inhibited by dipeptides in 10 mM concentration. The uptakes of [14C]-labelled MeAIB and glycine were not significantly different from that of sucrose. It is concluded that peptide formation effectively excludes the rapidly penetrating L-system amino acids, L-leucine and L-phenylalanine, from access to the L-system channel.


The Conservative Treatment of Some Hand and Carpal Fractures.
In recent decades, there has been a trend toward increased use of operative treatment of hand fractures. However, internal stabilization with wires or open reduction and internal fixation of the phalanges and the metacarpals carries a risk of surgical complications that can be avoided by using appropriate conservative treatment. In this article, some hand fractures that can be managed safely without surgery are discussed. In conclusion, when facing a fracture in the hand, the first consideration is whether the fracture can be treated nonoperatively and not which operative treatment is most appropriate. This applies to both displaced and undisplaced fractures.


The Role of the Dental Community in Oropharyngeal Cancer Prevention Through HPV Vaccine Advocacy.
As rates of human papillomavirus (HPV)-related oropharyngeal cancer (OPC) continue to rise, the dental community's role in primary prevention efforts related to HPV vaccination will become increasingly important. The aim of this study was to assess knowledge, beliefs, practices, and perceived barriers regarding HPV and HPV vaccine advocacy within the dental community. A sample of 266 dentists and dental hygienists completed an online survey, and responses were then analyzed using frequencies of responses, t tests, chi-square tests, and Spearman's correlations. Ninety percent of providers believe it is important to play an "active role" in their patients' general medical care, yet only 50% believe it is their responsibility to recommend the HPV vaccine. Only 50% feel knowledgeable enough to recommend the vaccine. 78.6% of providers rarely discuss HPV vaccination with their age-appropriate patients, and 82% rarely recommend the vaccine. The two most strongly agreed-upon barriers were "parent concerns about the safety or efficacy of the vaccine" (71.6%) and "parent opposition to HPV vaccination for philosophical or religious reasons" (72.6%). Dentists were more knowledgeable about HPV vaccination and more likely to recommend the vaccine than hygienists. Higher levels of HPV-related knowledge correlated positively with beliefs and practices that support HPV vaccine advocacy. We have identified multiple opportunities for intervention aimed at increasing vaccine advocacy among oral health providers. These include clarification of role from dental professional organizations, alleviation of concerns related to perceived parental objection to the vaccine, and educational interventions targeting knowledge deficits among oral health providers.


Neutrophil CD64 index as a good biomarker for early diagnosis of bacterial infection in pregnant women during the flu season.
Pregnant women are at high risk of developing febrile illness during the flu season. Early identification of a viral or bacterial infection is crucial in the management of febrile pregnant patients. Neutrophil CD64 (nCD64) has been shown to have more important diagnostic value in sepsis than traditional inflammatory indicators. The pregnant women enrolled were divided into three groups according to disease: influenza A infection, bacterial infection and healthy controls. Peripheral blood CD64, leukocyte, C-reactive protein (CRP), procalcitonin (PCT) and human Th1/Th2-related cytokines levels were routinely measured. The correlation between and diagnostic value of the nCD64 index and other biomarkers were evaluated using Spearman's correlation test and receiver operating characteristic (ROC) curve analysis. Pregnant women with bacterial infection had significantly elevated levels of leukocytes (8.4 vs. 5.95, 109/L; P = 0.004), CRP (89.70 vs. 50.05 mg/mL; P = 0.031), PCT (0.13 vs. 0.04 ng/mL; P = 0.010) and TNF-α (0.46 vs. 0.38 pg/mL; P = 0.012) and an elevated nCD64 index (12.16 vs. 0.81; P < 0.001) compared with those with influenza A infection. The area under the receiver operating characteristic (AUROC) curve of the nCD64 index to discriminate bacterial infection among pregnant women (AUROC = 0.9183, P < 0.0001) was the largest. The sensitivity and specificity of the nCD64 index at an optimal cut-off value of 3.16 were 84% and 100%, respectively, with a negative predictive value (NPV) of 94%. Our study demonstrates the clinical value of the nCD64 index in distinguishing between bacterial infection and influenza A in pregnant women.


Heterocyclic Compounds Bearing Triazine Scaffold and Their Biological Significance: A Review.
Benzene is a six-membered hydrocarbon ring system and if three carbon-hydrogen units of benzene ring are replaced by nitrogen atoms then triazine is formed. Triazines are present in three isomeric forms 1,2,3- triazine, 1,2,4-triazine, and 1,3,5-triazine according to the position of the nitrogen atom. These are weak bases having weaker resonance energy than benzene, so nucleophilic substitution is preferred than electrophilic substitution. Triazine is an interesting class of heterocyclic compounds in medicinal chemistry. Numerous synthetic derivatives of triazine have been prepared and evaluated for a wide spectrum of biological activities in different models with desired findings such as antibacterial, antifungal, anti-cancer, antiviral, antimalarial, antiinflammatory, antiulcer, anticonvulsant, antimicrobial, insecticidal and herbicidal agents. Triazine analogs have exposed potent pharmacological activity. So, triazine nucleus may be considered as an interesting core moiety for researchers for the development of future drugs.


Exploring the Lignin Catabolism Potential of Soil-Derived Lignocellulolytic Microbial Consortia by a Gene-Centric Metagenomic Approach.
An exploration of the ligninolytic potential of lignocellulolytic microbial consortia can improve our understanding of the eco-enzymology of lignin conversion in nature. In this study, we aimed to detect enriched lignin-transforming enzymes on metagenomes from three soil-derived microbial consortia that were cultivated on "pre-digested" plant biomass (wheat straw, WS1-M; switchgrass, SG-M; and corn stover, CS-M). Of 60 selected enzyme-encoding genes putatively involved in lignin catabolism, 20 genes were significantly abundant in WS1-M, CS-M, and/or SG-M consortia compared with the initial forest soil inoculum metagenome (FS1). These genes could be involved in lignin oxidation (e.g., superoxide dismutases), oxidative stress responses (e.g., catalase/peroxidases), generation of protocatechuate (e.g., vanAB genes), catabolism of gentisate, catechol and 3-phenylpropionic acid (e.g., gentisate 1,2-dioxygenases, muconate cycloisomerases, and hcaAB genes), the beta-ketoadipate pathway (e.g., pcaIJ genes), and tolerance to lignocellulose-derived inhibitors (e.g., thymidylate synthases). The taxonomic affiliation of 22 selected lignin-transforming enzymes from WS1-M and CS-M consortia metagenomes revealed that Pseudomonadaceae, Alcaligenaceae, Sphingomonadaceae, Caulobacteraceae, Comamonadaceae, and Xanthomonadaceae are the key bacterial families in the catabolism of lignin. A predictive "model" was sketched out, where each microbial population has the potential to metabolize an array of aromatic compounds through different pathways, suggesting that lignin catabolism can follow a "task division" strategy. Here, we have established an association between functions and taxonomy, allowing a better understanding of lignin transformations in soil-derived lignocellulolytic microbial consortia, and pinpointing some bacterial taxa and catabolic genes as ligninolytic trait-markers.


Spatial frequency channels derived from individual differences.
Contrast sensitivity functions differ from observer to observer. We propose that these differences arise because each observer has unique weights for the outputs of the neural channels that underlie the contrast sensitivity function. By applying principal components analysis to individual contrast sensitivity functions of 297 observers, estimates of the channel tuning curves were found. We find evidence for three broadly tuned bandpass channels with peaks at 4, 8, and 16c/deg and bandwidth near 1.3 octaves. These channel tuning curves were reproduced in a cross-validation study of 56 observers.


Phase II trial and pharmacokinetic assessment of intravenous melphalan in patients with advanced prostate cancer.
Alkylating agents have been reported to yield response rates of up to 20% in hormone-refractory prostate cancer. Melphalan was studied in four small trials in which the drug was given orally. In this phase II trial, melphalan (30 mg/m2) was given intravenously every 28 days to 27 patients with hormone-refractory prostate cancer. Pharmacokinetic sampling was performed so as to describe the clearance of melphalan in this population and in an attempt to carry out pharmacodynamic modeling for toxicity and response. Prostate-specific antigen (PSA) was also assessed prospectively. No objective responses to this regimen were documented. The median survival for patients on this trial was 11.5 months. There was no correlation between drug clearance and measured creatinine clearance and no relationship between systemic exposure and toxicity. A decrease of > 50% in serum PSA that was sustained for > 6 weeks was documented in two patients, most notably in one patient who has survived for more than 29 months. Intravenous melphalan is not an active agent in hormone-refractory prostate cancer.


A facultative ectomycorrhizal association is triggered by organic nitrogen.
Increasing nitrogen (N) deposition often tends to negatively impact the functions of belowground ectomycorrhizal networks, although the exact molecular mechanisms underlying this trait are still unclear. Here, we assess how the root-associated fungus Clitopilus hobsonii establishes an ectomycorrhiza-like association with its host tree Populus tomentosa and how this interaction is favored by organic N over mineral N. The establishment of a functional symbiosis in the presence of organic N promotes plant growth and the transfer of 15N from the fungus to above ground plant tissues. Genomic traits and in planta transcriptional signatures suggest that C. hobsonii may have a dual lifestyle with saprotrophic and mutualistic traits. For example, several genes involved in the digestion of cellulose and hemicellulose are highly expressed during the interaction, whereas the expression of multiple copies of pectin-digesting genes is tightly controlled. Conversely, the nutritional mutualism is dampened in the presence of ammonium (NH4+) or nitrate (NO3-). Increasing levels of NH4+ led to a higher expression of pectin-digesting genes and a continuous increase in hydrogen peroxide production in roots, whereas the presence of NO3- resulted in toxin production. In summary, our results suggest that C. hobsonii is a facultative ectomycorrhizal fungus. Access to various forms of N acts as an on/off switch for mutualism caused by large-scale fungal physiological remodeling. Furthermore, the abundance of pectin-degrading enzymes with distinct expression patterns during functional divergence after exposure to NH4+ or organic N is likely to be central to the transition from parasitism to mutualism.


Laparoscopic Treatment of Mixed Malignant Ovarian Germ Cell Tumor in a 16-Year-Old Female Adolescent.
Malignant ovarian germ cell tumors are rare entities, although they account for a large proportion of ovarian masses in young women. These tumors have traditionally been removed via laparotomy, because of their large size and solid nature. The use of laparoscopy for treatment of adnexal masses in adolescents has been heavily debated and poorly studied to date. A 16-year-old female patient presented with abdominal pain and an 11-cm adnexal mass on ultrasound. An emergent laparoscopic salpingo-oophorectomy was performed without complication. Pathology revealed a mixed malignant ovarian germ cell tumor. Laparoscopic fertility-sparing surgery offers many benefits over laparotomy, and should be considered in cases of young women with large adnexal masses, even if potential for malignancy exists.


Gene augmented nuclear-targeting sonodynamic therapy via Nrf2 pathway-based redox balance adjustment boosts peptide-based anti-PD-L1 therapy on colorectal cancer.
Colorectal cancer is known to be resistant to immune checkpoint blockade (ICB) therapy. Sonodynamic therapy (SDT) has been reported to improve the efficacy of immunotherapy by inducing immunogenic cell death (ICD) of cancer. However, the SDT efficacy is extremely limited by Nrf2-based natural redox balance regulation pathway in cancer cells in response to the increased contents of reactive oxygen species (ROS). Nuclear-targeting strategy has shown unique advantages in tumor therapy by directly destroying the DNA. Thus it can be seen that Nrf2-siRNA augmented nuclear-targeting SDT could boost ICB therapy against colorectal cancer. The nuclear-targeting delivery system TIR@siRNA (TIR was the abbreviation of assembled TAT-IR780) with great gene carrier capacity and smaller diameter (< 60 nm) was designed to achieve the gene augmented nuclear-targeting SDT facilitating the anti-PD-L1 (programmed cell death-ligand-1) therapy against colorectal cancer. In CT26 cells, TIR@siRNA successfully delivered IR780 (the fluorescent dye used as sonosensitizer) into cell nucleus and Nrf2-siRNA into cytoplasm. Under US (utrasound) irradiation, TIR@siRNA notably increased the cytotoxicity and apoptosis-inducing activity of SDT through down-regulating the Nrf2, directly damaging the DNA, activating mitochondrial apoptotic pathway while remarkably inducing ICD of CT26 cells. In CT26 tumor-bearing mice, TIR@siRNA mediated gene enhanced nuclear-targeting SDT greatly inhibited tumor growth, noticeably increased the T cell infiltration and boosted DPPA-1 peptide-based anti-PD-L1 therapy to ablate the primary CT26 tumors and suppress the intestinal metastases. All results demonstrate that TIR@siRNA under US irradiation can efficiently inhibit the tumor progression toward colorectal CT26 cancer in vitro and in vivo by its mediated gene augmented nuclear-targeting sonodynamic therapy. Through fully relieving the immunosuppressive microenvironment of colorectal cancer by this treatment, this nanoplatform provides a new synergistic strategy for enhancing the anti-PD-L1 therapy to ablate colorectal cancer and inhibit its metastasis.


The effects of apoptotic, deported human placental trophoblast on macrophages: possible consequences for pregnancy.
During pregnancy, trophoblasts are shed into maternal blood from the placenta as they die. Trophoblasts are fetal cells and are therefore immunologically foreign to the maternal immune system, but the effects of shed trophoblasts on the maternal immune system are poorly characterized. We have used an in vitro villous explant model to harvest shed trophoblasts. These shed trophoblasts consist of multinucleated syncytial knots as well as mononuclear cells, and approximately 90% are apoptotic as determined by immunostaining with antibodies recognizing activated caspase-3 and the M30 cytokeratin neoepitope. U937 cells phagocytosed the shed apoptotic trophoblasts and, subsequently, secretion of the anti-inflammatory cytokine IL-10 was increased. In contrast, secretion of the proinflammatory cytokine Il-1beta by U937 cells was decreased after phagocytosis of apoptotic trophoblasts and the changes in both IL-10 and IL-1beta secretion were blocked by co-incubation with the phagocytosis inhibitor cytochalasin B. Shed trophoblasts caused a significant increase also in expression of the, immunosuppressive, tryptophan-metabolizing enzyme indoleamine 2,3-dioxygenase. We speculate that the shedding of trophoblasts may not be simply a mechanism the fetus uses to dispose of aged trophoblasts but rather shed apoptotic trophoblasts may provide a chronic source of tolerizing paternally derived antigens to regulate maternal immune responses to the fetus.


Evaluation of antibacterial efficacy of garlic (Allium sativum) and ginger (Zingiber officinale) crude extract against multidrug-resistant (MDR) poultry pathogen.
The study is aimed to understand the antibacterial sensitivity of native and Indian varieties of garlic (Allium sativum) and ginger (Zingiber officinale) crude extracts against multidrug-resistant (MDR) poultry pathogen (Escherichia coli and Salmonella sp.). Thin layer chromatography (TLC) is used to identify the target spices' bioactive antibacterial compounds. MDR E. coli and Salmonella sp. were isolated from poultry. The TLC-Bioautography technique was applied to explore the antibacterial potentiality of garlic and ginger. Inhibitory activities of garlic were Zone of inhibition (ZI) = 14.03 ± 0.15 mm and 19.70 ± 0.36 mm, Minimum inhibitory concentration (MIC): 0.625 and 0.325 mg/ml, and ginger were ZI = 14.63 ± 0.30 mm and 11.56 ± 0.51mm, MIC: 9.0 mg/ml against E. coli and Salmonella sp., respectively. Two bands of garlic (Rf value = 0.31 and 0.50) and one band of ginger (Rf value = 0.71) showed inhibitory potential in TLC-Bioautography against both MDR isolates. Garlic and ginger were effective against MDR E. coli and Salmonella sp. These spices could be a suitable alternative during the antibiotic void.


Responses of soil microbial diversity, network complexity and multifunctionality to three land-use changes.
Land-use change is one of the greatest challenges for natural ecosystem services. Soil microbiomes are essential for modulating multiple ecosystem functions. However, little is known about the impact of land-use changes on soil microbial communities and their associated soil functions. In this study, 150 alpine soil samples representing conversion of forests to shrublands or grasslands, and of shrublands to grasslands were investigated for bacterial, fungal and protistan community diversity, co-occurrence network, as well as their relationships with soil multifunctionality via a sampling strategy of space-for-time substitution. The conversion of forest to grassland increased the diversity of fungi and bacteria, and altered the microbial community structures of bacteria, fungi and protists, resulting a greater impact on soil microbiome than other land-use conversions. Cross-trophic interaction analyses demonstrated this conversion increased microbial network complexity and robustness, whereas forest to shrubland had the opposite trend. The land-use induced changes in soil multifunctionality were related with microbial network modules, but were not always associated with variations of microbial diversity. Random forest modeling further suggested the significant role of microbial modules in explaining soil multifunctionality, together with environmental factors. These findings indicate divergent responses of belowground multitrophic organisms to land-use changes, and the potential role of microbial module in forecasting soil multifunctionality.


Seasonal changes and alternation of food plant preference in some mesophyll-feeding leafhoppers.
The life histories of two bivoltine mesophyll-feeding leafhoppers, Lindbergina aurovittata (Douglas) and Edwardsiana rosae (L.), in South Wales are described. L. aurovittata alternates between evergreen Rubus and Quercus in the first generation and various deciduous broad-leaved trees in the second. Experiments on nymphal food preferences and adult oviposition behaviour show that nymphs of both generations tend to prefer older to younger leaves, but show little species specificity. However, adults are discriminating in oviposition preferences, which change regularly and completely in alternating generations. Similar results were obtained with E. rosae. Such obligate seasonal changes in host preferences and utilization are well known amongst aphids, but previously have not been reported in leafhoppers.


Controversies in the Spleen: Histiocytic, Dendritic, and Stromal Cell Lesions.
Histiocytic, dendritic, and stromal cell lesions that occur in the spleen are challenging diagnostically, not well studied due to their rarity, and therefore somewhat controversial. New techniques for obtaining tissue samples also create challenges as splenectomy is no longer common and needle biopsy does not afford the same opportunity for examination of tissue. Characteristic primary splenic histiocytic, dendritic, and stromal cell lesions are presented in this paper with new molecular genetic findings in some entities that help differentiate these lesions from those occurring in non-splenic sites, such as soft tissue, and identify possible molecular markers for diagnosis.


Temperature-dependent growth and emergence of functional leaves: an adaptive mechanism in the seedlings of the western Himalayan plant Podophyllum hexandrum.
As an adaptive mechanism, hypocotyl dormancy delays emergence of functional leaf until favorable season of growth in Podophyllum hexandrum, an endangered medicinal plant of the western Himalayas. However, upon exposure of the freshly germinated seedlings to favorable temperature (25 degrees C), functional leaves emerged within 20 days. Therefore, we examined regulation mechanisms of growth and development of this alpine plant by temperature under laboratory conditions. The seedlings were exposed to (1) 25 degrees C (temperature prevailing at the time of maximum vegetative growth), (2) 4 degrees C (mean temperature at the onset of winter in its natural habitat), and (3) 10 degrees C (an intermediate temperature). Slackened growth at 4 degrees C was followed by senescence of aerial parts and quiescence of roots and predetermined leaf primordia. Rapid development of leaf primordia at 25 degrees C was associated with increased starch hydrolysis. This was evident from higher alpha-amylase activity and reducing sugars. These parameters decreased on sudden exposure to 4 degrees C. In contrast, the roots (perennating organs) showed a slight increase (1.36-fold) in alpha-amylase activity. Growth and development in seedlings growing at 10 degrees C (temperature less adverse than 4 degrees C) were comparatively faster. The content of reducing sugars and alpha-amylase activity were also higher in all the seedling parts at 10 degrees C as compared to 4 degrees C. This indicated larger requirements for sugar by the seedlings at 10 degrees C. Irrespective of temperature, maximum changes in nitrate and nitrate reductase occurred during the initial 10 days, i.e., when the readily available form of sugars (reducing sugar) was highest. This indicated that a temperature-dependent availability of carbon, but not temperature itself, was an important regulator of uptake and reduction of nitrogen.


Blood serum amyloid A as potential biomarker of pembrolizumab efficacy for patients affected by advanced non-small cell lung cancer overexpressing PD-L1: results of the exploratory "FoRECATT" study.
Identifying the patients who may benefit the most from immune checkpoints inhibitors remains a great challenge for clinicians. Here we investigate on blood serum amyloid A (SAA) as biomarker of response to upfront pembrolizumab in patients with advanced non-small-cell lung cancer (NSCLC). Patients with PD-L1 ≥ 50% receiving upfront pembrolizumab (P cohort) and with PD-L1 0-49% treated with chemotherapy (CT cohort) were evaluated for blood SAA and radiological response at baseline and every 9 weeks. Endpoints were response rate (RR) according to RECIST1.1, progression-free (PFS) and overall survival (OS). The most accurate SAA cut-off to predict response was established with ROC analysis in the P cohort. In the P Cohort (n = 42), the overall RR was 38%. After a median follow-up of 18.5 months (mo), baseline SAA ≤ the ROC-derived cut-off (29.9 mg/L; n = 28/42.67%) was significantly associated with higher RR (53.6 versus 7.1%; OR15, 95% CI 1.72-130.7, p = 0.009), longer PFS (17.4 versus 2.1 mo; p < 0.0001) and OS (not reached versus 7.2mo; p < 0.0001) compared with SAA > 29.9 mg/L. In multivariate analysis, low SAA positively affects PFS (p = 0.001) and OS (p = 0.048) irrespective of ECOG PS, number of metastatic sites and pleural effusion. SAA monitoring (n = 40) was also significantly associated with survival endpoints: median PFS 17.4 versus 2.1 mo and median OS not reached versus 7.2 mo when SAA remained low (n = 14) and high (n = 12), respectively. In the CT Cohort (n = 30), RR was not affected by SAA level (p > 0.05) while low SAA at baseline (n = 17) was associated with better PFS (HR 0.38, 95% CI 0.16-0.90, p = 0.006) and OS (HR 0.25, 95% CI 0.09-0.67, p < 0.001). Low SAA predicts good survival outcomes irrespective of treatment for advanced NSCLC patients and higher likelihood of response to upfront pembrolizumab only. The strong prognostic value might be exploited to easily identify patients most likely to benefit from immunotherapy. A further study (FoRECATT-2) is ongoing to confirm results in a larger sample size and to investigate the effect of SAA on immune response in vitro assays.


Price discounts on alcohol in a city in Northern England.
To describe the extent and nature of price discounts on alcohol in Newcastle upon Tyne, England. An observational survey in stores licensed for off-sales in December 2010 to January 2011. A total of 2018 price discounts in 29 stores led to a median saving of 25% and required a median purchase of 20 standard UK alcohol units. Median price per standard unit was £0.92 (US$1.49; €1.05) before discount and £0.68 (US$1.10; €0.78) after discount. Restriction of price discounting should be considered as a public health policy.


Complete Genome Sequence of an Alphacoronavirus from Common Vampire Bats in Peru.
Bats host diverse coronaviruses, including taxa capable of pandemic spread in humans. We report the genome of an alphacoronavirus from a neotropical bat species (Desmodus rotundus) in Peru, which contributes to our understanding of bat coronaviruses in nature.


Macrophage migration inhibitory factor has a permissive role in concanavalin A-induced cell death of human hepatoma cells through autophagy.
Concanavalin A (ConA) is a lectin and T-cell mitogen that can activate immune responses. In recent times, ConA-induced cell death of hepatoma cells through autophagy has been reported and its therapeutic effect was confirmed in a murine in situ hepatoma model. However, the molecular mechanism of ConA-induced autophagy is still unclear. As macrophage migration inhibitory factor (MIF), which is a proinflammatory cytokine, can trigger autophagy in human hepatoma cells, the possible involvement of MIF in ConA-induced autophagy was investigated in this study. We demonstrated that cell death is followed by an increment in MIF expression and secretion in the ConA-stimulated human hepatoma cell lines, HuH-7 and Hep G2. In addition, ConA-induced autophagy and cell death of hepatoma cells were blocked in the presence of an MIF inhibitor. Knockdown of endogenous MIF by small hairpin RNA confirmed that MIF is required for both ConA-induced autophagy and death of hepatoma cells. Furthermore, signal pathway studies demonstrated that ConA induces signal transducer and activator of transcription 3 (STAT3) phosphorylation to trigger MIF upregulation, which in turn promotes Bcl-2/adenovirus E1B 19 kDa-interacting protein 3 (BNIP3)-dependent autophagy. By using a murine in situ hepatoma model, we further demonstrated that MIF contributes to anti-hepatoma activity of ConA by regulating STAT3-MIF-BNIP3-dependent autophagy. In summary, our findings uncover a novel role of MIF in lectin-mediated anti-hepatoma activities by regulating autophagy.


Factors affecting the rate of purine ribonucleotide dephosphorylation in human erythrocytes.
Purine ribonucleotide dephosphorylation was measured in intact human erythrocytes in vitro to evaluate those factors which might regulate this process in vivo. It was found that purine nucleotides which exist predominantly in the triphosphate form (e.g. ATP and GTP) are protected from dephosphorylation while those nucleotides normally present as the monophosphate (e.g. IMP) are susceptible to dephosphorylation. This point was emphasised by studying an individual whose erythrocytes accumulated ITP rather than IMP; erythrocytes from this individual has a more stable pool of inosine phosphates than did erythrocytes from normal individuals. The concentration of intracellular phosphate was also shown to affect the rate of dephosphorylation. The dephosphorylation of IMP was inhibited at intracellular phosphate concentrations above approx. 3 mM. AMP dephosphorylation (in cells whose AMP concentration was increased by incubating them in the presence of 2-deoxyglucose) was inhibited by phosphate more strongly than was found for IMP. In contrast, the dephosphorylation of GMP did not appear to be affected by phosphate concentration. High oxygen tension was a powerful stimulator of IMP dephosphorylation while low oxygen tension protected IMP from dephosphorylation. This finding shows that human erythrocytes are similar to those of other mammals in this regard and points to a possible physiological determinant of purine turnover in these cells.


[Effects of breast massage on breast pain, breast-milk sodium, and newborn suckling in early postpartum mothers].
In this study the effects of breast massage on breast pain, breast-milk sodium, and newborn suckling in early postpartum mothers were investigated. The design was a non-synchronized nonequivalent control group pretest-posttest design. Sixty postpartum mothers who were admitted to a postpartum care center and had problems with breastfeeding were recruited. Of these mothers, 44 were assigned to the intervention group and received two 30-minute breast massages within 10 days of postpartum period. The others were assigned control group and received only routine care. Breast pain was measured using a numeric pain scale and number of times newborns suckled was observed throughout breastfeeding. Breast milk was self-collected to evaluate breast-milk sodium. Mean age of postpartum mothers was 30 years old. Compared to the control group, women in the intervention group reported significant decreases in breast pain (p<.001), increases in number of times newborns suckled after the first and second massage (p<.001), and a decrease in breast-milk sodium after the first massage (p=.034). Breast massage may have effects on relieving breast pain, decreasing breast-milk sodium, and improving newborn suckling. Breast massage can be used to solve breast problems. Further research is needed to validate our findings.


Continuous corneal intrastromal ring implantation for treatment of keratoconus in an Iranian population.
To evaluate the effect of mechanical implantation of a continuous intrastromal ring in keratoconus. Prospective, interventional, nonrandomized, case series. The MyoRing (Dioptex GmbH) was implanted after creation of an intrastromal pocket for 95 eyes of 95 patients with moderate and advanced keratoconus. All patients had at least 12 months of follow-up. Preoperative and postoperative visual acuity, keratometry, aberrometry, and refraction were the main outcome measures of the study. A significant improvement in uncorrected and corrected distance visual acuity was observed 1 month after surgery, which was consistent with the significant reduction in sphere (5.74 diopters [D]) and cylinder (3.02 D). No significant changes were detected in these parameters afterward. Furthermore, a significant corneal flattening of a mean value of 9.78 D was found. Both spherical myopia and astigmatism underwent reduction, but the reduction in myopia was more remarkable than astigmatism. Higher-order aberrations and coma-like aberrations decreased significantly, but spherical aberrations increased after surgery. No significant change in central corneal thickness was observed at any point after operation. There were no significant differences between 2 keratometry groups (higher or lower than 53 D) in visual gain after the procedure. There were no major complications during or after surgery. MyoRing explantation was performed in 4 eyes (4%). The refraction, visual acuity, and corneal topography returned to the preoperative status 1 month later for all 4 eyes. MyoRing implantation has an acceptable efficacy profile in moderate and advanced keratoconus.


T-cell receptor expressed on an autoreactive T-cell clone, clone 4. I. Induction of various T-receptor functions by anti-T idiotypic antibodies.
Three monoclonal antibodies (1G3, 2H11, and 3G12) specific for a syngeneic Ek-specific T-cell clone, clone 4, have been established. The antibodies specifically blocked not only the activation of the clone in response to the specific antigen Ek but also the activation by IL-2. Kinetic studies of the blocking activity revealed that the antibodies blocked activation not only through steric hindrance of the antigen-binding site of the receptor but also via inhibition of DNA synthesis. The antibodies induced unresponsiveness of the clone to the specific antigen Ek, but not to nonspecific activation by IL-2. The state of unresponsiveness induced by 1G3 continued for 14 days, the longest time so far examined. The recovery from the unresponsiveness (tolerance) was not observed unless the clone cells proliferated vigorously in response to IL-2. The idiotope recognised by 1G3 was different from that by 2H11 and/or 3G12. This might explain some functional differences elicited by the antibodies.


Design and Implementation of Competency Based Postgraduate Medical Education in Otorhinolaryngology: The Pilot Experience in India.
The worldwide call for a shift towards competency based postgraduate medical education has until recently gone largely unheeded in India, despite the Medical Council of India enshrining the principle in its regulations for postgraduate institutions. This paper details the first concrete attempt at establishing a CBME curriculum in Otorhinolaryngology in India. The design and implementation of the CBME curriculum was carried out in four phases, in a time-bound manner over a period of 6 months. Phase I consisted of an extensive literature review and a clarification of the major objectives of the program. Phase II involved the listing out of 20-30 entrustable professional activities (EPAs) for each specialty and the 13 core EPAs common to all incoming residents and the subsequent mapping of these EPAs to their respective domains of competence and year-wise levels of competence. This was followed by the development of milestones for each EPA and appropriate clinical vignettes. Phase III focused on development of 360° assessment strategies, including the in-house development of an e-portfolio. Phase IV was dedicated to the implementation of the CBME curriculum, and involved various sensitization and orientation programs for faculty and the new residents. This exercise in designing and implementing a CBME program showed the important role that intra-departmental and inter-institutional cross-communication and exchange of ideas vies-a-vie workshops and personal communication play in bridging the lapses in knowledge in this emerging area, reaching consensus to achieve project goals and for finding relevant solutions to common problems. Medical education in India presents its own peculiar set of logistical and cultural challenges. Keeping in line with the recommendations of the Medical Council of India regarding Postgraduate Medical Education, it is essential that medical colleges in India not fall behind the international paradigm shift towards CBME.


The Value of Current Laboratory Tests in Diagnosing Food, Venom, and Drug Allergies.
An accurate diagnosis of IgE-mediated allergies is necessary to inform risk management for severe allergic reactions including anaphylaxis for food, venom, and drug allergies. The most widely available laboratory test for allergy is serum-specific IgE testing, which is routinely used for food allergy and insect sting allergy. Testing for specific IgE is limited by high sensitivity and low specificity, resulting in concern regarding overdiagnosis. Testing of allergen components has led to improved diagnosis for some food and venom allergens. Additional options for laboratory tests, such as epitope analysis, basophil activation, and mast cell activation, are being investigated for their potential to optimize diagnosis and provide predictors for reaction severity and treatment response. In contrast, laboratory testing for drug allergy is more limited because to date, there are no well-validated commercial assays in the United States. Furthermore, it is important to diagnose delayed reactions to medications, because these also significantly affect decision-making regarding therapeutic options for infectious disorders. Reliable tests for both immediate and delayed drug hypersensitivity are much needed, because drug allergy labels can significantly limit treatment options for patients. Research in this area is emerging.


Interleukin-12 inhibits eosinophil differentiation from bone marrow stem cells in an interferon-gamma-dependent manner in a mouse model of asthma.
Intrapulmonary administration of IL-12 has been shown to inhibit the number of eosinophils in lung murine models of asthma, but the precise mechanism of this inhibition has not been reported. The purpose of this study was to examine whether IL-12 treatment inhibits bone marrow eosinophilopoiesis, and to elucidate the role of IFN-gamma in this process. To elucidate the in vivo and in vitro effects of IL-12 on eosinophil differentiation from murine bone marrow (BM) stem cells, and to examine the mechanistic role of IFN-gamma in this process. Allergen-sensitized BALB/c mice were administered low doses of intranasal IL-12 at the time of allergen challenge, and the number of eosinophils in BM was determined 3 days later. The direct actions of IL-12 on eosinophil differentiation from BM cells were determined in vitro. The mechanistic role of IFN-gamma was assessed by measuring IFN-gamma induction by IL-12 in BM cell cultures, and through the use of IFN-gamma KO mice. Treatment of allergic mice with intrapulmonary IL-12 (1 ng or 10 ng) reduced eosinophils in BM by 43%. Culture of BM cells from allergen-sensitized mice with IL-3 + IL-5 induced eosinophil differentiation in vitro. Addition of IL-12 to these cultures inhibited eosinophil differentiation, with maximal inhibition (45%) occurring at 10 ng/mL IL-12 concentration. IL-12 induced IFN-gamma production from BM cultures, and failed to inhibit eosinophil differentiation in IFN-gamma-knockout mice, indicating a critical mechanistic role for IFN-gamma. This study demonstrates that IL-12 selectively inhibits BM eosinophilopoiesis, and that this effect is mediated by IFN-gamma. Intrapulmonary IL-12 has suppressive effects on BM eosinophilopoiesis that may represent a novel mechanism contributing to the anti-eosinophilic effects of IL-12 in allergic airway disease.


Interstitial fluid of isolated perfused rat hearts: glucose and lactate concentration.
Interstitial fluid (IF) emerging at the ventricular surface of isolated perfused rat hearts was collected and assayed for rate of production, protein content, glucose and lactate concentration. The influence of four perfusion media was investigated during an experimental period of 150 min: Krebs-Henseleit solution (KH) containing glucose (5.5 mM); KH containing glucose and pyruvate (2 mM); Hypoxic KH (20% O2), substrates as 2; KH containing isoprenaline (8 X 10(-9) M), substrates as 2. Interstitial fluid was produced at a rate of 20 to 100 microliters/min/gww and contained proteins (0.5 to 3 g/l). Interstitial glucose concentration was lower than venous concentration by up to 50%. Interstitial lactate concentration was higher by up to 600%. Permeability X surface area products of glucose calculated from transcapillary concentration differences and transfer rates were different depending on experimental conditions, but were within the rather large range of P X S values for molecules of similar size obtained by other authors. Those of lactate were higher by a factor of 3 to 9 and can be interpreted to be influenced by the metabolic activity of the endothelium. The results demonstrate that interstitial substrate concentration can differ very markedly from intravascular concentration and cannot be estimated reliably without assay of the interstitial fluid. Capillary permeability seems to be variable under the experimental conditions of this study.


Application of sensitive enzymeimmunoassays for oxytocin and prolactin determination in blood plasma of yaks (Poephagus grunniens L.) during milk let down and cyclicity.
Highly sensitive and specific enzymeimmunoassays for oxytocin and prolactin determination in yak plasma using the biotin-streptavidin amplification system and the second antibody coating technique were validated and applied for determining their profiles during milk let down and cyclicity in yaks. Oxytocin EIA was conducted taking duplicate 200 microl of unknown plasma samples and standards per well. The lowest detection limit was 0.2 pg/well, which corresponded to 1pg/ml plasma. Prolactin EIA was carried out directly in 50 microl of yak plasma. The sensitivity of EIA procedure was 5 pg/well prolactin, which corresponded to 0.1 ng/ml plasma. Mean plasma prolactin concentrations although high at estrus were not statistically different (P > 0.05) from the hormone concentrations on other days. Mean plasma prolactin concentrations during non-breeding season were significantly higher (P < 0.001) than that recorded in breeding season. Oxytocin and prolactin profiles were also obtained in two yaks before, during and after milking. A sharp release of oxytocin and prolactin shortly after udder stimulation was observed. High levels of oxytocin and prolactin were maintained during milking, falling sharply thereafter.


Novel roles of complement in renal diseases and their therapeutic consequences.
The complement system functions as a part of the innate immune system. Inappropriate activation of the complement pathways has a deleterious effect on kidneys. Recent advances in complement research have provided new insights into the pathogenesis of glomerular and tubulointerstitial injury associated with complement activation. A new disease entity termed 'C3 glomerulopathy' has recently been proposed and is characterized by isolated C3 deposition in glomeruli without positive staining for immunoglobulins. Genetic and functional studies have demonstrated that several different mutations and disease variants, as well as the generation of autoantibodies, are potentially associated with its pathogenesis. The data from comprehensive analyses suggest that complement dysregulation can also be associated with hemolytic uremic syndrome and more common glomerular diseases, such as IgA nephropathy and diabetic kidney disease. In addition, animal studies utilizing genetically modified mice have begun to elucidate the molecular pathomechanisms associated with the complement system. From a diagnostic point of view, a noninvasive, MRI-based method for detecting C3 has recently been developed to serve as a novel tool for diagnosing complement-mediated kidney diseases. While novel therapeutic tools related to complement regulation are emerging, studies evaluating the precise roles of the complement system in kidney diseases will still be useful for developing new therapeutic approaches.


Adult movement of the native holly leafminer, Phytomyza ilicicola Loew (Diptera: Agromyzidae): consequences for host choice within and between habitats.
Phytomyza ilicicola is a specialist herbivore, the larvae of which mine, and the adults of which feed and oviposit on the developing leaves of American Holly, Ilex opaca. Adult Phytomyza have been shown to discriminate among host plants on the basis of several host and habitat factors. Such discrimination may explain, in part, the observed density variation of larval Phytomyza among individual trees and between forest and suburban habitats. In order for discrimination to influence density, adult Phytomyza must be sufficiently vagile to move among habitats and to encounter many hosts. We experimentally monitored the movement of adults to ask if this were so. In addition we asked whether inter-host movements were altered by resource concentration, and whether vagility offered an escape from parasitism. To answer these questions, three isolated holly trees to serve as colonization sources were selected in early spring, prior to adult emergence. Potted target trees were then placed singly and in groups of four at 3 compass directions around and at 3 different distances (10, 25, and 50 m) from each of the source trees. Six weeks after exposure to adult Phytomyza, the number of feeding punctures and first instar larvae were counted. Clones were then overwintered in cold frames and in early spring, just prior to adult emergence, all pupae were examined for parasitism by the dominant parasitoid Opius striatriventris. Feeding and parasitism were similar at all 3 distances from the source trees, indicating that Opius was at least as vagile as Phytomyza. However, first instar larvae decreased with distance. There was no evidence that resource concentration affected adult movement. Our results suggest that Phytomyza are sufficiently vagile to choose among hosts within and between habitats. However, movement does not translate into equally high oviposition at all distances from a source, nor does it provide an escape from parasitism by Opius.


Monitoring dynamic changes of glutathione redox state in subcellular compartments of human cells - an approach based on rxYFP biosensor.
The kinetic and spatial separation of redox systems renders redox biology studies a particularly challenging field. Genetically encoded biosensors including the glutathione-specific redox-sensitive yellow fluorescent protein (rxYFP) provide an alternative way to overcome the limitations of conventional glutathione/glutathione disulfide (GSH/GSSG) redox measurements. In this study, the plasmids expressing respectively cytosol-, nucleus-, and mitochondrial matrix- targeted rxYFP were created and introduced to human cervical carcinoma (HeLa) cells. The rxYFP redox states were monitored by direct assessment of the oxidized to reduced rxYFP ratio via redox protein extraction, redox Western blot and signal quantification. RxYFP proteins expressed in the cytosol, nucleus or mitochondrial matrix of HeLa cells were responsive to the intracellular redox state changes induced by reducing as well as oxidizing agents. Compartment-targeted rxYFP sensors were able to detect different steady-state redox conditions between the cytosol, nucleus and mitochondrial matrix. Furthermore, rxYFP sensors were able to sense dynamic and compartment-specific redox changes caused by 100µM hydrogen peroxide (H2O2). Mitochondrial matrix-targeted rxYFP displayed a greater dynamics of oxidation in response to a H2O2 challenge than the cytosol- and nucleus-targeted sensors, largely due to a more alkaline local pH environment. Our data provide direct evidence that mitochondrial glutathione redox state is maintained and regulated independently from that of the cytosol and nucleus. Complementary to existing redox sensors and conventional redox measurements, compartment-targeted rxYFP sensors provide a novel tool for examining mammalian cell redox homeostasis, permitting high resolution readout of steady glutathione state and dynamics of redox changes.


RNA sequencing role and application in clinical diagnostic.
Although whole-exome sequencing and whole-genome sequencing has tremendously improved our understanding of the genetic etiology of human disorders, about half of the patients still do not receive a molecular diagnosis. The high fraction of variants with uncertain significance and the challenges of interpretation of noncoding variants have urged scientists to implement RNA sequencing (RNA-seq) in the diagnostic approach as a high throughput assay to complement genomic data with functional evidence. RNA-seq data can be used to identify aberrantly spliced genes, detect allele-specific expression, and identify gene expression outliers. Amongst eight studies utilizing RNA-seq, a mean diagnostic uplift of 15% has been reported. Here, we provide an overview of how RNA-seq has been implemented to aid in identifying the causal variants of Mendelian disorders.


Formic acid burn--local and systemic effects. Report of a case.
Formic acid is an organic acid which has mainly been used in industry, but in recent years its use in agriculture has greatly increased in the Scandinavian countries. Formic acid is more caustic to the skin than acetic acid. However, reports on formic acid burns are scarce in the medical literature, and no serious burn has previously been described. A few reports on formic acid ingestion have been published. It is of great interest that formic acid has been shown to play the major role in the methanol poisoning syndrome. A case of severe burn with undiluted formic acid is reported. In the beginning, the dermal injury seemed to be of minor degree, but then the patient, a 15-year-old girl, developed signs of systemic formic acid intoxication, including metabolic acidosis, intravascular hemolysis and hemoglobinuria. These signs were similar to those described as a result of formic acid ingestion. The systemic effects were successfully treated without late sequelae, but the burn turned out mainly to be full-thickness, resulting in major scarring, as commonly seen after various chemical burns.


Iron uptake by ZIP8 and ZIP14 in human proximal tubular epithelial cells.
In patients with iron overload disorders, increasing number of reports of renal dysfunction and renal iron deposition support an association between increased iron exposure and renal injury. In systemic iron overload, elevated circulating levels of transferrin-bound (TBI) and non-transferrin-bound iron (NTBI) are filtered to the renal proximal tubules, where they may cause injury. However, the mechanisms of tubular iron handling remain elusive. To unravel molecular renal proximal tubular NTBI and TBI handling, human conditionally immortalized proximal tubular epithelial cells (ciPTECs) were incubated with 55Fe as NTBI and fluorescently labeled holo-transferrin as TBI. Ferrous iron importers ZIP8 and ZIP14 were localized in the ciPTEC plasma membrane. Whereas silencing of either ZIP8 or ZIP14 alone did not affect 55Fe uptake, combined silencing significantly reduced 55Fe uptake compared to control (p < 0.05). Furthermore, transferrin receptor 1 (TfR1) and ZIP14, but not ZIP8, colocalized with early endosome antigen 1 (EEA1). TfR1 and ZIP14 also colocalized with uptake of fluorescently labeled transferrin. Furthermore, ZIP14 silencing decreased 55Fe uptake after 55Fe-Transferrin exposure (p < 0.05), suggesting ZIP14 could be involved in early endosomal transport of TBI-derived iron into the cytosol. Our data suggest that human proximal tubular epithelial cells take up TBI and NTBI, where ZIP8 and ZIP14 are both involved in NTBI uptake, but ZIP14, not ZIP8, mediates TBI-derived iron uptake. This knowledge provides more insights in the mechanisms of renal iron handling and suggests that ZIP8 and ZIP14 could be potential targets for limiting renal iron reabsorption and enhancing urinary iron excretion in systemic iron overload disorders.


Changes in nuclear proteins of rat testis cells separated by velocity sedimentation.
The technique of velocity sedimentation at unit gravity has been used to separate rat testis cell suspensions into fractions enriched in particular cell types. Changes in the nuclear proteins from the various fractions have been characterized by polyacrylamide gel electrophoresis, and correlated with the changing morphology of the nucleus during spermatogenesis. The most striking alterations in both protein composition and nuclear morphology occur during spermatid maturation as both histone and non-histone proteins are replaced by highly basic, low molecular weight, spermatidal proteins. This replacement process is accompanied by a quantitative reduction in both histone and non-histone proteins. The synthesis of at least three basic proteins has been identified with late stage spermatids. One of these proteins is a highly basic sperm-specific protein containing high levels of cyst(e)ine and arginine. A second protein synthesized in late stage spermatids is lysine rich, while the third protein contains cyst(e)ine and co-migrates with histone F2a1 on acid-urea polyacrylamide gels. The changes in protein composition of rat testis nuclei after irradiation or hypophysectomy reflect the resulting changes in the cellular composition of the testis. After selective elimination of the germinal cells by irradiation, the electrophoretic pattern of acid-soluble proteins from the testis is very similar to that of somatic tissue. Thus, the cellular specificity of nuclear proteins demonstrated here using cell separation techniques is also apparent following treatments which selectively alter the cellular composition of the testis.


Concurrent validity and reliability of new application for 6-min walk test in healthy adults.
Evaluation assessments for physical performance, such as walking tests, are important for measuring a person's well-being. As of current, medical technology is primarily used to administer these assessments. However, medical devices are not easily accessible and are intended for research purposes only, and hence inconvenient for clinical use. Therefore, we aimed to develop a prototype physical performance assessor device with a mobile application and explored concurrent validity and reliability between the standard 6-min walk test (6MWT) and wearable sensor 6MWT using 6-min walk distance in healthy adults. Sixty healthy males and females, above 18 years of age, were required to attach a sensor to their dominant ankle while the standard protocol for 6MWT was performed. After completing the walking test, the distance from the wearable sensor 6MWT with a mobile application and the standard 6MWT were recorded and compared. There was no significant difference between the distance between the standard 6MWT (410.12 ± 74.03 m) and the distance obtained with the wearable sensor. Concurrent validity was found to be moderate, and Cronbach's alpha was 0.79, which indicated good internal consistency. The innovative prototype wearable walking sensor with a mobile application can effectively evaluate physical performance in healthy individuals.Clinical trial registration number: TCTR20220801002.


Structural insight into the recognition between Sufu and fused in the Hedgehog signal transduction pathway.
Hedgehog signaling plays a crucial role in embryogenesis and adult tissue homeostasis, and mutations of its key components such as Suppressor of fused (Sufu) are closely associated with human diseases. The Ser/Thr kinase Fused (Fu) promotes Hedgehog signaling by phosphorylating the Cubitus interruptus (Ci)/Glioma-associated oncogene homologue (Gli) family of transcription factors. Sufu associates with both Fu and Ci/Gli, but the recognition mechanism between Sufu and Fu remains obscure. Here, our structure of the N-terminal domain (NTD) of Drosophila Sufu (dSufu) in complex with the Sufu-binding site (SBS) of Fu reveals that both main-chain β sheet formation and side-chain hydrophobic interactions contribute to the recognition between Sufu and Fu, and point mutations of highly conserved interface residues eliminated their association. Structural comparison suggests that Fu and Ci/Gli bind on opposite sides of dSufu-NTD, allowing the formation of a Fu-dSufu-Ci ternary complex which facilitates the phosphorylation of Ci/Gli by Fu. Hence, our results provide insights into the Sufu-Fu recognition mechanism.


[Biomechanism of intracranial and spinal trauma of child born in occipitoanterior presentation as used in forensic examination].
The new conception of mechanism and sequence of damages typical for labor trauma is presented: injury of ligaments of atlas and axis and anterior atlanto-occipital membrane, ruptures of synchondrosises of cranial bones because of vertebral column pressing on occipital bone; tears of dura mater of brain upon basilar impression in the area of focal spot; subarachnoid hemorrhage and intracerebral bleeding in case of cerebral compression by bone borders where they overly override; ruptures of ligaments of cervico-occipital and intervertebral joints in case of excess flexion and overextension of head. Authors show that damage of sutures, bones of calvarium and partition parts of dura materialize later than injury of synchondrosises of base and they are absent if sinchondrosises are not damaged. Also iatrogenic damages of fetal in labor and the mechanism of their formation are described.


[Evaluation of the anterior chamber depth after cataract surgery with OCT Visante].
OCT Visante allows the visualisation of the anterior segment structures and particularly of the irido-corneal angle by a non-contact method. We made a prospective study on 14 patients to demonstrate by OCT Visante the changes of irido-corneal angle and depth of the anterior chamber after cataract surgery. 14 eyes of 14 patients have undergone a phacoemulsification of the cataract with posterior chamber lens implantation. All patients have had a complete ophthalmologic examination, an OCT Visante with angle measurement and an A-Scan for determination of the lens thickness. All these exams were repeated at 1 week and 1 month postoperatively. In all patients, we noticed an increase of the postoperative iridocorneal angle. The mean preoperative angle was 18.3 degrees +/- 11.81 SD, 38.65 degrees +/- 3.95 SD at 1 week postoperatively and 45.7 degrees +/- 8.72 SD at 1 month. Imagery by OCT Visante allowed us by a non-invasive method to quantify very precisely the opening of the irido-corneal angle and the increase of the anterior chamber depth after cataract surgery. OCT Visante is a method allowing the visualisation and quantification of anterior segment changes after cataract surgery.


Pachychoroid neovasculopathy has clinical properties that differ from conventional neovascular age-related macular degeneration.
To determine the clinical properties of pachychoroid neovasculopathy (PNV) that differ from conventional neovascular age-related macular degeneration (nAMD) and suggest that they are different clinical entities. To accomplish this, we reviewed the medical records of 100 consecutive patients diagnosed with nAMD. All of the patients were Japanese, and their mean age was 75.5 years. There were 72 men and 28 women. For the bilateral cases, only the right eye was analyzed. An eye was diagnosed with PNV when a macular neovascularization (MNV) was detected just above the dilated choroidal vessels. The Indocyanine green angiographic (ICGA) and en face optical coherence tomographic (OCT) images were used to assess the vertical symmetry of the medium and large choroidal vessels. The subfoveal choroidal thickness (SCT) was also measured manually in the OCT images. After reclassification, there were 29 (29%) patients with typical nAMD (25 with type 1 MNV, 4 with type 2 MNV), 43 (43%) with PNV, 21 (21%) with polypoidal choroidal vasculopathy, and 7 (7%) with retinal angiomatous proliferation. Of the 43 PNV, 17 (39.5%) had polypoidal lesions and 26 (60.5%) had no polypoidal lesions. The percentage of eyes with vertical asymmetry of the medium and large choroidal vessels was significantly greater in the 35 PNV (81.4%) than in the 16 non-PNV (28.1%; P < 0.01) cases. The mean SCT was significantly thicker in the PNV eyes than in the non-PNV eyes (298 ± 96 μm vs. 228 ± 82 μm; P < 0.01). The response of PNV to anti-vascular endothelial growth factor treatments was better than that of non-PNV eyes [higher dry macula rate after the loading period (90.9% vs. 59.1%), fewer total number of injections (11.0 ± 2.9 vs. 13.4 ± 3.2), and longer treatment intervals for the anti-VEGF therapy (8.4 ± 3.1 vs. 13.4 ± 3.2 weeks) at 2 years (all P < 0.01)]. These differences in the morphology and response to anti-VEGF treatments suggest that PNV is a separate clinical entity to conventional nAMD.


Incidence of human papillomavirus contamination of transvaginal probes in Japan and possible contamination prevention strategy.
To clarify the present status of human papillomavirus (HPV) contamination of transvaginal probes in Japan and propose a preventive method. This study was performed at three institutes: a tertiary center, secondary hospital, and primary facility. To identify contamination rates, probes were disinfected and covered with probe covers and condoms; the cover was changed for each patient. The probes were tested for HPV, and those with HPV detected were analyzed to identify the type of HPV. Next, nurses put on new gloves before covering the probe for each patient, and the probes were similarly tested for HPV. A total of 120 probes were tested, and HPV was detected from a total of five probes, a contamination rate of 4.2 % (5/120). HPV was detected in all three institutes. Importantly, high-risk HPV, i.e., HPV-52, 56, and 59, was detected. After the "glove change strategy" was implemented, HPV was not detected on any of 150 probes tested at any of the three institutions. In Japan, the HPV contamination rate of vaginal probes in routine practice was 4.2 %. There was no HPV contamination of probes after changing the gloves for cover exchange for each patient. This strategy may prevent HPV probe contamination.


DNA flow cytometry and morphometry as prognostic indicators in advanced ovarian cancer: a step forward in predicting the clinical outcome.
In 74 patients with advanced ovarian cancer, the value of DNA flow cytometry and morphometry was evaluated and compared with histological grading and clinical outcome. By means of both DNA flow cytometry and morphometry a clear distinction was found between a favorable group of 15 patients, with a median survival of more than 60 months, and the remaining patients, of whom the majority died during the same follow-up period. The levels of significance for DNA flow cytometry (P = 0.0002) and morphometry (P = 0.0001) with respect to survival of the favorable and unfavorable groups of patients were higher than for histological grading (P = 0.02). In a multivariate analysis it was demonstrated that morphometry proved to be the factor of most relevance for survival, although the effect of DNA flow cytometry was almost as good. Data such as presence of ascites, the size of the residual tumor mass, choice of chemotherapy, FIGO stage, and histological grade were not of additional prognostic relevance in this analysis. It is concluded that both morphometry and DNA flow cytometry are a step forward in identifying favorable and unfavorable groups of patients with advanced ovarian cancer. For the daily practice of management of ovarian cancer patients, morphometry may be an attractive support of visual grading.