Study of kefir drinks produced by backslopping method using kefir grains from Bosnia and Herzegovina: Microbial dynamics and volatilome profile.
Kefir is a well-known health-promoting beverage that can be produced by using kefir grains (traditional method) or by using natural starter cultures from kefir (backslopping method). The aim of this study was to elucidate the microbial dynamics and volatilome profile occurring during kefir production through traditional and backslopping methods by using five kefir grains that were collected in Bosnia and Herzegovina. The results from conventional pour plating techniques and amplicon-based sequencing were combined. The kefir drinks have also been characterized in terms of their physico-chemical and colorimetric parameters. A bacterial shift from Lactobacillus kefiranofaciens to Acetobacter syzygii, Lactococcus lactis and Leuconostoc pseudomesenteroides from kefir grains in traditional kefir to backslopped kefir was generally observed. Despite some differences within samples, the dominant mycobiota of backslopped kefir samples remained quite similar to that of the kefir grain samples. However, unlike the lactic acid and acetic acid bacteria, the yeast counts decreased progressively from the grains to the backslopped kefir. The backslopped kefir samples showed higher protein, lactose and ash content and lower ethanol content compared to traditional kefir samples, coupled with optimal pH values that contribute to a pleasant sensory profile. Concerning the volatilome, backslopped kefir samples were correlated with cheesy, buttery, floral and fermented odors, whereas the traditional kefir samples were correlated with alcoholic, fruity, fatty and acid odors. Overall, the data obtained in the present study provided evidence that different kefir production methods (traditional vs backslopping) affect the quality characteristics of the final product. Hence, the functional traits of backslopped kefir should be further investigated in order to verify the suitability of a potential scale-up methodology for backslopping.


Studies on Structural, Optical, Thermal and Electrical Properties of Perylene-Doped p-terphenyl Luminophors.
A simple solid state reaction technique was employed for the preparation of polycrystalline luminophors of p-terphenyl containing different amounts of perylene followed by spectral characterization techniques viz. XRD, SEM, TGA-DSC, UV-Visible spectroscopy, thermo-electrical conductivity, fluorescence spectroscopy, fluorescence life time spectroscopy and temperature dependent fluorescence. X-ray diffraction profiles of the doped p-terphenyl reveal well-defined and sharp peaks indicate homogeneity and crystallinity. The SEM micrograph of pure p-terphenyl exhibit flakes like grains and then compact and finally gets separately with perylene amounts. The observed results indicate that closed packed crystal structures of doped p-terphenyl during crystal formation. The band gaps estimated from UV-visible spectroscopy decreased from 5.20 to 4.10 eV, while thermo-electrical conductivity increases with perylene content. The fluorescence spectra showed partial quenching of p-terphenyl fluorescence and simultaneously sensitization of perylene fluorescence at the excitation wavelength of p-terphenyl (290 nm) due to excitation energy transfer from p-terphenyl to perylene. The observed sensitization results are in harmony with intense blue color seen in fluorescence microscopy images and has high demand in scintillation process.


Anesthesia in patients with Brugada syndrome: two case reports.
Brugada syndrome is a rare disease. It causes sudden cardiac arrest, which is a serious life-threatening event. Sudden cardiac death mostly results from coronary artery disease. However, patients with Brugada syndrome show normal cardiac anatomy and no evidence of ischemia or electrolyte imbalance. Anesthesia in patients with Brugada syndrome is challenging due to its unpredictable nature, and is worth our attention. We report two cases of Brugada syndrome during anesthesia. In case one, a 31-year-old Filipino laborer was scheduled for laparoscopic appendectomy. The patient denied any preexisting cardiac disease. The preoperative vital signs were stable, with mild fever of 37.9 °C. The operation was smooth. During the emergence period, the patient suffered from sudden onset of ventricular tachycardia. After resuscitation, the cardiac rhythm returned to normal. Later, he was confirmed to have a genetic trait of Brugada syndrome. In case two, a young Taiwanese patient with pre-diagnosed Brugada syndrome underwent an operation. The perioperative precautions were taken to prevent the occurrence of ventricular arrhythmia. The surgery was uneventful. Brugada syndrome, although rare, has the highest incidence in South East Asian healthy young males. It brings attention to possible fatal cardiac arrhythmia in this population. Careful preoperative evaluation and perioperative management can help reduce the harmful outcome of the disease and prevent any untoward events.


Analysis of Human Acupoint Biological Information and Neural Electric Activity Based on Ultrasonographic Image.
To find out the bioelectric signal discriminative features of different acupoints at the same time and at the same acupoint, and then analyze these differences to determine the type of acupuncture points and assist in judging the patient's condition, a hardware system was designed that can collect subcutaneous potential information of acupoints. Then, the changes of ultrasonic images were analyzed after acupuncture at Zu San Li, as well as the influence of acupuncture at Zu San Li on vagal nerve activity. The hardware system is composed of 4 parts: an analog circuit module, a digital circuit module, a power module, and a host personal computer. The power module adopted dual-module power supply mode, which was composed of MC78M05CT and LM2576 and their related peripheral circuits to ensure the stability of the circuit. The analog circuit module collected the electric signals from the 2 acupoints of Zu San Li and Shang Ju Xu of the volunteers' calves through a multistage electrode probe. In the amplifier module, Butterworth filter and 50 HZ notch are used to reduce the noise of electric signals. The filtered and amplified electric signals are converted into A/D converter in the digital circuit module. Serial communication is used to transmit data to the upper computer of the personal computer to compare the noise reduction effect of acupoint information, multiple sequences of different acupoints, and multiple sequences of the same acupoint. The influence of acupuncture at the Zu San Li acupoint on the muscle tension around the acupoint was investigated by shear wave acoustic elastic imaging technology. The influence of acupuncture at this acupoint was explored on vagal nerve activity. The results show that the wavelet noise reduction method can greatly filter out the interference signals. In the case of artificial stimulation, the signal waveform of each twist shows a big increase, and the comparison shows that there is no significant change in the frequency of other bands. Through ultrasonic image evaluation, it was found that acupuncture at Zu San Li can significantly affect the mean value of Young modulus. After acupuncture at Zu San Li, the vagal activity was increased and the balance ratio of sympathetic vagal activity was reduced. Part of the potential frequency of the acupoint is extremely active, which is the response potential of the subcutaneous induction tissue at the acupoint, which proved the feasibility of collecting electric signals around the acupoints in this kind of hardware system. It is of great significance to diagnose the disease according to the characteristics of acupoint electric signals.


111Indium-trastuzumab visualises myocardial human epidermal growth factor receptor 2 expression shortly after anthracycline treatment but not during heart failure: a clue to uncover the mechanisms of trastuzumab-related cardiotoxicity.
Trastuzumab can induce cardiotoxicity, particularly when combined with anthracyclines. Myocardial human epidermal growth factor receptor 2 (HER2) expression may be transiently upregulated by a compensatory mechanism following cardiac stress. 111In-DTPA-trastuzumab, scintigraphy can detect HER2 positive tumour lesions, however previously, we found myocardial uptake in only 1 of the 15 anthracycline-pre-treated patients with a median of 11 months after the last anthracycline administration. To evaluate whether myocardial HER2 expression is upregulated by anthracycline-induced cardiac stress or in case of heart failure by chronic pressure or volume overload, we performed 111In-DTPA-trastuzumab scans in patients shortly after anthracyclines and with non-anthracycline-related heart failure. Patients within 3 weeks after undergoing 4-6 cycles first-line anthracycline-based chemotherapy and patients with heart failure due to cardiac disease underwent gammacamera imaging 48 and 96 h after 111In-DTPA-trastuzumab intravenously. Myocardial 111In-DTPA-trastuzumab uptake was observed in 5 out of 10 anthracycline-treated patients, who all were without symptomatic cardiac dysfunction. None of the 10 heart failure patients showed myocardial uptake. Shortly after completion of anthracycline treatment, myocardial HER2 over-expression was detectable in 50% of the patients. 111In-DTPA-trastuzumab scintigraphy after anthracyclines prior to adjuvant trastuzumab potentially identifies patients susceptible for trastuzumab-related cardiotoxicity and thus may facilitate the optimal timing of trastuzumab therapy.


Bacterial crowd control with iron.
Bacterial biofilms account for more than 80% of human infections. Hergenrother and coworkers report in this issue that high concentrations of iron salts can block the formation of these bacterial communities; this represents an attractive new method for biofilm control.


Insulin hypoglycemia increases the levels of neuropeptide Y and calcitonin gene-related peptide, but not of chromogranins A and B, in rat chromaffin granules.
The levels and subcellular distribution of chromogranin A and B, of calcitonin gene-related peptide (CGRP) and of neuropeptide Y (NPY) were investigated in rat adrenals before and after insulin treatment. Six days after insulin-induced hypoglycemia the levels of chromogranin A and B were similar to controls, however those of NPY and CGRP were increased by a factor of 2.5 and 35, respectively. This treatment also elevated mRNA levels of NPY and CGRP, establishing an increased biosynthesis of these two neuropeptides. As shown by subcellular fractionation, all peptides were present in chromaffin granules after insulin treatment. Furthermore, immunostaining at the ultrastructural level demonstrated the co-localization of chromogranin A, NPY and CGRP within the same chromaffin granules. These results establish that insulin-induced hypoglycemia changes the levels of the secretory peptides in chromaffin granules leading to an altered composition of the secretory cocktail. Apparently, the biosynthesis of the secretory peptides and their storage organelles can be regulated in distinct patterns.


[Influences of food browning on nutrition. II. Effects of ingestion of non-dialyzable browning substances, dopamelanin and chlorogenic acid on growth and protein and lipid metabolism in rats (author's transl)].
In a previous paper, we reported an inhibitory effect on growth rate and plasma lipid composition in rats of browning substances obtained from ethanol extraction of minced egg plant. In order to know active principle(s) in browning substances, attempts were made to condense the substances and chlorogenic acid was separated. The ingestion of chlorogenic acid and other browning substances such as dopameranin and catechin were examined for their biological activity. Active principle(s) in the browning substances appeared to interfere with digestion and utilization of dietic protein. The principle resided in dialyzable fraction in enzymatically produced materials and in nondialyzable fraction in materials obtained by chemical oxidation. Ingestion of nondialyzable fraction caused an elevation of total serum cholesterol level, but little change in serum fatty acid composition.


An electron microscopic study on the lining mucosa of developing rat vocal cords.
To explain the developmental process of epithelial cells of true vocal cords, larynges obtained from 42 fetal and 36 developing young rats were examined using scanning and transmission electron microscopies. Although the membranous portion was indistinguishable from the arytenoid portion in the prenatal stage, there was no squamous cell epithelium on the glottis. The lining epithelium of vocal cords in 15, 16, and 17 day fetal rats was composed of cuboidal and columnar cells which had a primary cilium. Immediately after birth the membranous portion of vocal cords became recognizable, and ciliated cells present on vocal cords retrogressed rapidly while nonciliated squamous cells grew and extended. Results of the present study show that epithelial cells of ciliated columnar type covering vocal cords change remarkably to nonciliated squamous cells between prenatal and postnatal stages.


Congenitally corrected transposition of the great arteries.
Congenitally corrected transposition of the great arteries (cc-TGA, also known as L-transposition) is a rare condition that represents approximately 0.5% of congenital heart disease. It is characterized by atrioventricular and ventriculoarterial discordance. Patients with cc-TGA are at risk for cardiac arrhythmias due to abnormalities of the conduction system. We present an autopsy case of a 49-year-old man with cc-TGA who died unexpectedly, likely as a result of a fatal arrhythmia. This case illustrates the cardiac pathology characteristic of cc-TGA and describes how to grossly identify inversion of the atrioventricular valves.


A Phase II Trial of S-1 and Oxaliplatin in Patients with Metastatic Breast Cancer Previously Treated with Anthracycline and Taxane (KCSG-BR07-03).
This single-arm phase II trial investigate the efficacy and safety of S-1 plus oxaliplatin (SOX) in patients with metastatic breast cancer. Patients with metastatic breast cancer previously treated with anthracyclines and taxanes were enrolled. Patients received S-1 (40-60 mg depending on patient's body surface area, twice a day, day 1-14) and oxaliplatin (130 mg/m2, day 1) in 3 weeks cycle until disease progression or unacceptable toxicity. The primary endpoint was objective response rate (ORR) according to Response Evaluation Criteria in Solid Tumor 1.1. Secondary endpoints included time-to-progression (TTP), duration-of-response (DoR), overall survival (OS), and adverse events. A total of 87 patients were enrolled from 11 institutions in Korea. Hormone receptor was positive in 54 (62.1%) patients and six (6.9%) had human epidermal growth factor receptor 2-positive disease. Forty-eight patients (85.1%) had visceral metastasis and 74 (55.2%) had more than three sites of metastases. The ORR of SOX regimen was 38.5% (95% confidence interval [CI], 26.9 to 50.0) with a median TTP of 6.0 months (95% CI, 5.1 to 6.9). Median DoR and OS were 10.3 months (95% CI, 5.5 to 15.1) and 19.4 (95% CI, not estimated) months, respectively. Grade 3 or 4 neutropenia was reported in 28 patients (32.1%) and thrombocytopenia was observed in 23 patients (26.6%). This phase II study showed that SOX regimen is a reasonable option in metastatic breast cancer previously treated with anthracyclines and taxanes.


Sex and gender aspects on intensive care. A cohort study.
To investigate if patient sex affects intensive care unit (ICU) admission and discharge patterns. Specifically, we investigate if the patients sex affects length of stay (LOS) and probability of ICU discharge and 30- and 90-day mortality. In a retrospective cohort study with 8598 adult patients admitted between 2006 and 2016 to a university hospital ICU in Stockholm, Sweden, we analysed ICU-LOS using univariate and multivariable quantile regression, and performed a competing risk regression model to assess the association between probability of discharge and sex. Mortality was analysed using logistic regression. Of 8598 included patients, 37% were women. No differences in age or median severity illness scores were found. After excluding trauma patients men were older, had a higher Charlson Co-morbidity Index, higher median SAPS 3 and higher probability of mortality than women. Women had a higher probability of being discharged from the ICU. There were no differences in 30- or 90-day mortality. In this large cohort study of critically ill patients we have shown that women and men had an equal length of stay, but women had a significantly higher probability of being discharged. There was no difference in mortality between women and men.


A simple and sensitive liquid chromatography method for the determination of low dihydrocodeine concentrations in human plasma: its application in Chinese healthy volunteers.
A simple, sensitive and modified method was developed for determination of low dihydrocodeine (CAS 125-28-0) concentrations in human plasma by high performance-liquid chromatography (HPLC) with diode array detector. Measurement was performed on a Zorbax XDB-C18 analytical column together with a XDB-C18 precolumn at 40 degrees C after a simple one-step extraction. An isocratic mobile phase consisting of acetonitrile-0.1% trifluoroacetic acid (TFA)-water (12:40:48, v/v/v), was run at a flow rate of 1.0 mL/min. Good chromatographic separation was achieved in less than 6.2 min. This assay was linear over a concentration range of 2.50-100 ng/mL with a lower limit of quantification at 2.50 ng/mL. The intra- and inter-day precision (relative standard deviation) was less than 6.00 and 6.62%, respectively, at all concentration levels studied, while the intra- and inter-day accuracy was 1.50-3.73% and -1.35-1.92%, respectively. Recoveries were 76.10-83.81% with coefficients of variation of 1.86-6.93%. Stability of dihydrocodeine in plasma proved to be good. The validated method was successfully applied to a bioequivalence study of dihydrocodeine after a single oral administration of 20 mg dihydrocodeine tartrate in Chinese healthy male volunteers.


Corrigendum to "A Collaborative Brain-Computer Interface Framework for Enhancing Group Detection Performance of Dynamic Visual Targets".
[This corrects the article DOI: 10.1155/2022/4752450.].


Detection of active human cytomegalovirus by the promyelocytic leukemia body assay in cultures of PBMCs from patients undergoing hematopoietic stem cell transplantation.
A novel detection system was established previously for cells infected with the human cytomegalovirus (HCMV) in vitro that utilizes the unique IE1-dependent nuclear dispersion of promyelocytic leukemia (PML) bodies early in the HCMV replication cycle. This assay system, designated "the PML assay," makes use of the GFP-PML-expressing cell line SE/15, and allows real-time monitoring of infected cells by fluorescence microscopy without any staining procedures. A rapid and quantitative drug susceptibility testing was developed for low-titer clinical isolates propagated in fibroblasts in vitro. The present study sought to exploit the PML assay for evaluating in vivo status of HCMV without virus isolation. Progeny viruses were detected directly from peripheral blood mononuclear cells (PBMCs) infected in vivo obtained from hematopoietic stem cell transplantation recipients. The overall positivity of the PML assay tended to correlate with the levels of genomic DNA. Direct phenotypic susceptibility testing detected one ganciclovir (GCV)-resistant case among 19 samples, which was confirmed further by genomic and plaque reduction assays. However, in another patient with the sequence-proven mutant confirmed by sequencing, the progeny viruses exhibiting GCV-resistance were not detected. Studies on the isolated virus from the latter patient suggested the possibility that replication efficiency may differ between PBMCs and lesions infected in vivo, which may hamper the detection of GCV-resistant viruses by the PML assay, at least in this case. Taken together, the PML assay is sufficiently sensitive to monitor replication-competent HCMV directly from PBMCs infected in vivo, and provides a novel tool for comparing the characteristics of HCMV strains infected in vivo.


A new method for the evaluation of dental implant stability using an inductive sensor.
We developed a new method for the measurement of dental implant stability by analyzing the impulse response of the implant. The movement of the implant was measured by an inductive sensor with a dedicated adaptor. The large inductance of the adapter amplified the small displacement signal of the implant. The Periotest (Siemens, Bensheim, Germany) was used as a source of excitation force to acquire the impact response of the implant. Power spectrum analysis was applied to the impact response of the implant. The peak frequency of the spectrum was used as a measure of the implant stability. The performance of the system was tested and verified through simulation of the implant-bone interface in an in vitro model. Various implant-bone interfacial conditions were assessed. Holes of varying depth and diameter were drilled into a dental implantation model. Two types of impression materials (EXAMIXFINE, Regisil Rigid) with different degrees of hardness were used to fix the implant into the hole. The implant stability was also measured using the ISQ (implant stability quotient) by resonance frequency analysis on the Osstell Mentor (Integration Diagnostics AB, Goteborgsvagen, Sweden) for comparison. Linear regression analysis of the peak frequency as a stability parameter showed a linear relationship with both the depth and the diameter of the hole (p<0.05). When EXAMIXFINE was used, the peak frequency was linearly associated with the depth (R(2)=0.443) and diameter (R(2)=0.396) of the hole. When Regisil Rigid was used, the peak frequency also showed a linear relationship with the depth (R(2)=0.555) and diameter (R(2)=0.350) of the hole. The peak frequency also increased as the hardness of the impression material increased. Differentiability of the system was evaluated by an ANOVA test. A statistically significant difference (p<0.01) was found between all implantation conditions, except in one case using the Regisil Rigid material. In contrast, the ISQ value did not consistently differentiate under several implantation conditions. The developed method could differentiate the stability changes in simulated implantation conditions with a wider dynamic range and with higher resolution than the ISQ value.


MOF-derived LDH modified flame-retardant polyurethane sponge for high-performance oil-water separation: Interface engineering design based on bioinspiration.
Frequent petrochemical spill accidents and secondary fire hazards have threatened the ecological environment and environmental safety. The traditional purification technology has the problems of high energy consumption and secondary pollution, which also brings new challenges to spill disposal. Herein, we demonstrate a biomimetic structure-based flame-retardant polyurethane (PU) sponge (FPUF@MOF-LDH@HDTMS) for continuous oil-water separation. Inspired by desert beetle and lotus leaf, the biomimetic micro-nano composite structure was constructed by in-situ growth of metal-organic framework-derived layered double hydroxide (MOF-LDH) on the surface of the PU sponge. After grafting MOF-LDH with hexadecyltrimethoxysilane, FPUF@MOF-LDH@HDTMS showed excellent superhydrophobic/superoleophilic performance (water contact angle=153° and oil contact angle=0°). FPUF@MOF-LDH@HDTMS can easily and quickly adsorb oily liquids suspended/settled in the water thanks to the unique bionic structure. FPUF@MOF-LDH@HDTMS has excellent oil/organic solvents absorption capacity; even after 20 cycles of use still maintains high adsorption capacity. More importantly, the continuous oil-water separation through FPUF@MOF-LDH@HTMS has achieved a separation efficiency of up to 99.1%. In addition, the bionic superhydrophobic sponge has excellent flame retardancy, which reduces the possibility of secondary fire caused by PU sponges. Thus, the biomimetic micro-nano composite structure provides a new design strategy for the more high-performance oil-water separation sponges.


From the double-helix to novel approaches to the sequencing of large genomes.
Elucidation of the structure of DNA by Watson and Crick [Nature 171 (1953) 737-738] has led to many crucial molecular experiments, including studies on DNA replication, transcription, physical mapping, and most recently to serious attempts directed toward the sequencing of large genomes [Watson, Science 248 (1990) 44-49]. I am totally convinced of the great importance of the Human Genome Project, and toward achieving this goal I strongly favor 'top-down' approaches consisting of the physical mapping and preparation of contiguous 50-100-kb fragments directly from the genome, followed by their automated sequencing based on the rapid assembly of primers by hexamer ligation together with primer walking. Our 'top-down' procedures totally avoids conventional cloning, subcloning and random sequencing, which are the elements of the present 'bottom-up' procedures. Fragments of 50-100 kb are prepared in sufficient quantities either by in vitro excision with rare-cutting restriction systems (including Achilles' heel cleavage [AC] or the RecA-AC procedures of Koob et al. [Nucleic Acids Res. 20 (1992) 5831-5836]) or by in vivo excision and amplification using the yeast FRT/Flp system or the phage lambda att/Int system. Such fragments, when derived directly from the Escherichia coli genome, are arranged in consecutive order, so that 50 specially constructed strains of E. coli would supply 50 end-to-end arranged approx. 100-kb fragments, which will cover the entire approx. 5-Mb E. coli genome. For the 150-Mb Drosophila melanogaster genome, 1500 of such consecutive 100-kb fragments (supplied by 1500 strains) are required to cover the entire genome. The fragments will be sequenced by the SPEL-6 method involving hexamer ligation [Szybalski, Gene 90 (1990) 177-178; Fresenius J. Anal. Chem. 4 (1992) 343] and primer walking. The 18-mer primers are synthesized in only a few minutes from three contiguous hexamers annealed to the DNA strand to be sequenced when using an over 100-fold excess of hexamers and T4 DNA ligase at room temperature, preferably in the presence of the single-strand-binding (SSB) protein of E. coli. These 18-nt primers are immediately extended by the DNA polymerase, Sequenase 2.0, in the dideoxy sequencing reaction. Very high quality sequencing ladders are obtained for single-stranded DNA or denatured double-stranded approx. 50-kb fragments, as exemplified by phage lambda DNA.(ABSTRACT TRUNCATED AT 400 WORDS)


DIF-1 induces its own breakdown in Dictyostelium.
DIF-1 is a novel chlorinated alkyl phenone which induces differentiation of prestalk cells in Dictyostelium discoideum. It is broken down and inactivated by a cytoplasmic enzyme, DIF-1 3(5)-dechlorinase (hereafter referred to as DIF-1 dechlorinase), which is found only in prestalk cells. We show that DIF-1 dechlorinase levels are induced at least 50-fold when cells are treated with DIF-1. This response is rapid--enzyme activity doubles within 15 min and is fully induced within an hour--and occurs early in development, before other prestalk markers can be induced by DIF-1. Maximum inducibility is seen towards the end of aggregation, when DIF-1 dechlorinase is barely detectable in uninduced cells. The dose-dependence reveals a threshold concentration of DIF-1 (15 nM) below which almost no response is seen. Cyclic AMP, which is the chemoattractant during aggregation and plays a key role in later development, suppresses the induction of DIF-1 dechlorinase by DIF-1. We conclude that induction of DIF-1 dechlorinase is one of the first steps on the developmental pathway which leads to prestalk cell differentiation, and suggest that the resulting negative feedback on DIF-1 levels is an important part of the mechanism by which cells decide whether to become prestalk or prespore cells.


Metabolite Comparison between Serum and Follicular Fluid of Dairy Cows with Inactive Ovaries Postpartum.
Inactive ovaries (IO) accounts for 50% of ovarian disease in postpartum dairy cows, which seriously affects their reproductive efficiency. To investigate the metabolic changes in the serum and follicular fluid of dairy cows with IO during lactation, six estrus (E) cows and six IO cows at 50 to 55 days in milk were selected based on B ultrasonic detection and clinical manifestations. The differential metabolites in serum and follicular fluid between the E cows and IO cows were identified by ultra-high-pressure liquid chromatography-quadrupole time-of-flight mass spectrometry, combined with multidimensional statistical methods. The results showed that dairy cows with IO were in a subclinical ketosis status where beta-hydroxybutyrate (BHB) exceeded 1.20 mmol/L, 14 differential metabolites in the serum of IO cows included 10 increased metabolites and 4 decreased metabolites, and 14 differential metabolites in the follicular fluid of IO cows included 8 increased metabolites and 6 decreased metabolites. These differential metabolites mainly involved nine metabolic pathways. The common enrichment pathway of different metabolites in serum and follicular fluid were glycerophospholipid metabolism and pentose and glucuronate interconversions. In conclusion, there were significant differences in the differential metabolites and enrichment pathways between serum and follicular fluid of IO cows, implying that there were complex changes in blood metabolism and local follicular metabolism of IO cows, whose interactions need further investigation.


Prevalence, awareness, treatment, and control of hypertension in the Czech Republic. Results of two nationwide cross-sectional surveys in 1997/1998 and 2000/2001, Czech Post-MONICA Study.
Hypertension is an easily diagnosed and eminently modifiable risk factor for the development of all clinical manifestations of atherosclerosis. Despite the availability of a simple, non-invasive, and rather accurate method of measuring blood pressure (BP), and overwhelming evidence that reducing BP effectively prevents cardiovascular events, hypertension at the population level is not managed optimally. In 1997/1998 and 2000/2001, two surveys for cardiovascular risk factors were conducted in nine districts of the Czech Republic, involving a 1% population random sample aged 25-64 years in each district. In concordance with the MONICA Project, the present study confirms a high prevalence of hypertension in the Czech population, detecting an increase in prevalence for the male population over a period of 3 years (males from 38.8 in 1997/1998 to 42.3 in 2000/2001; P<0.05). Within the same period, there is also a significant increase in the awareness of hypertension in males (from 57.3 in 1997/1998 to 60.0 in 2000/2001; P<0.05), and an increase in the number of male hypertensives being treated by antihypertensive drugs (from 30.9 to 44.3; P<0.05). Control of hypertension did not change in either sex, being still suboptimal (males 16.4%, females 25.4% in the last survey in 2000/2001).


Stabilization of the non-canonical adenine-adeninium base pair by N(7) coordination of Zn(II).
A new zinc (II) compound with 9-ethyladenine (9-EtA) of formula [Zn(9-EtA-N7)Cl(3)](9-EtAH) has been synthesized and characterized by X-ray diffraction. Its X-structure consists of an Zn(II) anionic complex and 9-ethyladeninium as counteranion. The Zn(II) complex shows a distorted tetrahedral geometry in which three Cl and an 9-EtA coordinates through N(7) position are the ligands. An indirect chelation via intramolecular H-bond between N(6)H and an Cl ligand is present in the complex. The network of [Zn(9-EtA-N7)Cl(3)](9-EtAH) shows interesting features. Thus, self-association of coordinated adenine-adeninium takes place by H-bonding of N(6)-H...N(1) and N(6)-H...N(7), leading to a polymeric ribbon-like 1D supramolecular arrangement. Ab initio calculations have been applied in order to study the stability of the adenine-adeninium interaction due to the coordination of the Zn(II) to the N(7) position and to compare experimental and theoretical structural data.


Growth behavior of plexiform neurofibromas after surgery.
The aim of this study was to analyze growth rate and identify prognostic factors for progression of postoperative plexiform neurofibromas in patients with neurofibromatosis type 1. We measured postoperative tumor volume change per year on magnetic resonance imaging. Linear regression models were applied to identify risk factors for tumor progression. Fifty-two patients (mean age: 25.4 years (3.2-64.2 years)) with 56 plexiform neurofibromas were analyzed. Initial median tumor volume was 40.3 ml (SD: 1,552 (0-10,800)). Surgical indications included disfigurement (n = 21), pain (n = 20), and functional deficits (n = 16). Sixteen percent of all cases experienced acute surgical complication, and 13% showed late complication. Eight patients (19%; 6 children and 2 adults) with residual tumor had repeat surgery for tumor progression. Median tumor progression was 0.6% change per year (SD ± 27.4; range: -59.2 to 88.1) and 2.9% from baseline (SD ± 163.9; range -1,001.3 to 81.8). Patients aged 21 years and younger had the highest progression rate (P < 0.01). For every year of age, the mean growth rate decreased by -0.463 mean percent (P = 0.03). With age as a continuous variable, age, the site of the tumor, and depth were the only factors associated with tumor progression. Fourteen plexiform neurofibromas (10 nodular and 4 diffuse) in 13 patients (5 children and 8 adults) were completely resected (by visualization) and did not relapse during observation (mean: 2.9 years; range: 1.1-5.8 years). Age, tumor type, location, and depth are helpful to estimate the progression of plexiform neurofibromas after surgery. Patients benefit from elective surgery of small and completely removable plexiform neurofibromas.


Retrospective monitoring of minimal residual disease using hairpin-shaped clone specific primers in B-cell lymphoma affected dogs.
Lymphoma is one of the most common forms of cancer in dogs as it is in humans but, unlike humans, the cure rates in canines are still very low. Despite the fact that high grade B-cell lymphomas are considered to be chemotherapy responsive, almost all treated dogs ultimately relapse and die due to the residual malignant lymphocytes, namely minimal residual disease (MRD). It would be extremely valuable for clinicians to detect, monitor and quantify MRD for risk group stratification, effective treatment intervention and outcome prediction. The PCRs targeting the Ig gene rearrangements constitute one of the most reliable tools to this end. We have recently validated a method which exploits hairpin-shaped primers for quantifying MRD. In the present study, that method is conveniently used for retrospectively monitoring MRD in the peripheral blood of 8 dogs diagnosed with B-cell lymphoma who underwent chemotherapy. All dogs attained complete remission. The median disease-free interval was 254.5 days (range 63-774) while the median survival time was 313.5 days (range 143-817 days). At admission, all dogs, except one which had already been treated with prednisone, had circulating neoplastic cells. All dogs attained complete remission (CR) which was almost always matched with a complete MRD response. The persistence of MRD despite apparent CR indicated a worse prognosis and a short duration of CR. Finally, the relapse is consistently anticipated by the reappearance of MRD in the peripheral blood. The study confirmed the suitability of an MRD monitoring assay as a clinical decision-making tool.


Addiction history moderates the effect of prefrontal 10-Hz transcranial alternating current stimulation on habitual action selection.
Individuals with substance use disorders (SUDs) transition more quickly from goal-directed to habitual action-selection, but the neural mechanisms underlying this phenomenon remain unclear. Data from animal models suggest that drugs of abuse can modify the neurocircuits that regulate action-selection, enhancing circuits that drive inflexible, habit-based stimulus-response (S-R) action-selection and weakening circuits that drive flexible, goal-directed actions. Here, we tested the effect of bilateral 10-Hz transcranial alternating current stimulation (10Ηz-tACs) of the dorsolateral prefrontal cortex on action-selection in men and women with a SUD history and an age- and sex-matched control group. We tested the hypothesis that true 10Ηz-tACS versus active sham stimulation would reduce perseverative errors after changed response contingencies for well-learned S-R associations, reflecting reduced habit-based action-selection, specifically in the SUD group. We found that 10 Hz-tACS increased perseverative errors in the control group, but in the SUD group, 10 Hz-tACS effects on perseverative errors depended on substance abuse duration: a longer addiction history was associated with a greater reduction of perseverative errors. These results suggest that 10Ηz-tACs altered circuit level dynamics regulating behavioral flexibility, and provide a foundation for future studies to test stimulation site, frequency, and timing specificity. Moreover, these data suggest that chronic substance abuse is associated with altered circuit dynamics that are ameliorated by 10Ηz-tACs. Determining the generalizability of these effects and their duration merits investigation as a direction for novel therapeutic interventions. These findings are timely based on growing interest in transcranial stimulation methods for treating SUDs.NEW & NOTEWORTHY Treating the executive dysfunction associated with addiction is hampered by redundancies in pharmacological regulation of different behavioral control circuits. Thus, nonpharmacological interventions hold promise for addiction treatment. Here, we show that, among people with an addiction history, 10-Hz transcranial alternating current stimulation (10Hz-tACS) of the dorsolateral prefrontal cortex can reduce habitual actions. The fact that 10Hz-tACS can regulate behavioral flexibility suggests its possible utility in reducing harmful habitual actions.


Death by deamination: a novel host restriction system for HIV-1.
A human gene with potent antiretroviral activity has recently been found to encode a new member of the family of cytidine deaminases, previously known to be involved in mRNA editing, immunoglobulin gene class switching, and immunoglobulin gene hypermutation. The enzyme attacks viral DNA as it is synthesized in infected cells and prevents the formation of functional proviruses. The Vif gene of HIV-1 blocks this host restriction and so allows virus replication.


Leiomyosarcoma: One disease or distinct biologic entities based on site of origin?
Leiomyosarcoma (LMS) can originate from the retroperitoneum, uterus, extremity, and trunk. It is unclear whether tumors of different origin represent discrete entities. We compared clinicopathologic features and outcomes following surgical resection of LMS stratified by site of origin. Patients with LMS undergoing resection at a single institution were retrospectively reviewed. Clinicopathologic variables were compared across sites. Survival was calculated using the Kaplan-Meier method and compared using log-rank and Cox regression analyses. From 1983 to 2011, 138 patients underwent surgical resection for LMS. Retroperitoneal and uterine LMS were larger, higher grade, and more commonly associated with synchronous metastases. However, disease-specific survival, recurrence-free survival, and recurrence patterns were not significantly different across the four sites. Synchronous metastases (HR 3.20, P < 0.001), but not site of origin, size, grade, or margin status, were independently associated with worse DSS. A significant number of recurrences and disease-related deaths were noted beyond 5 years. Although larger and higher grade, retroperitoneal and uterine LMS share similar survival and recurrence patterns with their trunk and extremity counterparts. LMS of various anatomic sites may not represent distinct disease processes based on clinical outcomes. The presence of metastatic disease remains the most important prognostic factor for LMS.


Elderly trauma associated with high-risk recreational activity: A population-based study, United States, 2010 through 2016.
Understanding trends in prevalence and etiology is critical to public health strategies for prevention and management of injury related to high-risk recreation in elderly Americans. The National Emergency Department Sample from 2010 through 2016 was queried for patients with a principal diagnosis of trauma (ICD-9 codes 800.0-959.9) and who were 55 years and older. High-risk recreation was determined from e-codes a priori. Primary outcome measures were mortality and total hospital charges. Of the 29,491,352 patient cohort, 458,599 (1.56%) engaged in high-risk activity, including those age 85 and older. High-risk cases were younger (median age 61 vs 70) and majority male (71.87% vs 39.24%). The most frequent activities were pedal cycling (45.81%), motorcycling (29.08%), and off-road vehicles (9.13%). Brain injuries (8.82% vs 3.88%), rib/sternal fractures (13.35% vs 3.53%), and cardiopulmonary injury (5.25% vs 0.57%) were more common among high-risk cases. Mortality (0.75% vs 0.40%) and total median hospital charges ($3,360 vs $2,312) were also higher for high-risk admissions, where the odds of mortality increased exponentially per year of age (odds ratio, 1.06; 99.5% CI, 1.05-1.08). High-risk recreation was associated with more than $1 billion in total hospital charges and more than 100 deaths among elderly Americans per year. Morbidity, mortality, and resource utilization due to high-risk recreation extend into the ninth decade of life. The patterns of injury described here offer opportunities for targeted injury prevention education to minimize risk among this growing segment of the United States population.


Surgeon's garb and infection control: what's the evidence?
Surgical site infections are an important complication resulting from surgery. Before the 20th century a significant number of surgeries resulted in death from sepsis. One of the rituals resulting from efforts to reduce infection is the donning of surgical garb. A large body of literature has developed on the efficacy of these specialized garments. The available evidence for the efficacy of each individual garb at reducing surgical site infections will be explored. The relevance of this evidence to the outpatient dermatologic surgery setting will also be discussed.


Is there a role for endovascular stent implantation in the management of postoperative right ventricular outflow tract obstruction in the era of transcatheter valve implantation?
The optimal management pathway for the dysfunctional right ventricular outflow tract (RVOT) is uncertain. We evaluated the long-term outcomes and clinical impact of stent implantation for obstructed RVOTs in an era of rapidly progressing transcatheter pulmonary valve technology. Retrospective review of 151 children with a biventricular repair who underwent stenting of obstructed RVOT between 1991 and 2017. RVOT stenting resulted in significant changes in peak right ventricle (RV)-to-pulmonary artery (PA) gradient (39.4 ± 17.1-14.9 ± 8.3; p < 0.001) and RV-to-aortic pressure ratio (0.78 ± 0.22-0.49 ± 0.13; p < 0.001). Subsequent percutaneous reinterventions in 51 children to palliate recurrent stenosis were similarly effective. Ninety-nine (66%) children reached the primary outcome of subsequent pulmonary valve replacement (PVR). Freedom from PVR from the time of stent implantation was 91%, 51%, and 23% at 1, 5, and 10 years, respectively. Small balloon diameters for stent deployment were associated with shorter freedom from PVR. When additional children without stent palliation (with RV-to-PA conduits) were added to the stent cohort (total 506 children), the multistate analysis showed the longest freedom from PVR in those with stent palliation and subsequent catheter reintervention. Pulmonary regurgitation was well-tolerated clinically. Indexed RV dimensions and function estimated by echocardiography remained stable at last follow up or before primary outcome. Prolongation of conduit longevity with stent implant remains an important strategy to allow for somatic growth to optimize the risk-benefit profile for subsequent surgical or transcatheter pulmonary valve replacement performed at an older age.


Mechanisms underlying obesity associated with systemic and renal hemodynamics in essential hypertension.
Obesity and hypertension frequently coexist and are considered major "factors of risk" associated with coronary heart disease. This report identifies the systemic and renal hemodynamic alterations associated with obesity in normotensive and essential hypertensive patients. An expanded intravascular (plasma) volume is associated with obesity, which is related to an increased venous return to the heart, increased cardiac output, and increased blood flow to kidneys and other organs in normal and hypertensive patients. The cardiac consequences result in structural and functional alterations. Although recently postulated pathophysiologic mechanisms have been associated with obesity, none has yet been related to the hemodynamic alterations associated with obesity that coexist in patients with essential hypertension.


The Power of Sophisticated Phenotypic Screening and Modern Mechanism-of-Action Methods.
The enthusiasm for phenotypic screening as an approach for small-molecule discovery has increased dramatically over the last several years. The recent increase in phenotype-based discoveries is in part due to advancements in phenotypic readouts in improved disease models that recapitulate clinically relevant biology in cell culture. Of course, a major historical barrier to using phenotypic assays in chemical biology has been the challenge in determining the mechanism of action (MoA) for compounds of interest. With the combination of medically inspired phenotypic screening and the development of modern MoA methods, we can now start implementing this approach in chemical probe and drug discovery. In this Perspective, we highlight recent advances in phenotypic readouts and MoA determination by discussing several case studies in which both activities were required for understanding the chemical biology involved and, in some cases, advancing toward clinical development.


COVID-19 vaccine hesitancy among youth.
The purpose of this evaluation is to describe COVID-19 vaccine hesitancy among a sample of youth aged 12-15 and inform efforts to increase vaccination uptake among youth populations. We used data collected in May 2021 through a health education program for 9th graders (N = 345). We used Spearman correlations and chi-square tests to assess the statistical significance of bivariate relationships. Less than half of students (42%) reported they are not hesitant at all about getting a COVID-19 vaccine. The remainder reported they were "a little hesitant" (22%), "somewhat hesitant" (21%), or "very hesitant" (15%). There were no statistically significant differences across age, gender, race/ethnicity, parental education, self-reported grades, or hours playing video games during school days. There was a statistically significant relationship between COVID-19 vaccine hesitancy and hours of TV watched during school days. The prevalence of COVID-19 vaccine hesitancy in this sample is high, but more research is needed to estimate its prevalence for youth at the state or national level. Correlates of COVID-19 vaccine hesitancy among adult populations may not match those found in youth populations. Research which recognizes youth as agentic rather than passive participants in decision-making opens opportunities for developing age-appropriate health communication and interventions for vaccination.


Pulmonary alveolar proteinosis in a child: HRCT findings before and after bronchoalveolar lavage.
We demonstrate the high-resolution computed tomography (HRCT) findings of pulmonary alveolar proteinosis in a child before and after bronchoalveolar lavage. The CT pattern in our case differs from the pattern described in previous reports. We found a more homogeneous distribution of the pulmonary changes and a "crazy paving" pattern. High-resolution CT may be helpful in the differential diagnosis of this rare disease and in the follow-up of the pulmonary changes after bronchoalveolar lavage.


Neurophysiological findings in the Rett syndrome, II: Visual and auditory brainstem, middle and late evoked responses.
Nine girls with the Rett syndrome (RS) were investigated neurophysiologically using evoked potentials techniques. Visual- (VER) and auditory-evoked responses, including the early (ABR), middle (MLR) and late components (ACR), were recorded. There was evidence of variable, multilevel impairment of the nervous system. While ABR and MLR indicated lesions at the brainstem and midbrain levels, the late responses and VER pointed to an intra-cerebral/cortical defect. It is suggested that the perceptual mechanisms still functioned and some discrimination properties remained.


Brief Report: A Pilot Study of the Use of a Virtual Reality Headset in Autism Populations.
The application of virtual reality technologies (VRTs) for users with autism spectrum disorder (ASD) has been studied for decades. However, a gap remains in our understanding surrounding VRT head-mounted displays (HMDs). As newly designed HMDs have become commercially available (in this study the Oculus Rift™) the need to investigate newer devices is immediate. This study explored willingness, acceptance, sense of presence and immersion of ASD participants. Results revealed that all 29 participants (mean age = 32; 33 % with IQ < 70) were willing to wear the HMD. The majority of the participants reported an enjoyable experience, high levels of 'presence', and were likely to use HMDs again. IQ was found to be independent of the willingness to use HMDs and related VRT immersion experience.


Treating the physical symptoms of depression with second-generation antidepressants: a systematic review and metaanalysis.
Approximately two-thirds of patients with depression experience physical pain symptoms. Coexisting pain complicates the treatment of depression and is associated with worse depression outcomes. The authors reviewed the effect of newer antidepressants on pain in patients with depression. The authors searched systematically for trials of second-generation antidepressants that enrolled depression patients and reported pain outcomes, pooling changes on the pain visual-analog scale (VAS), using random-effects models. Eight trials were eligible. Pooled analysis of head-to-head trials showed no difference in VAS between duloxetine and paroxetine. Both drugs were superior to placebo. The authors found insufficient evidence to support the choice of one second-generation antidepressant over another in patients with pain accompanying depression.


Treatment of dermatofibroma sarcoma protuberans using modified Mohs micrographic surgery: no recurrences and smaller defects.
The purpose of this study was to evaluate the use of modified Mohs micrographic surgery (MMS) for dermatofibroma sarcoma protuberans (DFSP) in a single institution by a single surgeon. The authors conducted a retrospective analysis of 25 patients with DFSP who were treated with modified MMS over the past 19 years at Scripps Clinic. Of the 25 patients treated with modified MMS for DFSP, there were no identifiable recurrences. The mean follow-up time was 101 months. The defect sizes of DFSP lesions treated by modified MMS are smaller than if the lesions had been treated with the standard wide local excision (WLE) margins of 3 cm. This is a retrospective analysis using the data of one surgeon. Five of the 25 patients were not examined in our office. Not all patients had 5-year follow-up. There were no cases of the fibrosarcomatous variant. Our data support the growing literature that modified MMS achieves excellent local control for DFSP with a possible benefit of smaller defects when compared with treatment with WLE. The authors have indicated no significant interest with commercial supporters.


Machine Learning Identification of Immunotherapy Targets in Low-Grade Glioma Using RNA Sequencing Expression Data.
Immunotherapy has revolutionized cancer treatment in the past decade, but significant hurdles remain. Human studies with immune checkpoint inhibitors targeting programmed cell death protein have demonstrated suboptimal efficacy in the setting of low-grade gliomas (LGGs). Identification of mechanisms leading to inadequate anti-tumor immunity is paramount. The current study evaluates and validates barriers to immunotherapy using a novel machine learning algorithm. We utilized The Cancer Genome Atlas (TCGA) to generate expression levels of 28 immune genes related to known immunotherapeutic targets or lymphocyte cytolytic activity. We created training and testing groups and 3 machine learning models to determine the genes most highly correlated to cytolytic activity (CYT). The 3 models were run through multiple regression by exhaustive selection, LASSO, and random forest. We validated computational results by comparing expression of pertinent genes in patient-derived glioma samples. Our models demonstrated linearity, a low mean-squared error, and consistent results with respect to the most important variables. Expression of ICOS, IDO1, and CD40 were the most important variables in all models and demonstrated positive correlation with CYT. Other variables included TIGIT and CD137. Genetic analysis from 3 IDH-mutants (IDHm) and 3 IDH-wild type (IDHwt) patient-derived glioma samples validated TCGA data and demonstrated lower levels of CYT in IDHm gliomas compared with IDHwt. This novel methodology has elucidated 3 potential targets for immunotherapy development in LGGs. We also demonstrated a novel method of analyzing data using advanced statistical techniques that can be further used in developing treatments for other diseases as well.


The Novel Familial ST-Depression Syndrome - Current Knowledge and Perspectives.
Familial ST-depression syndrome represents a novel inherited disease characterized by nonischemic ST-segment depressions in multiple leads. The ECG phenotype appears to debut around puberty, while the typical onset of arrhythmias occurs around 50 years of age. Clinical manifestations include supraventricular arrhythmias, fast polymorphic ventricular tachycardia, sudden cardiac death, and left ventricular systolic dysfunction. The optimal treatment is unknown but asymptomatic individuals without red flags may not need treatment. In contrast, ICD implantation should be considered in patients with probable arrhythmic syncope and in those fulfilling general criteria for ICD treatment. Future research should focus on establishing the disease prevalence, optimizing risk stratification and treatment, and elucidating the underlying genetic etiology.


Induction of erythroid differentiation in Friend leukemia cells by bromodeoxyuridine.
Treatment of Friend leukemia cells with BrdU, the thymidine analog which interferes with DMSO induced differentiation in these cells as well as the expression of differentiated character in many other cell systems, is capable of inducing erythroid differentiation. Globin mRNA, as assayed by hybridization to globin cDNA, increases 2.5- to 30-fold after appropriate treatment with BrdU. This effect was observed with several different subclones of three independent Friend tumor cell lines. After BrdU treatment, globin mRNA content may reach up to 10-20% of the levels in DMSO induced cultures. The induction of erythroid differentiation is also apparent when accumulated heme content or the appearance of benzidine positive cells is monitored. One Friend cell line (745) we examined was not induced by BrdU although it incorporated an amount of BrdU into its DNA comparable to that incorporated by the other cell lines. In addition, BrdU did interfere with DMSO induction in this cell line. These results suggest that two different mechanisms may be operative in regulating erythroid differentiation in Friend leukemia cells. While BrdU interferes with the mechanism activated by DMSO treatment, this analog could independently activate an alternative mechanism.


Diseases of the nose and sinuses.
This article discusses the diagnosis and management of acute and chronic diseases of the nasal cavity and sinuses. Also discussed are the anatomy of the upper respiratory tract, guidelines for obtaining a thorough history and performing a complete physical examination, and various diagnostic procedures, such as rhinoscopy, culture, and serology.


Allantoic cyst: a prenatal clue to patent urachus.
A patent urachus, which is typically diagnosed as leakage from the umbilicus postnatally, can present as an allantoic cyst in the umbilical cord antenatally. We report a case of a patent urachus with an allantoic cyst diagnosed via fetal MR imaging at 24 weeks' gestation. Early detection allowed for appropriate counseling and prompt corrective surgery after birth.


The influence of reconstruction methods on food retention phenomenon in the remnant stomach after a subtotal gastrectomy.
Food retention and bile reflux has been frequently observed in gastric cancer patients following a subtotal gastrectomy. The aim of this study was to determine whether reconstruction methods after the distal subtotal gastrectomy influenced the degree of food residue and bile reflux. The prospectively collected data was reviewed retrospectively for 522 patients with early gastric cancer who had undertaken a follow-up endoscopic examination after a distal subtotal gastrectomy between 2003 and 2006. The incidence of food retention was 55.5%, 31.9%, and 20.9% at 3, 12, and 24 months after distal subtotal gastrectomy, respectively. The food residue score was higher in the Billroth I (stapling) group than the Billroth II (hand sewing) group at 3 months after surgery (P = 0.006). The incidence of bile reflux was higher in the Billroth II group than in the Billroth I group at 12 and 24 months after surgery (P < 0.001, P = 0.002, respectively). No significant association was found between the food retention and body weight changes. Food retention was detected in lots of patients after subtotal gastrectomy and the reconstructive methods after subtotal gastrectomy was not relevant to food retention.


The effect of white spot syndrome virus (WSSV) envelope protein VP28 on innate immunity and resistance to white spot syndrome virus in Cherax quadricarinatus.
VP28 is the most abundant membrane protein of WSSV, and the recombinant protein VP28 (VP26 or VP24) was constructed for the immune protection experiment in this study. Crayfish were immunized by intramuscular injection of recombinant protein V28 (VP26 or VP24) at a dose of 2 μg/g. The survival rate of crayfish immunized by VP28 showed a higher value than by VP26 or VP24 after WSSV challenge. Compared with the WSSV-positive control group, the VP28-immunized group could inhibit the replication of WSSV in crayfish, increasing the survival rate of crayfish to 66.67% after WSSV infection. The results of gene expression showed that VP28 treatment could enhance the expression of immune genes, mainly JAK and STAT genes. VP28 treatment also enhanced total hemocyte counts and enzyme activities including PO, SOD, and CAT in crayfish. VP28 treatment reduced the apoptosis of hemocytes in crayfish, as well as after WSSV infection. In conclusion, VP28 treatment can enhance the innate immunity of crayfish and has a significant effect on resistance to WSSV, and can be used as a preventive tool.


Synthesis of intramolecular cross-coupling analogues of forskolin.
A ring distortion strategy was applied to the synthesis of a series of intramolecular cross-coupled analogues of forskolin 1. Treatment with palladium acetate, forskolin underwent an intramolecular cross-coupling reaction to generate a novel cycloalkene ether 2 in 85% yield. Under the same conditions, a series of forskolin ester analogues 4a-4d were prepared from 1-OH ester derivatives of forskolin 3a-3d in 85-93% yields. Treating cycloalkene ether 2 with aryl iodides in the presence of a palladium catalyst afforded Z-isomers arylation products 5a-5e in a stereoselective manner in 70-85% yields.


Exposure to imidacloprid induce oxidative stress, mitochondrial dysfunction, inflammation, apoptosis and mitophagy via NF-kappaB/JNK pathway in grass carp hepatocytes.
Imidacloprid (IMI) is a neonicotinoid compound widely used in agriculture production, causing surface water pollution and threatening non-target organisms. The aim of this study was to analyze the effects of IMI on grass carp (Ctenopharyngodon idellus) liver cell (L8824) injury. The L8824 cells were exposed to different doses of IMI (65 mg/L, 130 mg/L and 260 mg/L) for 24 h. Our results demonstrated that exposure IMI significantly suppressed the activity of anti-oxidant enzymes (SOD, CAT and T-AOC) and accumulated oxidase (MDA) levels, and promoting reactive oxygen species (ROS) generation in L8824 cells. Additionally, mitochondrial membrane potential (ΔΨ m), mitochondria-derived ROS and ATP content and the MitoTracker Green indicated that IMI aggravated mitochondrial dysfunction, thereby inducing inflammation and enhancing pro-inflammatory genes (NF-kappaB, TNFα, IL-1β and IL-6) expressions. However, the addition of 2 mM N-acetyl-l-cysteine (NAC) can reverse these adverse effects of high-dose IMI- induced. Hence, ROS is the main factor of IMI-induced mitochondrial dysfunction and inflammation. We further found that exposure to IMI induced apoptosis, which is characterized by promoting release of cytochrome c (Cyt-C), and increasing the expression of Bcl-2-Associated X (BAX), cysteinyl aspartate specific proteinases (Caspase 9 and 3), decreasing Bcl-2 level. Immunofluorescent staining, qRT-PCR and Western Blot results indicated that IMI exposure also activated mitophagy, which was demonstrated by the expression of mitophagy-related genes (BNIP3, LC3B and P62). Conversely, scavenging JNK by SP600125(10 μM) alleviated the expression of mitochondrial apoptosis and mitophagy-related gene induced by high-dose IMI. Therefore, these results of study demonstrated that IMI-induced oxidative stress to regulate mitochondrial dysfunction, thus causing inflammation, mitochondrial apoptosis and mitophagy in grass carp hepatocytes through NF-kappaB/JNK pathway.


Update on the treatment of Duchenne muscular dystrophy.
Duchenne muscular dystrophy is the most severe childhood form of muscular dystrophy caused by mutations in the gene responsible for dystrophin production. There is no cure, and treatment is limited to glucocorticoids that prolong ambulation and drugs to treat the cardiomyopathy. Multiple treatment strategies are under investigation and have shown promise for Duchenne muscular dystrophy. Use of molecular-based therapies that replace or correct the missing or nonfunctional dystrophin protein has gained momentum. These strategies include gene replacement with adeno-associated virus, exon skipping with antisense oligonucleotides, and mutation suppression with compounds that "read through" stop codon mutations. Other strategies include cell therapy and surrogate gene products to compensate for the loss of dystrophin. All of these approaches are discussed in this review, with particular emphasis on the most recent advances made in each therapeutic discipline. The advantages of each approach and challenges in translation are outlined in detail. Individually or in combination, all of these therapeutic strategies hold great promise for treatment of this devastating childhood disease.


Simultaneous triple-parametric optical mapping of transmembrane potential, intracellular calcium and NADH for cardiac physiology assessment.
Investigation of the complex relationships and dependencies of multiple cellular processes that govern cardiac physiology and pathophysiology requires simultaneous dynamic assessment of multiple parameters. In this study, we introduce triple-parametric optical mapping to simultaneously image metabolism, electrical excitation, and calcium signaling from the same field of view and demonstrate its application in the field of drug testing and cardiovascular research. We applied this metabolism-excitation-contraction coupling (MECC) methodology to test the effects of blebbistatin, 4-aminopyridine and verapamil on cardiac physiology. While blebbistatin and 4-aminopyridine alter multiple aspects of cardiac function suggesting off-target effects, the effects of verapamil were on-target and it altered only one of ten tested parameters. Triple-parametric optical mapping was also applied during ischemia and reperfusion; and we identified that metabolic changes precede the effects of ischemia on cardiac electrophysiology.


Amyloid beta deregulates astroglial mGluR5-mediated calcium signaling via calcineurin and Nf-kB.
The amyloid hypothesis of Alzheimer's disease (AD) suggests that soluble amyloid β (Aβ) is an initiator of a cascade of events eventually leading to neurodegeneration. Recently, we reported that Aβ deranged Ca(2+) homeostasis specifically in hippocampal astrocytes by targeting key elements of Ca(2+) signaling, such as mGluR5 and IP3 R1. In the present study, we dissect a cascade of signaling events by which Aβ deregulates glial Ca(2+) : (i) 100 nM Aβ leads to an increase in cytosolic calcium after 4-6 h of treatment; (ii) mGluR5 is increased after 24 h of treatment; (iii) this increase is blocked by inhibitors of calcineurin (CaN) and NF-kB. Furthermore, we show that Aβ treatment of glial cells leads to de-phosphorylation of Bcl10 and an increased CaN-Bcl10 interaction. Last, mGluR5 staining is augmented in hippocampal astrocytes of AD patients in proximity of Aβ plaques and co-localizes with nuclear accumulation of the p65 NF-kB subunit and increased staining of CaNAα. Taken together our data suggest that nanomolar [Aβ] deregulates Ca(2+) homeostasis via CaN and its downstream target NF-kB, possibly via the cross-talk of Bcl10 in hippocampal astrocytes.


Emodin attenuates TNF-α-induced apoptosis and autophagy in mouse C2C12 myoblasts though the phosphorylation of Akt.
Emodin, a major component of Rheum palmatum, has been reported to significantly protect neural tissue against apoptosis and autophagy. However, the effects and underlying mechanisms of action of emodin in muscle atrophy are still poorly defined. In this study, we investigated the protective effects and the underlying mechanisms by which emodin acts on tumor necrosis factor alpha (TNF-α)-induced apoptosis and autophagy in mouse C2C12 myoblasts. Emodin, at various concentrations, decreased TNF-α-induced apoptosis in C2C12 myoblasts, which were analyzed by Hoechst 33342 staining and annexin V/PI analysis. Emodin also inhibited the collapse of the mitochondrial membrane potential and the generation of reactive oxygen species in TNF-α-stimulated C2C12 myoblasts. Consistent with these results, the expression of Bcl-2 was increased, whereas the expression of Bax, cleaved-caspase 3 and cleaved-PARP was decreased after emodin treatment. These data demonstrate that emodin attenuated apoptosis in TNF-α-stimulated C2C12 myoblasts through mitochondrial signaling pathways. In addition, emodin inhibited autophagy in TNF-α-stimulated C2C12 myoblasts by suppressing the expression of LC3-II, Beclin-1 and Atg7. Emodin also resulted in the upregulation of the phosphorylated forms of Akt. Taken together, these results suggest that emodin inhibited apoptosis and autophagy in TNF-α-induced C2C12 myoblasts, possibly through the activation of phosphorylated Akt. Our findings suggest that emodin could be a potential therapeutic agent in the treatment of muscle atrophy.


Plasma levels of glucose, alanine, lactate, and beta-hydroxybutyrate in the unfed spiny dogfish shark (Squalus acanthias) after surgery and following mammalian insulin infusion.
Serial blood samples were collected from the unfed spiny dogfish shark (Squalus acanthias) by use of a cannula chronically implanted in the dorsal aorta. Plasma glucose, alanine, lactate, and beta-hydroxybutyrate levels were estimated by standard enzymatic procedures. The metabolite levels were estimated immediately after the surgery, for an additional 3 to 5 postoperative days before experimental use, and for 7 to 9 days after a single infusion of mammalian insulin (50, 100, and 250 IU/kg body wt) or the control solution. Plasma glucose, alanine, and lactate levels declined for 1 or 2 days after the surgery, but beta-hydroxybutyrate levels were essentially unchanged. Plasma glucose levels remained relatively stable for the remainder of the pretreatment period, and in the control animals during the subsequent treatment period. In contrast, plasma alanine and beta-hydroxybutyrate levels increased during both the pretreatment period and the treatment period in the control animals. Plasma lactate levels increased in the control animals after Day 3 of the treatment period. Insulin infusions resulted in severe, prolonged depressions of plasma glucose and alanine levels. Plasma lactate levels also fell after insulin infusion, but the depressed levels were interrupted by precipitous, large increases, followed by sharp declines, that were evident for one to three sampling periods and whose onset varied between Days 3 and 5 in individual animals. The levels increased again in some animals between Days 6 and 9. Plasma beta-hydroxybutyrate levels were comparatively resistant to insulin. After an early decline, plasma beta-hydroxybutyrate rose to levels that were near the control levels by Days 2 through 4, but fell to near the initial levels by Day 5 and thereafter. The results suggest that the spiny dogfish, a species that consumes reliable and abundant amounts of protein and lipid, but little carbohydrate, and that may feed as seldom as every 2 weeks, uses ketone bodies (beta-hydroxybutyrate and presumably acetoacetate) as primary fuels. Glucose is apparently synthesized by gluconeogenesis, alanine and lactate are probable substrates, and ketone bodies are the likely sources of energy. The evidence supports the hypothesis that glucose is synthesized to maintain muscle glycogen reserves that are used for anaerobic glycolysis when the animal is engaged in predation, escape, or other circumstances that dictate vigorous swimming. If other tissues and organs require glucose as a fuel source, the amounts must be exceptionally small.


Interaction of apolipoprotein A-II with recombinant HDL containing egg phosphatidylcholine, unesterified cholesterol and apolipoprotein A-I.
The preparation of discoidal, recombinant HDL (r-HDL) containing various phospholipids, apolipoproteins and a range of concentrations of unesterified cholesterol has been reported by several investigators. The present study describes the preparation of r-HDL containing both apolipoprotein (apo) A-I and apo A-II. r-HDL with 100:1 (mol:mol) egg PC.apo A-I and 0 (Series I), 5 (Series II) or 10 (Series III) mol% unesterified cholesterol were prepared by the cholate dialysis method. The resulting complexes had a Stokes' radius of 4.7 nm and contained two molecules of apo A-I per particle. When the r-HDL (2.0 mg apo A-I) were supplemented with 1.0 mg of apo A-II, one of the apo A-I molecules was replaced by two molecules of apo A-II. This modification was not accompanied by a loss of phospholipid, nor by major change in particle size. The addition of 2.5 or 4.0 mg of apo A-II resulted in the displacement of both apo A-I molecules from a proportion of the r-HDL and the formation of smaller particles (Stokes' radius 3.9 nm), which contained half the original number of egg PC molecules and three molecules of apo A-II. The amount of apo A-I displaced was dependent on the concentration of unesterified cholesterol in the r-HDL: when 2.5 mg of apo A-II was added to the Series I, II and III r-HDL, 44, 60 and 70%, respectively, of the apo A-I was displaced. Addition of 4.0 mg of apo A-II did not promote further displacement of apo A-I from any of the r-HDL. By contrast, the association of apo A-II with r-HDL was independent of the concentration of unesterified cholesterol and was a linear function of the amount of apo A-II which had been added. It is concluded that (1), the structural integrity of egg PC.unesterified cholesterol.apo A-I r-HDL, which contain two molecules of apo A-I, is not affected when one of the apo A-I molecules is replaced by two molecules of apo A-II; (2), when both apo A-I molecules are replaced by apo A-II, small particles which contain three molecules of apo A-II are formed; and (3), the displacement of apo A-I from r-HDL is facilitated by the presence of unesterified cholesterol in the particles.


Cloning and characterization of human Golgi phosphoprotein 2 gene (GOLPH2/GP73/GOLM1) promoter.
Human Golgi phosphoprotein 2 gene (also known as GOLPH2, GP73 or GOLM1) encodes an epithelial-specific Golgi membrane protein which can be induced by virus infection. It is also overexpressed in a number of tumors and is currently considered as an early diagnosis marker for hepatocellular carcinoma. However, little is known about how GOLPH2 is dysregulated in these disease conditions and the functional implications of its overexpression. The aim of this study is to investigate human GOLPH2 regulation mechanisms. We cloned a 2599 bp promoter fragment of GOLPH2 and found it maintained epithelial specificity. By deletion analysis, a repressive region (-864 to -734 bp), a positive regulatory region (-734 to -421 bp) and a core promoter region (-421 to -79 bp) were identified. Sequence analysis revealed that GOLPH2 core promoter was devoid of canonical TATA element and classified as a TATA-less promoter. Adenoviral early region 1A (E1A) was able to activate GOLPH2 and the CtBP interaction domain of E1A was sufficient but not required for activation. A GC-box motif (-89 to -83 bp) in GOLPH2 core promoter region partly mediated E1A transactivation. These results delineated regulatory regions and functional element in GOLPH2 promoter, elucidated adenoviral E1A stimulation mechanisms and provided insight into GOLPH2 functions.


Mesenchymal stem cells from periapical lesions modulate differentiation and functional properties of monocyte-derived dendritic cells.
Immunoregulatory mechanisms within periapical lesions (PLs) are as of yet unexplored. Considering the crucial role of DCs in controlling the immune response within PLs, the immunomodulatory properties of mesenchymal stem cells (MSCs), and the colocalization of MSCs and DCs in situ, we wondered whether MSCs from PLs modulate the development and functions of DCs. Using a model of monocyte-derived DCs, we showed that PL-MSCs inhibited differentiation of DCs via soluble factors, of which IL-6 had a minor effect, but did not impair their subsequent maturation induced by pro-inflammatory cytokines. However, upon maturation such DCs favored the production of Th2/Th17 cytokines by allogenic CD4(+) lymphocytes in coculture, compared with mature DCs differentiated without PL-MSCs. PL-MSC-differentiated DCs, cultivated with pro-inflammatory cytokines and PL-MSCs, although phenotypically mature, exhibited poor allostimulatory activity, induced anergy, Th2 polarization, differentiation of suppressive CD4(+) CD25(high) CD39(+) Treg-cell subsets via IDO-1-, ILT-3-, and ILT-4-dependent mechanisms, and increased production of TGF-β in the coculture. In contrast, DCs cultivated with PL-MSCs only during maturation stimulated proliferation and Th1 polarization of CD4(+) T cells in an IL-12-independent manner. In conclusion, PL-MSCs significantly modulate the development and functions of DCs, depending on the phase of DCs development during which the interaction occurs.


Early (stage A) prostatic cancer. VI. A critical look at the follow-up.
The 4 year follow-up of an original series of 100 patients treated by subtotal prostatectomy and analysed on histopathological grounds is presented. 87 out of 100 were traced and were in the following groups: 37/45 A1, 27/29 A2, 10/12 A3, 13/14 benign prostatic hyperplasia. No therapy was performed. All three patients who died of prostatic carcinoma fitted into substage A3, all three patients living with metastases fitted into A1. The progression observed is significant when related to the brief interval of time, the size of the prostatic microcarcinoma and the histological grade (well differentiated tubular carcinomas in 5 out of 6 cases). Prostatectomy with capsulectomy is strongly recommended in order to prevent progression.


Skin Cancer.
Skin cancer accounts for most malignancies across the globe. They are primarily divided into melanoma and nonmelanoma skin malignancies. Nonmelanoma skin cancer includes basal cell carcinoma and squamous cell carcinoma. Fair skin and chronic ultraviolet B exposure are the most important risk factors. Primary prevention is achieved by avoiding sun exposure and tanning beds.


Gallium-67 imaging in the diagnosis of blastomycosis.
67Ga imaging was used in a group of 13 untreated patients with documented South American blastomycosis in an attempt to obtain additional information on the site and extent of the lesions. All diagnoses were confirmed by demonstration of the fungi in the lesions or body fluids. Serologic tests were performed on all patients. 67Ga imaging was correlated with X-ray in all patients, and with bone imaging when bony lesions were suspected. 67Ga imaging was positive for all lesions detected on chest X-ray and by clinical evaluation. More importantly, lesions clinically unsuspected and missed by other diagnostic techniques were shown on 67Ga imaging: bony and hepatic lesions in three patients; a brain lesion in one patient and parotid gland involvement in five patients. There was good agreement between 67Ga imaging and clinical and laboratory data, scintigraphy being more sensitive. Follow-up studies after specific therapy demonstrated decreased uptake in the lungs and other sites. 67Ga imaging is a useful tool for detection of suspected and unsuspected lesions of blastomycosis and to evaluate the response of the disease to specific therapy.


Reduced expression of CD47 during murine red blood cell (RBC) senescence and its role in RBC clearance from the circulation.
Almost 2 percent of murine blood red blood cells (RBCs) are destroyed each day and are replaced by fresh RBCs generated through the process of erythropoiesis. RBCs to be destroyed are phagocytosed by macrophages in the reticuloendothelial system, especially in the spleen. CD47 molecules on RBCs may regulate the susceptibility of RBC to destruction by phagocytosis because its recognition by inhibitory receptor (signal regulatory protein alpha) on macrophages sends a negative signal, which if sufficiently strong, may abort the phagocytic response altogether. The aim of this study was to investigate whether age-dependent changes in CD47 expression on circulating RBCs have a role in destruction of senescent RBCs by macrophages. A two-step in vivo biotinylation method for labeling mouse RBCs in vivo was used to track the CD47 expression levels as well as the turnover of circulating RBCs of defined age groups. Our results indicate that CD47 expression levels decrease on circulating RBCs throughout their life span in circulation. The oldest RBCs in circulation have 30 percent lower mean expression of CD47 than the youngest RBCs. Depletion of macrophages by administration of clodronate-loaded liposomes resulted in a significant decrease in the mean expression of CD47 on RBCs of all age groups and a significant accumulation of senescent RBCs in blood and spleen. A decrease in mean expression of CD47 and accumulation of senescent RBCs in macrophage-depleted mice were significantly higher for spleen RBCs compared to blood RBCs. Our results provide supportive evidence for a role of decreasing CD47 expression on aging circulating RBCs in their destruction by macrophages.


Effect of cimetidine on the pharmacokinetics of temafloxacin.
Cimetidine, a widely prescribed histamine H2-receptor antagonist, is known to interact with a variety of drugs; consequently, it is important to determine its potential for interaction with any new drug. The interaction between cimetidine and a new quinolone, temafloxacin, has been examined in an open randomised 2-period crossover study in 12 healthy adults. Half the volunteers received cimetidine 400mg 3 times daily for 8 days. On day 5, a single temafloxacin 400mg dose was administered. No other drugs were allowed during this period. The other volunteers did not receive cimetidine (or any other drugs) for an 8-day period except for temafloxacin 400mg on day 5. Blood and urine were sampled serially after temafloxacin administration and daily thereafter until the last day of the study. A 2-week washout period preceded crossover. Pharmacokinetic analyses showed that cimetidine did not affect the rate or extent of temafloxacin absorption, as evidenced by unchanged peak plasma concentration, time to peak plasma concentration, and terminal-phase volume of distribution. However, renal and total clearance values for temafloxacin were reduced by 19%, and elimination half-life and area under the concentration vs time curve were increased in the presence of cimetidine. The most likely mechanism underlying these effects is inhibition by cimetidine of tubular secretion of temafloxacin in the kidney. The lack of clinically significant adverse effects and the small magnitude of the reduction in temafloxacin clearance suggest that the interaction is of little clinical consequence.


Angiographic and embryologic considerations in five cases of middle cerebral artery fenestration.
Five cases of unilateral middle cerebral artery fenestration were observed during the prospective evaluation of 1466 consecutive cerebral angiograms (0.43% of 1170 patients) between January 1999 and July 2001. In each case, an early branching temporopolar artery was seen to arise from the inferior limb of the fenestrated segment. This finding suggests that early branching temporopolar arteries may participate in the formation of middle cerebral artery fenestration by interfering with the normal fetal development of the middle cerebral artery.


Ocular trauma and other catastrophes.
Trauma is one of the leading causes of monocular blindness worldwide. Recognizing, evaluating, and managing vision-threatening injuries, such as globe rupture, retrobulbar hemorrhage, intraocular foreign bodies, and hyphemas, are paramount for emergency physicians to reduce the risk of vision loss and other complications. Other emergent ocular complaints, such as acute-angle closure glaucoma, and retinal detachment also require temporizing emergency management that can reduce complications while awaiting definitive specialty care. Understanding these conditions and evidence-based management will enable the emergency physician to provide the most up-to-date standard of care.


Ranitidine: A Proposed Mechanistic Rationale for NDMA Formation and a Potential Control Strategy.
The formation of N-nitrosodimethylamine (NDMA) in ranitidine hydrochloride drug substance (DS) and drug products has attracted considerable attention over the last few years. The drug structure is unusual in that it contains a vinyl nitro moiety. Although a variety of studies have been carried out to understand how NDMA is formed in the DS solids, a mechanistic description of NDMA formation has remained elusive. A new mechanistic view of NDMA formation is detailed here. Autoxidation of ranitidine can rationalize nitrite ion and dimethylamine liberation from ranitidine. The subsequent nitrosation is argued to be due to conversion of nitrite ion to the gas phase nitrosating agent nitrosyl chloride, NOCl. Oxygen scavenging packaging systems should be able to stop the autoxidation, and thus shut down the nitrite release from ranitidine. Without nitrite release NDMA cannot form. This may provide a practical means to stabilize ranitidine DS and solid dosage formulations against NDMA formation.


Hemoglobin Cochin-Port-Royal: consequences of the replacement of the beta chain C-terminal by an arginine.
Hemoglobin Cochin Port-Royal beta 146 (HC3) His yields Arg is the second example in which the beta C-terminal residue is replaced. Owing to the known importance of His beta 146 in the co-operative effects of hemoglobin, the functional properties of this variant were carefully studied. It had a normal Hill coefficient but a reduced alkaline Bohr effect. However, the reduction in Bohr effect is less than the halving predicted from previous mutants and modified hemoglobins.


Enhanced cycle life of starter lighting ignition (SLI) type lead-acid batteries with electrolyte modified by ionic liquid.
The aim of the presented work was to improve the lifetime of lead-acid SLI (starting, lighting and ignition) batteries through electrolyte modification with ionic liquids. The conducted research included the synthesis and determination of the influence of di(hexadecyldimethylammonium) and di(octadecyldimethylammonium) sulphates on the basic parameters (capacity, cranking performance) of the starter battery as well as parameters affecting its lifetime (dynamic charge acceptance, corrosion, water consumption). It has been shown that the addition of these compounds increases corrosion resistance and reduces water consumption, resulting in an increase in cyclic durability by up to 36%. The improvement is associated with the absorption of ionic liquid molecules into the mass of lead(ii) sulphate, which was confirmed by physicochemical and electrochemical studies.


Formalization of gene regulation knowledge using ontologies and gene ontology causal activity models.
Gene regulation computational research requires handling and integrating large amounts of heterogeneous data. The Gene Ontology has demonstrated that ontologies play a fundamental role in biological data interoperability and integration. Ontologies help to express data and knowledge in a machine processable way, which enables complex querying and advanced exploitation of distributed data. Contributing to improve data interoperability in gene regulation is a major objective of the GREEKC Consortium, which aims to develop a standardized gene regulation knowledge commons. GREEKC proposes the use of ontologies and semantic tools for developing interoperable gene regulation knowledge models, which should support data annotation. In this work, we study how such knowledge models can be generated from cartoons of gene regulation scenarios. The proposed method consists of generating descriptions in natural language of the cartoons; extracting the entities from the texts; finding those entities in existing ontologies to reuse as much content as possible, especially from well known and maintained ontologies such as the Gene Ontology, the Sequence Ontology, the Relations Ontology and ChEBI; and implementation of the knowledge models. The models have been implemented using Protégé, a general ontology editor, and Noctua, the tool developed by the Gene Ontology Consortium for the development of causal activity models to capture more comprehensive annotations of genes and link their activities in a causal framework for Gene Ontology Annotations. We applied the method to two gene regulation scenarios and illustrate how to apply the models generated to support the annotation of data from research articles.


Functional characterization of the sea urchin sns chromatin insulator in erythroid cells.
Chromatin insulators are regulatory elements that determine domains of genetic functions. We have previously described the characterization of a 265 bp insulator element, termed sns, localized at the 3' end of the early histone H2A gene of the sea urchin Paracentrotus lividus. This sequence contains three cis-acting elements (Box A, Box B, and Box C + T) all needed for the enhancer-blocking activity in both sea urchin and human cells. The goal of this study was to further characterize the sea urchin sns insulator in the erythroid environment. We employed colony assays in human (K562) and mouse (MEL) erythroid cell lines. We tested the capability of sns to interfere with the communication between the 5'HS2 enhancer of the human beta-globin LCR and the gamma-globin promoter. We found that the sns sequence displays directional enhancer-blocking activity. By the use of antibodies against known DNA binding proteins, in electrophoretic mobility shift assays, we demonstrated the binding of the erythroid-specific GATA-1 and the ubiquitous Oct-1 and Sp1 transcription factors. These factors bind to Box A, Box B, and Box C + T, respectively, in both K562 and MEL nuclear extracts. These results may have significant implications for the conservation of insulator function in evolutionary distant organisms and may prove to be of practical benefit in gene transfer applications for erythroid disorders such as hemoglobinopathies and thalassemias.


Progress and Prospects of Inorganic Solid-State Electrolyte-Based All-Solid-State Pouch Cells.
All-solid-state batteries have piqued global research interest because of their unprecedented safety and high energy density. Significant advances have been made in achieving high room-temperature ionic conductivity and good air stability of solid-state electrolytes (SSEs), mitigating the challenges at the electrode-electrolyte interface, and developing feasible manufacturing processes. Along with the advances in fundamental study, all-solid-state pouch cells using inorganic SSEs have been widely demonstrated, revealing their immense potential for industrialization. This review provides an overview of inorganic all-solid-state pouch cells, focusing on ultrathin SSE membranes, sheet-type thick solid-state electrodes, and bipolar stacking. Moreover, several critical parameters directly influencing the energy density of all-solid-state Li-ion and lithium-sulfur pouch cells are outlined. Finally, perspectives on all-solid-state pouch cells are provided and specific metrics to meet certain energy density targets are specified. This review looks to facilitate the development of inorganic all-solid-state pouch cells with high energy density and excellent safety.


Clinical Characteristics and Analysis of Lung Cancer- Associated Acute Ischemic Stroke.
The incidence of lung cancer and acute ischemic stroke remains high in recent years, both of which occur mostly in people over 60 years old. In the present study, we aimed to further clarify the pathogenesis of lung cancer-associated acute ischemic stroke (LCA-AIS) by comparing and analyzing clinical characteristics of stroke patients with or without lung cancer. A total of 51 patients with lung cancer were selected as the case group (LCSG), and 78 patients without cancer history were adopted as the control group (SG). The data collected in this study included sex, age, traditional cerebrovascular disease risk factors (TCDRFs), blood test index, imaging findings, etiological typing, and prognosis evaluation. SPSS21.0 software was used for statistical analysis. Normally distributed data were analyzed by t-test, and count data were analyzed by chi-square test or exact probability method. P < 0.05 was considered statistically significant. In the case group, the levels of plasma D-dimer, fibrinogen degradation products (FDPs) and NIHSS, as well as the mRS score and mortality of patients, were higher, while the levels of RBC, Hb and Hcy were lower compared with the control group. Imaging findings showed that multivessel involvement was more common in the case group, and the infarcts were more likely to be multiple and involved in both the anterior and posterior circulations. The TOAST classification of LCSG was dominated by stroke of undetermined etiology (SUE) and stroke of other determined etiology (SOE). Statistical analysis showed that the patients were more likely to suffer from acute ischemic stroke within 1 year after the diagnosis of lung cancer (41 cases, 80.39%). Hypercoagulability and acute multiple brain infarcts were more common in patients with LCA-AIS, and hypoproteinemia and hyponatremia were more likely to occur in these patients, leading to worse prognosis. Patients were most likely to have a stroke within 1 year after the diagnosis of lung cancer.


Porokeratotic eccrine ostial and dermal duct nevus. Report of a case of adult onset.
We describe the case of a 38-year-old woman with adult-onset porokeratotic eccrine ostial and dermal duct nevus. The patient had a 9-year history of multiple keratotic papules and comedo-like pits on the medial border of the left foot. Light- and electron microscopic studies showed multiple cornoid lamella-like parakeratotic columns, which invariably were associated with hyperplastic eccrine ostia and distal sweat ducts. It is concluded that this entity, the first reported case of adult-onset porokeratotic eccrine ostial and dermal duct nevus, is a variant of the congenital form described previously by Abell and Read and by Aloi and Pippione.


Small renal caliceal calculi as a cause of pain.
We report 7 cases of long-standing lumbar or flank pain associated with small renal calculi located peripherally in a calix and no obvious obstruction. After stone removal by percutaneous puncture the patients were relieved of the pain.


Viability of brown trout embryos positively linked to melanin-based but negatively to carotenoid-based colours of their fathers.
'Good-genes' models of sexual selection predict significant additive genetic variation for fitness-correlated traits within populations to be revealed by phenotypic traits. To test this prediction, we sampled brown trout (Salmo trutta) from their natural spawning place, analysed their carotenoid-based red and melanin-based dark skin colours and tested whether these colours can be used to predict offspring viability. We produced half-sib families by in vitro fertilization, reared the resulting embryos under standardized conditions, released the hatchlings into a streamlet and identified the surviving juveniles 20 months later with microsatellite markers. Embryo viability was revealed by the sires' dark pigmentation: darker males sired more viable offspring. However, the sires' red coloration correlated negatively with embryo survival. Our study demonstrates that genetic variation for fitness-correlated traits is revealed by male colour traits in our study population, but contrary to predictions from other studies, intense red colours do not signal good genes.


Workload for radiologists during on-call hours: dramatic increase in the past 15 years.
The objective of this study is to investigate the workload for radiologists during on-call hours and to quantify the 15-year trend in a large general hospital in Western Europe. Data regarding the number of X-ray, ultrasound and computed tomography (CT) studies during on-call hours (weekdays between 6.00 p.m. and 7.00 a.m., weekends, and national holidays) between 2006 and 2020 were extracted from the picture archiving and communication system. All studies were converted into relative value units (RVUs) to estimate the on-call workload. The Mann-Kendall test was performed to assess the temporal trend. The total RVUs during on-call hours showed a significant increase between 2006 and 2020 (Kendall's tau-b = 0.657, p = 0.001). The overall workload in terms of RVUs during on-call hours has quadrupled. The number of X-ray studies significantly decreased (Kendall's tau-b = - 0.433, p = 0.026), whereas the number of CT studies significantly increased (Kendall's tau-b = 0.875, p < 0.001) between 2006 and 2020. CT studies which increased by more than 500% between 2006 and 2020 are CT for head trauma, brain CTA, brain CTV, chest CT (for suspected pulmonary embolism), spinal CT, neck CT, pelvic CT, and CT for suspected aortic dissection. The number of ultrasound studies did not change significantly (Kendall's tau-b = 0.202, p = 0.298). The workload for radiologists during on-call hours increased dramatically in the past 15 years. The growing amount of CT studies is responsible for this increase. Radiologist and technician workforce should be matched to this ongoing increasing trend to avoid potential burn-out and to maintain quality and safety of radiological care.


IFI16 restoration in hepatocellular carcinoma induces tumour inhibition via activation of p53 signals and inflammasome.
Interferon-inducible 16 (IFI16) is known to involve in p53-dependent tumour suppression and also the formation of inflammasome, which function, however, remains controversy during carcinogenesis as a pattern recognition receptor for tumour death-derived free DNA. In this study, we investigated the anti-tumour role of IFI16 in hepatocellular carcinoma (HCC). Hepatocellular carcinoma tissues (n = 20) and corresponding non-neoplastic tissues (n = 20) were collected to determine the expression of IFI16. After the transfection of pcDNA3.1-IFI16 into Huh7 and SMMC7721 cells in vitro, the influence of IFI16 overexpression on cell vitality, colony formation, apoptosis and migration were analysed. The role effect of IFI16 in vivo was further investigated. The expression of IFI16 was significantly decreased in tumour tissues and cell lines. Overexpression of IFI16 induced decrease of cell vitality, colony formation and increased apoptosis with impaired ability of migration. Mechanistically, IFI16 could activate p53 at Ser15 to up-regulate the p21WAF1/CIP1 level to inhibit tumour growth and migration, which was restored by the p53 inhibitor Pifithrin-α (20 μmol/L). Moreover, IFI16-induced tumour cell death promoted the recruitment of inflammasome complex to enhance tumour inhibition, but the caspase-1 inhibitor Ac-YVAD-CMK (50 μmol/L) could suppress this process in HCC. The results in vivo indicated that restored expression of IFI16 in tumour cells effectively promote tumour regression, which could be partly abrogated by the inhibition of activation of p53 signals or induced inflammasome. IFI16 is a tumour suppressor in HCC via activation of p53 signals and inflammasome.