Repeated liver stiffness measurement compared with paired liver biopsy in patients with non-alcoholic fatty liver disease.
The value of repeated liver stiffness measurement (LSM) in non-alcoholic fatty liver disease (NAFLD) has not been shown before. A longitudinal study of biopsy-proven NAFLD patients was conducted at the Asian tertiary hospital from November 2012 to January 2017. Patients with paired liver biopsies and LSM were followed prospectively for liver-related and non-liver related complications, and survival. The data for 113 biopsy-proven NAFLD patients (mean age 51.3 ± 10.6 years, male 50%) were analyzed. At baseline, advanced fibrosis based on histology and LSM was observed in 22 and 46%, respectively. Paired liver biopsy and LSM at 1-year interval was available in 71 and 80% of patients, respectively. High-risk cases (defined as patients with advanced fibrosis at baseline who had no fibrosis improvement, and patients who developed advanced fibrosis on repeat assessment) were seen in 23 and 53% of patients, based on paired liver biopsy and LSM, respectively. Type 2 diabetes mellitus was independently associated with high-risk cases. The median follow-up was 37 months with a total follow-up of 328 person-years. High-risk cases based on paired liver biopsy had significantly higher rates of liver-related complications (p = 0.002) but no difference in other outcomes. High-risk patients based on paired LSM had a significantly higher rate of liver-related complications (p = 0.046), cardiovascular events (p = 0.025) and composite outcomes (p = 0.006). Repeat LSM can predict liver-related complications, similar to paired liver biopsy, and may be useful in identifying patients who may be at an increased risk of cardiovascular events. Further studies in a larger cohort and with a longer follow-up should be carried out to confirm these observations.


Central nervous system infection due to Cryptococcus gattii sensu lato in India: Analysis of clinical features, molecular profile and antifungal susceptibility.
Cryptococcus gattii species complex has evolved as a pathogen in the last two decades causing infection among both immunocompetent and immunocompromised hosts. We aimed to analyse the clinical features of CNS infection caused by C. gattii sensu lato, molecular and antifungal susceptibility profile of this pathogen. Cases diagnosed to have CNS cryptococcosis were included in the study. Cryptococcus recovered from patient's specimen was identified by standard protocol. Species confirmation, mating type and molecular type determination were performed by PCR based methods. Antifungal susceptibility was tested in VITEK2C to amphotericin B, 5-flucytosine, fluconazole and voriconazole. Among 199 cases, 20 (10%) were due to C. gattii, comprising of 75% cryptococcal meningitis and 25% cryptococcoma cases. Young adult males were commonly affected. Headache and vomiting were prominent symptoms and 50% were immunocompromised. Among the isolates, 75%, 20% and 5% were C. tetragattii, C. gattii sensu stricto and C. bacillisporus respectively and all had mating type α. Four (20%) isolates of C. tetragattii and the only isolate of C. bacillisporus were resistant to fluconazole. The most common species isolated from south India is C. tetragattii. The study contributes to the epidemiology of C. gattii and reiterates the need for genotyping and antifungal susceptibility testing.


[Short-segment pedicle screw fixation combined with vertebroplasty for the treatment of Kümmell disease with kyphosis].
To explore the clinical effects of posterior short-segment pedicle screw internal fixation combined with vertebroplasty for the treatment of Kümmell disease with kyphosis. Twenty-four patients with Kümmell disease complicated with kyphosis treated by posterior short-segment pedicle screw internal fixation combined with vertebroplasty from January 2016 to December 2018 were retrospectively analyzed, including 6 males and 18 females, aged 63 to 85 (73.1±6.5) years old. The clinical effect was evaluate by visual analogue scale (VAS), Oswestry Disability Index (ODI), the anterior height of injured vertebral body, and the sagittal Cobb angle of the affected segment beforeoperation, at 3 days and final follow up after operation. And the surgical complications were observed. All 24 patients were followed up from 12 to 24 months with an average of (15.5±3.2) months. The VAS score was decreased from 5.21±1.06 preoperatively to 2.38±0.58 at 3 days postoperatively and 1.71±0.75 at final follow-up;ODI was decreased from (50.4±13.5)% preoperatively to (20.9±8.0)% at 3 days postoperatively and (16.7±9.6)% at final follow-up;the anterior height of injured vertebral body was restored from (8.0±4.2) mm before surgery to (18.1±5.0) mm at 3 days after surgery and (16.8±5.1) mm at final follow up;the sagittal Cobb angle of affected segment was decreased from (19.5±6.3)° preoperatively to (7.6±2.1)° at 3 days after surgery and(8.4±1.7)° at final follow-up. VAS, ODI, anterior height of injured vertebral body, and sagittal Cobb angle of affected segment were significantly improved at 3 days after operation and at final follow-up (P<0.05). Two patients had complications, including asymptomaticcement leakage in 1 patient and superficial wound infection in 1 patient. Posterior short-segment pedicle screw internal fixation combined with vertebroplasty for the treatment of Kümmell disease with kyphosis has relatively small surgical trauma, excellent clinical results, good vertebral height recovery, satisfactory correction of kyphotic angle, and fewer complications, etc. It is a safe and effective surgical method to treat Kümmell disease with kyphosis.


Co-evolutionary analysis of insulin/insulin like growth factor 1 signal pathway in vertebrate species.
Insulin/insulin like growth factor 1 signaling pathway plays an essential role on the regulation of variant, interrelated and fundamental processes, such as metabolism, growth, reproduction and aging. This pathway is highly conserved during evolution. We analyzed eight vertebrate species' insulin/insulin like growth factor 1 pathway and examined the co-evolutionary relationship between different protein components by quantifying similarity of phylogenetic trees. The collected species include Takifugu rubripes, Danio rerio, Xenopus laevis, Gallus gallus, Mus musculus, Rattous norvegicus, Pan troglodytes and Homo sapiens. Results show that interacting proteins in this pathway share highly co-evolutionary relationship while contrarily the proteins without interaction have low co-evolutionary relationship. We also predict some receptor and ligand partners enjoy highly binding potential in corresponding species' pathway.


Pharmacoproteomic investigation into antidepressant response in two mouse inbred strains.
In this study, we present a pharmacoproteomic investigation of response to antidepressants two inbred strains. Our aim was to uncover molecular mechanisms underlying antidepressant action and identify new biomarkers to determine therapeutic response to two antidepressants with proven efficacy in the treatment of depression but divergent mechanisms of action. Mice were treated with the pro-noradrenergic drug nortriptyline, the pro-serotonergic drug escitalopram or saline. Quantitative proteomic analyses were undertaken on hippocampal tissue from a study design that used two inbred mouse strains, two depressogenic protocols and a control condition, (maternal separation, chronic mild stress, control), two antidepressant drugs and two dosing protocols. The proteomic analysis was aimed at the identification of specific drug-response markers. Complementary approaches, 2DE and isobaric tandem mass tagging (TMT), were applied to the selected experimental groups. To investigate the relationship between proteomic profiles, depressogenic protocols and drug response, 2DE and TMT data sets were analysed using multivariate methods. The results highlighted significant strain- and stress-related differences across both 2DE and TMT data sets and identified the three gene products involved in serotonergic (PXBD5, YHWAB, SLC25A4) and one in noradrenergic antidepressant action (PXBD6).


[Indication and use of adjuvant analgesics on chronic pain].
Adjuvant analgesics including antidepressant and anticonvulsant are highly useful toward intractable chronic pains, especially neuropathic pain. In recent years, selection of the first line treatment is done by balancing between efficacies of candidate drugs and their potential adverse effects. While evaluating the efficacy of a drug is easy when it is used alone; however, it may not be always so if multiple drugs are used concurrently.


Transcriptional Regulation of Human UDP-Glucuronosyltransferase 2B10 by Farnesoid X Receptor in Human Hepatoma HepG2 Cells.
Little is known about transcriptional regulators of UDP-glucuronosyltransferase 2B10 (UGT2B10), an enzyme known to glucuronidate many chemicals and drugs such as nicotine and tricyclic antidepressants. Here, we uncovered that UGT2B10 was transcriptionally regulated by farnesoid X receptor (FXR), the bile acid sensing nuclear receptor. GW4064 and chenodeoxycholic acid (two specific FXR agonists) treatment of HepG2 cells led to a significant increase in the mRNA level of UGT2B10. The treated cells also showed enhanced glucuronidation activities toward amitriptyline (an UGT2B10 probe substrate). In reporter gene assays, the extent of UGT2B10 activation by the FXR agonists was positively correlated with the amount of cotransfected FXR. Consistently, knockdown of FXR by shRNA attenuated the induction effect on UGT2B10 expression. Furthermore, a combination of electrophoretic mobility shift assay and chromatin immunoprecipitation showed that the FXR receptor trans-activated UGT2B10 through its specific binding to the -209- to -197-bp region (an IR1 element) of the UGT2B10 promoter. In summary, our results for the first time established FXR as a transcriptional regulator of human UGT2B10.


An improved method for measurement of testosterone in human plasma and saliva by ultra-performance liquid chromatography-tandem mass spectrometry.
The aim of the study was to develop and validate a practical assay of clinically relevant testosterone levels in human plasma and saliva. We performed ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis on Atlantis dC18 steel column using a mobile phase of 2-mM ammonium acetate and acetonitrile (20:80, v: v) that was delivered at 0.3 ml/min. After adding d3-testosterone as an internal standard (IS), we extracted plasma and salivary samples with methyl tert-butyl ether. Mass spectrometry was performed in electrospray positive-ion mode. Targeted ion transitions were examined at m/z 289.18 → 97.04 and 292.24 → 97.04 for testosterone and IS, respectively. We validated the method according to the US Food and Drug Administration guidelines. Elution times for testosterone and IS were both around 1.35 min. Testosterone level was linearly associated (r 2 = 0.9975 and 0.9958) with peak area ratio of testosterone to IS between 0.5-50 ng/ml and 10-400 pg/ml in plasma and saliva, respectively. The coefficient of variation and bias were ≤12.6% and ≤±12.1% in plasma and ≤10.2% and ≤±5.3% in saliva. The extraction recovery of testosterone was ≥92% from plasma and ≥94% from saliva. Testosterone was stable (≥91%) for 24 h at room temperature and for 8 weeks at -20°C in both plasma and salivary samples. We report a simple, validated, UPLC-MS/MS assay that can be used to determine clinically relevant levels of testosterone in human plasma and saliva.


Nature of heavy quasiparticles in magnetically ordered heavy fermions UPd(2)Al(3) and UPt(3).
The optical conductivity of the heavy fermions UPd(2)Al(3) and UPt(3) has been measured in the energy range from 0.04 to 5 meV. In both compounds a well pronounced pseudogap of less than 1 meV develops in the optical response at low temperatures; we relate this to the antiferromagnetic ordering. From the energy dependence of the effective mass and scattering rate we conclude that the enhancement of the mass mainly occurs below the energy which is related to magnetic correlations between the local magnetic moments and the itinerant electrons. This implies that the magnetic order in these compounds is the prerequisite to the formation of the heavy quasiparticles and eventually to superconductivity.


Note: Portable rare-earth element analyzer using pyroelectric crystal.
We report a portable rare-earth element analyzer with a palm-top size chamber including the electron source of a pyroelectric crystal and the sample stage utilizing cathodoluminescence (CL) phenomenon. The portable rare-earth element analyzer utilizing CL phenomenon is the smallest reported so far. The portable rare-earth element analyzer detected the rare-earth elements Dy, Tb, Er, and Sm of ppm order in zircon, which were not detected by scanning electron microscopy-energy dispersive X-ray spectroscopy analysis. We also performed an elemental mapping of rare-earth elements by capturing a CL image using CCD camera.


Transporter protein and drug resistance of Trypanosoma.
Trypanosoma infection is one of the most important infections in livestock and humans. One of the main problems of its therapeutic control and treatment is the resurgence of drug resistance. One of the most studied causes of such resistance is the function of its adenosine transporter gene. A trypanosomal gene TbAT1 from Trypanosoma brucei has been cloned in yeast to demonstrate its function in the transport of adenosine and trypanocidal agents. Drug resistant trypanosomes showed a defective TbAT1 variant; furthermore, deletion of the gene and set point mutations in the transporter gene has been demonstrated from isolates from relapse patients. The molecular understanding of the mechanism of action trypanocidal agents and function of transporter gene can lead to control of drug resistance of Trypanosomes.


Antimicrobial activity of antiproteinases.
Low-molecular-mass neutrophil elastase inhibitors have been shown to be important in the control of lung inflammation. In addition to inhibiting the enzyme neutrophil elastase, these low-molecular-mass compounds (10 kDa) have been shown to have other activities. For example, secretory leucocyte proteinase inhibitor (SLPI) and elastase-specific inhibitor/SKALP (skin-derived antileucoproteinase)/elafin have also been shown to have "defensin"-like antimicrobial activities. Indeed, these inhibitors have antimicrobial properties in vitro against bacteria, fungi and, potentially, HIV. In addition, we have shown, using an adenovirus-mediated gene transfer overexpression strategy, that elafin is also active against Pseudomonas aeruginosa infection in mice in vivo. The mechanism of action is currently under investigation. In addition to these direct or indirect effects on microbes, it has been shown that lipopolysaccharide is able to up-regulate SPLI production in macrophages in vitro, and that the addition of recombinant SLPI to human monocytes or the transfection of macrophages with SPLI can down-regulate pro-inflammatory mediators such as tumour necrosis factor, presumably to limit self-damaging excessive inflammation. Using viral gene transfer vectors, we are currently investigating the potential of these inhibitors in various models of inflammation in vivo.


Optic nerve sheath diameter and lumbar puncture opening pressure in nontrauma patients suspected of elevated intracranial pressure.
The purpose of this study was to determine if patients with nontraumatic causes of elevated intracranial pressure (ICP) could be identified by ultrasound measurement of optic nerve sheath diameter (ONSD). It was hypothesized that an ONSD greater than or equal to 5 mm would identify patients with elevated ICP. This was a prospective observational trial comparing ONSD with ICP measured by opening pressure manometry on lumbar puncture (LP). The cohort consisted of a convenience sample of adult patients presenting to the emergency department, requiring LP. The ONSD measurement was performed before computed tomography and LP. The physician performing the LP was blinded to the result of the ONSD measurement. An opening pressure on manometry of greater than or equal to 20 cm H2O and an ONSD greater than or equal to 5 mm were considered elevated. Fifty-one patients were included in our study, 24 (47%) with ICP greater than or equal to 20 cm H2O and 27 (53%) with ICP less than 20 cm H2O. The sensitivity of ONSD greater than or equal to 5 for identifying elevated ICP was 75% (95% confidence interval, 53%-90%) with specificity of 44% (25%-65%). The area under the receiver operator characteristic curve was 0.69 (0.54-0.84), suggesting a relationship between ONSD and ICP. An ONSD greater than or equal to 5 mm was associated with elevated ICP in nontraumatic causes of elevated ICP. Although a relationship exists, a sensitivity of 75% does not make ONSD measurement an adequate screening examination for elevated ICP in this patient population.


Barriers and Facilitators of Telemedicine Among Physicians at a University Hospital.
Background and aims This study explored the perceived barriers and facilitators to the implementation of telemedicine among physicians and estimated and compared the prevalence of telemedicine use among physicians before and during coronavirus disease 2019 (COVID-19). Methods This cross-sectional study was conducted at King Saud University in Riyadh. A convenience sample of 163 physicians working at King Khalid University Hospital (KKUH) completed an online survey. Demographic data, patterns of use, and perceived barriers and facilitators of telemedicine were collected using a 5-point Likert scale. Results Our research showed that 61.3% (n = 100) of the physicians surveyed had used telemedicine in their careers. The prevalence of telemedicine before the onset of COVID-19 was 18.4%, whereas during COVID-19, it increased to 59.5%, which is an increase of 330% (P < .001). Most of the respondents (50.9%; n = 83) used it weekly (27%) or occasionally (23.9%). The most prevalent perceived barrier was technical difficulties (68.7%; n = 112), and the most prevalent perceived facilitator was that telemedicine can reduce unnecessary face-to-face appointments (86.5%; n = 141). Conclusion The use of telemedicine to provide health care is on the rise, especially in the case of emergencies. Different specialties face different facilitators and barriers, and the potential of telemedicine implementation depends on the work environment and the needs of the specialty. Several obstacles need to be overcome before telemedicine becomes a consistently used method for providing health care.


Bis(1-tert-butyl-1H-imidazole-κN3)dichloridocobalt(II).
In the crystal structure of the title compound, [CoCl(2)(C(7)H(12)N(2))(2)], molecular units are formed by coordination of the unsubstituted N atoms of two tert-butyl-substituted imidazole molecules and two chloride ligands, which distinguishes the complex from structures of imidazolium-based dications with tetrachloridocobaltate dianions. There are two crystallographically independent molecules in the asymmetric unit, related by a noncrystallographic inversion centre.


Enrichment of Biscuits with Matcha Green Tea Powder: Its Impact on Consumer Acceptability and Acute Metabolic Response.
Matcha green tea powder (MGTP) is made with finely ground green tea leaves that are rich in phytochemicals, most particularly catechins. Shortbread biscuits were enriched with MGTP and evaluated for consumer acceptability and potential functional health properties. Baking decreased the content of total catechins by 19% compared to dough, although epimerization increased the amount of (+)-gallocatechin gallate at the expense of other catechins such as (-)-epigallocatechin gallate. Consumer acceptability tests using a 9-point hedonic scale showed that consumers preferred enriched biscuits with low content of MGTP (2 g of MGTP 100 g-1 of flour), and an increase of sugar content did not significantly improve the acceptability of MGTP-enriched biscuits. Overall, enrichment of biscuits with MGTP did not significantly affect the postprandial glucose or triglyceride response (area under curve) compared to non-enriched biscuits consumed with water or MGTP drink. Enriching biscuits with Matcha green tea is acceptable to consumers, but may not bring significant postprandial effects.


Adipose derived delivery vehicle for encapsulated adipogenic factors.
Hydrogels derived from adipose tissue extracellular matrix (AdECM) have shown potential in the ability to generate new adipose tissue in vivo. To further enhance adipogenesis, a composite adipose derived delivery system (CADDS) containing single- and double-walled dexamethasone encapsulated microspheres (SW and DW Dex MS) has been developed. Previously, our laboratory has published the use of Dex MS as an additive to enhance adipogenesis and angiogenesis in adipose tissue grafts. In the current work, AdECM and CADDS are extensively characterized, in addition to conducting in vitro cell culture analysis. Study results indicate the AdECM used for the CADDS has minimal cellular and lipid content allowing for gelation of its collagen structure under physiological conditions. Adipose-derived stem cell (ASC) culture studies confirmed biocompatibility with the CADDS, and adipogenesis was increased in experimental groups containing the hydrogel scaffold. In vitro studies of AdECM hydrogel containing microspheres demonstrated a controlled release of dexamethasone from SW and DW formulations. The delivery of Dex MS via an injectable hydrogel scaffold combines two biologically responsive components to develop a minimally, invasive, off-the-shelf biomaterial for adipose tissue engineering. Scientists and doctors have yet to develop an off-the-shelf product for patients with soft tissue defects. Recently, the use of adipose derived extracellular matrix (adECM) to generate new adipose tissue in vivo has shown great promise but individually, adECM still has limitations in terms of volume and consistency. The current work introduces a novel composite off-the-shelf construct comprised of an adECM-based hydrogel and dexamethasone encapsulated microspheres (Dex MS). The hydrogel construct serves not only as an injectable protein-rich scaffold but also a delivery system for the Dex MS for non-invasive application to the defect site. The methods and results presented are a progressive step forward in the field of adipose tissue engineering.


Proton ordering in (NH4)3H(SO4)2 at low-temperature phase transitions.
Single-crystal neutron diffraction was used to investigate the H-atom disorder in triammonium hydrogen disulfate (TAHS), (NH4)3H(SO4)2, below room temperature. Crystal structure analysis of the monoclinic phase III shows an increase of proton ordering with decreasing temperature in the (SO4)H(SO4) dimer. Moreover, the NH4(+) groups on a general position begin ordering in this phase. The monoclinic unit cell of TAHS-IV doubles in the b direction and a slight distortion of SO4(2-) and NH4(+) tetrahedra is observed. The order parameter introduced by Landau was determined for the second-order II/III and III/IV phase transitions from the intensities of the superstructure reflections. TAHS-V has a triclinic space group and the crystal structure seems to be completely ordered according to a structure analysis by single-crystal X-ray diffraction measurements. In addition, the decisive role of the dynamical disorder of different ammonium groups on successive phase transitions is discussed. Additional peaks were observed by X-ray powder diffraction measurements at ∼ 70 K on cooling, which refers to the V/VII phase transition. These additional peaks remained up to ∼ 85 K on heating. They were described with a doubling of the unit cell along all three principal crystallographic directions.


Conserved expression of the glutamate NMDA receptor 1 subunit splice variants during the development of the Siberian hamster suprachiasmatic nucleus.
Glutamate neurotransmission and the N-methyl-D-aspartate receptor (NMDAR) are central to photic signaling to the master circadian pacemaker located in the hypothalamic suprachiasmatic nucleus (SCN). NMDARs also play important roles in brain development including visual input circuits. The functional NMDAR is comprised of multiple subunits, but each requiring the NR1 subunit for normal activity. The NR1 can be alternatively spliced to produce isoforms that confer different functional properties on the NMDAR. The SCN undergoes extensive developmental changes during postnatal life, including synaptogenesis and acquisition of photic signaling. These changes are especially important in the highly photoperiodic Siberian hamster, in which development of sensitivity to photic cues within the SCN could impact early physiological programming. In this study we examined the expression of NR1 isoforms in the hamster at different developmental ages. Gene expression in the forebrain was quantified by in situ hybridization using oligonucleotide probes specific to alternatively spliced regions of the NR1 heteronuclear mRNA, including examination of anterior hypothalamus, piriform cortex, caudate-putamen, thalamus and hippocampus. Gene expression analysis within the SCN revealed the absence of the N1 cassette, the presence of the C2 cassette alone and the combined absence of C1 and C2 cassettes, indicating that the dominant splice variants are NR1-2a and NR1-4a. Whilst we observe changes at different developmental ages in levels of NR1 isoform probe hybridization in various forebrain structures, we find no significant changes within the SCN. This suggests that a switch in NR1 isoform does not underlie or is not produced by developmental changes within the hamster SCN. Consistency of the NR1 isoforms would ensure that the response of the SCN cells to photic signals remains stable throughout life, an important aspect of the function of the SCN as a responder to environmental changes in quality/quantity of light over the circadian day and annual cycle.


Perovskite-Coated Thermochromic Transparent Wood: A Novel Material for Smart Windows in Energy-Efficient and Sustainable Buildings.
Transparent wood (TW) has emerged as a sustainable alternative to conventional glass as an energy-efficient window glazing material owing to its exceptional optical transparency and superior mechanical and thermal performances. However, it is challenging to develop the TW-based color-switching smart windows with both high optical performance and mechanical strengths. In this work, an optically switchable and mechanically robust perovskite-coated thermochromic transparent wood (PTTW) is developed for use as smart windows to achieve an effective solar modulation and thermal management. PTTW exhibits a substantial solar modulation ability Δτsol of 21.6% and a high clear-state luminous transmittance τlum of 78.0%, which enable an efficient thermal regulation while ensuring high visual clarity. PTTW also offers enhanced mechanical properties (i.e., tensile strength σtens = 71.4 MPa and flexural strength σflex = 93.1 MPa) and improved thermal properties [i.e., thermal conductivity K = 0.247 W/(m·K) and heat capacity C = 1.69 J/(g·°C)] compared to glass-based smart windows, as well as excellent performance stability (i.e., 200 heating-cooling cycles), manifesting its applicability in real building scenarios. In addition, PTTW also demonstrates a remarkable thermal-regulating performance (i.e., 5.44 °C indoor air temperature regulation) and an energy-saving potential (i.e., 12.9% heating, ventilation, and air conditioning energy savings) in Hong Kong. Overall, this study contributes to the progression toward energy-efficient and sustainable buildings.


An experimental field approach to parasitism and immune defence in voles.
The fox tapeworm Echinococcus multilocularis is typically perpetuated in a cycle with red foxes as definitive hosts and various rodent species as intermediate hosts. In this study, foxes were baited with a highly efficient drug against cestodes (praziquantel) in 5 blocks of 1 km2. Voles, Arvicola terrestris, the most abundant intermediate host species, were trapped in the 5 baited blocks and in 5 non-baited control blocks. Baiting the foxes reduced the prevalence of E. multilocularis in fox faecal samples in the baited blocks, but voles trapped in the two blocks did not differ in their infection rates. However, voles from the baited blocks had significantly smaller spleen masses and were more likely to be infested with mites than those from the control blocks, possibly reflecting different immunological activities. Our study suggests that the environmental contamination with E. multilocularis eggs, and perhaps those of other tapeworms, influences the immune system of the intermediate host species A. terrestris in the wild.


The Influence of Weather Variation, Urban Design and Built Environment on Objectively Measured Sedentary Behaviour in Children.
With emerging evidence indicating that independent of physical activity, sedentary behaviour (SB) can be detrimental to health, researchers are increasingly aiming to understand the influence of multiple contexts such as urban design and built environment on SB. However, weather variation, a factor that continuously interacts with all other environmental variables, has been consistently underexplored. This study investigated the influence of diverse environmental exposures (including weather variation, urban design and built environment) on SB in children. This cross-sectional observational study is part of an active living research initiative set in the Canadian prairie city of Saskatoon. Saskatoon's neighbourhoods were classified based on urban street design into grid-pattern, fractured grid-pattern and curvilinear types of neighbourhoods. Diverse environmental exposures were measured including, neighbourhood built environment, and neighbourhood and household socioeconomic environment. Actical accelerometers were deployed between April and June 2010 (spring-summer) to derive SB of 331 10-14 year old children in 25 one week cycles. Each cycle of accelerometry was conducted on a different cohort of children within the total sample. Accelerometer data were matched with localized weather patterns derived from Environment Canada weather data. Multilevel modeling using Hierarchical Linear and Non-linear Modeling software was conducted by factoring in weather variation to depict the influence of diverse environmental exposures on SB. Both weather variation and urban design played a significant role in SB. After factoring in weather variation, it was observed that children living in grid-pattern neighbourhoods closer to the city centre (with higher diversity of destinations) were less likely to be sedentary. This study demonstrates a methodology that could be replicated to integrate geography-specific weather patterns with existing cross-sectional accelerometry data to understand the influence of urban design and built environment on SB in children.


Two-step model of stop codon recognition by eukaryotic release factor eRF1.
Release factor eRF1 plays a key role in the termination of protein synthesis in eukaryotes. The eRF1 consists of three domains (N, M and C) that perform unique roles in termination. Previous studies of eRF1 point mutants and standard/variant code eRF1 chimeras unequivocally demonstrated a direct involvement of the highly conserved N-domain motifs (NIKS, YxCxxxF and GTx) in stop codon recognition. In the current study, we extend this work by investigating the role of the 41 invariant and conserved N-domain residues in stop codon decoding by human eRF1. Using a combination of the conservative and non-conservative amino acid substitutions, we measured the functional activity of >80 mutant eRF1s in an in vitro reconstituted eukaryotic translation system and selected 15 amino acid residues essential for recognition of different stop codon nucleotides. Furthermore, toe-print analyses provide evidence of a conformational rearrangement of ribosomal complexes that occurs during binding of eRF1 to messenger RNA and reflects stop codon decoding activity of eRF1. Based on our experimental data and molecular modelling of the N-domain at the ribosomal A site, we propose a two-step model of stop codon decoding in the eukaryotic ribosome.


Major histocompatibility complex-restricted and unrestricted T helper cells recognizing minor histocompatibility antigens of B cell surfaces.
The present experiments analyze the functional properties of helper T cells specific for "minor" histocompatibility antigens. T cells from C3H/HeJ mice, primed in vivo and highly enriched in vitro for reactivity to membrane antigens of C3H/Tif B cells, specifically proliferate, and provide polyclonal help to splenic B cells from strains carving a variety of different H-2 haplotypes on C3H or BALB backgrounds, while failing to respond to cells carrying the same H-2 haplotypes on C57BL or A backgrounds. Since it has been previously demonstrated (A.A. Augustin and A. Coutinho, J. Exp. Med. 1980. 151: 587) and B cell activation in this system strictly requires direct, specific recognition of B cell surface antigens by helper cells and does not result from the production of soluble "mitogenic" or "nonspecific helper factors", it is concluded that this phenomenon represents specific, major histocompatibility complex (MHC)-unrestricted T cell help. In addition, it has now been found that expression of helper activity requires viability of the helper cells and is partially radiation-sensitive. Lack of MHC restriction is not a general property of specific helper cells which directly recognize B cell "minor" antigens, since BALB.C3H anti-C3H/Tif T cells appear to be restricted by H-2 in their polyclonal helper activity. The helper activity mediated by specific anti-"minor", H-2 restricted helper cells could not be inhibited by anti-VH antibodies, and the inhibition obtained with anti-Ia antibodies appeared to operate at the level of B cell induction, rather than at the level of helper cell activation.


On interrater reliability for Leonhard's classification of endogenous psychoses.
Twenty-two patients with a diagnosis of schizophrenia according to DSM-III-R and ICD-10 criteria, for whom the long-term courses of illness were all well documented, were classified by 4 independent investigators using Leonhard's classification of endogenous psychoses. With regard to the large nosological groups of cycloid psychoses (including the subtypes of anxiety-happiness psychosis, confusional psychosis and motility psychosis), of unsystematic schizophrenias (including the subtypes of affect-laden paraphrenia, cataphasia and periodic catatonia), and of systematic schizophrenias (divided into systematic catatonias, systematic paraphrenias, and hebephrenias), is was possible to reach a high level of agreement in the diagnosis, representing a Cohen kappa coefficient of 0.82 and 0.89, respectively. In only 2 out of 22 patients were discrepancies observed in the assignment to the above-mentioned groups. This clearly shows the high reliability of Leonhard's classification, which allows a differentiated diagnostic and prognostic judgement of schizophrenic psychoses according to the DSM-III-R and ICD-10.


[Psychoanalysis and psychoanalysts in France between 1939 and 1945].
France was affected by a deep psychoanalytic silence, both in theoretical and clinical fields, throughout the four years of German occupation. The psychoanalytic Institute closed its doors and the Revue française de psychanalyse interrupted its publication as soon as the armistice of 1940. Some people, e.g. Rudolf Loewenstein or Princess Marie Bonaparte, emigrated, others fought, e.g. Sacha Nacht or Paul Schiff. Daniel Lagache went on with his researches within the works of the University of Strasburg, which had sought shelter in Clermont-Ferrand. René Laforgue cooperated with the German occupant; after the Liberation of France in 1944 he was discarded from a group in which, a few years before, he may have entertained the hope of playing an outstanding part. Quite a few of those he analyzed remained faithful to him--which will have an important bearing on the evolution of the psychoanalytic movement in France after 1945. In 1945, indeed, the new generation emerging from the last convulsions of the war, will gather around the most influential potential leaders: Sacha Nacht and Jacques Lacan. Only after the latency of those silent years, only after those years of violent struggles and cowardice, will the psychoanalytic elaborations appear, and also the institutional feuds to which they gave vent--in a conflictual climate amounting to the schism of 1953.


Epithelial glycoprotein-330 mediates endocytosis of plasminogen activator-plasminogen activator inhibitor type-1 complexes.
Epithelial glycoprotein 330 (gp330) is structurally similar to the multifunctional alpha 2-macroglobulin receptor/low density lipoprotein receptor-related protein (alpha 2MR/LRP), gp330 and alpha 2MR/LRP bind Ca2+ with high affinity, and both receptors bind and mediate endocytosis of alpha 2MR-associated protein (RAP). In the present report, we describe that affinity-purified gp330 from rabbit renal cortex binds plasminogen activator inhibitor type-1 (PAI-1) complexed with urokinase-type plasminogen activator (uPA). alpha 2M-methylamine, which binds with high affinity to alpha 2MR/LRP, did not bind to gp330. The apparent Kd for binding of uPA.PAI-1 complexes was about 0.8 nM at 4 degrees C. The binding was calcium-dependent and inhibited by recombinant RAP (rRAP) and tissue type plasminogen activator-PAI-1 complexes. Thin sections of rabbit renal proximal tubules bound 125I-labeled uPA.PAI-1 and rRAP in the apical part of proximal tubules corresponding to the localization of gp330. The binding of 125I-uPA.PAI-1 complexes in tubules was abolished by excess unlabeled rRAP, and a rRAP-inhibitable endocytosis and degradation of labeled uPA.PAI-1 complexes was demonstrated by perfusion of isolated rabbit proximal tubules. The results establish an endocytotic function of gp330 and suggest that gp330 is an important component of the fibrinolytic system in gp330-containing epithelial as found in, for example, kidney and lung.


In vitro mutations in dihydrofolate reductase that confer resistance to methotrexate: potential for clinical application.
Mammalian cells cultured in the presence of the chemotherapeutic agent, methotrexate, develop resistance to this drug. Sometimes this is due to mutations in the gene for dihydrofolate reductase, the primary target of methotrexate. However, it has not been possible to link such polymorphism to resistance of neoplastic disease to therapy with methotrexate. Nevertheless, interest in this possibility lead to the introduction of many mutations into the cDNA for human DHFR by mutagenesis. Most of the corresponding enzyme variants have been expressed in Escherichia coli and characterized. Many mutations in codons for hydrophobic residues at the active site greatly decrease inhibition by methotrexate, and by the related substrate analogue, trimetrexate, while allowing the retention of considerable catalytic efficiency. Introduction of some of these mutants into mammalian cells by retroviral transfer provides substantial protection from toxic effects of the inhibitors, and has promise for the myeloprotection of patients receiving therapy with methotrexate or trimetrexate. Another potential use is in therapy for inherited disorders of hematopoiesis, where genetic modification of enough cells is a perennial problem. After transplantation of bone marrow that has been transduced with a bicistronic vector encoding both the mutant DHFR and a therapeutic gene, subsequent administration of methotrexate or trimetrexate should permit selection and enrichment of genetically modified hematopoietic cells.


Antagonistic effect of atipamezole on xylazine-induced sedation, bradycardia, and ruminal atony in calves.
Three doses of an alpha 2-adrenoreceptor antagonist, atipamezole, were administered to reverse xylazine-induced sedation, bradycardia, and ruminal atony in calves. Once a week for 4 weeks, each of 6 calves was administered IV 1 treatment of: 0.3 mg of xylazine/kg of body weight, followed in 10 minutes by 1 ml of 0.9% NaCl; 0.3 mg of xylazine/kg, followed in 10 minutes by 3 micrograms of atipamezole/kg; 0.3 mg of xylazine/kg, followed in 10 minutes by 10 micrograms of atipamezole/kg; or 0.3 mg of xylazine/kg, followed in 10 minutes by 30 micrograms of atipamezole/kg. The order of the 4 treatments in each calf was selected at random. Xylazine alone caused lateral recumbency for 33.6 +/- 7.1 minutes (mean +/- SEM). Atipamezole administered at dosages of 3, 10, and 30 micrograms/kg shortened xylazine-induced lateral recumbency to 20.5 +/- 3.0, 10.2 +/- 0.2, and 9.3 +/- 0.5 minutes, respectively. Calves given xylazine alone stood at greater than 60 minutes after the onset of recumbency. Atipamezole given at 3, 10, and 30 micrograms/kg shortened the time from onset of lateral recumbency to standing to 40.2 +/- 6.9, 12.8 +/- 1.1, and 10.0 +/- 0.7 minutes, respectively. Drowsiness was found in calves given the lowest dosage of atipamezole (3 micrograms/kg) after the calves stood. Atipamezole given at dosages of 10 and 30 micrograms/kg reversed xylazine-induced ruminal atony in a dose-dependent manner. In addition, 30 micrograms of atipamezole/kg reversed xylazine-induced bradycardia, but the lower dosages of this antagonist did not. Results indicated that 30 micrograms of atipamezole/kg should be a useful antidote for xylazine overdose in cattle.


Changes of several acute phase factors in broiler chickens in response to infectious bronchitis virus infection.
This study was performed to clarify the acute phase response following infectious bronchitis virus inoculation. Ninety clinically healthy 1-d-old Ross chicks were randomly assigned into 2 groups: control (n = 20) and infected group (n = 70). At the age of 20 d, all birds in the infected group were challenged intranasally with allantoic fluid containing 10(5) embryo lethal dose (ELD50)/0.1 mL of the infectious bronchitis virus. Blood samples were collected from 20 clinically healthy and 70 infected chicks at prior and 1, 2, 3, 5, 7, 11, 13, 15, and 20 d postinoculation. On d 1, 7, and 11 postinoculation 4 chickens from the experimental group and 2 chickens from the control group were randomly selected. Their trachea, lungs, and cecal tonsil were collected for virus detection and quantitation by real-time reverse-transcription PCR assay. In the serum the acute phase proteins (haptoglobin and serum amyloid A), pro-inflammatory cytokines (interferon-γ and tumor necrosis factor-α), and serum sialic acid (total, TSA; lipid-bound, LBSA; and protein-bound, PBSA) concentrations were measured using validated standard procedures. All variables were significantly higher in the infected birds after virus inoculation compared with the healthy group (P < 0.05). There were positive correlations between all variables in the infected group. Correlation coefficients were significantly positive between haptoglobin and interferon-γ, LBSA and TSA, and TSA and LBSA (P < 0.05). There were positive correlations among viral RNA and all studied variables; however, these correlations were not statistically significant (P > 0.05).


PROTAC chemical probes for histone deacetylase enzymes.
Over the past three decades, we have witnessed the progression of small molecule chemical probes designed to inhibit the catalytic active site of histone deacetylase (HDAC) enzymes into FDA approved drugs. However, it is only in the past five years we have witnessed the emergence of proteolysis targeting chimeras (PROTACs) capable of promoting the proteasome mediated degradation of HDACs. This is a field still in its infancy, however given the current progress of PROTACs in clinical trials and the fact that FDA approved HDAC drugs are already in the clinic, there is significant potential in developing PROTACs to target HDACs as therapeutics. Beyond therapeutics, PROTACs also serve important applications as chemical probes to interrogate fundamental biology related to HDACs via their unique degradation mode of action. In this review, we highlight some of the key findings to date in the discovery of PROTACs targeting HDACs by HDAC class and HDAC isoenzyme, current gaps in PROTACs to target HDACs and future outlooks.


Neuropsychiatric complications of corticosteroids in older people.
The neuropsychiatric effects of corticosteroids are well described in the literature but relatively little is known specifically about their effects among older people. As the population ages, and increasing numbers of older people receive assertive medical treatment for physical conditions, more patients are likely to present to psychiatric services suffering from adverse neuropsychiatric effects associated with corticosteroid use. A review of the effects of corticosteroids among older people was carried out through Medline and PubMed searches and a review of article references. Very little has been specifically written about steroid use and the effects on older people. This paper briefly outlines the current level of knowledge of neuropsychiatric effects of steroids, and relates this to general treatment considerations for older subjects. Recommendations for further research and early identification of complications are made.


Theoretical studies on the mobility-shift assay of protein-DNA complexes.
The theory of mass transport coupled to reversible macromolecular interactions under chemical kinetic control forms the basis for computer simulation of the electrophoretic mobility-shift behavior of protein-DNA complexes. Model systems include (i) specific binding of a univalent protein molecule to a single site on the DNA molecule; (ii) the putative cage effect; (iii) cooperative binding to multiple sites; (iv) formation of looped complexes of 1:1 and 2:1 stoichiometry; (v) noncooperative and cooperative, nonspecific binding modes; and (vi) binding of dimerizing transcriptional factors to response elements of target genes. Favorable comparison of simulated with experimental mobility-shift behavior indicates that the phenomenological mechanisms, whereby observed mobility-shift patterns are generated during electrophoresis, are embodied in the theory. These studies have provided guidelines for definitive interpretation of mobility-shift assays and for the design of experiments to develop a detailed understanding of the particular system under investigation.


Planning and realisation of complex intentions in patients with Parkinson's disease.
There is some evidence that patients with Parkinson's disease may impaired in prospective memory performance (planning and self initiated realisation of delayed intentions). Little is known about the effect of the disease on distinct phases of prospective memory and the potential mechanisms underlying these effects. To investigate intention formation, intention retention, intention initiation, and intention execution of patients with Parkinson's disease and test for the mediating influence of working memory, inhibition, short term retrospective memory, and divided attention. 16 patients with Parkinson's disease and 16 age and education matched normal controls were given a complex event based prospective memory task which differentiates four phases of prospective remembering. In addition, participants completed tasks assessing potential cognitive mediators. On the prospective remembering task, Parkinson patients were impaired in the intention formation phase and showed a trend towards impairment in the intention initiation. In contrast, there were no impairments of retrospective intention retention or the fidelity with which the patients executed their previously developed plan. The group effects were related to interindividual differences in working memory span. The results suggest that the planning phase of prospective remembering is specifically impaired in Parkinson's disease, and that the impairment is related to working memory deficit. In contrast, even when complex intentions have to be remembered, the retrospective storage of intentions to be performed is not impaired.


Soluble Platelet Release Factors as Biomarkers for Cardiovascular Disease.
Platelets are the main players in thrombotic diseases, where activated platelets not only mediate thrombus formation but also are involved in multiple interactions with vascular cells, inflammatory components, and the coagulation system. Although in vitro reactivity of platelets provides information on the function of circulating platelets, it is not a full reflection of the in vivo activation state, which may be relevant for thrombotic risk assessment in various disease conditions. Therefore, studying release markers of activated platelets in plasma is of interest. While this type of study has been done for decades, there are several new discoveries that highlight the need for a critical assessment of the available tests and indications for platelet release products. First, new insights have shown that platelets are not only prominent players in arterial vascular disease, but also in venous thromboembolism and atrial fibrillation. Second, knowledge of the platelet proteome has dramatically expanded over the past years, which contributed to an increasing array of tests for proteins released and shed from platelets upon activation. Identification of changes in the level of plasma biomarkers associated with upcoming thromboembolic events allows timely and individualized adjustment of the treatment strategy to prevent disease aggravation. Therefore, biomarkers of platelet activation may become a valuable instrument for acute event prognosis. In this narrative review based on a systematic search of the literature, we summarize the process of platelet activation and release products, discuss the clinical context in which platelet release products have been measured as well as the potential clinical relevance.


Cohesive communities in dynamic brain functional networks.
In large-scale brain network dynamics, brain nodes switching between modules has been found to correlate with cognition. However, how the brain nodes engage in this kind of reorganization of modules is unclear. Based on a functional magnetic resonance imaging dataset, we construct dynamic brain functional networks and investigate nodal module temporal dynamic behavior by applying the multilayer network analysis approach. We reveal three cohesive communities that are groups of brain nodes linked in the same community during brain module dynamic reorganization. We show that the cohesive communities have higher clustering coefficients and lower characteristic path lengths than the controlled community, indicating cohesive communities are the parts of brain networks with high information processing efficiency. The smaller sample entropy of functional connectivity in cohesive communities also proves their property of being more "static" compared with the controlled community in brain dynamics. Specifically, compared with the controlled community, the functional connectivity of cohesive communities is restricted strictly by structure connectivity and shows more similarity to structure connectivity. More importantly, we find that the cohesive communities are stable not only in the resting state but also when processing cognitive tasks. Our results not only show that cohesive communities may be the fundamental community organization to support brain network dynamics but also provide insights into the intrinsic structural relationship between the resting state and task states of the brain.


Molecular dissection of mesenchymal-epithelial interactions in the hair follicle.
De novo hair follicle formation in embryonic skin and new hair growth in adult skin are initiated when specialized mesenchymal dermal papilla (DP) cells send cues to multipotent epithelial stem cells. Subsequently, DP cells are enveloped by epithelial stem cell progeny and other cell types to form a niche orchestrating hair growth. Understanding the general biological principles that govern the mesenchymal-epithelial interactions within the DP niche, however, has been hampered so far by the lack of systematic approaches to dissect the complete molecular make-up of this complex tissue. Here, we take a novel multicolor labeling approach, using cell type-specific transgenic expression of red and green fluorescent proteins in combination with immunolabeling of specific antigens, to isolate pure populations of DP and four of its surrounding cell types: dermal fibroblasts, melanocytes, and two different populations of epithelial progenitors (matrix and outer root sheath cells). By defining their transcriptional profiles, we develop molecular signatures characteristic for the DP and its niche. Validating the functional importance of these signatures is a group of genes linked to hair disorders that have been largely unexplored. Additionally, the DP signature reveals novel signaling and transcription regulators that distinguish them from other cell types. The mesenchymal-epithelial signatures include key factors previously implicated in ectodermal-neural fate determination, as well as a myriad of regulators of bone morphogenetic protein signaling. These findings establish a foundation for future functional analyses of the roles of these genes in hair development. Overall, our strategy illustrates how knowledge of the genes uniquely expressed by each cell type residing in a complex niche can reveal important new insights into the biology of the tissue and its associated disease states.


Dual effects of Haemophilus influenzae on guinea pig tracheal beta-adrenergic receptor function: involvement of oxygen-centered radicals from pulmonary macrophages.
The Gram-negative bacterium Haemophilus influenzae has been shown to cause a deterioration of the guinea pig pulmonary beta-adrenergic receptor system. In the present study we investigated further the mechanisms behind this effect. To this extent we evaluated the involvement of pulmonary macrophages (PM). Treatment of guinea pigs with killed H. influenzae bacteria resulted in the accumulation of a factor in the serum which could specifically stimulate PM. Thus stimulated, PM from nontreated animals caused a decrease of tracheal beta-adrenergic receptor function in vitro. This effect was evident by a decrease of the maximal response of the dose-response curves to isoprenaline, whereas the EC50 values did not change. Catalase and thiourea abolished the PM-induced effects, whereas superoxide dismutase did not, indicating that oxygen-centered radicals, in particular the highly reactive hydroxyl radical, may be responsible for the observed effects. In addition, dexamethasone also inhibited the decrease of tracheal beta-adrenergic receptor function. When activated PM, taken from animals that had been pretreated with killed H. influenzae bacteria 4 days beforehand, were stimulated with serum from a H. influenzae-treated animal, a potentiation of tracheal beta-adrenergic receptor function was observed.


Saccharin drinking rather than open field behaviour predicts initial ethanol acceptance in Wistar rats.
This study examined the relationship between saccharin drinking, open field behaviour and ethanol drinking in Wistar rats. Correlational analysis revealed that both absolute saccharin drinking and an increase in total fluid intake in the presence of saccharin positively correlated with the initial acceptance of increasing ethanol concentrations in a two-bottle choice situation (2-8% v/v ethanol vs water). This relationship disappeared, however, during further weeks of ethanol drinking when ethanol was available in a three-bottle choice situation (8% ethanol vs 16% ethanol vs water). In contrast, none of the behavioural parameters measured in the open field test (forward locomotion, rearings, central entries, time in central area) correlated with subsequent ethanol consumption. These results indicate that saccharin drinking, rather than open field parameters, may predict subsequent ethanol intake during the initial period of exposure to low ethanol concentrations.


First report of meningococcal ciprofloxacin resistance in Greece due to invasive isolates of the sequence type ST-3129.
A local outbreak caused by Neisseria meningitidis occurred in the migration camp in the Greek island of Lesbos during January-February 2020 (4 of 5 cases). In total, 5 samples positive for N. meningitidis were further investigated for sero-/genogroup, PorA, and WGS analysis. MenB was found among 3 cases, while in two cases, MenY was identified. WGS analysis and antibiotic susceptibility testing on the 2 culture positive MenB samples showed the new ST-3129, ciprofloxacin-resistant clone was circulating among the immigrants in the aforementioned camp. This is the first report of ciprofloxacin resistance in Greece.


Regularized inverse holographic volume reconstruction for 3D particle tracking.
The key limitations of digital inline holography (DIH) for particle tracking applications are poor longitudinal resolution, particle concentration limits, and case-specific processing. We utilize an inverse problem method with fused lasso regularization to perform full volumetric reconstructions of particle fields. By exploiting data sparsity in the solution and utilizing GPU processing, we dramatically reduce the computational cost usually associated with inverse reconstruction approaches. We demonstrate the accuracy of the proposed method using synthetic and experimental holograms. Finally, we present two practical applications (high concentration microorganism swimming and microfiber rotation) to extend the capabilities of DIH beyond what was possible using prior methods.


The t(11;16)(q23;p13) translocation in myelodysplastic syndrome fuses the MLL gene to the CBP gene.
The recurrent translocation t(11;16)(q23;p13) has been reported to be associated with therapy-related acute leukemia. The MLL gene involved in other 11q23 abnormalities was also rearranged by this translocation. We analyzed two patients with myelodysplastic syndrome with t(11;16) and showed that the MLL gene on 11q23 was fused with CREB-binding protein (CBP) gene on 16p13 in these patients. The CBP gene encodes a transcriptional adaptor/coactivator protein and it is mutated in patients with Rubinstein-Taybi syndrome. The CBP gene is also involved in acute myeloid leukemia (AML) with t(8;16)(p11;p13). In-frame MLL-CBP fusion transcripts combine the MLL AT-hook motifs and DNA methyltransferase homology region with a largely intact CBP. Our results combined with the finding of the MOZ-CBP fusion in t(8;16)-AML suggest that the CBP gene may be associated with leukemogenesis through translocations.


Gender differences in cerebral aneurysm location.
A limited number of studies consisting predominantly of ruptured aneurysms have looked at differences in anatomical distribution of aneurysms between male and females. Unlike all other causes of stroke, subarachnoid hemorrhages (SAH) occur more often in women and are thought to be a result of both hormonal influences and variation in wall shear stress. This paper retrospectively looks at a cohort of largely unruptured intracranial aneurysms to determine if there exists a gender discrepancy in the anatomic distribution of cerebral aneurysms. A retrospective review of consecutive patients with ruptured and unruptured intradural saccular cerebral aneurysms treated endovascularly was performed. Six hundred eighty-two aneurysms were treated. Seventy-two percentage of the patients were women and 27% of patients presented with SAH. Among women, most aneurysms were located along the ICA (54%) while men the ACA (29%, compared to 15% in women), a discrepancy evident in both unruptured and ruptured groups. Females tended to present later in life (59 vs. 55 years), with multiple aneurysms (11 vs. 6% in men), and with SAH (28 vs. 23% in men) - the majority of these ruptured aneurysms were located at the ICA (42%), while men at the ACA (47%). Additionally, the majority (68%) of ruptured ICA aneurysms were PCOM. Understanding the natural history of aneurysms is imperative in treating incidentally found aneurysms. Significant differences exist between the genders in relation to aneurysm location, the most pronounced at the ICA and ACA. Previously described hormonal and hemodynamic theories behind cerebral aneurysm pathogenesis seem like plausible reasons to explain these differences.


Hormone-specific regulation of the kidney androgen-regulated gene promoter in cultured mouse renal proximal-tubule cells.
The kidney androgen-regulated protein (KAP) is specifically expressed and differentially regulated by androgens and tri-iodothyronine (T(3)) in intact mouse early (PCT) and late (PR) proximal-tubule cells. Until now, detailed characterization of the molecular elements mediating androgen-responsive gene expression in the kidney has been hampered by the lack of appropriate cultured cell systems suitable for DNA transfection studies. In the present study we have analysed the hormone-dependent transactivation of the KAP gene promoter in immortalized differentiated PCT and PR proximal-tubule cells derived from L-PK/Tag1 transgenic mice. Transient transfection studies with different KAP promoter constructs indicated that a 224 bp-truncated fragment was sufficient to mediate cell-specific expression of the KAP promoter. Dihydrotestosterone (DHT) stimulated in an androgen-dependent manner the transactivation of KAP in PCT and PR cells, while mutation of a putative androgen-response element (ARE) sequence located at -39 bp from the transcription initiation site abolished the transactivation induced by DHT. Furthermore, insulin-like growth factor 1 (IGF-1), but not T(3), enhanced the androgen-dependent transactivation of KAP in cultured PCT cells. These results demonstrate that the short 224 bp fragment of the KAP promoter is sufficient to drive the proximal-tubule androgen-specific regulated expression of KAP and reveal synergistic interactions between IGF-1 and androgens for KAP regulation in PCT cells.


Risk of occupational allergy to stored grain arthropods and false pest-risk perception in Czech grain stores.
Arthropods are a documented cause of occupational allergy in cereal stores. Since the current allergenic risk of various arthropods in grain stores is not known, we evaluated its extent using data from the Czech Republic (CZ). We surveyed 514 grain storage units for pest composition and density. Recalculating literature data we established 4 density dependent classes of pooled mite "allergy-risk level" (ARL) in stored grain: (i) safe-ARL: 0 mites.g(-1) grain, (ii) low-ARL: up to 1 mite.g(-1) grain, (iii) high-ARL: from 1 to 5 mites.g(-1) grain, (iv) danger-acute asthma-ARL: higher than 5 mites.g(-1) grain. Farmers (15) were questioned for pest taxon-related pesticide treatments. Mites constituted the largest group of collected pests (92%) followed by psocids (5%), beetles (3%) and moths (0%). 60% of mites belonged to known allergen producing species; the most abundant were Acarus siro, A. faris, Tyrophagus putrescentiae and Lepidoglyphus destructor. Grain samples belonged to the established ARL classes as follows: (i) safe-ARL: 37% (ii) low-ARL: 53%; (iii) high-ARL: 6%; (iv) danger-acute asthma-ARL: 4%. The enquiry among farmers revealed that almost no pesticides were targeted solely to control mites. This study suggests that mites represent, due to their allergenic potential, density and frequency, the most serious source of allergens in stored grain in CZ. However, the medical aspect of pest control--such as allergy avoidance strategy--is overlooked since grain feeding insects were mostly chemically controlled, regardless of their relatively low density and allergen production in comparison with mites.


Survival of primary lung cancer patients in Brunei Darussalam, 1987-2012.
In 2009 to 2011, the commonest cause of death was by cancer in Brunei Darussalam, and 16.5% and 19.5% of cancer deaths were due to lung cancer in 2004 and 2011 respectively. This study was to investigate the survival of lung cancer patients in Brunei Darussalam. This retrospective cohort study was conducted in 2013 & 2014 for those who were diagnosed as primary lung cancer in the period of 1987 to 2012. Data were retrieved from patients' medical records and death certificates using pretested data collection form. Survival analyses namely Kaplan-Meier method, and log-rank test to compare survival between groups. We retrieved 630 primary lung cancer records. Majority was diagnosed at the late stages, 42.5% at Stage IV & 33.4% at Stage III. The overall median survival time was 6.1 months whereas 2.6 and 79.1 months for Stage IV and Stage I respectively. The overall 6-month, 1-, 3- and 5-year survival rates were 50.2%, 32.1%, 14.5% and 8.8% respectively. Survival duration had significantly improved from 1987-1999 to 2000-2012 (P=0.001) although significant higher proportion of Stage IV was diagnosed in the second period (P=0.008). Overall survival rates and duration of primary lung cancer in Brunei Darussalam were comparable with some developed countries. However, through effective public intervention such as increase awareness, early case detection, and effective anti-smoking strategies, survival of lung cancer patients can certainly be improved and the burden of disease can be reduced.


Acoustic phonetic study of the Sora vowel system.
This paper is an acoustic phonetic study of vowels in Sora, a Munda language of the Austroasiatic language family. Descriptions here illustrate that the Sora vowel system has six vowels and provide evidence that Sora disyllables have prominence on the second syllable. While the acoustic categorization of vowels is based on formant frequencies, the presence of prominence on the second syllable is shown through temporal features of vowels, including duration, intensity, and fundamental frequency. Additionally, this paper demonstrates that acoustic categorization of vowels in Sora is better in the prominent syllable than in the non-prominent syllable, providing evidence that syllable prominence and vowel quality are correlated in Sora. These acoustic properties of Sora vowels are discussed in relation to the existing debates on vowels and patterns of syllable prominence in Munda languages of India. In this regard, it is noteworthy that Munda languages, in general, lack instrumental studies, and therefore this paper presents significant findings that are undocumented in other Munda languages. These acoustic studies are supported by exploratory statistical modeling and statistical classification methods.


Food safety regulation: reforming the Delaney Clause.
The safety of food has been an age-old concern. Early civilizations adopted laws that punished sellers of tainted food. In this country, before food safety became a responsibility of the federal government every state had enacted laws prohibiting the sale of food that contained poisonous substances. The modern scientific and legal instruments available to the US Food and Drug Administration and allied agencies have improved regulation and advances in food preparation, preservation, and storage have contributed to a safer food supply. Even so, some observers believe that contemporary threats to food safety have grown more serious, and they surely excite intense public concern. For nearly two decades Congress has been debating the adequacy of current laws governing food safety. In the closing months of the 104th Congress, both parties finally agreed on the first significant legislative change in over a generation. This chapter examines the origins of the issues that were the focus of this extended debate and analyzes the implications of their resolution.


Temperature effect on the shear-induced cell damage in biofabrication.
Biofabrication that incorporates living cells to manufacture various bioproducts is often carried out at different temperatures as the process demands. In the process, cells are subjected to mechanical forces, which may damage cells if the forces reach a certain level. Previous studies have shown that the cell damage is mainly caused by shear stress; however, none of them looked at the temperature effect on cell damage. In the present work, the influence of temperature on shear-induced cell damage was investigated experimentally by using a cone-and-plate rheometer, and based on the experimental results, a cell damage law was established to quantitatively describe the relationship between the cell damage percent and temperature. The so-established cell damage law was then applied to the modeling of the cell damage percent that occurs in the biofabrication process in which pressurized air was applied to dispense Schwann cells suspended in the alginate solution at different temperatures. The agreement between the model predictions and the experimental results suggests that the method presented in this article is effective for use in the investigation of the temperature effect, thereby providing a cue to preserve cell viability in the biofabrication processes.


Implementation of the ETAR method for 3D inhomogeneity correction using FFT.
The equivalent tissue-air-ratio (ETAR) method employs three-dimensional (3D) CT pixel information to approximate scatter dose contribution for inhomogeneity correction. In general, the calculation provides better agreement with measurements than the one-dimensional (1D) methods typically used in commercial treatment planning systems. In its original implementation, the 3D formulation of the ETAR method is modified empirically as a 2D calculation in order to reduce computation time. The modification compromises the use of the method in several treatment geometries. An examination of the ETAR formulation shows that the calculation can be expressed as a convolution and thus can be performed in 3D using fast Fourier transform (FFT) techniques. The algorithm has been implemented as a 3D FFT convolution. Making use of the symmetric properties of the FFT, the new approach shows significant savings in computation time without excessive memory requirement. Despite its fundamental limitations when applied to regions of electronic disequilibrium, the ETAR method offers a practical solution to improving current dose calculation in 3D treatment planning, particularly when the more advanced scatter ray-tracing dose calculation algorithms remain impractical for clinical use. Recent work to extend the method to approximate electron transport is also encouraging.


Upregulation of connexin 43 and apoptosis‑associated protein expression by high glucose in H9c2 cells was improved by resveratrol via the autophagy signaling pathway.
The expression of connexin43 (Cx43) protein and the apoptotic rate of cardiomyocytes may be regulated by autophagy and associated with diabetic cardiomyopathy. It is possible that the beneficial effect of resveratrol on diabetic cardiomyocytes occurs via the autophagy pathway. However, it remains to be elucidated whether resveratrol treatment may attenuate the hyperglycemia‑induced remodeling of Cx43 and apoptosis through the regulation of autophagy. H9c2 cardiac cells were incubated with 5.5 and 25 mM glucose, 25 mM glucose with chloroquine (50 µM), and 25 mM glucose with or without resveratrol (10, 25 µM) for 24 h. H9c2 cells were also incubated with 25 µM resveratrol in the presence of chloroquine (50 µM). Cell viability was determined using an MTT cell survival assay. Cytotoxicity was determined by quantification of the release of lactate dehydrogenase. The expression of Cx43, autophagic maker proteins [Beclin‑1, p62 and microtubule‑associated protein 1 light chain 3 (LC3)], apoptosis maker proteins (B‑cell lymphoma‑2 and Bcl‑2 associated X protein), AMP‑activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) were determined using western blotting. Resveratrol treatment led to reduced Cx43 expression levels compared with the 25 mM glucose treatment and significantly reduced the expression of apoptosis‑associated proteins in H9c2 cells under hyperglycemic conditions. Autophagy was increased as indicated by the upregulation of Beclin‑1 and p62 expression and the reduced LC3‑II/LC3‑I ratio. AMPK expression was increased, whereas mTOR expression was reduced in the resveratrol treatment groups. Treatment with chloroquine reversed effect of resveratrol. In conclusion, administration resveratrol may protect H9c2 cells against hyperglycemia‑induced Cx43 upregulation and apoptosis, which may be mediated through the induction of the autophagy signaling pathway.


Combined medical and surgical procedure in acute myocardial infarction--a preliminary report.
The purpose of this investigation was to evaluate a new therapeutic approach, by which acutely ischemic human myocardium could be reperfused. The procedure was as follows: intracoronary application of nitroglycerine, intraluminal recanalization by catheters and intracoronary application of streptokinase. Before and after these interventions coronary angiograms were made. Surgical coronary revascularization was performed 1--64 days after reestablishment of flow through the occluded vessels. Seven patients with acute myocardial infarction were treated in this manner. The mean preoperation ejection fraction was 48.6%, the mean enddiastolic pressure was 17.5 mmHg and the mean maximal CPK activity was 616 units/liter (u/l). By the conservative regimen outlined, reopening of the occluded vessels was achieved in all cases. Critical stenoses of 80--85%, however, persisted. But, reperfusion appeared sufficient since in none of the patients hemorrhagic myocarditis was produced intra-operatively. There was no hospital mortality. The mean post-operative ejection fraction was 60% and the mean enddiastolic pressure was 10.6 mmHg. It is concluded that ischemic myocardium can be reperfused by the active conservative regime tested. Emergency revascularization performed after induced reperfusion appears to carry a low operative risk.


An Intradiscal Granuloma Due to a Retained Wooden Foreign Body.
We report a patient with a wooden foreign body granuloma in the intervertebral disc space being symptomatic 17 years after a paraspinal penetrant trauma. According to the our result of the search for wooden foreign body granulomas, this is the first case suffered from a wooden foreign body granuloma in the intervertebral disc space that reported in the literature. In this report, we emphasized the importance of rigorous examination and follow up in paraspinal wooden penetrant traumas.


A Frameshift Mutation in KIT is Associated with White Spotting in the Arabian Camel.
While the typical Arabian camel is characterized by a single colored coat, there are rare populations with white spotting patterns. White spotting coat patterns are found in virtually all domesticated species, but are rare in wild species. Theories suggest that white spotting is linked to the domestication process, and is occasionally associated with health disorders. Though mutations have been found in a diverse array of species, fewer than 30 genes have been associated with spotting patterns, thus providing a key set of candidate genes for the Arabian camel. We obtained 26 spotted camels and 24 solid controls for candidate gene analysis. One spotted and eight solid camels were whole genome sequenced as part of a separate project. The spotted camel was heterozygous for a frameshift deletion in KIT (c.1842delG, named KITW1 for White spotting 1), whereas all other camels were wild-type (KIT+/KIT+). No additional mutations unique to the spotted camel were detected in the EDNRB, EDN3, SOX10, KITLG, PDGFRA, MITF, and PAX3 candidate white spotting genes. Sanger sequencing of the study population identified an additional five kITW1/KIT+ spotted camels. The frameshift results in a premature stop codon five amino acids downstream, thus terminating KIT at the tyrosine kinase domain. An additional 13 spotted camels tested KIT+/KIT+, but due to phenotypic differences when compared to the KITW1/KIT+ camels, they likely represent an independent mutation. Our study suggests that there are at least two causes of white spotting in the Arabian camel, the newly described KITW1 allele and an uncharacterized mutation.


ART in Europe, 2019: results generated from European registries by ESHRE†.
What are the data and trends on ART and IUI cycle numbers and their outcomes, and on fertility preservation (FP) interventions, reported in 2019 as compared to previous years? The 23rd ESHRE report highlights the rising ART treatment cycles and children born, alongside a decline in twin deliveries owing to decreasing multiple embryo transfers; fresh IVF or ICSI cycles exhibited higher delivery rates, whereas frozen embryo transfers (FET) showed higher pregnancy rates (PRs), and reported IUI cycles decreased while maintaining stable outcomes. ART aggregated data generated by national registries, clinics, or professional societies have been gathered and analyzed by the European IVF-Monitoring (EIM) Consortium since 1997 and reported in a total of 22 manuscripts published in Human Reproduction and Human Reproduction Open. Data on medically assisted reproduction (MAR) from European countries are collected by EIM for ESHRE each year. The data on treatment cycles performed between 1 January and 31 December 2019 were provided by either national registries or registries based on initiatives of medical associations and scientific organizations or committed persons in one of the 44 countries that are members of the EIM Consortium. Overall, 1487 clinics offering ART services in 40 countries reported, for the second time, a total of more than 1 million (1 077 813) treatment cycles, including 160 782 with IVF, 427 980 with ICSI, 335 744 with FET, 64 089 with preimplantation genetic testing (PGT), 82 373 with egg donation (ED), 546 with IVM of oocytes, and 6299 cycles with frozen oocyte replacement (FOR). A total of 1169 institutions reported data on IUI cycles using either husband/partner's semen (IUI-H; n = 147 711) or donor semen (IUI-D; n = 51 651) in 33 and 24 countries, respectively. Eighteen countries reported 24 139 interventions in pre- and post-pubertal patients for FP, including oocyte, ovarian tissue, semen, and testicular tissue banking. In 21 countries (21 in 2018) in which all ART clinics reported to the registry 476 760 treatment cycles were registered for a total population of approximately 300 million inhabitants, allowing the best estimate of a mean of 1581 cycles performed per million inhabitants (range: 437-3621). Among the reporting countries, for IVF the clinical PRs per aspiration slightly decreased while they remained similar per transfer compared to 2018 (21.8% and 34.6% versus 25.5% and 34.1%, respectively). In ICSI, the corresponding PRs showed similar trends compared to 2018 (20.2% and 33.5%, versus 22.5% and 32.1%) When freeze-all cycles were not considered for the calculations, the clinical PRs per aspiration were 28.5% (28.8% in 2018) and 26.2% (27.3% in 2018) for IVF and ICSI, respectively. After FET with embryos originating from own eggs, the PR per thawing was at 35.1% (versus 33.4% in 2018), and with embryos originating from donated eggs at 43.0% (41.8% in 2018). After ED, the PR per fresh embryo transfer was 50.5% (49.6% in 2018) and per FOR 44.8% (44.9% in 2018). In IVF and ICSI together, the trend toward the transfer of fewer embryos continues with the transfer of 1, 2, 3, and ≥4 embryos in 55.4%, 39.9%, 2.6%, and 0.2% of all treatments, respectively (corresponding to 50.7%, 45.1%, 3.9%, and 0.3% in 2018). This resulted in a reduced proportion of twin delivery rates (DRs) of 11.9% (12.4% in 2018) and a similar triplet DR of 0.3%. Treatments with FET in 2019 resulted in twin and triplet DR of 8.9% and 0.1%, respectively (versus 9.4% and 0.1% in 2018). After IUI, the DRs remained similar at 8.7% after IUI-H (8.8% in 2018) and at 12.1% after IUI-D (12.6% in 2018). Twin and triplet DRs after IUI-H were 8.7% and 0.4% (in 2018: 8.4% and 0.3%) and 6.2% and 0.2% after IUI-D (in 2018: 6.4% and 0.2%), respectively. Eighteen countries (16 in 2018) provided data on FP in a total number of 24 139 interventions (20 994 in 2018). Cryopreservation of ejaculated sperm (n = 11 592 versus n = 10 503 in 2018) and cryopreservation of oocytes (n = 10 784 versus n = 9123 in 2018) were most frequently reported. Caution with the interpretation of results should remain as data collection systems and completeness of reporting vary among European countries. Some countries were unable to deliver data about the number of initiated cycles and/or deliveries. The 23rd ESHRE data collection on ART, IUI, and FP interventions shows a continuous increase of reported treatment numbers and MAR-derived livebirths in Europe. Although it is the largest data collection on MAR in Europe, further efforts toward optimization of both the collection and the reporting, from the perspective of improving surveillance and vigilance in the field of reproductive medicine, are awaited. The study has received no external funding and all costs are covered by ESHRE. There are no competing interests.


Reliability of ground reaction forces in the aquatic environment.
The aim of this study was to verify the reliability of the kinetic parameters of gait using an underwater force platform. A total of 49 healthy participants with a median age of 21years were included. The kinetic gait data were collected using a 0.6×0.6×0.1m aquatic force plate (Bertec®), set in a pool (15×13×1.30m) with a water depth of 1.20m and water temperature of 32.5°C. Participants walked 10m before reaching the platform, which was fixed to the ground. Participants were instructed to step onto the platform with their preferred limb and data from three valid attempts were used to calculate the average values. A 48-h interval between tests was used for the test-retest reliability. Data were analyzed using interclass correlation coefficients (ICC) and results demonstrated that reliability ranged from poor to excellent, with ICC scores of between 0.24 and 0.87 and mean differences between (d¯)=-0.01 and 0.002. The highest reliability values were found for the vertical (Fz) and the lowest for the mediolateral components (Fy). In conclusion, the force platform is reliable for assessing the vertical and anteroposterior components of power production rates in water, however, caution should be applied when using this instrument to evaluate the mediolateral component in this environment.


Integrated health and care systems in England: can they help prevent disease?
Over the past 12 months, there has been increasing policy rhetoric regarding the role of the National Health Service (NHS) in preventing disease and improving population health. In particular, the NHS Long Term Plan sees integrated care systems (ICSs) and sustainability and transformation partnerships (STPs) as routes to improving disease prevention. Here, we place current NHS England integrated care plans in their historical context and review evidence on the relationship between integrated care and prevention. We ask how the NHS Long Term Plan may help prevent disease and explore the role of the 2019 ICS and STP plans in delivering this change. We reviewed the evidence underlying the relationship between integrated care and disease prevention, and analysed 2016 STP plans for content relating to disease prevention and population health. The evidence of more integrated care leading to better disease prevention is weak. Although nearly all 2016 STP plans included a prevention or population health strategy, fewer than half specified how they will work with local government public health teams, and there was incomplete coverage across plans about how they would meet NHS England prevention priorities. Plans broadly focused on individual-level approaches to disease prevention, with few describing interventions addressing social determinants of health. For ICSs and STPs to meaningfully prevent disease and improve population health, they need to look beyond their 2016 plans and fill the gaps in the Long Term Plan on social determinants.


The contralateral foot in children with unilateral clubfoot: a study of pressures and forces involved in gait.
The unaffected contralateral side of patients with unilateral clubfeet has sometimes been taken as control in foot pressure measurement studies. However, it has never been shown that the pressure pattern under the contralateral foot is similar to a normal foot. Sixteen patients with unilateral clubfoot and 110 normal subjects took part in this study. All participants were aged from 4 to 8 years. Studies comprised clinical examination and foot pressure measurements in barefoot walking. Evaluations of the measurements were completed by the calculation of forces acting under 10 anatomical foot areas normalized to ground reaction force at foot-flat and push-off. Significant differences in peak pressure were observed between the control group and the contralateral foot of patients under the heel and the metatarsals 1 to 4. Significant differences in the forces at foot-flat and push-off are seen especially in the midfoot. Both sides of the patients with unilateral clubfeet exert significantly less ground reaction force than normal subjects. Different pressure and force distributions of clubfoot children on their contralateral side compared with the normal feet of the control group may indicate differences in the general control pattern generated by the central nervous system. Differences can still be observed after normalization of the forces. We therefore conclude that when foot pressure measurements are made with clubfeet, it might be advisable to use a collective of normal feet, instead of the contralateral foot, for comparison.


Rotational Acetabular Osteotomy.
Hip dysplasia is the most common cause of secondary osteoarthritis (OA). To prevent the early onset of secondary OA, Nishio's transposition osteotomy, Steel's triple osteotomy, Eppright's dial osteotomy, Wagner's spherical acetabular osteotomy, Tagawa's rotational acetabular osteotomy (RAO), and Ganz' periacetabular osteotomy (PAO) have been proposed. PAO and RAO are now commonly used in surgical treatment of symptomatic acetabular dysplasia in Europe, North America, and Asia. The aim of this paper is to present the followings: the patient selection criteria for RAO; the surgical technique of RAO; the long-term outcome of RAO; and the future perspectives.


Review of the Immune Checkpoint Inhibitors in the Context of Cancer Treatment.
Checkpoint proteins are an integral part of the immune system and are used by the tumor cells to evade immune response, which helps them grow uncontrollably. By blocking these proteins, immune checkpoint inhibitors can restore the capability of the immune system to attack cancer cells and stop their growth. These findings are backed by adequate clinical trial data and presently, several FDA-approved immune checkpoint inhibitors exist in the market for treating various types of cancers, including melanoma, hepatocellular, endometrial, lung, kidney and others. Their mode of action is inhibition by targeting the checkpoint proteins CTLA-4, PD-1, PD-L1, etc. They can be used alone as well as in amalgamation with other cancer treatments, like surgery, radiation or chemotherapy. Since these drugs target only specific immune system proteins, their side effects are reduced in comparison with the traditional chemotherapy drugs, but may still cause a few affects like fatigue, skin rashes, and fever. In rare cases, these inhibitors are known to have caused more serious side effects, such as cardiotoxicity, and inflammation in the intestines or lungs. Herein, we provide an overview of these inhibitors and their role as biomarkers, immune-related adverse outcomes and clinical studies in the treatment of various cancers, as well as present some future perspectives.


Biomarkers of immune checkpoint inhibitor efficacy in cancer.
Immune checkpoint inhibitor-based therapies that target ctla-4, PD-1, or the PD-1 ligand PD-L1 have received approval in Canada and many parts of the world for the treatment of melanoma, renal cell cancer, urothelial cancer, classical Hodgkin lymphoma, and non-small-cell lung cancer. However only a small proportion of patients derive long-term clinical benefit. Here, we describe the biomarkers associated with the complex relationship between tumour-related immune stimulus, T cell-mediated immune response, and immune modulation of the microenvironment that can help to predict improved patient outcomes.


Ultrastructural and immunocytochemical characterization of an immortalized human breast epithelial cell line, MCF-10.
The human breast epithelial cell line MCF-10 was derived from s.c. mastectomy tissue from a 36-year-old, parous, premenopausal woman with fibrocystic disease. It was initiated as a mortal cell line (MCF-10M), which senesces when transferred serially in 1.05 mM calcium. These cells spontaneously gave origin to two immortal sublines, MCF-10A, or attached cells, and MCF-10F, or floating cells, which have proliferated for more than 4 years in Dulbecco's modified essential medium and Ham's F-12 either with the customary calcium concentration of 1.05 mM (DMEM-H) or in medium containing 0.04 mM calcium or low calcium. Studies reported here indicate that MCF-10 is a mammary epithelial cell line. Electron microscopy showed that both MCF-10A and MCF-10F have characteristics of luminal ductal cells, but not of myoepithelial cells. When grown for more than 1200 days in Dulbecco's modified essential medium-Ham's F-12 and low calcium media, respectively, they maintained their epithelial characteristics, although the concentration of calcium exerted a powerful influence on cell morphology. Cells grown in Dulbecco's modified Eagle's medium-Ham's F12 medium are low cuboidal with numerous desmosomes and short microvilli, whereas those grown in low calcium medium have significantly reduced number of desmosomes, are more spherical, and have greater numbers of microvilli which are longer than those of cells grown in DMEM-H. The breast epithelial orgin of these cells was confirmed by immunocytochemical detection of epithelial sialomucins and keratins. The monoclonal antibodies MFA-breast and MC5 and the polyclonal antibody epithelial membrane antigen were used to detect the epithelial sialomucins. Keratins were characterized by using KA-4, K-14, AE1/AE3, and K-19 specific antibodies. It was concluded that MCF-10A and MCF-10F cells are breast epithelial cells and that they represent an important tool for studies of the basic processes of growth and carcinogenesis.


Natal origin and age-specific egress of Pacific bluefin tuna from coastal nurseries revealed with geochemical markers.
Geochemical chronologies were constructed from otoliths of adult Pacific bluefin tuna (PBT) to investigate the timing of age-specific egress of juveniles from coastal nurseries in the East China Sea or Sea of Japan to offshore waters of the Pacific Ocean. Element:Ca chronologies were developed for otolith Li, Mg, Mn, Zn, Sr, and Ba, and our assessment focused on the section of the otolith corresponding to the age-0 to age-1 + interval. Next, we applied a common time-series approach to geochemical profiles to identify divergences presumably linked to inshore-offshore migrations. Conspicuous geochemical shifts were detected during the juvenile interval for Mg:Ca, Mn:Ca, and Sr:Ca that were indicative of coastal-offshore transitions or egress generally occurring for individuals approximately 4-6 mo. old, with later departures (6 mo. or older) linked to overwintering being more limited. Changepoints in otolith Ba:Ca profiles were most common in the early age-1 period (ca. 12-16 mo.) and appear associated with entry into upwelling areas such as the California Current Large Marine Ecosystem following trans-Pacific migrations. Natal origin of PBT was also predicted using the early life portion of geochemical profile in relation to a baseline sample comprised of age-0 PBT from the two primary spawning areas in the East China Sea and Sea of Japan. Mixed-stock analysis indicated that the majority (66%) of adult PBT in our sample originated from the East China Sea, but individuals of Sea of Japan origin were also detected in the Ryukyu Archipelago.


Management of adenoid cystic carcinoma of the head and neck: a single-institute study with over 25-year follow-up.
Adenoid cystic carcinoma is a rare malignant tumor arising from exocrine glands such as the major and minor salivary glands of the paranasal sinuses or the external auditory canal. Although multiple retrospective clinical studies of ACC have been reported to date, clinical questions, such as 1) long-term prognosis beyond 20 years, 2) usefulness and suitability for treatment of therapeutic interventions, 3) therapeutic goal to aim for, and 4) prognosis by recurrence sites, are still unclear. To improve understanding and management of adenoid cystic carcinoma of the head and neck (ACC), a retrospective study with 58 new ACC cases between 1991 and 2016 was performed. The median observation period was 66.8 months (range 3-316 months). The overall clinical stages were as follows: I, 6.9%; II, 25.9%; III, 19.0%; and IV, 48.2%. Histology was cribriform/tubular type (C-T type) in 62.0% and solid type in 27.5%. The main treatment strategy was definitive surgery, which was performed in 75.2% of cases. Overall 10-year, 20-year, and 25-year survivals were 63.7, 27.3, and 20.0%, respectively. Similarly, disease-specific survival (DSSs) was 65.7, 51.2, and 38.4%, respectively, and disease-free survival was 25.2, 9.4, and 9.4%, respectively. Conducting surgery (HR: 0.19, 95% CI: 0.06-0.61, p = 0.005) and C-T type (HR: 0.32, 95% CI: 0.11-0.93, p = 0.036) were independent prognostic predictors of DSS. DSS was significantly prolonged after salvage surgery for both locoregional recurrence (p = 0.004) and lung metastatic recurrence (p = 0.012, vs best supportive care). In ACC cases, both initial surgical treatment and repetitive surgical resection of resectable recurrent lesions, including both locoregional and lung metastases, resulted in longer survival. The major goal of treatment for ACC may be long-term survival including cancer-bearing survival, resulting in either natural death or intercurrent-disease death, since judging cure of ACC is almost impossible. Retrospectively registered.


Diagnostic Pearls and Clinical Implications of Prekallikrein Deficiency.
Prekallikrein (PK) deficiency is extremely rare, and manifestations are not well characterized due to a small number of cases reported and the lack of scientific clarity about its role in clot formation in vivo. Here, we report a case of a 64-year-old male, with no known history of abnormal bleeding, who scheduled to undergo deep brain stimulator placement for control of his Parkinson's disease. During pre-procedure testing, activated partial thromboplastin time (PTT) was found to be prolonged at 146 seconds. Mixing studies were suggestive of a coagulation factor deficiency. His PTT characteristically became shorter with prolonged incubation, providing a clue at testing for PK levels, which were found to be severely low. He, subsequently, underwent surgery without any complications. Our case further highlights the clinical pearls for diagnosis and further endorses that these patients can safely undergo surgical procedures without the need for plasma transfusions or factor concentrate usage.


Effect of a Fusion Peptide by Covalent Conjugation of a Mitochondrial Cell-Penetrating Peptide and a Glutathione Analog Peptide.
Previously, we designed and synthesized a library of mitochondrial antioxidative cell-penetrating peptides (mtCPPs) superior to the parent peptide, SS31, to protect mitochondria from oxidative damage. A library of antioxidative glutathione analogs called glutathione peptides (UPFs), exceptional in hydroxyl radical elimination compared with glutathione, were also designed and synthesized. Here, a follow-up study is described, investigating the effects of the most promising members from both libraries on reactive oxidative species scavenging ability. None of the peptides influenced cell viability at the concentrations used. Fluorescence microscopy studies showed that the fluorescein-mtCPP1-UPF25 (mtgCPP) internalized into cells, and spectrofluorometric analysis determined the presence and extent of peptide into different cell compartments. mtgCPP has superior antioxidative activity compared with mtCPP1 and UPF25 against H2O2 insult, preventing ROS formation by 2- and 3-fold, respectively. Moreover, we neither observed effects on mitochondrial membrane potential nor production of ATP. These data indicate that mtgCPP is targeting mitochondria, protecting them from oxidative damage, while also being present in the cytosol. Our hypothesis is based on a synergistic effect resulting from the fused peptide. The mitochondrial peptide segment is targeting mitochondria, whereas the glutathione analog peptide segment is active in the cytosol, resulting in increased scavenging ability.