[Cells on intraocular lenses. Comparison of specular and slit-lamp microscopy findings].
Examination with a specular microscope facilitates diagnostic evaluation of cells on the surface of intraocular lenses. Similarly, slit-lamp examination makes specular microscopic studies possible. However, image resolution and magnification are not achieved by using this method, though the amount of time and instrumental effort is minimal. Various types of cells can be differentiated. Fibroblast-like cells can only be demonstrated readily in the specular areas, whereas multinucleate giant cells can usually also be seen by using focal illumination. It is difficult to differentiate the smaller, mononucleated cells.


Potassium channel openers as potential therapeutic weapons in ion channel disease.
The opening of potassium (K+) channels, causing hyperpolarization of the cell membrane, is a physiological means of decreasing cell excitability. Thus, drugs with this property will demonstrate a broad clinical potential. The identification of synthetic molecules that evoke physiological responses (for example smooth muscle relaxation) by the opening of K+ channels led to a new direction in the pharmacology of ion channels. The term "potassium channel openers" was initially associated with a group of chemically diverse agents (for example, cromakalim, pinacidil, nicorandil) that evoke K+ efflux through adenosine 5'-triphosphate (ATP)-sensitive K+ channels (KATP). This finding initiated a search to identify molecules that specifically open other K+ channel subtypes (for example large conductance calcium-activated K+ channels [BKCa]). K+ channel opening properties have been demonstrated in a diverse range of synthetic chemical structures and endogenous substances. Second generation KATP channel openers (KATPCOs) demonstrate heterogeneous pharmacology indicative of independent sites of action for the different agents. Successful cloning of the KATP channel has shed light on the heterogeneity of the structure targeted by KATPCOs. Expression of the actions of KATPCOs involves three isoforms of the sulfonylurea (SUR) receptor (which forms the beta subunit of the KATP channel). The distribution of the SUR isoforms (and potential of identifying new isoforms) provides unique targets for the development of selective KATPCOs giving focused therapeutic approaches to clinical conditions for example cardiac ischemia, urinary incontinence, neurodegeneration, obesity and autoimmune diseases. BKCa channels are found in a diverse array of tissues and due to voltage and Ca sensitivity may work as a negative feedback process. A variety of small synthetic molecules (for example, NS004, fenamates) and natural product-derived compounds (DHS-I, maxikdiol) have been identified as selective BKCa channel openers which should have a profound impact in controlling diseases. The discovery of numerous variants of the alpha subunit (ion conductance pore) and beta subunit (contributes biophysical and pharmacological properties) complex of the BKCa channel gives potential to target specific tissues with selective openers. Little is known, however, about the site(s) of interaction of openers of these channels. The discovery of K+ channel subtype-specific openers and their evaluation in different diseases will determine the degree to which these channels (KATP, BKCa), or their isoforms, represent realistic therapeutic targets. Drugs already marketed that open K+ channels were discovered empirically, and most have serious safety and efficacy problems. New scientific methods, utilizing molecular insight, are implicating K+ channel dysfunction in numerous disease states and are identifying new targets for the future generation of K+ channel opening drugs.


[Strengths and challenges in connection to the clinical education in anaesthesiology:].
We performed a qualitative interview study regarding perceived strengths and challenges in connection to the clinical curriculum in anaesthesiology. Medical students and the medical specialists in charge of supervising the clinical curriculum pointed out that the primary strengths were that the staff was committed to tutoring, good general structure and possibility to explore basic and more advanced clinical procedures. Challenges were seen in the current lack of cooperation between hospitals and the general lack of knowledge regarding medical students' curriculum. Future processes for better practice sharing between hospitals should be explored.


Impact of 12-Week Moderate-Intensity Aerobic Training on Inflammasome Complex Activation in Elderly Women.
Aging often associates with a chronic low-grade inflammatory status that can be consequent to the activation of Toll-like receptors (TLRs) and the downstream NLR family pyrin domain containing 3 (NLRP3) inflammasome and causes a chronic secretion of pro-inflammatory cytokines. Since exercise has known anti-inflammatory effects, we investigated the effect of Nordic walking training on inflammasome activation and downstream effectors in elderly women. A population of elderly women was divided into EXP (n = 29) that completed 12 weeks of the moderate-intensity aerobic training program and CTRL (n = 29), performing no activity. Blood samples were taken before and after the first (T1-pre and T1-post, respectively) and last (T2-pre and T2-post, respectively) exercise unit. Inflammasome activation status was assessed by whole blood NLRP3 and TLR4 expression by RT-qPCR. Serum levels of IL-1β, IL-6, TNFα, and IL-18 cytokines were assayed by multiplex fluorescent beads-based immunoassays or ELISA. NLRP3 and TLR4 levels were reduced 2 folds between T1-pre and T2-pre and induced at T2-post, compared to T2-pre, by 2.6- and 2.9-fold, respectively. A single exercise bout elicited a 1. 38-, 1. 5-, and 1.36-fold rise of IL-1β, TNFα, and IL-6 concentration, respectively, although not significant, at the beginning of the training (T1-pre vs. T1-post), a 1.4-fold decrease for IL-1β and TNFα at the end of the training (T1-pre vs. T2-pre), and a 2-, 1.8- and 1.26-fold increase after the last exercise session (T2-pre vs. T2-post) for the three cytokines. When stratifying the population based on BMI in normal weight (NW) and overweight (OW), NLRP3 and TLR4 expression was affected only in NW. As for inflammatory cytokines, IL-1β was modulated in NW at the beginning of the training, whereas in OW at the end of the training; for TNFα, this time-dependent modulation was significant only in OW. Applied aerobic training affected the resting expression of inflammasome constituents (NLRP3 and TLR4) and levels of downstream effectors (IL-1β, TNFα, and IL-6). However, at the end of the program, participants acquire an acute inflammatory response to exercise that was absent at baseline. Future studies would have to define the molecular mechanisms associated with, and how to potentiate, the exercise-associated inflammatory response.


Proteomic Analysis of Stage-II Breast Cancer from Formalin-Fixed Paraffin-Embedded Tissues.
Breast cancer is the most frequently occurring disease among women worldwide. The early stage of breast cancer identification is the key challenge in cancer control and prevention procedures. Although gene expression profiling helps to understand the molecular mechanism of diseases or disorder in the living system, gene expression pattern alone is not sufficient to predict the exact mechanisms. Current proteomics tools hold great application for analysis of cancerous conditions. Hence, the generation of differential protein expression profiles has been optimized for breast cancer and normal tissue samples in our organization. Normal and tumor tissues were collected from 20 people from a local hospital. Proteins from the diseased and normal tissues have been investigated by 2D gel electrophoresis and MALDI-TOF-MS. The peptide mass fingerprint data were fed into various public domains like Mascot, MS-Fit, and Pept-ident against Swiss-Prot protein database and the proteins of interest were identified. Some of the differentially expressed proteins identified were human annexin, glutathione S-transferase, vimentin, enolase-1, dihydrolipoamide dehydrogenase, glutamate dehydrogenase, Cyclin A1, hormone sensitive lipase, beta catenin, and so forth. Many types of proteins were identified as fundamental steps for developing molecular markers for diagnosis of human breast cancer as well as making a new proteomic database for future research.


Danger assessment of a spring-gun device (muzzleloader) loaded only with black powder.
We report a case involving a spring-gun device (muzzleloader) loaded solely with gunpowder, installed next to shoes to prevent the neighbors' puppy from removing them. The booby trap was triggered by the 15-year-old dog-owners son when he tried to put the shoes out of the reach of the puppy. The boy suffered second degree superficial burns located mainly at the dorsal side of the right hand and fingers. To estimate the danger of the used weapon, several tests were undertaken on soap blocks from different distances and with different loads of black powder. The particle density per mm and the depth of black powder tattooing in the soap was compared with the boy's injuries, and found conclusive with the gun-owners statement regarding the loading of the weapon. Furthermore, our experiments indicated that the gunpowder load involved was not able to inflict permanent damage, not even to the eyes, at the here estimated firing distance.


Biomarker potential of the LEF1/TCF family members in breast cancer: Bioinformatic investigation on expression and clinical significance.
The LEF1/TCF transcription factor family is related to the development of diverse tissue types, including the mammary tissue, and dysregulation of its expression and function has been described to favor breast tumorigenesis. However, the clinical and biological relevance of this gene family in breast cancer is still poorly understood. Here, we used bioinformatics approaches aiming to reduce this gap. We investigated its expression patterns in molecular and immune breast cancer subtypes; its correlation with immune cell infiltration, and its prognostic values in predicting outcomes. Also, through regulons construction, we determined the genes whose expression is influenced by these transcription factors, and the pathways in which they are involved. We found that LEF1 and TCF3 are over-expressed in breast tumors regarding non-tumor samples, while TCF4 and TCF7 are down-expressed, with the gene's methylation status being associated with its expression dysregulation. All four transcription factors presented significance at the diagnostic and prognostic levels. LEF1, TCF4, and TCF7 presented a significant correlation with immune cell infiltration, being associated with the immune subtypes of less favorable outcomes. Altogether, this research contributes to a more accurate understanding of the expression and clinical and biomarker significance of the LEF1/TCF transcription factors in breast cancer.


Identification of a Small Molecule That Enhances Ferroptosis via Inhibition of Ferroptosis Suppressor Protein 1 (FSP1).
Glutathione peroxidase 4 (GPX4) is an intracellular enzyme that oxidizes glutathione while reducing lipid peroxides and is a promising target for cancer therapy. To date, several GPX4 inhibitors have been reported to exhibit cytotoxicity against cancer cells. However, some cancer cells are less sensitive to the known GPX4 inhibitors. This study aimed to explore compounds showing synergistic effects with GPX4 inhibitors. We screened a chemical library and identified a compound named NPD4928, whose cytotoxicity was enhanced in the presence of a GPX4 inhibitor. Furthermore, we identified ferroptosis suppressor protein 1 as its target protein. The results indicate that NPD4928 enhanced the sensitivity of various cancer cells to GPX4 inhibitors, suggesting that the combination might have therapeutic potential via the induction of ferroptosis.


Three-Year Outcomes of Second-generation Trabecular Micro-bypass Stents (iStent inject) With Phacoemulsification in Various Glaucoma Subtypes and Severities.
This longitudinal case series provides valuable long-term data on the safety and efficacy of iStent inject with concomitant cataract surgery. Patients experienced significant and durable reductions in both intraocular pressure (IOP) and medication burden through 3 years postoperative. The purpose of this study was to examine the 3-year effectiveness and safety of iStent inject second-generation trabecular micro-bypass stent implantation with concomitant phacoemulsification cataract surgery in eyes with various glaucoma subtypes and severities. This was a single-center consecutive case series. Three-year outcomes included mean IOP and medications, the proportions of eyes with IOP ≤18, ≤15, and ≤12 mm Hg, and success (absence of secondary glaucoma interventions). Safety included best-corrected visual acuity (BCVA), cup-to-disc ratio, visual field (VF) mean deviation, retinal nerve fiber layer (RNFL), and ganglion cell-inner plexiform layer (GCIPL) thickness, and adverse events. A total of 124 eyes with different glaucoma subtypes and severities were included. At 3 years postoperative, mean IOP reduced from 16.90±3.85 mm Hg preoperatively to 13.17±2.83 mm Hg (22% reduction, P<0.001) and mean medication burden decreased from 2.38±1.29 medications preoperatively to 1.16±1.22 medications (51% reduction, P<0.001). At 3 years, 96% of eyes achieved IOP ≤18 mm Hg (vs. 69% preoperatively), 80% of eyes achieved IOP ≤15 mm Hg (vs. 40% preoperatively), and 42% of eyes achieved IOP ≤12 mm Hg (vs. 7% preoperatively) with 76% of eyes eliminating ≥1 medication and 37% of eyes eliminating ≥2 medications versus preoperative medication burden. The 3-year cumulative survival rate was 74%. Postphacoemulsification BCVA improvement was preserved, and cup-to-disc ratio, VF mean deviation, and RNFL and GCIPL thickness remained stable. A favorable safety profile included no intraoperative complications and few, transient, postoperative adverse events. Significant and sustained IOP and medication reductions were achieved through 3 years after iStent inject implantation with cataract surgery in a real-world clinical population with mild-to-severe glaucoma, along with favorable safety including stable BCVA, VF, and RNFL and GCIPL thickness.


Effects of base polymer hydrophobicity and end-group modification on polymeric gene delivery.
A new 320-member polymer library of end-modified poly(β-amino ester)s was synthesized. This library was chosen such that small differences to the structures of component backbone, side-chain, and end-group monomers could be systematically and simultaneously evaluated. The in vitro transfection efficacy and cytotoxicity of DNA nanoparticles formed from this library were assessed. This library approach not only enabled us to synthesize and test a large variety of structures rapidly but also provided us with a robust data set to analyze for the effect of small structural permutations to polymer chain structure. Small changes to the side chains, backbones, and end groups within this polymer library produced dramatic results, with transfection efficacy of CMV-Luc varying over 4 orders in a 96-well plate format. Increasing hydrophobicity of the base polymer backbone and side chain tended to increase transfection efficacy, but the most hydrophobic side chains and backbones showed the least requirement for a hydrophobic pair. Optimal PBAE formulations were superior to commercially available nonviral alternatives FuGENE HD and Lipofectamine 2000, enabling ~3-fold increased luminescence (2.2 × 10(6) RLU/well vs 8.1 × 10(5) RLU/well) and 2-fold increased transfection percentage (76.7% vs 42.9%) as measured by flow cytometry with comparable or reduced toxicity.


Indomethacin sustained-release suppositories containing sugar ester in polyethylene glycol base.
Indomethacin (IM) sustained-release suppositories were prepared by the fusion method using sugar ester and polyethylene glycol 4000 (PEG). The suppositories were evaluated by in vitro release testing, X-ray analysis and in vivo absorption testing in rabbits. X-ray analysis showed that IM was amorphous in PEG-base suppositories. In a release test, slow-release was obtained when the sugar ester content of a suppository was 60%. The IM plasma level following the administration of the suppository was well sustained in the absorption test. The main slow-release mechanism is considered to be the release of IM from the matrix composed of sugar ester and PEG, which is represented by the Higuchi equation. A good correlation between the release test and the absorption test was obtained. It is considered that the amorphous state of IM in this type of sustained-release suppository would enhance the release and absorption of IM in the rectum of the rabbit, whose rectal fluid volume is small.


[The clinical efficacy of epristeride in the treatment of benign prostatic hyperplasia].
To estimate the clinical safety, efficacy of Epristeride in the treatment of benign prostatic hyperplasia (BPH). One hundred and fourty-one BPH patients, aged from 50 to 83 years old [(66.18 +/- 6.41) yr], were given Epristeride tablets 5 mg p.o., twice daily for 120 days. To synthetically estimate the efficacy by the changing rate of international prostate symptom score (IPSS), maximum flow rate (Qmax), the volume of prostate (V) and the residual urine (Ru). The total efficacy was 81.29%. The adverse reactions include 1 case (0.72%) of exanthem, 1 case (0.72%) of nausea and vomiting, 1 case (0.72%) of insomnia, 1 case (0.72%) of tinnitus and 3 cases (2.16%) of erectile dysfunction. Two patients discontinued the treatment due to the side effects (1.44%). There was no clinical significantly abnormal laboratory index during the treatment. Epristeride tablets was safe and effective in the treatment of BPH.


Two new species of Willowsia (Collembola: Entomobryidae) from Guizhou Province, China.
The genus Willowsia Shoebotham, 1917 belongs to the family Entomobryidae. Within the family it is characterised by having 4-segmented antennae, 8+8 eyes, spines and scales absent from dens, claw with unpaired inner teeth, bidentate mucro and scales on body. Twenty-six species have been described in the genus worldwide including eight species from China. Two new species, Willowsia pseudoshi sp. nov. and W. variabilis sp. nov., from China are described here. The former species has pale yellow antennae except for distal part of Antenna IV which has some blue pigment and 4 macrochaetae are present on Abdomen I. The latter species has the whole antenna pigmented blue and 7-8 macrochaetae present on Abdomen I. A key to Chinese species of Willowsia is given.


Development and application of a rapid and sensitive LC-MS/MS method for simultaneous quantification of six diterpene lactones in rats after oral administration of Rhizoma Dioscoreae Bulbiferae extract.
Diterpene lactones have been considered as the main therapeutic and hepatotoxic constituents of Rhizoma Dioscoreae Bulbiferae in recent years. In this work, a simple, rapid and accurate LC-MS/MS method was established and validated to determine six diterpene lactones in rat plasma simultaneously, including Diosbulbin B (DIOB), Diosbulbin C (DIOC), Diosbulbin D (DIOD), Diosbulbin G (DIOG), Diosbulbin J (DIOJ) and Diosbulbin L (DIOL), after oral administration of Rhizoma Dioscoreae Bulbiferae extract. The six diterpene lactones, with the inclusion of two pairs of isomer (DIOB & D, DIOC & L), and Buspirone (internal standard, IS) were successfully separated using an XDB-C18 column with the gradient elution, consisting of water with 0.1% (v/v) FA and methanol with 0.1% (v/v) FA, under a flow rate of 0.50 mL/min in 5.8 min. Precursor-product ion transitions were optimized to be m/z 362.1 → 317.1, 363.1 → 207.1, 345.0 → 299.2, 364.3 → 347.0, 396.3 → 379.3, 363.2 → 345.1 and 386.3 → 122.2 for DIOB, DIOC, DIOD, DIOG, DIOJ, DIOL and buspirone at positive ion mode with an electrospray ionization source (ESI), respectively. The linearity ranges of this present method were 0.50 to 500 μg/L for DIOB, 20.0 to 20,000 μg/L for DIOC and 2.00 to 2000 μg/L for DIOD, DIOG, DIOJ and DIOL, respectively. And the LLOQs were as low as 0.20 μg/L for DIOB, 20.0 μg/L for DIOC and 2.00 μg/L for DIOB, D, G, J and L. The accuracy of each analyte was within the range of 95.8% to 101.0% and the precision was <11.3%. No matrix effect and carry over was observed, and the recovery of the six analytes ranged from 87.3% to 109% with the RSD <11.4% within the concentrations range. The validated method was further applied to the pharmacokinetics investigation of DIOB, DIOC, DIOD, DIOG, DIOJ and DIOL successfully after oral administration of Rhizoma Dioscoreae Bulbiferae extract at 1.53 g/kg in rats.


[Bilateral keratoconus: a case report].
Among corneal dystrophies, the keratoconus is one of the most frequently observed among young adults. A clinico pathological case is reported in a 13-year-old-girl of African origin. The diagnosis of bilateral keratoconus was established based on the obvious changes of the corneal curvature and thickness. After an unsuccessful attempt to improve vision with contact lenses, a keratoplasty was finally performed on one side to remove the pathological cornea. Its histopathological study found the characteristic changes of keratoconus: breaks of Bowman's layer and corneal thinning.


Hydatid disease at Royal North Shore Hospital: results of surgical treatment.
A study of the results of surgery for hydatid disease was undertaken at Royal North Shore Hospital for the period 1966-83. There were 51 patients who underwent surgery for this disease and follow-up was obtained in all but eight cases. Comparison of cumulative survivals for this group with that of a matched Australian population showed that patients undergoing surgical treatment of their hydatid can anticipate a normal life expectancy. There was no evidence that patients with extrahepatic disease fared any worse than those with disease confined to the liver. Documented recurrence of hydatid disease occurred in eight cases following surgery and only one patient actually died from hydatid disease. This was a case in which spillage occurred at the initial procedure. This paper is presented as a reminder that hydatid disease still occurs with sufficient frequency to pose therapeutic problems for surgeons. Whilst total eradication of the parasite will only occur with enforcement of public health measures, surgery would still appear to be effective therapy for established hydatid disease so long as care is taken to avoid cyst rupture.


Current Prophylaxis and Treatment Approaches for Acute Graft-Versus-Host Disease in Haematopoietic Stem Cell Transplantation for Children With Acute Lymphoblastic Leukaemia.
Acute graft-versus-host disease (aGvHD) continues to be a leading cause of morbidity and mortality following allogeneic haematopoietic stem cell transplantation (HSCT). However, higher event-free survival (EFS) was observed in patients with acute lymphoblastic leukaemia (ALL) and grade II aGvHD vs. patients with no or grade I GvHD in the randomised, controlled, open-label, international, multicentre Phase III For Omitting Radiation Under Majority age (FORUM) trial. This finding suggests that moderate-severity aGvHD is associated with a graft-versus-leukaemia effect which protects against leukaemia recurrence. In order to optimise the benefits of HSCT for leukaemia patients, reduction of non-relapse mortality-which is predominantly caused by severe GvHD-is of utmost importance. Herein, we review contemporary prophylaxis and treatment options for aGvHD in children with ALL and the key challenges of aGvHD management, focusing on maintaining the graft-versus-leukaemia effect without increasing the severity of GvHD.


Associations of e-cigarette experimentation with support for tobacco control policies in the European Union, 2012-2014.
There are limited data on the potential effects of e-cigarette experimentation on support for tobacco control policies. To bridge this gap, we assessed associations between e-cigarette experimentation and support for tobacco control policies in the European Union 2012-2014. We also investigated variations across tobacco-use status, e-cigarette experimentation and socio-demographic characteristics. Datasets were used from the Special Eurobarometer for Tobacco surveys performed in 2012 (n=26 751) and 2014 (n=27 801). Tobacco control policies assessed were: banning advertising, policies to keep tobacco out of sight, banning online sales, banning flavors, standardized packaging, tax increases, and policies to reduce illicit trade in tobacco. We use multilevel logistic regression models to assess variations in socio-demographics and tobacco/e-cigarette use with support for these policies in 2014, and examined changes in support for these policies, between 2012 and 2014, separately by tobacco-use status (never, current, and former smokers). Population support for tobacco control policies was high in 2014: policies to reduce illicit trade had the highest level of support at 70.1%, while tax increases were the least likely measure to be supported with 52.3% support. Among never and former smokers, experimentation with e-cigarettes was associated with reduced support for all tobacco control policies assessed. For example, never smokers who had experimented with e-cigarettes were less likely to support either tobacco advertising bans (adjusted odds ratio aOR=0.57, 95% confidence interval 0.46-0.71) or standardized packaging for tobacco (aOR=0.58, 95% CI: 0.47-0.71). Former smokers who had experimented with e-cigarettes were less likely to either support standardized packaging for tobacco (aOR=0.70, 95% CI: 0.60-0.82) or keeping tobacco out of sight (aOR=0.77, 95% CI: 0.65-0.90). Among current smokers, e-cigarette experimentation was not associated with support for the tobacco control policies assessed. E-cigarette experimentation was consistently associated with reduced support for tobacco control policies among never and former smokers but not among current smokers. The implications of these findings for tobacco control are unknown, but the data support concerns that e-cigarette experimentation may affect public support for established tobacco control policies within specific subgroups. Further research is needed to assess potential long-term impacts on tobacco control policies.


Detection of an infrared near-field optical signal by attaching an infrared-excitable phosphor to the end of a photocantilever.
To improve the signal-to-noise ratio of near-field scanning optical microscopy, we propose attaching an infrared-excitable phosphor (IEP) to a photocantilever. One source of noise is the light scattered from locations on the sample surface other than that of the probe tip. By detecting only the light scattered from the tip, we can obtain a near-field optical signal without noise. We attached an IEP particle to a photocantilever to convert infrared light to visible light and we used 1550-nm infrared illumination, so the light scattered from the sample was only infrared. The silicon photodiode of the photocantilever is 10(6) times less sensitive to infrared light than to visible light. As a result, only the converted visible light from the IEP particle, i.e. the signal containing the near-field optical information from the tip, was detected. We verified that the photocantilever detected the signal in the evanescent light produced by infrared illumination and that the detected signal was the light converted by the IEP. The experimental results show the feasibility of detecting infrared light and not the background light through the use of the IEP.


LUMBAR syndrome: A case manifesting as cutaneous infantile hemangiomas of the lower extremity, perineum and gluteal region, and a review of published work.
We herein report a rare case of LUMBAR syndrome. A 1-month-old female infant presented with extensive segmental hemangiomas on the left lower extremity, left perineum and gluteal region with ulceration. Bilateral labia minoras were asymmetrical. Both legs were asymmetrical with left leg atrophy, and the intergluteal cleft was deviated. A dark red pustule and a sacrococcygeal dimple could be seen in the lumbosacral region. Lipomyelomeningocele, tethered cord and sacrum dysplasia were noted by magnetic resonance imaging. The patient was found to have an absent left kidney at 32 weeks of pregnancy. Eventually, we draw the diagnosis of LUMBAR syndrome. In addition, we discuss the clinical manifestation, diagnosis, treatment and pathogenesis by a review of published work.


Population health of Fallow deer (Dama dama) on Little St. Simons Island, Georgia, USA.
Fallow deer (Dama dama) were introduced to Little St. Simons Island, Georgia, USA in the 1920s and thrive at high population densities, to the exclusion of white-tailed deer (Odocoileus virginina). The presence of introduced pathogens and parasites as a result of their introduction is currently unknown, as is the impact of native disease on the exotic fallow deer. Hunter-killed fallow deer from 2003-2005 were necropsied and surveyed for evidence of infectious disease, parasitic agents, and toxicologic parameters. Fallow deer were positive for antibodies to bovine virus diarrhea virus I and II, bluetongue virus, and bovine adenovirus. Twenty species of bacteria were isolated from the internal organs, and 14 species of parasites were recovered including one abomasal nematode, Spiculopteragia asymmetrica, which is not known to occur in native North American ungulates. Concentrations of liver and copper were low, while lead, zinc, and iron were considered within normal levels. No clinical signs of disease were noted, and the overall health of the insular fallow deer was considered good.


Genome-wide comparison of four MRSA clinical isolates from Germany and Hungary.
Staphylococcus aureus is a drug-resistant pathogen, capable of colonizing diverse ecological niches and causing a broad spectrum of infections related to a community and healthcare. In this study, we choose four methicillin-resistant S. aureus (MRSA) clinical isolates from Germany and Hungary based on our previous polyphasic characterization finding. We assumed that the selected strains have a different genetic background in terms of the presence of resistance and virulence genes, prophages, plasmids, and secondary metabolite biosynthesis genes that may play a crucial role in niche adaptation and pathogenesis. To clarify these assumptions, we performed a comparative genome analysis of these strains and observed many differences in their genomic compositions. The Hungarian isolates (SA H27 and SA H32) with ST22-SCCmec type IVa have fewer genes for multiple-drug resistance, virulence, and prophages reported in Germany isolates. Germany isolate, SA G6 acquires aminoglycoside (ant(6)-Ia and aph(3')-III) and nucleoside (sat-4) resistance genes via phage transduction and may determine its pathogenic potential. The comparative genome study allowed the segregation of isolates of geographical origin and differentiation of the clinical isolates from the commensal isolates. This study suggested that Germany and Hungarian isolates are genetically diverse and showing variation among them due to the gain or loss of mobile genetic elements (MGEs). An interesting finding is the addition of SA G6 genome responsible for the drastic decline of the core/pan-genome ratio curve and causing the pan-genome to open wider. Functional characterizations revealed that S. aureus isolates survival are maintained by the amino acids catabolism and favor adaptation to growing in a protein-rich medium. The dispersible and singleton genes content of S. aureus genomes allows us to understand the genetic variation among the CC5 and CC22 groups. The strains with the same genetic background were clustered together, which suggests that these strains are highly alike; however, comparative genome analysis exposed that the acquisition of phage elements, and plasmids through the events of MGEs transfer contribute to differences in their phenotypic characters. This comparative genome analysis would improve the knowledge about the pathogenic S. aureus strain's characterization, and responsible for clinically important phenotypic differences among the S. aureus strains.


Maltol Promotes Mitophagy and Inhibits Oxidative Stress via the Nrf2/PINK1/Parkin Pathway after Spinal Cord Injury.
Spinal cord injury (SCI), a fatal disease in the central nervous system, is characteristic of weak neuronal regeneration ability and complex pathological progress. Activation of oxidative stress (OS) and apoptosis-mediated cell death significantly contributes to the progression of SCI. Current evidence suggests that maltol exerts natural antioxidative properties via obstructing OS and apoptosis. However, the significant effect of maltol on SCI treatment has never been evaluated yet. In our current study, we explored maltol administration that could trigger the expression of Nrf2 and promote the retranslocation of Nrf2 from the cytosol to the nucleus, which can subsequently obstruct OS signal and apoptosis-mediated neuronal cell death after SCI. Furthermore, we found that maltol treatment enhances PINK1/Parkin-mediated mitophagy in PC12 cells, facilitating the recovery of mitochondrial functions. Our findings propose that maltol could be a promising therapeutic candidate for the treatment and management of SCI.


Excited-state switching by per-fluorination of para-oligophenylenes.
Fluorination has become a versatile route to tune the electronic and optical properties of organic conjugated materials. Herein we report a new phenomenon, excited-state switching by per-fluorination of para-oligophenylenes, placing a low intensity 1(1)B(2) state below the 1(1)B(1) state, giving rise to large Stokes shifts. The switching is attributed to the specific impact of fluorine on the delocalized and localized frontier orbitals as elucidated by quantum-chemical calculations. The sterical demands of the fluorine atom additionally diminish efficient conjugation along the chain, leading to hypsochromic shifts with respect to the unsubstituted counterparts and to a weak chain length dependence of the absorption and unstructured emission spectra and enhanced internal conversion.


The present role of staphylococci in development of surgical hospital infection.
At present the problem of staphylococcal infection appears to be highly important. The research aim was to establish the role and significance of Staphylococci, particularly methicillin resistant in development of complications in modern surgery, sources and ways of Staphylococci appearance and spread in today surgical departments. Particularly seasonal chronorythms of microflora carriage by the medical personnel and students was studied. Most of the strains were resistant to traditional antibacterial remedies. Since the begin of the half-century antibiotic era Staphylococci are still a problem in Surgery. Adequate microbiologic monitoring of stuff and students is recommended to prevent a lot of complications.


Viability of hospital-based radiography programs.
To identify and address risks threatening hospital-based radiography programs and to report conditions that contribute to their survival. Program directors of the 201 hospital-sponsored radiography programs accredited by the Joint Review Committee on Education in Radiologic Technology were surveyed via questionnaire. Program demographic data are reported for comparison. More than 90% of the survey respondents believe their programs will remain viable over the next 5 years, indicating a possible plateau in program closures. A majority of the program directors cite financial concerns as a risk factor. The study results suggest a need for additional inquiry covering inter-related groups.


Tumor-induced osteomalacia: a single center experience.
To describe the clinical presentation, localization modalities, and management of patients with tumor-induced osteomalacia (TIO). We performed a retrospective analysis of case records of patients diagnosed with TIO between January 1996 and March 2010 at our institution in Mumbai, India. Nine patients (6 female and 3 male) with a mean age of 37.5 ± 17.5 years with biochemical and imaging evidence of TIO were included in the study. Overall, patients presented with proximal muscle weakness and pain. Three patients had neurofibromatosis 1, one had isolated schwannoma, and one had epidermal nevus syndrome. The mean delay in diagnosis was 7.67 years. Biochemical studies revealed normal serum calcium (mean, 9.2 ± 0.8 mg/dL), low serum phosphorus (mean, 1.36 ± 0.54 mg/dL), and low maximal tubular reabsorption of phosphorus factored for glomerular filtration rate (mean, 0.94 ± 0.49 mg/dL). Fibroblast growth factor-23 was increased in 3 of the patients without neurofibromatosis but was normal or near-normal in all the patients with neurofibromas. A fludeoxyglucose F 18 positron emission tomography (FDG PET) scan helped to localize the tumors in 4 of the 5 patients with diagnoses other than neurofibromatosis. Three patients underwent surgical excision and were cured. One patient underwent biopsy and partial excision. Histopathologic findings were suggestive of phosphaturic mesenchymal tumor, benign fibrous histiocytoma, nonossifying fibroma, and sciatic nerve schwannoma. There is a well-known delay in the diagnosis of TIO. FDG PET is a useful modality for localization of tumors. Preoperative localization increases the odds for cure after surgical excision.


Rapid adherence to collagen IV enriches for tumour initiating cells in oral cancer.
Although several approaches for identification and isolation of carcinoma cells with tumour initiating properties have been established, enrichment of a population of pure and viable tumour-initiating cells (TICs) is still problematic. This study investigated possibilities to isolate a population of cancer cells with tumour initiating properties based on their adherence properties, rather than expression of defined markers or clonogenicity. Several human cell lines derived from oral dysplasia and oral squamous cell carcinoma (OSCC), as well as primary cells derived from patients with OSCC were allowed to adhere to collagen IV-coated dishes sequentially. Rapid adherent cells (RAC), middle adherent cells (MAC) and late adherent cells (LAC) were then harvested and further investigated for their morphology, stem cell-like properties and molecular profile while grown in vitro and tongue xenotransplantation in NOD-SCID mice at serial dilutions. RAC showed significantly higher colony forming efficiency (p<0.05), sphere forming ability, greater migration ability (p<0.05), exhibited longer G2 phase and displayed higher expression of integrin β1 and other stem-cell related molecules as compared to MAC and LAC. RAC induced tongue tumours in NOD-SCID mice with the highest incidence. These tumours were also bigger and metastasised more frequently in loco-regional lymph nodes than MAC and LAC. These findings prove for the first time that OSCC cells with tumour initiating properties can be enriched based on their rapid adhesiveness to collagen IV. This separation procedure provides a potentially useful tool for isolating TICs in OSCC for further studies on understanding their characteristics and drug-resistant behaviour.


Aortic valve repair and replacement after balloon aortic valvuloplasty in children.
Little is known about the incidence, indications, and results of surgical repair or replacement of the aortic valve after balloon aortic valvuloplasty (BAV) for congenital aortic stenosis in children. This study was designed to evaluate patterns of failure requiring operation after BAV for congenital aortic stenosis and to review our experience with successful repair, rather than replacement, of selected aortic valves after BAV. From March 1986 to June 1995, 60 patients with congenital aortic stenosis aged 1 day to 27 years (mean +/- standard deviation, 7.3 +/- 6 years) underwent BAV. Twenty-three patients (38%) required operation a mean of 44 +/- 37 months (range, 1 to 110 months) after BAV, because of severe aortic insufficiency in 13 patients and recurrent or residual aortic stenosis in 10 patients. Severe aortic insufficiency was invariably due to avulsion of a cusp from the annulus, with resulting cusp prolapse and insufficiency. Operative intervention consisted of valve replacement in 14 patients and valve repair in 9 patients. Repair techniques included reattachment of an avulsed cusp to the aortic annulus, relief of commissural fusion, and debridement of thickened cusps. Actuarial freedom from surgical intervention after BAV was 88% +/- 4% at 1 year, 70% +/- 6% at 5 years, and 51% +/- 12% at 9 years. The need for aortic valve operation was unrelated to age at the time of BAV, indication for operation (aortic insufficiency versus aortic stenosis), age of operation, or preoperative gradient. All patients survived aortic valve operation; there was one late death at an average follow-up of 27 +/- 20 months (range, 2 to 61 months) after aortic valve operation. Stenosis was well relieved in all patients undergoing valve replacement. The 9 valve repair patients have been followed for 22 +/- 14 months (range, 1 to 47 months). Echocardiographic follow-up of the valve repair patients revealed a mean residual aortic stenosis peak instantaneous gradient of 32 mm Hg and mild aortic insufficiency or less in all patients. Aortic valve operation is required in 5% to 7% of patients yearly after BAV. The need for operation appears to be unrelated to age at the time of BAV; aortic insufficiency predominates over aortic stenosis as an indication for operative intervention. Valve repair can be applied in some patients after BAV with good intermediate-term results and may delay the need for aortic valve replacement.


Cellular composition of the initial fracture hematoma compared to a muscle hematoma: a study in sheep.
Bone fracture leads to a cycle of inflammation, cellular migration, and proliferation to restore tissue integrity. Immune cells at the site of injury are involved especially in the early phase of the healing process, but little is known about the cells present in the initial fracture hematoma. The hypothesis of this study was that the cellular composition in a fracture hematoma differs from that found in a muscle hematoma and that these divergences get more pronounced over time. By using a reproducible osteotomy model and muscle trauma in sheep the distributions of the immune cell subpopulations were evaluated 1 and 4 h after surgery. The cell amount within the first 4 h increased in both hematoma. The number of dead cells was higher in the muscle hematoma. One hour postoperatively the initial fracture hematoma revealed a lower granulocyte percentage compared to the muscle hematoma. The ratio of T helper to cytotoxic T cells was higher in the fracture hematoma compared to the muscle hematoma at both investigated time points. B cell percentage increased in the fracture but not in the muscle hematoma from 1 to 4 h. This is the first study that compares the immune cell subpopulations of a fracture and muscle hematoma.


Alternate mechanism for amino acid entry into Neurospora crassa: extracellular deamination and subsequent keto acid transport.
The growth of the pm nbg mutant strain of Neurospora crassa was inhibited by the amino acid analog para-fluorophenylalanine despite the fact that none of the three constitutive amino acid permeases is functional in this strain. This observation led to the detection of both a deaminase which was released into the growth medium in response to para-fluorophenylalanine and a keto acid transport system which allowed entry of the resulting keto acid into the cell. The transported keto acid was recovered in cellular protein, suggesting its regeneration as the amino acid. The cooperative activity of these two systems represents an additional mechanism for the intracellular accumulation of amino acids, which is distinct from the known amino acid permeases.


Evaluating a Novel Simulation Course for Prehospital Provider Resuscitation Training in Botswana.
In 2012, Botswana embarked on an organized public approach to prehospital medicine. One goal of the Ministry of Health (MOH) was to improve provider education regarding patient stabilization and resuscitation. Simulation-based instruction is an effective educational strategy particularly for high-risk, low-frequency events. In collaboration with partners in the United States, the team created a short, simulation-based course to teach and update prehospital providers on common field responses in this resource-limited setting. The objective of this study was to evaluate an educational program for Botswanan prehospital providers via written and simulation-based examinations. We developed a two-day course based on a formal needs assessment and MOH leadership input. The subject matter of the simulation scenarios represented common calls to the prehospital system in Botswana. Didactic lectures and facilitated skills training were conducted by U.S. practitioners who also served as instructors for a rapid-cycle, deliberate practice simulation education model and simulation-based testing scenarios. Three courses, held in three cities in Botswana, were offered to off-duty MOH prehospital providers, and the participants were evaluated using written multiple-choice tests, videotaped traditional simulation scenarios, and self-efficacy surveys. Collectively, 31 prehospital providers participated in the three courses. The mean scores on the written pretest were 67% (standard deviation [SD], 10) and 85% (SD, 7) on the post-test (p < 0.001). The mean scores for the simulation were 42% (SD, 14.2) on the pretest and 75% (SD, 11.3) on the post-test (p < 0.001). Moreover, the intraclass correlation coefficient scores between reviewers were highly correlated at 0.64 for single measures and 0.78 for average measures (p < 0.001 for both). Twenty-one participants (68%) considered the course "extremely useful." Botswanan prehospital providers who participated in this course significantly improved in both written and simulation-based performance testing. General feedback from the participants indicated that the simulation scenarios were the most useful and enjoyable aspects of the course. These results suggest that this curriculum can be a useful educational tool for teaching and reinforcing prehospital care concepts in Botswana and may be adapted for use in other resource-limited settings.


[Extrarenal nodular nephrogenic blastema in the inguinal canal: report of two cases].
Two observations of extrarenal nephrogenic nodule in inguinal canal are reported. We underline the difficulties in differential diagnostic with true Wilms' tumor, and in sparing these pediatric cases from more aggressive treatment. The main morphological arguments in favor of non-tumoral nephrogenic nodular remnants are: the size of the lesion, the association with a congenital inguinal hernia suggesting a congenital abnormality, and the organoid organization of blastemic tissue surrounding the cavities lined with "hobnail" epithelium evoking a mesonephrogenic origin.


Accord No. 8 of 19 August 1988.
The objective of this Accord is to facilitate the entry into Honduras of foreigners who desire to invest in agricultural, agro-industrial, industrial, tourism, and export projects. The Accord establishes an investment visa scheme for 1) foreign investors and business persons and their spouses and economic dependents and 2) technicians and specialist experts and workers and their spouses and economic dependents. The visas are free and are good 1) for three months' residence in Honduras with three-month extensions possible up to a period of one year and 2) for one month, renewable in one-month periods up to six months. Further provisions of the Accord set forth procedures, documentary requirements, and registration requirements, among other things.


Functional and Structural Organization of Chlorophyll in the Developing Photosynthetic Membranes of Euglena gracilis Z: II. CHARACTERISTICS OF THE LATE FORMATION OF ACTIVE PHOTOSYSTEM II REACTION CENTERS DURING FIRST STAGES OF GREENING.
During light-induced greening of dark-grown, nondividing Euglena gracilis Z, there is a delay of about 10 hours in the formation of active photosystem II (PSII) reaction centers compared to chlorophyll synthesis. Experiments with greening under different light intensities rule out the possibility that this delay results from a late induction of active PSII reaction center formation when a definite amount of chlorophyll is attained in the early greened cells. Experiments on greening after preillumination show that this delay does not originate in a long, light-induced formation of specific synthesizing machinery for reaction center components. Experiments with greening in the presence of streptomycin show that, when this inhibitor of protein synthesis by chloroplastic ribosomes is added to dark-grown, preilluminated cells or to cells already greened for 24 hours, the formation of active PSII reaction centers is inhibited after a time which depends on the light intensity used for greening. Under very low light intensity (150 lux), the addition of streptomycin to 24-hour greened cells does not prevent further development of functional chloroplasts. These observations lead to the conclusion that streptomycin-insensitive chloro-plast development occurs due to syntheses of cytoplasmic origin and of light-induced pools of components synthesized early by chloroplastic ribo-somes. Conformational changes requiring time may allow the insertion of components necessary for the reorganization of PSII reaction centers in the developing thylakoid after synthesis. This hypothesis accounts for the observed delay in PSII reaction center formation compared to chlorophyll synthesis.


Prevention of Acute Proximal Junctional Fractures After Long Thoracolumbar Posterior Fusions for Adult Spinal Deformity Using 2-level Cement Augmentation at the Upper Instrumented Vertebra and the Vertebra 1 Level Proximal to the Upper Instrumented Vertebra.
Retrospective cohort analysis. To evaluate efficacy of proximal junction fracture (PJF) prevention in adult spinal deformity (ASD) using 2-level cement augmentation at the construct's proximal extent. Prevention of PJF after thoracolumbar fusions is critical because they may result in neurological injury. Cement augmentation of constructs' proximal vertebrae is postulated to decrease PJF. Patients with ASD after PSF from pelvis to thoracolumbar junction with 6 months or more follow-up were retrospectively studied. Demographics, deformity radiographical parameters, and health-related quality of life outcomes (HRQoL) scores were compared with patients with no cement, 2-level cement augmentation at upper instrumented vertebra (UIV) and vertebra 1 level proximal to UIV (UIV+1), and cement at another location ("Other"). Revision surgery for PJF was primary outcome. Univariable and multivariable logistic regression analyses were used for statistical analysis. 51 patients [female-29; male-22; average age: 65 yr (33-82)] met inclusion criteria (2-level-19; no-cement-23; "Other"-9). Average follow-up (mo) was longer for no-cement (25 ± 15) and "Other" (20 ± 16) than 2-level (15 ± 8) (P = 0.06). All perioperative radiographical parameters were similar, save first postoperative thoracic kyphosis and lumbopelvic mismatch. Compared with 2-level cement, non-2-level cement had significantly more revisions for PJF (0% vs. 19%; P = 0.02). After UIV adjustment, risks of PJF revision surgery were 13.1 times higher for "Other" (95% CI: 0.5-346.5, P = 0.12) and 9.2 times higher (95% CI: 0.4-239.1, P = 0.18) for no-cement. All HRQoL scores improved in 2-level cement; only back/leg pain significantly improved in non-2-level cement. Postoperative Oswestry Disability Index was significantly less in 2-level cement. The use of 2-level cement augmentation (UIV and UIV+1) in PSF from pelvis to thoracolumbar junction for ASD is associated with a decreased rate of acute proximal junctional fractures and associated revision surgeries. As only associations can be demonstrated from this study's design, prospective investigations with larger, consecutive cohorts should be performed to explore causal relationships. 3.


Synthesis and characterization of self-assembled polyaniline nanotubes/silica nanocomposites.
Self-assembled semiconducting, paramagnetic polyaniline nanotubes have been synthesized by the oxidative polymerization of aniline with ammonium peroxydisulfate in aqueous medium in the presence of colloidal silica particles of an average diameter approximately 12 nm, without added acid. The electrical conductivity of polyaniline nanotubes/silica nanocomposites is in the range (3.3-4.0)x10(-3) S cm(-1). The presence of paramagnetic polaronic emeraldine salt form of polyaniline and phenazine units in nanocomposites was proved by FTIR, Raman, and EPR spectroscopies. The influence of the initial silica/aniline weight ratio on the morphology of polyaniline/silica nanocomposites was studied by scanning and transmission electron microscopies. Nanocomposites synthesized by using the initial weight ratio silica/aniline<or=0.2 contain polyaniline nanotubes which have a typical outer diameter of 100-250 nm and an inner diameter of 10-80 nm, and nanorods with a diameter of 60-100 nm, accompanied with polyaniline/silica nanogranules, while the nanocomposite synthesized at weight ratio silica/aniline approximately 2 contains polyaniline/silica nanogranules with an average diameter of 35-70 nm. The evolution of molecular and supramolecular structure of polyaniline in the presence of colloidal silica is discussed.


Lipid A and related compounds. XXVII. An efficient synthesis of D-galactosamine-4-phosphate analogs of lipid A via a novel key intermediate.
A new methodology for chemical differentiation of one amino and four hydroxyl groups of D-galactosamine derivatives and its application for the synthesis of D-galactosamine-4-phosphate analogs of lipid A are described. Preliminary examination of biological activity revealed that the synthetic monosaccharides show mitogenic activity.


Prevalence of ectoparasitic arthropods on wild animals and cattle in the Las Merindades area (Burgos, Spain).
This paper reports the prevalence of ectoparasitic arthropods in sampled groups of wild (n = 128; 16 species) and domestic (n = 69; 3 species) animals in the Las Merindades area of the Province of Burgos, Spain. The study revealed that wild animals were more infested and with a wider variety of ectoparasites than domestic animals. The parasitic prevalence was 67% for wild animals and 48% for livestock. In this way, 39% of animals were infected by ticks. Ixodes ricinus and Ixodes hexagonus were the most prevalent species whereas Dermacentor reticulatus showed affinity for the fox and wolf. The overall prevalence of parasitisation by fleas was 27%. Ctenophthalmus spp. showed the wider range host in wild animals, while Pulex irritans was the most frequent specie found. The parasitic prevalences by lice (Trichodectes melis, Trichodectes canis and Trichodectes mustelae) and by mite (Neotrombicula spp., Laelaps agilis and Sarcoptes scabiei) were 4% and 12%, respectively. In both cases only wild animals were found parasited.


Calculation of the volume of an HPGe crystal.
An important characteristic of a high-purity germanium detector for gamma spectrometry is the volume of the crystal, which relates to the detection efficiency and to the measured background level. The crystal dimensions provided by the detector manufacturer were used to calculate the crystal volume. The calculation method accounts for the various sub-volumes of the crystal, in particular the contact hole in the back face and the rounding of the 90° right edge in the front face. The calculation was corroborated using the INVENTOR(®) 3D software. The crystal volume was calculated to be 419.7 cm(3) compared with the 413 cm(3) stated by the manufacturer.


[In vitro study of the antimutagenic activity of alphahederin].
Authors have studied the antimutagenic power of alphahederin (a saponin extracted from Hedera helix) versus a clastogenic agent, doxorubicin and an aneugenic agent, carbendazim. We have applied a protocol of incorporation of alphahederin (pretreatment, simultaneous treatment and post treatment) to determine a mechanism of action. According to this protocol, alphahederin induces a significant diminution of the rate of micronuclei wathever the phases of the protocol. These results demonstrate the antimutagenic activity of alphahederin with a mechanism of action, both desmutagenic and bioantimutagenic.


[Relation of the cyclic nucleotide system with the functional state of the cholinoreceptors].
It has been demonstrated in rat experiments that the concentration of cAMP and cGMP in the rat liver increased 15-90 min after administration of the cholinomimetic carbamylcholine (Cr) or the cholinolytic chlorosyl (Ch) at doses of 1 and 2 mg/kg, respectively. Administration of Ch 5 min before Cr blocked the signs of parasympathetic nervous system excitation and made the content of cGMP return to normal. The concentration of cAMP in the liver of these rats as was the case in administration of Ch alone was greatly enhanced, reaching 400% of the control after 60 min. Transcriptional activity of hepatocytes following separate or combined administration of Ch and Cr was increased in 6 hours. Preliminary administration of Ch at a dose of 0.025 mg/kg prevented the excitation of the parasympathetic nervous system and stimulation of hepatocyte transcriptional activity, bringing about a dramatic fall in the liver cAMP and cGMP content. The existence of the relationship between cholinoreceptor function and the pool of cyclic nucleotides in the target cells is postulated.


Tonic transmitter release in a graded potential synapse.
1. We studied graded synaptic transmission in the fly photoreceptor-interneuron synapse by using intracellular in situ recordings from pre- and postsynaptic cells. 2. A large presynaptic hyperpolarization after light adaptation, caused by the activation of the electrogenic Na+/K+ pump, drastically reduced the conspicuous postsynaptic dark noise. At the same time, the postsynaptic neurons depolarized, with an increase of input resistance of 5-10 M omega. 3. The spectral characteristics of the postsynaptic membrane noise in dark and during noise reduction, together with the other results, suggested that the transmitter release decreased dramatically approximately 12 mV below the resting potential of the presynaptic photoreceptors. 4. During the postsynaptic noise reduction, the saturated and subsaturated first-order visual interneuron responses were increased up to 9 mV with a time constant of recovery of approximately 10 s. This increase was shown to be caused by the negative shift of the reversal potential of the transmitter-gated (mainly Cl-) conductance, caused apparently by the reduced transmitter input. 5. The results strongly suggest that the photoreceptor transmitter release in fly is tonic, even in dark, and further support the modulation of the synaptic voltage transfer by postsynaptic Cl- extrusion.


Gender-Related Differences in Pelvic Morphometrics of the Retriever Dog Breed.
This study presents the results from a morphometric analysis of 52 dry Retriever dog pelvic bones (30 male, 22 female). A total of 20 parameters were measured using an osteometric board and digital vernier caliper. Six parameters were found to be significantly higher (P < 0.05) in males than in females, while one parameter was significantly higher (P < 0.05) in females than in males. However, none of the measured parameters demonstrated clear cut-off values with no intersect between males and females. Therefore, we generated a stepwise discriminant analysis from all 20 parameters in order to develop a possible working equation to discriminate gender from a dog pelvic bone. Stepwise discriminant analysis was used to create a discrimination function: Y = [82.1*PS/AII] - [50.72*LIS/LI] - [23.09*OTD/SP] + [7.69*SP/IE] + [6.52*IC/OW] + [7.67*ISA/OW] + [20.77*AII/PS] + [504.71*OW/ISA] - [90.84*PS/ISA] - [148.95], which showed an accuracy rate of 86.27%. This is the first study presenting an equation/function for use in discriminating gender from a dog's pelvic measurements. The results can be used in veterinary forensic anthropology and also show that a dog's pelvis presents sexual dimorphisms, as in humans.


Isoflurane and postoperative respiratory depression following laparoscopic surgery: A retrospective propensity-matched analysis.
Episodes of respiratory depression during the immediate postoperative recovery period (Phase I post-anesthesia recovery) have been associated with respiratory complications during Phase II recovery. Using multivariable analyses in several surgical cohorts, we previously identified potential associations between patient and perioperative factors and increased risk for Phase I respiratory depression. The aim of this study is to use the propensity-matched analysis to specifically assess for a potential association between the use of isoflurane and episodes of Phase I respiratory depression after laparoscopic operations. The electronic medical records of 8567 patients who underwent laparoscopic operations between January 1, 2010 and July 31, 2014, lasting ≥90 minutes, were retrospectively analyzed. Propensity-matched patients anesthetized without isoflurane were identified for 3403 patients anesthetized with isoflurane. Compared to the use of desflurane, sevoflurane or propofol infusion, maintenance of anesthesia with isoflurane was associated with an increased likelihood of Phase I respiratory depression (OR 95% CI, 1.32, 1.15-1.50, p < 0.001) and longer Phase I recovery (126 vs. 110 minutes, p < 0.001). The use of isoflurane was associated with increased rates of postoperative respiratory depression and postoperative recovery when compared to sevoflurane, desflurane, or propofol infusion.


Actor-observer differences in realism in confidence and frequency judgments.
Taking a social psychological approach to metacognitive judgments, this study analyzed the difference in realism (validity) in confidence and frequency judgments (i.e., estimates of overall accuracy) between one's own and another person's answers to general knowledge questions. Experiment 1 showed that when judging their own answers, compared with another's answers, the participants exhibited higher overconfidence, better ability to discriminate correct from incorrect answers, lower accuracy, and lower confidence. However, the overconfidence effect could be attributable to the lowest level of confidence. Furthermore, when heeding additional information about another's answers the participants showed higher confidence and better discrimination ability. The overconfidence effect of Experiment 1 was not found in Experiment 2. However, the results of Experiment 2 were consistent with Experiment 1 in terms of discrimination ability, confidence, and accuracy. Finally, in both experiments the participants gave lower frequency judgments of their own overall accuracy compared with their frequency judgments of another person's overall accuracy.


Rumination and Excessive Reassurance Seeking: Investigation of the Vulnerability Model and Specificity to Depression.
Recent research investigating cognitive and interpersonal models of depression have conceptualized rumination and excessive reassurance seeking (ERS) as vulnerabilities for the etiology of depression. However, research testing the vulnerability hypothesis for these constructs among youth is lacking. Additionally, the specificity of rumination and excessive reassurance seeking to depression is unclear. Two studies investigated associations between these constructs to depression and anxiety. In Study 1, concurrent associations between these constructs and depressive and anxious arousal were examined among 194 minority, underserved youth. Study 2 examined concurrent and longitudinal associations between both constructs and depressive and general anxiety symptoms in a community sample of 402 youth. Results from both studies supported rumination as a specific vulnerability for depressive symptoms, whereas ERS appeared to be a concomitant of negative affect common to depressive and anxiety symptoms. Findings inform cognitive and interpersonal theories of depression by advancing knowledge of how rumination and ERS relate to aspects of internalizing problems among diverse populations of youth.


The tight junction protein ZO-2 mediates proliferation of vascular smooth muscle cells via regulation of Stat1.
Recent evidence suggests that the zonula occludens protein 2 (ZO-2) might have additional cellular functions, beyond regulation of paracellular permeability of epithelial and endothelial cells. Deregulation of ZO-2 in response to ischaemia, hypertensive stress, and vascular injury implies its involvement in cardiovascular disorders, most likely via regulating the functional behaviour of vascular smooth muscle cells (VSMC). However, a role of ZO-2 in VSMC biology has yet to be established. Our study was designed to understand the specific functions of ZO-2 in human VSMC. The expression of ZO-2 and Stat1 upon vascular injury was studied using ex vivo organ culture of coronary arteries combined with immunohistochemistry. ZO-2 silencing in human primary VSMC was achieved by means of lentiviral gene transfer. Cell proliferation was assessed by analysing DNA synthesis and by cell counting. Stat1 expression was examined using immunoblotting, immunocytochemistry, TaqMan, and fluorescence activated cell sorting (FACS) analysis. Functional relevance of Stat1 up-regulation was studied using a Stat1 promoter-luciferase reporter assay and intracellular microinjections of a Stat1 specific antibody. ZO-2 was highly expressed in the media and neointima of dilated but not of control arteries, whereas expression of the transcription factor Stat1 was inversely regulated upon injury. Analysis of VSMC with down-regulated ZO-2 revealed increased expression of Stat1 in these cells, whereas Stat1 phosphorylation was not affected. Stat1 up-regulation in VSMC with ZO-2 silencing resulted in a coordinate activation of Stat1-specific genes and consequently led to inhibition of cell proliferation. This effect was restored by microinjection of a Stat1 neutralising antibody. Our data suggest that the tight junction protein ZO-2 is involved in regulation of VSMC growth control upon vascular injury that is mediated by the transcription factor Stat1. Our findings point to a novel function of ZO-2 in VSMC and implicate ZO-2 as a novel important molecular target in pathological states of vascular remodelling in cardiovascular diseases.


Effects of Cigarette Smoking on Preimplantation Embryo Development.
In this chapter, we first gave a brief introduction to the detriments of cigarette smoking, with an emphasis on its adverse effects on female reproductive health. Then, we outlined recent advances about the impacts of cigarette smoke on preimplantation embryo development. Additionally, toxicities of cadmium and benzo(a)pyrene (BaP) at this specific developmental window were also discussed, to illustrate the potential mechanisms involved in cigarette smoke-associated embryotoxicity. Finally, we provide an overview of the issues to be solved in the future research. Further studies about the molecular mechanism of cigarette smoking-associated female infertility may provide vital insights into developing new interventions for the women smokers and thus improving their reproductive outcomes.


Gold nanoparticles bifunctionalized by chemiluminescence reagent and catalyst metal complexes: synthesis and unique chemiluminescence property.
Despite much progress in functionalized gold nanomaterial (GNMs), chemiluminescent (CL) functionalized GNMs with high CL efficiency are far from fully developed. In this work, we report a general strategy for the synthesis of gold nanoparticles (GNPs) bifunctionalized by CL reagent and catalyst metal complexes (BF-GNPs) by taking N-(aminobutyl)-N-(ethylisoluminol) (ABEI) as a model of CL reagents. The complexes of 2-[bis[2-[carboxymethyl-[2-oxo-2-(2-sulfanylethylamino)ethyl]amino]ethyl]amino]acetic acid (DTDTPA) with various metal ions, including Co(2+), Cu(2+), Pb(2+), Ni(2+), Hg(2+), Cr(3+), Eu(3+), La(3+), Gd(3+), Sm(3+), Er(3+), Dy(3+), Ce(4+), and Ce(3+), were grafted on the surface of ABEI functionalized GNPs (ABEI-GNPs) to form a series of BF-GNPs. These BF-GNPs exhibited excellent CL activity. In particular, the CL intensity of DTDTPA/Co(2+)-ABEI-GNPs was over 3 orders of magnitude higher than ABEI-GNPs. This work demonstrates for the first time that metal complexes grafted on the surface of GNPs have unique catalytic activity on the CL reaction, superior to that in the liquid phase. Such BF-GNPs may find future applications in bioassays, microchips, and molecular/cellular imaging.


Urinary volatile compounds as biomarkers for lung cancer: a proof of principle study using odor signatures in mouse models of lung cancer.
A potential strategy for diagnosing lung cancer, the leading cause of cancer-related death, is to identify metabolic signatures (biomarkers) of the disease. Although data supports the hypothesis that volatile compounds can be detected in the breath of lung cancer patients by the sense of smell or through bioanalytical techniques, analysis of breath samples is cumbersome and technically challenging, thus limiting its applicability. The hypothesis explored here is that variations in small molecular weight volatile organic compounds ("odorants") in urine could be used as biomarkers for lung cancer. To demonstrate the presence and chemical structures of volatile biomarkers, we studied mouse olfactory-guided behavior and metabolomics of volatile constituents of urine. Sensor mice could be trained to discriminate between odors of mice with and without experimental tumors demonstrating that volatile odorants are sufficient to identify tumor-bearing mice. Consistent with this result, chemical analyses of urinary volatiles demonstrated that the amounts of several compounds were dramatically different between tumor and control mice. Using principal component analysis and supervised machine-learning, we accurately discriminated between tumor and control groups, a result that was cross validated with novel test groups. Although there were shared differences between experimental and control animals in the two tumor models, we also found chemical differences between these models, demonstrating tumor-based specificity. The success of these studies provides a novel proof-of-principle demonstration of lung tumor diagnosis through urinary volatile odorants. This work should provide an impetus for similar searches for volatile diagnostic biomarkers in the urine of human lung cancer patients.


Salt and blood pressure. Conventional wisdom reconsidered.
The salt hypothesis is that higher levels of salt in the diet lead to higher levels of blood pressure, increasing the risk of cardiovascular disease. Intersalt, a cross-sectional study of salt levels and blood pressures in 52 populations, is often cited to support the salt hypothesis, but the data are somewhat contradictory. Four of the populations (Kenya, Papua, and 2 Indian tribes in Brazil) do have low levels of salt and blood pressure. Across the other 48 populations, however, blood pressures go down as salt levels go up, contradicting the hypothesis. Experimental evidence suggests that the effect of a large reduction in salt intake on blood pressure is modest, and health consequences remain to be determined. Funding agencies and medical journals have taken a stronger position favoring the salt hypothesis than is warranted, raising questions about the interaction between the policy process and science.


Magnesium and regulation of carbohydrate metabolism at the molecular level.
Enzymatic reactions involving the transfer of high-energy phosphate require a divalent metal ion as a participant. This is normally Mg2+ in biological situations, as its level within the cell is greater than other divalent metal ions. Although available methods for determining free cellular Mg2+ have thus far given divergent results, there is a growing body of evidence that its concentration is relatively low, approximately 0.5 mM. This low level permits control of important parts of carbohydrate metabolism, especially glycolysis, by Mg2+. The role of Mg2+ in regulating the phosphoryl-transfer enzymes of this pathway is described. Also described briefly is a series of computer models of this metabolism in heart and pancreatic islets. These models indicate considerable changes in Mg2+ level, and consequent control, especially when there are sharp changes in the glycolytic rate.


Direct demonstration of discrete Ca2+ microdomains associated with different isoforms of adenylyl cyclase.
Ca(2+)-sensitive adenylyl cyclases (ACs) orchestrate dynamic interplay between Ca(2+) and cAMP that is a crucial feature of cellular homeostasis. Significantly, these ACs are highly selective for capacitative Ca(2+) entry (CCE) over other modes of Ca(2+) increase. To directly address the possibility that these ACs reside in discrete Ca(2+) microdomains, we tethered a Ca(2+) sensor, GCaMP2, to the N-terminus of Ca(2+)-stimulated AC8. GCaMP2-AC8 measurements were compared with global, plasma membrane (PM)-targeted or Ca(2+)-insensitive AC2-targeted GCaMP2. In intact cells, GCaMP2-AC8 responded rapidly to CCE, but was largely unresponsive to other types of Ca(2+) rise. The global GCaMP2, PM-targeted GCaMP2 and GCaMP2-AC2 sensors reported large Ca(2+) fluxes during Ca(2+) mobilization and non-specific Ca(2+) entry, but were less responsive to CCE than GCaMP2-AC8. Our data reveal that different AC isoforms localize to distinct Ca(2+)-microdomains within the plasma membrane. AC2, which is regulated via protein kinase C, resides in a microdomain that is exposed to a range of widespread Ca(2+) signals seen throughout the cytosol. By contrast, a unique Ca(2+) microdomain surrounds AC8 that promotes selectivity for Ca(2+) signals arising from CCE, and optimizes CCE-mediated cAMP synthesis. This direct demonstration of discrete compartmentalized Ca(2+) signals associated with specific signalling proteins provides a remarkable insight into the functional organization of signalling microdomains.


Complexity, centralization, and fragility in economic networks.
Trade networks, across which countries distribute their products, are crucial components of the globalized world economy. Their structure affects the mechanism of propagation of shocks from country to country, as observed in a very sharp way in the past decade, characterized by economic uncertainty in many parts of the world. Such trade structures are strongly heterogeneous across products, given the different features of the countries which buy and sell goods. By using a diversified pool of indicators from network science and product complexity theory, we quantitatively demonstrate that, overall, products with higher complexity-i.e., with larger technological content and/or number of components-are traded through more centralized networks-i.e., with a smaller number of countries concentrating most of the export flow. Since centralized networks are known to be more vulnerable, we argue that the current composition of production and trading is associated to high fragility at the level of the most complex-thus strategic-products.


Affordable IgY-based antiviral prophylaxis for resource-limited settings to address epidemic and pandemic risks.
The COVID-19 pandemic caused by SARS-CoV-2 exposed a global problem, as highly effective vaccines are challenging to produce and distribute, particularly in regions with limited resources and funding. As an alternative, immunoglobulins produced in eggs of immunized hens (IgY) can be a simple and inexpensive source for a topical and temporary prophylaxis. Here, we developed a method to extract and purify IgY antibodies from egg yolks of hens immunized against viral pathogen-derived proteins using low-cost, readily available materials, for use in resource-limited settings. Existing protocols for IgY purification and equipment were modified, including extraction from yolks and separation of water-soluble IgY using common household reagents and tools. A replacement for a commercial centrifuge was developed, using a home food processor equipped with a 3D printed adapter to enable IgY precipitation. IgY purification was verified using standard gel electrophoresis and Western blot analyses. We developed a step-by-step protocol for IgY purification for two settings in low- and middle-income countries (LMIC): a local laboratory, where commercial centrifuges are available, or a more rural setting, where an alternative for expensive centrifuges can be used. Gel electrophoresis and Western blot analyses confirmed that the method produced highly enriched IgY preparation; each commercial egg produced ~ 90 mg of IgY. We also designed a kit for IgY production in these two settings and provided a cost estimate of the kit. IgY purified from eggs of immunized local hens can offer a fast and affordable prophylaxis, provided that purification can be performed in a resource-limited setting. Here, we created a low-cost method that can be used anywhere where electricity is available using inexpensive, readily available materials in place of costly, specialized laboratory equipment and chemicals. This procedure can readily be used now to make an anti-SARS-CoV-2 prophylaxis in areas where vaccines are unavailable, and can be modified to combat future threats from viral epidemics and pandemics.


Chronic neurosteroid treatment prevents the development of morphine tolerance and attenuates abstinence behavior in mice.
The effect of neurosteroids on the development of morphine tolerance and dependence was examined in mice. Development of tolerance to the antinociceptive effect of morphine sulfate (10 mg/kg, twice daily for 9 days) was measured in the tail-flick test and dependence was assessed from naloxone (2 mg/kg)-precipitated withdrawal jumps on day 10 of testing. Concomitant chronic administration of neurosteroids, allopregnanolone (0.5 mg/kg), pregnenolone sulfate (2 and 5 mg/kg) or dehydroepiandrosterone sulfate (2 and 5 mg/kg), followed by morphine (10 mg/kg) prevented the development of tolerance to the antinociceptive effect of morphine and suppressed the naloxone-precipitated withdrawal jumps. In contrast, dehydroepiandrosterone acetate (5 mg/kg) failed to modulate the morphine tolerance and dependence. The inhibitory effect was also seen upon concomitant administration of a neurosteroid precursor, progesterone (1-10 mg/kg), and a mitochondrial diazepam binding inhibitor receptor agonist, 4'-chlordiazepam (0.25-1 mg/kg), while an adrenocorticosteroid, hydrocortisone (1 and 10 mg/kg), failed to do so. However, acute treatment with these neurosteroids was not associated with any decrease in withdrawal jumping behavior in morphine-dependent mice. Neurosteroids themselves, at doses employed in the study, did not exert any effects on antinociception. These results support a role for neurosteroids in the development of tolerance to and dependence on morphine and suggest the potential utility of specific neuroactive steroids in its treatment.


Importance of NADPH supply for improved L-valine formation in Corynebacterium glutamicum.
Cofactor recycling is known to be crucial for amino acid synthesis. Hence, cofactor supply was now analyzed for L-valine to identify new targets for an improvement of production. The central carbon metabolism was analyzed by stoichiometric modeling to estimate the influence of cofactors and to quantify the theoretical yield of L-valine on glucose. Three different optimal routes for L-valine biosynthesis were identified by elementary mode (EM) analysis. The modes differed mainly in the manner of NADPH regeneration, substantiating that the cofactor supply may be crucial for efficient L-valine production. Although the isocitrate dehydrogenase as an NADPH source within the tricarboxylic acid cycle only enables an L-valine yield of Y(Val/Glc) = 0.5 mol L-valine/mol glucose (mol Val/mol Glc), the pentose phosphate pathway seems to be the most promising NADPH source. Based on the theoretical calculation of EMs, the gene encoding phosphoglucoisomerase (PGI) was deleted to achieve this EM with a theoretical yield Y(Val/Glc) = 0.86 mol Val/mol Glc during the production phase. The intracellular NADPH concentration was significantly increased in the PGI-deficient mutant. L-Valine yield increased from 0.49 +/- 0.13 to 0.67 +/- 0.03 mol Val/mol Glc, and, concomitantly, the formation of by-products such as pyruvate was reduced.


In vitro and in vivo effectiveness of a novel injectable calcitonin-loaded collagen/ceramic bone substitute.
This study aimed to evaluate the effectiveness of a novel calcitonin-loaded calcium phosphate composite bone cement in vitro and in vivo. The novel composite bone cements were composed of NuROs injectable bone graft substitute, type I collagen, and/or salmon calcitonin. The setting time, porosity, wettability, compressive strength, compressive modulus, and crystallographic structures of cement specimens were determined. Degradation rate, calcitonin release rate, and osteoinductivity were assessed in vitro. In addition, osteogenic effect was examined in a rabbit model of femoral defect. The results revealed that addition of collagen/calcitonin did not substantially alter physical properties and degradation rate of bone cement specimens. Calcitonin was released into culture medium in a two-phase manner. Osteogenic effect of conditioned medium derived from calcitonin containing bone cement was observed. Finally, de novo bone growth and bone mineralization across the bone defect area were observed in rabbits after implantation of composite bone cement specimens. In conclusion, this novel calcitonin-loaded composite calcium phosphate bone cement exhibits biocompatibility, bioresorbability, osteoinductivity, and osteoconductivity, which may be suitable for clinical use.


Gold nanoparticles cellular toxicity and recovery: effect of size, concentration and exposure time.
Gold nanoparticles (AuNPs) are used in many applications; however, their interactions with cells and potential health risk(s) are not fully known. In this manuscript, we describe the interactions of AuNPs with human dermal fibroblasts and show that they can penetrate the plasma membrane and accumulate in large vacuoles. We also demonstrate that the uptake of the AuNPs is a function of time, their size and concentration. Specifically, we demonstrate that 45 nm AuNPs penetrate cells via clathrin-mediated endocytosis, while the smaller 13 nm enter mostly via phagocytosis. Furthermore, we provide evidence of cytoskeleton filament disruption as a result of AuNPs exposure and reconstitution during recovery (following AuNP removal), despite no changes in actin or beta-tubulin protein levels. In contrast, the expression of the extracellular matrix (ECM) proteins, collagen and fibronectin, was diminished in the cells exposed to AuNPs. We also examined the proliferation rates of cells exposed to AuNPs and show that its diminution is a function of apoptosis and speculate that apoptosis results from the number of vacuoles present in the cells, which is probably the main factor that disrupts the cytoskeleton causing cell area contraction and decreases in motility. Lastly, we also present data that indicates that AuNPs' damage to cells is not permanent and that the cells can completely recover as a function of AuNPs' size, concentration and exposure time. Taken together, our data suggest that AuNPs exert detrimental effects on cell function that could reverse following AuNPs removal.


Aquaporin-4 autoimmunity.
Neuromyelitis optica (NMO) and a related spectrum of inflammatory CNS disorders are unified by detection of a serum autoantibody specific for the aquaporin-4 (AQP4) water channel, which is abundant in astrocytic foot processes. The classic clinical manifestations of NMO are optic neuritis and longitudinally extensive transverse myelitis. Newly recognized manifestations of AQP4 autoimmunity include lesions of circumventricular organs and skeletal muscle. NMO is commonly relapsing, is frequently accompanied by other autoimmune disorders, and sometimes occurs in a paraneoplastic context. The goals of treatment are to minimize neurologic disability in the acute attack and thereafter to prevent relapses and cumulative disability. The disease specificity of AQP4 immunoglobulin (Ig) G approaches 100% using optimized molecular-based detection assays. Clinical, immunohistopathologic, and in vitro evidence support this antibody being central to NMO pathogenesis. Current animal models yield limited histopathologic characteristics of NMO, with no clinical deficits to date. Recent descriptions of a myelin oligodendrocyte glycoprotein autoantibody in a minority of patients with NMO spectrum phenotype who lack AQP4-IgG predict serologic delineation of additional distinctive disease entities.


Emerging therapeutic targets in breast cancer bone metastasis.
In the past decade, our understanding of the molecular mechanisms that underlie breast cancer pathology and progression has dramatically improved. Using this knowledge, we have identified additional targets and developed novel therapeutic interventions in breast cancer. Together, these translational research efforts are helping to usher us into an age of personalized cancer therapy. Metastasis to bone is a common and devastating consequence of breast cancer. Bisphosphonates, which represent the current gold standard in bone metastasis therapies, are being improved with newer and more efficacious generations of these compounds being developed. Breast cancer growth in the bone requires activation of various signaling pathways in both cancer cells and stromal cells, including those that are stimulated by TGF-beta and RANKL, and mediated through the Src tyrosine kinase. Bone cells and cancer cells alike express promising targets for therapeutic intervention, including Cathepsin K, CXCR4 and GPNMB. In this article we discuss the molecular mechanisms behind these pro-metastatic molecules and review the most recent findings in the clinical development of their associated targeted therapies.


The Multicenter Isradipine/Diuretic Atherosclerosis Study: a study of the antiatherogenic properties of isradipine in hypertensive patients. MIDAS Research Group.
Hypertension is a risk factor for the development of atherosclerosis and its complications, which are among the major causes of morbidity and mortality. Although recent clinical trials indicate that antihypertensive treatment reduces morbidity and mortality associated with stroke, congestive heart failure, and renal insufficiency, questions remain as to whether such treatment also prevents coronary heart disease (CHD) mortality. The observed reduction in CHD mortality from pooled clinical trial data was 10-14% and was much less than the expected 20-25% reduction for a 5-6 mm Hg reduction in diastolic pressure. One explanation may be that subtle adverse metabolic effects of treatment may have blunted the beneficial effects. Isradipine, a dihydropyridine calcium antagonist, is a potent antihypertensive drug with antiatherogenic properties in animal models. Therefore, we hypothesized that isradipine may be appropriate for testing the efficacy of antihypertensive treatment in retarding the progression of atherosclerosis in humans. The Multicenter Isradipine/Diuretic Atherosclerosis Study (MIDAS) is a clinical trial designed to compare the efficacy of isradipine (2.5 or 5 mg b.i.d.) with hydrochlorothiazide (12.5 or 25 mg b.i.d.) in retarding the progression of early carotid atherosclerosis as monitored by high-resolution B-mode ultrasonography.


Delay of Gratification as Reputation Management.
Although delay-of-gratification tasks have long been used as measures of self-control, recent evidence suggests that performance on these tasks is also driven by rational decision processes. The present research examined whether the effects of rational decision processes extend beyond costs and benefits embedded in the task itself to include anticipated consequences for the child's reputation. Across two studies, 3- and 4-year-olds from China (N = 273) were assigned to a standard delay-of-gratification condition or to a reputation condition in which they were told that their teacher or a peer would find out how long they had waited. Children waited longer in the reputation conditions and longer in the teacher condition than in the peer condition. This is the first evidence that children's performance on a delay-of-gratification task is sensitive to reputational concerns and to the identity of potential evaluators of their behavior.


Anionic contribution for fibrous maturation of protofibrillar assemblies of the human tau repeat domain in a fluoroalcohol solution.
Tau protein forms fibrous aggregates in the brain of patients with Alzheimer's disease. This type of aggregation in vitro is promoted efficiently by polyanions and anionic micelles. Here, we report another cosolvent system that induces the fibrous aggregation of human tau four-repeat domain (tau4RD). The protein aggregation was primarily achieved by a nonanionic agent 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), while the ionic condition was modified by inorganic salts. The aggregation analysis by three spectroscopic methods revealed a two-phase kinetics of the aggregation of tau4RD in the presence of HFIP at approximately 4-6%. Large increases in the light-scattering, the thioflavin-binding, and the secondary structure content of tau4RD have progressed within a few minutes at 37 degrees C, which was followed by another slower aggregation phase. Electron microscopic analysis demonstrated that the amorphous granules are formed in the faster step, which acquired a fibrous shape in the slower step in the solution containing NaCl. In the absence of the salt, however, the fibrous maturation was inhibited. Examination of various salt species in place of NaCl demonstrated that binding of anions to the precursor aggregates was essential for the fibrous maturation. On the basis of the results, we proposed an aggregation scheme of tau in which the formation of a thioflavin-binding intermediate occurred ahead of its fibrous maturation. The anionic environment was suggested to play a crucial role in the fibrous maturation and, therefore, could be an in vivo determinant of the morphology of the aggregates of tau.


Depression following myocardial infarction--an overseen complication with prognostic importance.
Myocardial infarction (MI) is a severe life event that is accompanied by an increased risk of depression. Mounting evidence suggests that post-MI depression is associated with adverse outcomes, but the underlying mechanisms of this association remain unclear, and no previous studies have examined whether the mental burden of MI is so heavy that it increases the risk of suicide. Although post-MI depression is common and burdensome, the condition remains under-recognised and under-treated. The development of new strategies to improve the quality of care for people with post-MI depression requires thorough understanding of the mechanisms that influence the prognosis as well as knowledge of the present care provided. The purpose of this PhD thesis is accordingly subdivided into four specific aims: 1. To estimate the prevalence of depression in people with MI after three months, and to estimate the provided hospital-based psychosocial rehabilitation (Paper I); 2. To examine GPs' practice of screening for depression in people with MI, and to analyse whether the screening rate varied among subgroups of people with a particularly high risk of post-MI depression (Paper II); 3. To examine the association between post-MI depression and new cardiovascular events or death, taking potential mediators into account (Paper III); 4. To examine the association between MI and suicide (Paper IV). Two different study designs were employed: a population-based cohort study using data obtained from registers and questionnaires sent to MI patients and their GPs (Paper I-III); a nationwide population-based matched case-control study using data obtained from registers (Paper IV). Three months after having suffered MI, about one fifth of the patients in our study had depression according to the Hospital Anxiety and Depression Scale (HADS). Upwards of half of the patients had participated in some rehabilitation, thirty per cent had participated in psychosocial support and three per cent reported that they had been examined for depression by questionnaire during the hospital-based rehabilitation. The rate of participation in psychosocial support and examination for depression was the same for patients with or without depression at three months following MI. A good fourth of the patients' GPs stated that they had screened the patient for depression within the first year after the patient suffered an MI. Three months after the MI, the screening rate was higher among patients with a history of mental illness and among patients with anxiety or depression than among patients without these conditions. Most of the GPs who performed screening followed guidelines by asking about specific depressive symptoms. According to the HADS, depression three months after MI was associated with an increased risk of a new cardiovascular event or death. This association was partly explained by the underlying cardiac disease severity and physical inactivity, but depression remained an independent prognostic risk factor after adjusting for these potential confounders. We found a tendency towards a weaker association with increasing physical activity and among users of antidepressants. MI was strongly associated with an increased risk of suicide. This risk was particularly high immediately after the MI, but remained high for more than five years after the MI. The association between MI and suicide remained stable throughout the study period (1981-2006), although many other factors changed (e.g. suicide rate, treatment of MI). This thesis demonstrated that post-MI depression is common, under-recognized and has a strong prognostic impact. About one in five patients have depression three months after MI. Guidelines recommend screening for depression, but the guidelines have not been systematically implemented either in the hospital-based rehabilitation or in Danish general practice. In the absence of systematic screening, we found that a significant part of those who had depression were not recognized. MI increases the risk of suicide, and depression following MI impairs the overall prognosis. The thesis indicates that physical activity and antidepressants modify the adverse prognosis in patients with post-MI depression, but larger studies are needed to clarify the impact of these potential modifiers and to evaluate how they may be catered for in the treatment of post-MI patients with depression.


The martyrdom of Doctor Andreas Vesalius.
The development of modern medicine began in the 16th century when Dr. Andreas Vesalius overthrew the previously uncontested medical dogma of the Greek physician Galen. Medical progress had been hindered for more than a millennium by strict adherence to Galen's authority. Flemish-born Vesalius challenged the political and societal forces of the time. At the University of Padua, he studied and later taught human anatomy by performing dissections. His discoveries were published in 1543 in his monumental De Humani Corporis Fabrica. Controversy led to his resignation from the University of Padua. His magnum opus was interpreted as a challenge to both academia and the church. He went to Spain, where he served as personal physician to Emperor Charles V. After almost 20 years in Spain, he became involved in an unfortunate incident that incurred the condemnation of the Inquisition. The royal court's intervention saved Vesalius from being burned at the stake, however, and he was ordered to make a pilgrimage to Jerusalem to atone for his error. On the return voyage, he became ill and died on the Greek island of Zante.


A distinct Cu(4)-thiolate cluster of human metallothionein-3 is located in the N-terminal domain.
Metallothionein-3 (MT-3), also known as neuronal growth inhibitory factor, is a metalloprotein expressed almost exclusively in the brain. Isolated MT-3 contains four Cu(I) and three Zn(II) ions organized in homometallic metal-thiolate clusters located in two independent protein domains. In this work a Cu(I) binding to metal-free MT-3 has been studied, aiming at the better understanding of the domain specificity for this metal ion. The cluster formation was followed by electronic absorption, circular dichroism, and by luminescence spectroscopy at room temperature and 77 K. The stepwise incorporation of Cu(I) into recombinant human apo-MT-3 revealed the cooperative formation of two Cu(4)S(9) clusters in succession, formed in both protein domains, i.e. Cu(4)- and Cu(8)-MT-3. Further binding of four Cu(I) caused an expansion of these Cu(I) cores, leading to fully metal-loaded Cu(12)-MT-3 containing Cu(6)S(9) and Cu(6)S(11) clusters in the beta- and alpha-domains of the protein, respectively. The location of the preferentially formed Cu(4) cluster in the protein was established by immunochemistry. Using domain-specific antibodies, in combination with limited tryptic digestion of a partially metal-occupied Cu(4)-MT-3, we could demonstrate that the Cu(4)S(9) cluster is located in the N-terminal beta-domain of the protein that contains a total of nine cysteine ligands.