Novel Assay to Detect RNA Polymerase I Activity In Vivo.
This report develops an analytically validated chromogenic in situ hybridization (CISH) assay using branched DNA signal amplification (RNAscope) for detecting the expression of the 5' external transcribed spacer (ETS) of the 45S ribosomal (r) RNA precursor in formalin-fixed and paraffin-embedded (FFPE) human tissues. 5'ETS/45S CISH was performed on standard clinical specimens and tissue microarrays (TMA) from untreated prostate carcinomas, high-grade prostatic intraepithelial neoplasia (PIN), and matched benign prostatic tissues. Signals were quantified using image analysis software. The 5'ETS rRNA signal was restricted to the nucleolus. The signal was markedly attenuated in cell lines and in prostate tissue slices after pharmacologic inhibition of RNA polymerase I (Pol I) using BMH-21 or actinomycin D, and by RNAi depletion of Pol I, demonstrating validity as a measure of Pol I activity. Clinical human prostate FFPE tissue sections and TMAs showed a marked increase in the signal in the presumptive precursor lesion (high-grade PIN) and invasive adenocarcinoma lesions (P = 0.0001 and P = 0.0001, respectively) compared with non-neoplastic luminal epithelium. The increase in 5'ETS rRNA signal was present throughout all Gleason scores and pathologic stages at radical prostatectomy, with no marked difference among these. This precursor rRNA assay has potential utility for detection of increased rRNA production in various tumor types and as a novel companion diagnostic for clinical trials involving Pol I inhibition.Implications: Increased rRNA production, a possible therapeutic target for multiple cancers, can be detected with a new, validated assay that also serves as a pharmacodynamic marker for Pol I inhibitors. Mol Cancer Res; 15(5); 577-84. ©2017 AACR.


Microscopic imaging and spectroscopy with scattered light.
Optical contrast based on elastic scattering interactions between light and matter can be used to probe cellular structure, cellular dynamics, and image tissue architecture. The quantitative nature and high sensitivity of light scattering signals to subtle alterations in tissue morphology, as well as the ability to visualize unstained tissue in vivo, has recently generated significant interest in optical-scatter-based biosensing and imaging. Here we review the fundamental methodologies used to acquire and interpret optical scatter data. We report on recent findings in this field and present current advances in optical scatter techniques and computational methods. Cellular and tissue data enabled by current advances in optical scatter spectroscopy and imaging stand to impact a variety of biomedical applications including clinical tissue diagnosis, in vivo imaging, drug discovery, and basic cell biology.


Transferring skills in quality collaboratives focused on improving patient logistics.
A quality improvement collaborative, often used by the Institute for Healthcare Improvement, is used to educate healthcare professionals and improve healthcare at the same time. However, no prior research has been done on the knowledge and skills healthcare professionals need to achieve improvements or the extent to which quality improvement collaboratives help enhance both knowledge and skills. Our research focused on quality improvement collaboratives aiming to improve patient logistics and tried to identify which knowledge and skills are required and to what extent these were enhanced during the QIC. We defined skills important for logistic improvements in a three-phase Delphi study. Based on the Delphi results we made a questionnaire. We surveyed participants in a national quality improvement collaborative to assess the skills rated as 1) important, 2) available and 3) improved during the collaborative. At two sense-making meetings, experts reflected on our findings and hypothesized on how to improve (logistics) collaboratives. The Delphi study found 18 skills relevant for reducing patient access time and 21 for reducing throughput time. All skills retrieved from the Delphi study were scored as 'important' in the survey. Teams especially lacked soft skills connected to project and change management. Analytical skills increased the most, while more reflexive skills needed for the primary goal of the collaborative (reduce access and throughput times) increased modestly. At two sense-making meetings, attendees suggested four improvements for a quality improvement collaborative: 1) shift the focus to project- and change management skills; 2) focus more on knowledge transfer to colleagues; 3) teach participants to adapt the taught principles to their own situations; and 4) foster intra-project reflexive learning to translate gained insights to other projects (inter-project learning). Our findings seem to suggest that Quality collaboratives could benefit if more attention is paid to the transfer of 'soft skills' (e.g. change, project management and communication skills) and reflexive skills (e.g. adjusting logistics principles to specific situations and inter-project translation of experiences).


Self-Medication Practices and Associated Factors Among Health-Care Professionals in Selected Hospitals of Western Ethiopia.
Even though the type, extent and reasons for self-medication practice (SMP) vary, globally self-medication (SM) is rising to relieve burdens on health services. However, inappropriate SMP results in economic wastes, damage of vital organs, incorrect therapy selection, risk of adverse drug reactions and development of antimicrobial-resistant pathogens. These consequences have severe implications including legal, ethical and quality of health-care delivery. Temporal increment and high prevalence of SM among health professionals is also a major bottleneck for Ethiopia. Hence, the study aimed to assess the SM among health-care professionals (HCPs) in selected governmental hospitals of Western Ethiopia. An instiution-based cross-sectional study was conducted among 338 HCPs using a pre-tested and self-adminstered questionnaries from March 1 to 25, 2018. Simple random sampling was used to select study participants and SMP (yes or no) was the outcome of the study variable. Data were entered and analyzed using SPSS version 20. Crude and adjusted odds ratios (95% CI) were calculated and all results were deemed to be statistically significant when p < 0.05. Among the 338 participants, 184 (54.4%) were females and the mean age of the study participants was 25±3.23 years. About 154 (45.6%) of them had work experience of less than 5 years and 49.7% were nurses by profession. The prevalence of SM was 73.4% with 3 months of recall for SM. Familiarity with medicines and ailments (46.8%) and mildness of illness (40.7%) were the most common reasons to self-medicate. The most frequently reported ailments were headache (37.1%) and gastric pain (29.8%). Analgesics (44.4%) and antibiotics (42.7%) were the most commonly used self-medicated categories of drugs. Female sex (Adjusted odds ratio [AOR] =2.13, 95% CI: 1.43-8.66), age 20-29 years (AOR=4.53, 95% CI: 1.01-14.45) and work experience of <5 years (AOR= 3.01, 95% CI: 1.32-11.71) were significantly associated with SMPs. The study revealed a high prevalence of SMP among HCPs. Sex, age, and work experience were significantly associated with SMPs. Hence, the use of prescription drugs without prescription should be discouraged and appropriate health education should be provided by all concerned bodies on the proper use of drugs.


QM assisted ML for 19F NMR chemical shift prediction.
Ligand-observed 19F NMR detection is an efficient method for screening libraries of fluorinated molecules in fragment-based drug design campaigns. Screening fluorinated molecules in large mixtures makes 19F NMR a high-throughput method. Typically, these mixtures are generated from pools of well-characterized fragments. By predicting 19F NMR chemical shift, mixtures could be generated for arbitrary fluorinated molecules facilitating for example focused screens. In a previous publication, we introduced a method to predict 19F NMR chemical shift using rooted fluorine fingerprints and machine learning (ML) methods. Having observed that the quality of the prediction depends on similarity to the training set, we here propose to assist the prediction with quantum mechanics (QM) based methods in cases where compounds are not well covered by a training set. Beyond similarity, the performance of ML methods could be associated with individual features in compounds. A combination of both could be used as a procedure to split input data sets into those that could be predicted by ML and those that required QM processing. We could show on a proprietary fluorinated fragment library, known as LEF (Local Environment of Fluorine), and a public Enamine data set of 19F NMR chemical shifts that ML and QM methods could synergize to outperform either method individually. Models built on Enamine data, as well as model building and QM workflow tools, can be found at https://github.com/PatrickPenner/lefshift and https://github.com/PatrickPenner/lefqm .


Time to Move Beyond a "One-Size Fits All" Approach to Inspiratory Muscle Training.
Inspiratory muscle training (IMT) has been studied as a rehabilitation tool and ergogenic aid in clinical, athletic, and healthy populations. This technique aims to improve respiratory muscle strength and endurance, which has been seen to enhance respiratory pressure generation, respiratory muscle weakness, exercise capacity, and quality of life. However, the effects of IMT have been discrepant between populations, with some studies showing improvements with IMT and others not. This may be due to the use of standardized IMT protocols which are uniformly applied to all study participants without considering individual characteristics and training needs. As such, we suggest that research on IMT veer away from a standardized, one-size-fits-all intervention, and instead utilize specific IMT training protocols. In particular, a more personalized approach to an individual's training prescription based upon goals, needs, and desired outcomes of the patient or athlete. In order for the coach or practitioner to adjust and personalize a given IMT prescription for an individual, factors, such as frequency, duration, and modality will be influenced, thus inevitably affecting overall training load and adaptations for a projected outcome. Therefore, by integrating specific methods based on optimization, periodization, and personalization, further studies may overcome previous discrepancies within IMT research.


Soy isoflavonoids and cancer prevention. Underlying biochemical and pharmacological issues.
The isoflavonoids in soy, genistein and daidzein, have been proposed to contribute an important part of the anti-cancer effect of soy. Although there have been many interesting studies on the effects of isoflavones on biochemical targets in tissue culture experiments, in most cases the concentrations used by investigators have exceeded 10 microM. However, based on simple pharmacokinetic calculations involving daily intake of isoflavones, absorption from the gut, distribution to peripheral tissues, and excretion, it is unlikely that blood isoflavone concentrations even in high soy consumers could be greater than 1-5 microM. Experiments designed to evaluate these pharmacological principles were carried out in anesthetized rats with indwelling biliary catheters and in human volunteers consuming soy beverages. The data from these experiments indicate that genistein is efficiently absorbed from the gut, taken up by the liver and excreted in the bile as its 7-O-beta-glucuronide. Re-infused genistein 7-O-beta-glucuronide was also well absorbed from the gut, although this occurred in the distal small intestine. In human subjects fed a soy beverage for a period of two weeks, plasma levels of genistein and daidzein, determined by HPLC-mass spectrometry, ranged from 0.55-0.86 microM, mostly as glucuronide and sulfate conjugates. In summary, genistein is well absorbed from the small intestine and undergoes an enterohepatic circulation. Although the plasma genistein levels achievable with soy food feeding are unlikely to be sufficient to inhibit the growth of mature, established breast cancer cells by chemotherapeutic-like mechanisms, these levels are sufficient to regulate the proliferation of epithelial cells in the breast and thereby may cause a chemopreventive effect.


Developing a project-oriented introduction to neuroscience lab at hope college.
The Introduction to Neuroscience course at Hope College includes a three-hour laboratory period each week. Seven of the fifteen weeks of the lab are used for a lab project that is focused on understanding the effects of gonadal hormones on brain and behavior. Students perform ovariectomies and implant sham, estradiol, or testosterone capsules in rats and then carry out five experiments: 1) Sexual Behavior, 2) Spatial Learning using the Morris Water Maze, 3) The Size of the Sexually Dimorphic Nucleus, 4) Phosphorylation of NMDA Receptors, and 5) Long Term Potentiation in Hippocampal Slices. The experiments are designed to provide the students with experiences at different levels of neuroscience, while improving their skills in statistics, using the primary literature, and scientific writing. The students generate interesting and statistically significant data which they summarize in a journal style lab reports. Using a Self Assessment of Learning Gains tool, we learned that students perceive the lab project improves their ability to A) pose questions from more than one disciplinary perspective that can be addressed by collecting and evaluating scientific evidence, B) learn about complex science problems that require insight from more than one discipline, C) extract main points from a scientific article and develop a coherent summary, and D) write reports using scientific data as evidence. Based on our results, we believe an extended lab project in an introductory neuroscience course can be used to engage students in neuroscience topics and help them develop the skills and habits of neuroscientists.


Determining the optimal number of patient rooms for an acute care unit.
With nearly $200 billion of healthcare construction expected by 2015, nurse leaders must expand their knowledge and capabilities in healthcare design. This bimonthly department, edited by Jaynelle F. Stichler, DNS, RN, FACHE, prepares nurse leaders to use the evidence-based design process to ensure that new, expanded, and renovated hospitals facilitate optimal patient outcomes, enhance the work environment for healthcare providers, and improve organizational performance. The authors of this article discuss issues to be considered in determining the optimal number of patient rooms to design for an acute care patient care unit.


Paludicola gen. nov. and Revision of the Species Formerly in Batrachospermum Section Turfosa (Batrachospermales, Rhodophyta).
Since the first phylogenetic study of the order Batrachospermales, Batrachospermum was shown to be paraphyletic. Subsequently, sections of the genus have been methodically investigated using DNA sequences and morphology in order to propose new genera and delineate species. Batrachospermum section Turfosa is the last section with multiple species yet to be examined. New sequence data of specimens from Europe and the United States were combined with the sparse sequence data already available. Phylogenetic analyses using rbcL and COI-5P sequences showed this section to be a well-supported clade, distinct from Batrachospermum section Batrachospermum and its segregate genera. Section Turfosa is raised to the generic rank as Paludicola gen. nov. Substantial genetic variation within the genus was discovered and 12 species are recognized based on DNA sequence data as well as morphological characters and geographic distribution. The following morphological characters were applied to distinguish species: branching pattern (pseudodichotomous or irregular), whorl size (reduced or well developed), primary fascicles (curved or straight), spermatangia origin (primary or secondary fascicles), and carposporophyte arrangement (loose or dense). Previously published species were transferred to the new genus: P. turfosa, P. keratophyta, P. orthosticha, P. phangiae, and P. periploca. Seven new species are proposed as follows: P. groenbladii from Europe; P. communis, P. johnhallii, and P. leafensis from North America; and P. aquanigra, P. diamantinensis, and P. turfosiformis from Brazil. In addition, three unsequenced species in the section, P. bakarensis, P. gombakensis, and P. tapirensis, were transferred to the new genus.


Aerosolized bexarotene inhibits lung tumorigenesis without increasing plasma triglyceride and cholesterol levels in mice.
Prior studies have shown the retinoid X receptor (RXR) agonist bexarotene has preventive efficacy in rodent models of mammary and lung tumorigenesis albeit causing hypertriglyceridemia and hypercholesterolemia. We reasoned that bexarotene delivered by inhalation may provide sufficient dose directly to the respiratory tract to achieve efficacy while avoiding these side effects. In this study, the chemopreventive activity of aerosolized bexarotene was investigated in the benzo(a)pyrene [B(a)P]-induced mouse lung tumor model as assessed by tumor multiplicity and tumor load. Aerosolized bexarotene significantly decreased tumor multiplicity and tumor load by 43% and 74%, respectively. Our data showed that bexarotene can both inhibit proliferation and promote apoptosis in vivo. Our data also show that aerosolized bexarotene did not increase plasma total cholesterol and triglyceride level compared with diet group. These results indicate that aerosolization may be a safe and effective route of administering bexarotene for chemoprevention of lung cancer.


Diverse effects of naturally occurring base lesions on the structure and stability of the human telomere DNA quadruplex.
Various base lesions continuously form in cellular nucleic acids and the unrepaired lesions are promutagenic and procarcinogenic. Though natural base lesions have been extensively studied in double-stranded DNA models, these studies are only less than a decade old for non-canonical DNA models, such as quadruplexes. Here we present a report on the effects of three frequently occurring natural lesions that can form in the TTA loops on the structure of the human telomere quadruplex d[AG3(TTAG3)3]. We compared the effect of the abasic site and 8-oxoadenine replacing adenine and 5-hydroxymethyluracil substituting for thymine. The results showed that the three lesions impacted the stability and quadruplex folding in markedly different ways. The effects depended on the type of lesion and the position in the sequence. Analogous lesions of guanine in the G-tetrads extensively destabilized the quadruplex and the effects depended more on the position than on the type of lesion. The distinct effects of the loop substitutions as well as comparison of the modifications of the loops and the quadruplex tetrads are discussed in this communication.


The Clinical Effect of Repetitive Transcranial Magnetic Stimulation on the Disturbance of Consciousness in Patients in a Vegetative State.
To explore the effect of combining repetitive transcranial magnetic stimulation (rTMS) and conventional rehabilitation on the recovery of consciousness in patients in a persistent vegetative state (PVS). A total of 48 patients in a PVS were randomly divided into a treatment and control group. Patients in the treatment group were treated with rTMS to stimulate the dorsolateral prefrontal cortex, and patients in the control group were treated with false stimulation. All patients were evaluated using scales and neuroelectrophysiological assessment before treatment, after 30 days of treatment, and following 60 days of treatment. Based on the Coma Recovery Scale-Revised (CRS-R) and electroencephalogram (EEG) grading indexes, the treatment group was significantly higher than those of the control group after 30 and 60 days of treatment. The average difference in the three measurements between the two groups before treatment, at 30 days, and 60 days was 0.04, 1.54, and 2.09 for CRS-R and 0.08, -0.83, and -0.62 for EEG indexes, respectively. The latency periods of each wave of the brainstem auditory evoked potentials (BAEPs) in the treatment group were shorter than those in the control group after 30 and 60 days of treatment. In both groups, the BAEP scores after 30 days of treatment were significantly higher than the scores before treatment, and the scores after 60 days of treatment were higher than the scores after 30 days. In patients in a PVS, rTMS assists in the recovery of consciousness function.


[Walking does not require continuity of the medullar "locomotion strip"].
Bilateral lesions interrupting the "locomotor strips" were produced in the medulla of mesencephalic cats. After that locomotion could be evoked by stimulation of the rostral and caudal part of the "strip". However, in some experiments the locomotion threshold rose after the lesion. The relationship between the "locomotor strips" and other brain stem structures and spinal cord is under discussion.


Hypoxia and trophoblast differentiation: a key role for PPARγ.
Tissue oxygen tension regulates differentiation of multiple types of stem cells. In the placenta, hypoxia has been associated with abnormal trophoblast differentiation and placental insufficiency syndromes of preeclampsia (PE) and intrauterine growth restriction (IUGR). Peroxisome proliferator-activated receptor-γ (PPARγ) is a ligand-activated transcription factor involved in many cellular processes, including differentiation. We have previously shown that PPARγ-null trophoblast stem (TS) cells show a defect in differentiation to labyrinthine trophoblast, instead differentiating preferentially to trophoblast giant cells (TGC). Since PPARγ is known to be regulated by hypoxia in adipose tissue, we hypothesized that there may be a link between oxygen tension, PPARγ expression, and trophoblast differentiation. We found that hypoxia reduced PPARγ expression by a mechanism independent of both hypoxia-inducible factor (HIF) and histone deacetylases (HDACs). In addition, PPARγ partially rescued hypoxia-induced inhibition of labyrinthine differentiation in wild-type TS cells but was not required for hypoxia-induced inhibition of TGC differentiation. Finally, we show that induction of labyrinthine trophoblast differentiation by HDAC inhibitor treatment is independent of both PPARγ and Gcm1. We propose a model with two pathways for labyrinthine trophoblast differentiation of TS cells, one of which is dependent on PPARγ and inhibited by hypoxia. Since hypoxia is associated with PE and IUGR, we propose that PPARγ may at least partially mediate hypoxia-induced placental insufficiency and as such may be a promising therapeutic target for these disorders.


Nicotinic acetylcholine receptor (CHRN) expression and function in cultured human adult fungiform (HBO) taste cells.
In rodents, CHRNs are involved in bitter taste transduction of nicotine and ethanol. Currently, it is not clear if CHRNs are expressed in human taste cells and if they play a role in transducing the bitter taste of nicotine and ethanol or in the synthesis and release of neurohumoral peptides. Accordingly, we investigated the expression and functional role of CHRNs in HBO cells. Using molecular techniques, we demonstrate that a subset of HBO cells express CHRNs that also co-express TRPM5, T1R3 or T2R38. Exposing HBO cells to nicotine or ethanol acutely or to nicotine chronically induced a differential increase in the expression of CHRN mRNA and protein in a dose- and time-dependent manner. Acutely exposing HBO cells to a mixture containing nicotine plus ethanol induced a smaller increase in CHRN mRNAs relative to nicotine or ethanol treatment alone. A subset of HBO cells responded to nicotine, acetylcholine and ATP with a transient increase in [Ca2+]i. Nicotine effects on [Ca2+]i were mecamylamine sensitive. Brain-derived neurotrophic factor (BDNF) protein was detected in HBO cells using ELISA. Acute nicotine exposure decreased BDNF in HBO cells and increased BDNF release in the medium. CHRNs were also detected in HEK293 cells by RT-PCR. Unlike HBO cells, CHRNs were localized in most of HEK293 cells and majority of HEK293 cells responded to nicotine and ethanol stimulation with a transient increase in [Ca2+]i. BDNF levels in HEK293 cells were significantly higher than in HBO cells but the nicotine induced release of BDNF in the media was a fraction of the BDNF cellular content. We conclude that CHRNs are expressed in TRPM5 positive HBO cells. CHRN mRNA expression is modulated by exposure to nicotine and ethanol in a dose- and time-dependent manner. Nicotine induces the synthesis and release of BDNF in HBO cells.


Targeted delivery of Doxorubicin by folic acid-decorated dual functional nanocarrier.
Doxorubicin (DOX) is one of the most commonly used antineoplastic agents, but its clinical application is oftentimes coupled with severe side effects. Selective delivery of DOX to tumors via nanosized drug carrier represents an attractive approach to this problem. Previously, we developed a dual functional nanomicellar carrier, PEG5K-embelin2 (PEG5K-EB2), which was able to deliver paclitaxel (PTX) selectively to tumors and to achieve an enhanced therapeutic effect. In the present study, we examined the utility of PEG5K-EB2 to deliver DOX to tumors. In addition, folic acid (FA) was coupled to the surface of the PEG5K-EB2 micelles (FA-PEG5K-EB2) to further improve the selective targetability of the system. DOX-loaded PEG5K-EB2 micelles were uniformly spherical particles with a diameter of approximately 20 nm. Incorporation of FA had minimal effect on the size of the particles. The DOX loading efficiency was as high as 91.7% and 93.5% for PEG5K-EB2 and FA-PEG5K-EB2, respectively. DOX formulated in PEG5K-EB2 micelles (with or without FA decoration) demonstrated sustained kinetics of DOX release compared to free DOX. FA-PEG5K-EB2 significantly facilitated the intracellular uptake of DOX over free DOX and PEGylated liposomal DOX (Doxil) in breast cancer cells, 4T1.2, and drug resistant cells, NCI/ADR-RES. P-gp ATPase assay showed that PEG5K-EB2 significantly inhibited the function of the P-gp efflux pump. The maximum tolerated dose of DOX-loaded PEG5K-EB2 micelles was 15 mg/kg in mice, which was 1.5-fold greater than that for free DOX. Pharmacokinetics (PK) and biodistribution studies showed that both types of DOX-loaded micelles, especially FA-PEG5K-EB2, were able to significantly prolong the blood circulation time of DOX and facilitate its preferential accumulation at the tumor tissue. Finally, DOX/PEG5K-EB2 mixed micelles demonstrated significantly enhanced tumor growth inhibitory effect with minimal toxicity in comparison to free DOX and Doxil and the antitumor activity was further enhanced after the decoration by folic acid. Our data suggest that FA-PEG5K-EB2 micelles represent a promising DOX delivery system that warrants more study in the future.


Hepatitis C Virus, Inflammation, and Cellular Aging: Turning Back Time.
There is evidence that hepatitis C virus (HCV) infection, like HIV infection, may be associated with chronic inflammation, immune activation, and immune senescence, which contribute to increased risks for cardiometabolic or other diseases outside the liver, as well as to ongoing damage in the liver. These effects may persist after a sustained virologic response (SVR) is achieved with HCV therapy. Such findings support initiation of treatment for HCV-infected individuals before damage to the liver is apparent and monitoring of individuals for complications even after an SVR is achieved. Fibrosis is not always reversible after SVR is achieved, and this should serve as an argument against waiting until fibrosis develops before initiating treatment for HCV-infected individuals. This article summarizes a presentation by Susanna Naggie, MD, MHS, at the IAS-USA continuing education program, Management of Hepatitis C Virus in the New Era: Small Molecules Bring Big Changes, in New York, New York, in September 2015.


Promoting community practitioners' use of evidence-based approaches to increase breast cancer screening.
Many women do not get mammography screenings at the intervals recommended for early detection and treatment of breast cancer. The Guide to Community Preventive Services (Community Guide) recommends a range of evidence-based strategies to improve mammography rates. However, nurses and others working in community-based settings make only limited use of these strategies. We report on a dissemination intervention that partnered the University of North Carolina with the Susan G. Komen Triangle Affiliate to disseminate Community Guide breast cancer screening strategies to community organizations. The intervention was guided by social marketing and diffusion of innovation theory and was designed to provide evidence and support via Komen's existing relationships with grantee organizations. The present study reports the findings from a formative evaluation of the intervention, which included a content analysis of 46 grant applications pre- and post intervention and focus groups with 20 grant recipients.


A Single-Domain Antibody Targeting Complement Component C5 Acts as a Selective Inhibitor of the Terminal Pathway of the Complement System and Thus Functionally Mimicks the C-Terminal Domain of the Staphylococcus aureus SSL7 Protein.
The complement system is an efficient anti-microbial effector mechanism. On the other hand abnormal complement activation is involved in the pathogenesis of multiple inflammatory and hemolytic diseases. As general inhibition of the complement system may jeopardize patient health due to increased susceptibility to infections, the development of pathway-specific complement therapeutics has been a long-lasting goal over the last decades. In particular, pathogen mimicry has been considered as a promising approach for the design of selective anti-complement drugs. The C-terminal domain of staphylococcal superantigen-like protein 7 (SSL7), a protein secreted by Staphylococcus aureus, was recently found to be a specific inhibitor of the terminal pathway of the complement system, providing selective inhibition of cell lysis mediated by the membrane attack complex (MAC). We describe here the selection by phage display of a humanized single-domain antibody (sdAb) mimicking the C-terminal domain of SSL7. The antibody, called sdAb_E4, binds complement C5 with an affinity in the low micromolar range. Furthermore, sdAb_E4 induces selective inhibition of MAC-mediated lysis, allowing inhibition of red blood cell hemolysis and inhibition of complement deposition on apopto-necrotic cells, while maintaining efficient bactericidal activity of the complement terminal pathway. Finally, we present preliminary results indicating that sdAb_E4 may also be efficient in inhibiting hemolysis of erythrocytes from patients with paroxysmal nocturnal hemoglobinuria. Our data provide a proof of concept for the design of a selective MAC inhibitor capable of retaining complement bacteriolytic activity and this study opens up promising perspectives for the development of an sdAb_E4-derived therapeutics with application in the treatment of complement-mediated hemolytic disorders.


Detection and toxin production of Staphylococcus aureus in sudden infant death cases in Hungary.
The potential role of microbial agents was investigated in 13 cases of Sudden Infant Death Syndrome and in 9 non-SIDS cases in Budapest between September 1996 and May 1998. Autopsy, histological examination and microbiological tests were performed on samples of blood, cerebrospinal fluid, pharyngeal samples and lung tissue from infants under one year died suddenly, without previous diseases. The multifactorial pathomechanism of SIDS was suggested by the isolation of toxin producing Staphylococcus aureus-, Enterobacteriaceae and Candida albicans strains in large number and by the detection of Parainfluenza Type 2 virus antigen. S. aureus proved the predominant bacteria in the SIDS cases. Nasopharyngeal microbial flora and S. aureus carrier of 100 age matched healthy infants were tested during the same period. S. aureus was isolated from 54% of SIDS cases and 37% from healthy infants /OR = 1.986 (95% Confidence interval = 0.55-7.33), p = 0243/. The enterotoxin and TSST-1 toxin producing activity of S. aureus showed the characteristic difference. The toxigenic S. aureus was detected in 46% of SIDS cases and 16% of healthy infants /OR = 4.5 (95% CI = 1.15-17.72), p = 0.010/. The distribution of toxigenic and nontoxigenic isolates was 86% in SIDS cases and 43% in healthy infants /OR = 7.875 (CI = 0.78-191.89), p = 0.041/.


Colloidal properties of nanoparticular biogenic selenium govern environmental fate and bioremediation effectiveness.
Microbial selenium (Se) bioremediation is based on conversion of water soluble, toxic Se oxyanions to water insoluble, elemental Se. Formed biogenic elemental Se is of nanometer size, hampering straightforward separation from the aqueous phase. This study represents the first systematic investigation on colloidal properties of pure biogenic Se suspensions, linking electrophoretic mobility (ζ-potential) to column settling behavior. It was demonstrated that circumneutral pH, commonly applied in bioremediation, is not appropriate for gravitational separation due to the negative ζ-potential preventing agglomeration. Mono/di/trivalent counter cations and acidity (protons) were used to screen efficiently the intrinsic negative charge of biogenic Se suspensions at circumneutral pH. Fast settling was induced by La(3+) addition in the micromolar range (86.2 ± 3.5% within 0.5 h), whereas considerably higher concentrations were needed when Ca(2+) or Na(+) was used. Colloidal stability was furthermore studied in different model waters. It was demonstrated that surface waters as such represent a fragile system regarding colloidal stability of biogenic Se suspensions (ζ-potential ∼ -30 mV), whereas dissolved organic matter increases colloidal stability. In marine waters, biogenic Se is colloidally destabilized and is thus expected to settle, representing a potential sink for Se during transport in the aquatic environment.


Dance Your Heart Out: A Community's Approach to Addressing Cardiovascular Health by Using a Logic Model.
Cardiovascular health has been identified as a prioritized community health need according to recent community health needs assessment data. While the Affordable Care Act mandates that nonprofit hospitals conduct a community health needs assessment, little guidance exists on how to address the identified needs. Logic models provide systematic structure and necessary direction in how communities can start to address their identified health needs. Completing logic models in a nonlinear fashion is encouraged to employ a strengths-based approach and verify the logic. This article provides an application of logic models as one strategy to generate a community-based program theory to improve cardiovascular health.


Cognitive decline and education in mild dementia.
Recent studies suggested that education may modify the clinical expression of dementia and Alzheimer's disease through its association with a brain reserve capacity. We studied whether education would be related to degree of cognitive decline in mild dementia. Equations to estimate premorbid cognitive ability were derived from a representative normative sample of 83 community-dwelling African Americans using age, education, and gender as independent variables and Word List Learning (WLL) and Animal Fluency (AF) scores from the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) neuropsychological test battery as dependent variables. These equations were applied to a second sample of 131 African Americans (22 with dementia, 109 healthy) who completed CERAD test batteries as part of an epidemiologic study of dementia in the community. Differences between obtained and estimated premorbid WLL and AF test scores were calculated and then analyzed in a 2 (Education) x 2 (Diagnosis) ANOVA. A significant interaction association between Education and Diagnosis on WLL scores and a borderline significant interaction on AF scores showed that the high-education demented group had a greater cognitive decline from estimated premorbid levels than the low-education demented group. Thus, at comparable levels of clinical dementia severity, greater cognitive decline occurred in highly educated patients than in low-educated patients.


Early 11C-PIB frames and 18F-FDG PET measures are comparable: a study validated in a cohort of AD and FTLD patients.
The availability of new PET ligands offers the potential to measure fibrillar β-amyloid in the brain. Nevertheless, physiologic information in the form of perfusion or metabolism may still be useful in differentiating causes of dementia during life. In this study, we investigated whether early (11)C-Pittsburgh compound B ((11)C-PIB) PET frames (perfusion (11)C-PIB [pPIB]) could provide information equivalent to blood flow and metabolism. First, we assessed the similarity of pPIB and (18)F-FDG PET images in a test cohort with various clinical diagnoses (n = 10), and then we validated the results in a cohort of patients with Alzheimer disease (AD) (n = 42; mean age ± SD, 66.6 ± 10.6 y; mean Mini-Mental State Examination [MMSE] score ± SD, 22.2 ± 6.0) or frontotemporal lobar degeneration (FTLD) (n = 31; age ± SD, 63.9 ± 7.1 y, mean MMSE score ± SD, 23.8 ± 6.7). To identify the (11)C-PIB frames best representing perfusion, we ran on a test cohort an iterative algorithm, including generating normalized (cerebellar reference) perfusion pPIB images across variable frame ranges and calculating Pearson R values of the sum of these pPIB frames with the sum of all (18)F-FDG frames (cerebellar normalized) for all brain tissue voxels. Once this perfusion frame range was determined on the test cohort, it was then validated on an extended cohort and the power of pPIB in differential diagnosis was compared with (18)F-FDG by performing a logistic regression of regions-of-interest tracer measure (pPIB or (18)F-FDG) versus diagnosis. A 7-min window, corresponding to minutes 1-8 (frames 5-15), produced the highest voxelwise correlation between (18)F-FDG and pPIB (R = 0.78 ± 0.05). This pPIB frame range was further validated on the extended AD and FTLD cohort across 12 regions of interest (R = 0.91 ± 0.09). A logistic model using pPIB was able to classify 90.5% of the AD and 83.9% of the FTLD patients correctly. Using (18)F-FDG, we correctly classified 88.1% of AD and 83.9% of FTLD patients. The temporal pole and temporal neocortex were significant discriminators (P < 0.05) in both models, whereas in the model with pPIB the frontal region was also significant. The high correlation between pPIB and (18)F-FDG measures and their comparable performance in differential diagnosis are promising in providing functional information using (11)C-PIB PET data. This approach could be useful, obviating (18)F-FDG scans when longer-lived amyloid imaging agents become available.


Reduced ability to recover from spindle disruption and loss of kinetochore spindle assembly checkpoint proteins in oocytes from aged mice.
Currently, maternal aging in women, based on mouse models, is thought to raise oocyte aneuploidy rates, because chromosome cohesion deteriorates during prophase arrest, and Sgo2, a protector of centromeric cohesion, is lost. Here we show that the most common mouse strain, C57Bl6/J, is resistant to maternal aging, showing little increase in aneuploidy or Sgo2 loss. Instead it demonstrates significant kinetochore-associated loss in the spindle assembly checkpoint protein Mad2 and phosphorylated Aurora C, which is involved in microtubule-kinetochore error correction. Their loss affects the fidelity of bivalent segregation but only when spindle organization is impaired during oocyte maturation. These findings have an impact clinically regarding the handling of human oocytes ex vivo during assisted reproductive techniques and suggest there is a genetic basis to aneuploidy susceptibility.


Seating and Wheeled Mobility Clinicians Contribute to the Wound Care Team.
Many know that the wound care team consists of physicians and nurses with specialized training. Many may not know of physical therapists (PT) or occupational therapists (OT) with training in seating and wheeled mobility who address skin injuries in people who are full-time wheelchair users. PTs/OTs address the fit and use of their wheelchair to their body and look at their daily function while looking for causes of skin injury otherwise not seen and can help prevent them in the future. Therefore, this makes PTs and OTs with this expertise a valuable part of the wound care team.


A phase II study of 5-fluorouracil/L-leucovorin/oxaliplatin (mFOLFOX6) in Japanese patients with metastatic or unresectable small bowel adenocarcinoma.
Several studies have suggested that chemotherapy prolonged survival in patients with metastatic or recurrent small bowel adenocarcinoma (SBA); however, there is still no standard chemotherapy regimen. Here, we evaluated the efficacy and safety of a 5-fluorouracil (5-FU)/L-leucovorin (l-LV)/oxaliplatin (mFOLFOX6) protocol as a first-line therapy for patients with SBA. This was a multicenter, single-arm, open-label phase II study. Eligibility criteria included histologically confirmed adenocarcinoma, age 20-80 years, and an Eastern Cooperative Oncology Group performance status (PS) of 0-2. The primary endpoint was 1-year progression-free survival (PFS). The secondary endpoints included overall response rate (ORR), overall survival (OS), overall PFS, and safety. Between April 2010 and November 2012, 24 patients were enrolled from 12 institutions. The median age of the patients was 63 years (range 31-79) and there was a male/female ratio of 18/6. The number of PS 0/1 patients was 17/7 and locally advanced/metastatic disease was seen in 2/22 patients, respectively. The primary tumor site was the duodenum in 14 patients (58%) and jejunum in 10 patients (42%). The median follow-up time was 14.7 months (3.7-40.3). The 1-year PFS was 23.3%. The ORR was 9/20 (45%). The median PFS and OS times were 5.9 months (95% confidence interval [CI] 3.0-10.2) and 17.3 months (95% CI 11.7-19.0), respectively. Major grade 3/4 toxicities were neutropenia (38%), anemia/peripheral neuropathy (25%), and stenosis (17%). There were no treatment-related deaths. Although the primary endpoint was not met, mFOLFOX6 showed effective and good tolerance as a first-line treatment for SBA.


Cancer mortality among Brazilian dentists.
Previous studies have variably shown excess risks of elected cancers among dentists. National Brazilian mortality data were used to obtain mortality patterns among dentists between 1996 and 2004. Cancer mortality odds ratios (MORs) and cancer proportional mortality ratios for all cancer sites were calculated, using the general population and physicians and lawyers as comparison groups. Female dentists from both age strata showed higher risks for breast, colon-rectum, lung, brain, and non-Hodgkin lymphoma. Compared to physicians and lawyers, higher MOR estimates were observed for brain cancer among female dentists 20-49 yr. Among male dentists, higher cancer mortality was found for colon-rectum, pancreas, lung, melanoma, and non-Hodgkin lymphoma. Higher risk estimates for liver, prostate, bladder, brain, multiple myeloma and leukemia were observed among 50-79 yr old male dentists. If confirmed, these results indicate the need for limiting occupational exposures among dentists in addition to establishing screening programs to achieve early detection of selected malignant tumors.


Crystallization and preliminary crystallographic study of rBmKalphaIT1, a recombinant alpha-insect toxin from the scorpion Buthus martensii Karsch.
Alpha-insect scorpion toxins are a distinct group of scorpion neurotoxins for which no crystal structures are yet available. A novel alpha-insect toxin named BmKalphaIT1 from the scorpion Buthus martensii Karsch (BmK) has been expressed as an inclusion body in Escherichia coli and purified by chromatography after renaturation. Recombinant BmKalphaIT1 (rBmKaIT1) was crystallized using the vapour-diffusion technique in hanging drops at 296 K. The crystals, which were grown in sodium phosphate, belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 30.24 (1), b = 36.51 (3), c = 57.08 (2) A. Diffraction data were collected to 2.1 A resolution using synchrotron radiation. There appears to be one rBmKalphaIT1 molecule in the asymmetric unit.


[Characteristics of microsporogenesis in the Siberian larch growing under the conditions of technogenic load].
Studies of microsporogenesis in the Siberian larch growing in Krasnoyarsk and its suburbs have shown that meiosis starts in October. Microsporocytes winter at prophase I (leptoneme, diploneme). Reduction divisions in male generative buds are resumed and terminated in spring, in March. However, in the case of thaws during the autumn-winter period, meiotic division proceeds in the larch buds and this leads to the formation of degrading tetrads and pollen. Hence, the organic quiescence is absent in the larch in winter. It was shown that in the larch growing in the city, meiosis proceed more asynchronously than in the background tree stands. An increase of chromosomal aberrations during the reduction division was noted under the conditions of technogenic pollution.


A Bibliometric Study on Charcot Foot Deformity.
In this study, we investigated the literature's publication trends related to the Charcot foot deformity. Using bibliometric analysis to examine the data of origin, this analysis was carried out by conducting an electronic search of the Web of Science database for research articles between 1970 and March 2023. We used the following search term in the search bar: TI = (Charcot foot OR Charcot foot deformity OR Charcot's foot OR Charcot Osteopathic Arthropathy) with English language and article-format filtering for documents. The bibliometric analysis was carried out using R's "Bibliometrix" package program. A total of 437 articles were found in the electronic search. A total number of 1513 authors from around the world contributed to the Charcot foot literature, with the most articles published (42.1%) originating in the United States. The United States had the highest proportion of citations (3332 citations). The highest number of articles (n = 245) on Charcot foot deformity was in the last decade. 2021 was the year with the most articles (n = 34). The authors from the United States and the United Kingdom had the highest number of international collaborations. The study offers researchers a current overview of essential data and may help direct future research by summarizing the main points and research trends on the topic of Charcot foot deformity.


Side-effects associated with ketamine use in depression: a systematic review.
This is the first systematic review of the safety of ketamine in the treatment of depression after single and repeated doses. We searched MEDLINE, PubMed, PsycINFO, and Cochrane Databases and identified 288 articles, 60 of which met the inclusion criteria. After acute dosing, psychiatric, psychotomimetic, cardiovascular, neurological, and other side-effects were more frequently reported after ketamine treatment than after placebo in patients with depresssion. Our findings suggest a selective reporting bias with limited assessment of long-term use and safety and after repeated dosing, despite these being reported in other patient groups exposed to ketamine (eg, those with chronic pain) and in recreational users. We recommend large-scale clinical trials that include multiple doses of ketamine and long-term follow up to assess the safety of long-term regular use.


Physiological and molecular characterization of aristolochic acid transport by the kidney.
Consumption of herbal medicines derived from Aristolochia plants is associated with a progressive tubulointerstitial disease known as aristolochic acid (AA) nephropathy. The nephrotoxin produced naturally by these plants is AA-I, a nitrophenanthrene carboxylic acid that selectively targets the proximal tubule. This nephron segment is prone to toxic injury because of its role in secretory elimination of drugs and other xenobiotics. Here, we characterize the handling of AA-I by membrane transporters involved in renal organic anion transport. Uptake assays in heterologous expression systems identified murine organic anion transporters (mOat1, mOat2, and mOat3) as capable of mediating transport of AA-I. Kinetic analyses showed that all three transporters have an affinity for AA-I in the submicromolar range and thus are likely to operate at toxicologically relevant concentrations in vivo. Structure-activity relationships revealed that the carboxyl group is critical for high-affinity interaction of AA-I with mOat1, mOat2, and mOat3, whereas the nitro group is required only by mOat1. Furthermore, the 8-methoxy group, although essential for toxicity, was not requisite for transport. Mouse renal cortical slices avidly accumulated AA-I, achieving slice-to-medium concentration ratios >10. Uptake by slices was sensitive to known mOat1 and mOat3 substrates and the organic anion transport inhibitor probenecid, which also blocked the production of DNA adducts formed with reactive intracellular metabolites of AA-I. Taken together, these findings indicate that OAT family members mediate high-affinity transport of AA-I and may be involved in the site-selective toxicity and renal elimination of this nephrotoxin.


Accelerated thrombolysis in a rabbit model of carotid artery thrombosis with liposome-encapsulated and microencapsulated streptokinase.
The present study compares the efficacy of two formulations of encapsulated streptokinase to streptokinase in a rabbit model of carotid artery thrombosis. Arterial thrombosis followed the injection of thrombin mixed with autologous whole blood into a carotid artery of New Zealand white rabbits. Thirty minutes after the confirmation of an occlusive thrombus, one of four streptokinase formulations was infused at a dosage of 6000 IU/kg into the jugular vein. Free streptokinase (FREE SK) was compared to identical dosages of streptokinase encapsulated in a liposome (LESK), streptokinase entrapped in a water-soluble polymer (MESK), and free streptokinase admixed with blank microparticles (FREE SK + BLANK). Carotid arterial blood flow was determined by pulsed Doppler flowmetry to confirm clot formation and reperfusion. Two hours after drug infusion, the rabbits were killed and the residual thrombus mass was determined. Compared to FREE SK (74.5 +/- 16.9 min; mean +/- SEM), LESK demonstrated significantly reduced reperfusion times (19.3 +/- 4.6 min) while MESK exhibited even greater improvement (7.3 +/- 1.6 min). FREE SK + BLANK showed no statistical improvement versus FREE SK. LESK and MESK also resulted in reduced residual clot mass and greater return of arterial blood flow. These studies suggest that encapsulation of streptokinase offers a potential method of improved fibrinolytic treatment for patients with clot-based disorders. MESK performed slightly better than LESK with improved production and storage characteristics.


[Frankfurt Vaccination Intervention Study--Friva].
The success of the recommendation of vaccinations on the occasion of school-beginner examinations is assessed. The feedback rate of the intervention group amounts to 65%. The recommended vaccinations were carried out at 60% (measles-mumps-rubella) up to 90% (tetanus).


The cost-effectiveness of government actions to reduce sodium intake through salt substitutes in Vietnam.
Dietary sodium reduction is recommended to reduce the burden of cardiovascular disease. In Vietnam food products including salt, fish sauce and bot canh contribute to ~ 70% of dietary sodium intake. Reduced sodium versions of these products can be produced by replacing some of the sodium chloride with potassium chloride. We aimed to assess the cost-effectiveness of three alternative approaches to introducing reduced sodium products onto the market with a view to lowering population sodium intake in Vietnam. The three salt substitution strategies included voluntary, subsidised and regulatory approaches targeting salt, fish sauce and bot canh products. Costs were modelled using the WHO-CHOICE methodology. A Markov cohort model was developed to evaluate the cost-effectiveness of each strategy versus no intervention from the government perspective. The model linked each intervention strategy to assumed changes in levels of sodium intake and then to systolic blood pressure. Changes in SBP were linked to a probability of ischaemic heart disease or stroke. The model followed people over their lifetime to assess average costs and quality adjusted life years (QALYs) gained for each strategy. The voluntary salt substitution strategy was assumed to require no investment by government. Following ramp up (years 6+), the average annual costs for the subsidised and regulatory strategies were 21,808,968,902 ₫ (US$ 977,354) and 12,949,953,247 ₫ (US$ 580,410) respectively. Relative to no intervention, all three salt substitution strategies were found to be cost-effective. Cost savings were driven by reductions in strokes (32,595; 768,384; 2,366,480) and ischaemic heart disease (IHD) events (22,830; 537,157; 1,648,590) for the voluntary, subsidised & regulatory strategies, respectively. The voluntary strategy was least cost-effective (- 3445 ₫ US$ -0.15; 0.009 QALYs gained) followed by the subsidised strategy (- 43,189 ₫ US$ -1.86; 0.022 QALYs gained) and the regulatory strategy delivered the highest cost savings and health gains (- 243,530 ₫ US$ -10.49; 0.074 QALYs gained). This research shows that all three modelled salt substitution strategies would be good value for money relative to no intervention in Vietnam. The subsidised alternative would require the highest level of government investment; however the implementation costs will be exceeded by healthcare savings assuming a reasonable time horizon is considered.


Splice Site Variants in the KCNQ1 and SCN5A Genes: Transcript Analysis as a Tool in Supporting Pathogenicity.
Approximately 75% of clinically definite long QT syndrome (LQTS) cases are caused by mutations in the KCNQ1, KCNH2 and SCN5A genes. Of these mutations, a small proportion (3.2-9.2%) are predicted to affect splicing. These mutations present a particular challenge in ascribing pathogenicity. Here we report an analysis of the transcriptional consequences of two mutations, one in the KCNQ1 gene (c.781_782delinsTC) and one in the SCN5A gene (c.2437-5C>A), which are predicted to affect splicing. We isolated RNA from lymphocytes and used a directed PCR amplification strategy of cDNA to show mis-spliced transcripts in mutation-positive patients. The loss of an exon in each mis-spliced transcript had no deduced effect on the translational reading frame. The clinical phenotype corresponded closely with genotypic status in family members carrying the KCNQ1 splice variant, but not in family members with the SCN5A splice variant. These results are put in the context of a literature review, where only 20% of all splice variants reported in the KCNQ1, KCNH2 and SCN5A gene entries in the HGMDPro 2015.4 database have been evaluated using transcriptional assays. Prediction programmes play a strong role in most diagnostic laboratories in classifying variants located at splice sites; however, transcriptional analysis should be considered critical to confirm mis-splicing. Critically, this study shows that genuine mis- splicing may not always imply clinical significance, and genotype/phenotype cosegregation remains important even when mis-splicing is confirmed.


A Monolithic Gimbal Micro-Mirror Fabricated and Remotely Tuned with a Femtosecond Laser.
With the advent of ultrafast lasers, new manufacturing techniques have come into existence. In micromachining, the use of femtosecond lasers not only offers the possibility for three-dimensional monolithic fabrication inside a single optically transparent material, but also a means for remotely, and arbitrarily, deforming substrates with nanometer resolution. Exploiting this principle and combining it with flexure design, we demonstrate a monolithic micro-mirror entirely made with a femtosecond laser and whose orientation is tuned in a non-contact manner by exposing some part of the device to low energy femtosecond pulses. Given the non-contact nature of the process, the alignment can be very precisely controlled with a resolution that is many orders of magnitude better than conventional techniques based on mechanical positioners.


Long head of the biceps tendon pain: differential diagnosis and treatment.
Though the role of the long head of the biceps tendon (LHBT) in shoulder pathology has been extensively investigated, it remains controversial. Historically, there have been large shifts in opinions on LHBT function, ranging from being a vestigial structure to playing a critical role in shoulder stability. Today, despite incomplete understanding of its clinical or biomechanical involvement, most investigators would agree that LHBT pathology can be a significant cause of anterior shoulder pain. When the biceps tendon is determined to be a significant contributor to a patient's symptoms, the treatment options include various conservative interventions and possible surgical procedures, such as tenotomy, transfer, or tenodesis. The ultimate treatment decision is based upon a variety of factors, including the patient's overall medical condition, severity, and duration of symptoms, expectations, associated shoulder pathology, and surgeon preference. The purpose of this manuscript is to review current anatomic, functional, and clinical information regarding the LHBT, including conservative treatment, surgical treatment, and postsurgical rehabilitation regimens. Level 5.


Hypoxia, but not reoxygenation, induces interleukin 6 gene expression through NF-kappa B activation.
Interleukin (IL) 6 is one of major mediators of inflammation, and IL-6 gene activation during hypoxia/reoxygenation has been implicated in the pathogenesis of ischemia/reperfusion injury. However, molecular events involved in IL-6 gene expression during hypoxia/reoxygenation remain to be identified. We have previously shown that NF-kappa B plays an essential and indispensable role in the transcriptional activation of the IL-6 gene induced by various stimuli, including IL-1 and tumor necrosis factor-alpha. We show here that hypoxia, but not reoxygenation, induces the activation of NF-kappa B through the degradation of a major inhibitor of NF-kappa B, I kappa B alpha. This hypoxia-induced NF-kappa B activation resulted in the kappa B-dependent transcriptional activation of the IL-6 gene. Interestingly, the time course of hypoxia-induced NF-kappa B activation was rather slow as compared with those of NF-kappa B activation induced by other stimuli, such as IL-1: a significant NF-kappa B activation was not observed before 1 hr of hypoxia treatment and persisted for up to 7 hr of hypoxia treatment. However, hypoxia-induced NF-kappa B activation was not inhibited by cycloheximide, which indicates that hypoxia directly triggers NF-kappa B activation. Furthermore, while hypoxia is unlikely to generate reactive oxygen intermediates, pretreatment of cells with antioxidants such as N-acetyl cysteine and alpha-tocopherol inhibited NF-kappa B activation induced by hypoxia. Thus, we discuss possible implications of these results for a postulated role of reactive oxygen intermediates in NF-kappa B activation.


Risk of regional recurrence in triple-negative breast cancer patients: a Dutch cohort study.
Triple-negative breast cancer is associated with early recurrence and low survival rates. Several trials investigate the safety of a more conservative approach of axillary treatment in clinically T1-2N0 breast cancer. Triple-negative breast cancer comprises only 15 % of newly diagnosed breast cancers, which might result in insufficient power for representative results for this subgroup. We aimed to provide a nationwide overview on the occurrence of (regional) recurrences in triple-negative breast cancer patients with a clinically T1-2N0 status. For this cohort study, 2548 women diagnosed between 2005 and 2008 with clinically T1-2N0 triple-negative breast cancer were selected from the Netherlands Cancer Registry. Follow-up data until 2014 were analyzed using Kaplan-Meier. Sentinel lymph node biopsy was performed in 2486 patients, and (completion) axillary lymph node dissection in 562 patients. Final pathologic nodal status was pN0 in 78.5 %, pN1mi in 4.5 %, pN1 in 12.3 %, pN2-3 in 3.6 %, and pNx in 1.1 %. During a follow-up of 5 years, regional recurrence occurred in 2.9 %, local recurrence in 4.2 % and distant recurrence in 12.2 %. Five-year disease-free survival was 78.7 %, distant disease-free survival 80.5 %, and 5-year overall survival 82.3 %. Triple-negative clinically T1-2N0 breast cancer patients rarely develop a regional recurrence. Their disease-free survival is more threatened by distant recurrence, affecting their overall survival. Consequently, it seems justified to include triple-negative breast cancer patients in randomized controlled trials investigating the safety of minimizing axillary staging and treatment.


[Experimental study of the pathogenesis of dysentery].
It was shown that experimental dysentery could be produced against the background of various pathologic conditions of experimental animals (avitaminosis, concurrent diseases, etc). The studies in vivo and in vitro showed that intraepithelial parasitising of Shigella occurred in dysentery but did not play the leading role in the pathogenesis of the disease. Macrophages played a more important role, particularly in formation of the bacterial carrier state. In order to resolve some disputable problems of the pathogenesis of dysentery it is necessary to take into account the biological properties of the causative agent which change under the influence ov various effects, as well as the state of reponsiveness of the host.


Multidrug resistance in a human small cell lung cancer cell line selected in adriamycin.
A multidrug resistant variant (H69AR) of the human small cell lung cancer cell line NCI-H69 was obtained by culturing these cells in gradually increasing doses of Adriamycin up to 0.8 microM after a total of 14 months. H69AR expresses the multidrug resistant phenotype because it is cross-resistant to anthracycline analogues including daunomycin, epirubicin, menogaril, and mitoxantrone as well as to acivicin, etoposide, gramicidin D, colchicine, and the Vinca alkaloids, vincristine and vinblastine. H69AR is also similar to other multidrug resistant cell lines in that it displays little or no cross-resistance to bleomycin, 5-fluorouracil, and carboplatin. It has a slight collateral sensitivity to 1-dehydrotestosterone and lidocaine. H69AR has increased cell-cell adhesiveness compared to H69, but a similar growth rate in vitro and tumorigenicity in nude mice. When cultured in the absence of Adriamycin, there is a 40% decrease in resistance by 35 days of culture, compared to cells in continuous culture in drug, but no further decrease in resistance up to 181 days. Monoclonal antibodies to P-glycoprotein have no detectable reactivity with H69AR cells as determined by enzyme-linked immunosorbent assay and immunoblotting techniques. Thus, unlike most multidrug resistant cell lines, H69AR does not appear to express enhanced levels of P-glycoprotein. H69AR will provide a useful model for the study of multidrug resistance in human small cell lung cancer.


The effects of the 2004 tsunami on a coastal aquifer in Sri Lanka.
On December 26, 2004, the earthquake off the southern coast of Sumatra in the Indian Ocean generated far-reaching tsunami waves, resulting in severe disruption of the coastal aquifers in many countries of the region. The objective of this study was to examine the impact of the tsunami on groundwater in coastal areas. Field investigations on the east coast of Sri Lanka were carried out along a transect located perpendicular to the coastline on a 2.4 km wide sand stretch bounded by the sea and a lagoon. Measurements of groundwater table elevation and electrical conductivity (EC) of the groundwater were carried out monthly from October 2005 to August 2007. The aquifer system and tsunami saltwater intrusion were modeled using the variable-density flow and solute transport code HST3D to understand the tsunami plume behavior and estimate the aquifer recovery time. EC values reduced as a result of the monsoonal rainfall following the tsunami with a decline in reduction rate during the dry season. The upper part of the saturated zone (down to 2.5 m) returned to freshwater conditions (EC < 1000 µS/cm) 1 to 1.5 years after the tsunami, according to field observations. On the basis of model simulations, it may take more than 15 years for the entire aquifer (down to 28 m) to recover completely, although the top 6 m of the aquifer may become fresh in about 5 years.


Peptic ulcer and gastric acid secretion in patients with insulinoma.
We evaluated the incidence of associated peptic ulcers and the gastric acid secretion in our 20 series of insulinoma, and obtained the following results: 1) Four out of 20 patients (20%) with insulinoma were associated with peptic ulcers. 2) Except for the maximum acid concentration, the preoperative gastric secretion tests, such as the basal acid output and maximum acid output, failed to indicate hyperacidity in those patients. And patients with a short duration of illness by insulinoma neither showed hyperacidity nor were associated with peptic ulcers. 3) A tendency of reduced gastric acid secretion and the amelioration of peptic ulcers was observed in those who underwent the extirpation of insulinoma. 4) In the most recent case received gastric analysis with continuous monitoring of blood glucose level, an increase of basal acid output was observed during the hypoglycemic period in which the blood glucose levels reduced to around 30 mg/100 ml. These results indicate that the low blood glucose level and the high gastric acid secretion produced under the condition on an empty stomach, may be important factors related to the high association rates of the peptic ulcers in patients with insulinoma.


An assessment of maturity from anthropometric measurements.
The range of variability between individuals of the same chronological age (CA) in somatic and biological maturity is large and especially accentuated around the adolescent growth spurt. Maturity assessment is an important consideration when dealing with adolescents, from both a research perspective and youth sports stratification. A noninvasive, practical method predicting years from peak height velocity (a maturity offset value) by using anthropometric variables is developed in one sample and cross-validated in two different samples. Gender specific multiple regression equations were calculated on a sample of 152 Canadian children aged 8-16 yr (79 boys; 73 girls) who were followed through adolescence from 1991 to 1997. The equations included three somatic dimensions (height, sitting height, and leg length), CA, and their interactions. The equations were cross-validated on a combined sample of Canadian (71 boys, 40 girls measured from 1964 through 1973) and Flemish children (50 boys, 48 girls measured from 1985 through 1999). The coefficient of determination (R2) for the boys' model was 0.92 and for the girls' model 0.91; the SEEs were 0.49 and 0.50, respectively. Mean difference between actual and predicted maturity offset for the verification samples was 0.24 (SD 0.65) yr in boys and 0.001 (SD 0.68) yr in girls. Although the cross-validation meets statistical standards for acceptance, caution is warranted with regard to implementation. It is recommended that maturity offset be considered as a categorical rather than a continuous assessment. Nevertheless, the equations presented are a reliable, noninvasive and a practical solution for the measure of biological maturity for matching adolescent athletes


Evidence-Based Consensus Recommendations for Colposcopy Practice for Cervical Cancer Prevention in the United States.
The American Society for Colposcopy and Cervical Pathology (ASCCP) Colposcopy Standards recommendations address the role of colposcopy and directed biopsy for cervical cancer prevention in the United States (US). The recommendations were developed by an expert working group appointed by ASCCP's Board of Directors. An extensive literature review was conducted and supplemented by a systematic review and meta-analysis of unpublished data. In addition, a survey of practicing colposcopists was conducted to assess current colposcopy practice in the US. Recommendations were approved by the working group members, and the final revisions were made based on comments received from the public. The recommendations cover terminology, risk-based colposcopy, colposcopy procedures, and colposcopy adjuncts. The ASCCP Colposcopy Standards recommendations are an important step toward raising the standard of colposcopy services delivered to women in the US. Because cervical cancer screening programs are currently undergoing important changes that may affect colposcopy performance, updates to some of the current recommendations may be necessary in the future.


PROFILES AND PREDICTORS OF SUBJECTIVE WELL-BEING AND HEALTH-SEEKING BEHAVIOUR AMONG OLDER PERSONS WITH MULTIMORBIDITY IN A RURAL PRIMARY CARE ENVIRONMENT IN SOUTH-EASTERN NIGERIA.
Subjective well-being(SWB) and health-seeking behaviour(HSB) are important medical channels to gauge the health status of older persons with multimorbidity particularly in resource-poor environment with limited socio-medical and health infrastructure. The aim of the study was to determine the profiles and predictors of SWB and HSB among older persons with multimorbidity in a rural primary care environment in South-Eastern Nigeria. A cross-sectional study was conducted on 214 older persons with multimorbidity from January to December 2022 in South-Eastern Nigeria. Data collection was done using structured, pretested and interviewer guided questionnaire. Subjective well-being was determine using single-item self-rated health status questionnaire. Multimorbidity was determine using Multimorbidity Assessment Questionnaire for Primary Care(MAQ-PC). The ages of study participants were 60-99 years(x=68±4.5 years). There were 123(57.5%) females. Ninety-three (43.5%) of the study participants had good SWB while 86(40.2%) had appropriate HSB. The most common multimorbidities were aggregate disorders of cardiovascular, musculoskeletal and gastrointestinal systems. The independent predictors of poor SWB were advanced old age(ORs=3.101; 95%CI(1.526-6.019); P=.024), low education(ORs=4.056; 95%CI(2.512-8.509); P=.025) and non-household family structure(ORs=2.225; 95%CI(1.078-6.192); P= .001). Inappropriate HSB were predicted by males(ORs=2.428; 95%CI(1.107-6.379); P=.039), low education(ORs=3.007; 95%CI(1.387-7.001); P=.013) and retirees with no secondary occupation (ORs=4.761;95%CI(1.351- 8.072); P= .001). The study has shown the SWB and HSB among the study participants. The independent predictors of poor SWB were advanced old age, low education and non-household family structure. Inappropriate HSB werepredicted by males, low education and retirees with no secondary occupation. Subjective well-being and HSB should be explored during clinical consultations with older persons with multimorbidity particularly in the sub-region with double whammy of non-communicable and communicable diseases.


[Attention shifting and dissociated vertical squint].
Some patients with dissociated vertical deviation show an increasing upward drift of their squinting eye when reading small numbers at a distance of 5 m, making a special effort to shift attention to the non-squinting eye. This finding is at variance with the view that the manifestation of dissociated vertical deviation may be induced by visual inattention. The phenomenon could be explained by analogy to the occlusion test, by a binocular imbalance in visual information processing. The phenomenon may be found frequently in patients who require surgery for an especially disfiguring intermittent vertical squint. Looking for the phenomenon may be useful when deciding whether to operate for a dissociated vertical deviation: If increasing vertical deviation is observed during binocular acuity testing, surgical treatment should be considered.


Morphological substrates of cognitive decline in nonagenarians and centenarians: a new paradigm?
Brain aging is characterized by the formation of neurofibrillary tangles (NFT) and senile plaques (SP) in both cognitively intact individuals and patients with Alzheimer's disease (AD). The ubiquitous presence of these lesions and the steady increase of the prevalence of dementia up to 85 years have strongly supported a continuum between normal brain aging and AD. In this context, the study of nonagenarians and centenarians could provide key informations about the characteristics of extreme aging. We provide here a detailed review of currently available neuropathological data in very old individuals and critically discuss the patterns of NFT, SP and neuronal loss distribution as a function of age. In younger cohorts, NFTs are usually restricted to hippocampal formation, whereas clinical signs of dementia appear when temporal neocortex is involved. SPs would not be a specific marker of cognitive impairment as no correlation was found between their quantitative distribution and AD severity. The low rate of AD lesions even in severe AD as well as the weakness of clinicopathological correlations reported in the oldest-old indicate that AD pathology is not a mandatory phenomenon of increasing chronological age. Our recent stereological observations of hippocampal microvasculature in oldest-old cases challenge the traditional lesional model by revealing that mean capillary diameters is an important structural determinant of cognition in this age group.


Homeopathy and the new fundamentalism: a critique of the critics.
Though in use for over 200 years, and still benefiting millions of people worldwide today, homeopathy is currently under continuous attacks for being "unscientific." The reasons for this can be understood in terms of what might be called a "New Fundamentalism," emanating particularly but not exclusively from within biomedicine, and supported in some sections of the media. Possible reasons for this are discussed. New Fundamentalism's hallmarks include the denial of evidence for the efficacy of any therapeutic modality that cannot be consistently "proven" using double-blind, randomized controlled trials. It excludes explanations of homeopathy's efficacy; ignores, excoriates, or considers current research data supporting those explanations incomprehensible, particularly from outside biomedicine: it is also not averse to using experimental bias, hearsay, and innuendo in order to discredit homeopathy. Thus, New Fundamentalism is itself unscientific. This may have consequences in the future for how practitioners, researchers, and patients of homeopathy/complementary and alternative medicine engage and negotiate with primary health care systems.


Nerve grafting. Functional results after primary vs delayed repair.
The finding of heightened metabolic activity in nerve cell bodies after axonal injury has led to speculation that delaying motor nerve repair would be beneficial. Using a sciatic nerve model, 100 rats were grouped based on nerve autograft donor and recipient site variables. Animals were subjected to electrophysiologic testing at 3, 6, 9, and 12 weeks after grafting. Twitch force and nerve compound action potential parameters were calculated and compared using two-way analysis of variance. Previous studies have suggested interaction between donor and recipient variables. We cannot support these findings. Our data revealed no significant difference between immediate or delayed nerve repair or between fresh or predegenerated nerve grafts.


The lipid raft microdomain-associated protein reggie-1/flotillin-2 is expressed in human B cells and localized at the plasma membrane and centrosome in PBMCs.
Reggie-1/flotillin-2 is a plasma membrane-associated cytoplasmic protein, which defines non-caveolar raft microdomains. Reggie-1/flotillin-2 is enriched in detergent insoluble (TX100) membrane fractions (DIG), co-localizes with activated GPI-linked proteins and the fyn-kinase in neurons and T cells, and thus apparently participates in the assembly of protein complexes essential for signal transduction. In T cells activated by crosslinking the GPI-linked protein Thy-1 or by crosslinking the ganglioside GM1, reggie-1/flotillin-2 co-localizes with the T cell receptor. To determine whether reggie-1/flotillin-2 is also expressed in B cells, primary B cells from human blood and cell lines representing the developmental stages of pro, pre, mature and plasma B cells were analyzed by Western blotting, RT-PCR and immunofluorescence. Here, we show that reggie-1/flotillin-2 is expressed throughout B cell development, as well as in primary B cells, purified by cell sorting. On non-activated mature B cell Raji cell line we found reggie-1/flotillin-2 are exclusively in the detergent (TX100) insoluble membrane fractions that are staining positive for the raft marker GM1. Immunofluorescence microscopy showed that reggie-1/flotillin-2 is localized at the plasma membrane and marks intracellular spots in PBMCs. Confocal co-localization studies showed that reggie-1/flotillin-2 is associated with the plasma membrane, and the centrosomes (microtubule organizing centers) in these PBMCs. Comparison of reggie-1/flotillin-2 cDNA sequences with the genomic sequence database allowed us to determine the exon/intron structures in mouse and human. The gene organizations are highly conserved suggesting an important function of reggie-1/flotillin-2. Since reggie/flotillin proteins co-cluster with the T cell receptor and fyn kinases upon T cell stimulation, our findings of reggie-1/flotillin-2 in B cells suggest a similar role in B cell function.


Mortality during the two years after coronary artery bypass grafting in relation to perioperative factors and urgency of operation.
The purpose of this study was to describe mortality during the 2-year-period after coronary artery bypass surgery (CABG) in relation to perioperative risk factors and urgency of operation. All the patients in western Sweden were included in whom CABG was performed between June 1988 and June 1991, without concomitant procedures or re-operations. The study was prospective in design. In all, 2000 patients were operated upon and 186 (9.3%) of the operations were acute. There was a significant relationship between the urgency of the operation and mortality. Early mortality was 2.4% in elective operations and 5.4-62.5% in urgent to emergency operations. The 30-day to 2-year mortality was 4.2%. The perioperative risk indicators independently associated with early mortality were neurologic complications, serum-aspartate aminotransferase (S-ASAT) more than 2.0 microkat/l, urgency of operation, the use of circulatory assist devices, re-operation and ventilator time more than 24 h. The risk indicators for mortality after 30 days were pneumothorax, longer intensive care unit (ICU) time, the use of inotropic drugs and neurologic complications. In conclusion, the multivariate analysis reveals the urgency of the operation as a predictor of early mortality after CABG, but no significant association with mortality was found after 30 days. When excluding death within 30 days, three additional independent predictors of mortality were identified.


Biomimetic Soft Polymer Microstructures and Piezoresistive Graphene MEMS Sensors Using Sacrificial Metal 3D Printing.
Recent advances in 3D printing technology have enabled unprecedented design freedom across an ever-expanding portfolio of materials. However, direct 3D printing of soft polymeric materials such as polydimethylsiloxane (PDMS) is challenging, especially for structural complexities such as high-aspect ratio (>20) structures, 3D microfluidic channels (∼150 μm diameter), and biomimetic microstructures. This work presents a novel processing method entailing 3D printing of a thin-walled sacrificial metallic mold, soft polymer casting, and acidic etching of the mold. The proposed workflow enables the facile fabrication of various complex, bioinspired PDMS structures (e.g., 3D double helical microfluidic channels embedded inside high-aspect ratio pillars) that are difficult or impossible to fabricate using currently available techniques. The microfluidic channels are further infused with conductive graphene nanoplatelet ink to realize two flexible piezoresistive microelectromechanical (MEMS) sensors (a bioinspired flow/tactile sensor and a dome-like force sensor) with embedded sensing elements. The MEMS force sensor is integrated into a Philips 9000 series electric shaver to demonstrate its application in "smart" consumer products in the future. Aided by current trends in industrialization and miniaturization in metal 3D printing, the proposed workflow shows promise as a low-temperature, scalable, and cleanroom-free technique of fabricating complex, soft polymeric, biomimetic structures, and embedded MEMS sensors.


[Prevention of tumor implantation during surgery of cervical and endometrial carcinoma].
The careful cleansing of the vagina with physiologic saline and 96% ethyl alcohol, the insert of a big, dry swab for two minutes in front of the portio vaginalis cervicis and the cleansing once more again with a dry swab are an effective prophylaxis against tumor implantation from a carcinoma of the endometrium or of the cervix. This could be confirmed not only by cytology prior to operation and by cytology antecedent to cleansing the vagina immediately prior to the removal of the uterus from the vagina but also by the follow-up study of 517 surgically treated cancers of the endometrium and 241 cervical cancers.


Orbital fibrous histiocytoma: case report and literature review.
Fibrous histiocytomas are a diverse group of soft tissue tumors classified histiologically as benign, locally aggressive, and malignant. These tumors are found throughout the body, but seem to have an affinity for the periorbital area. They account for one percent of all ocular masses and are the most common primary mesenchymal tumor of the orbit. Associated ocular signs and symptoms include decreased visual acuity, proptosis, diplopia, pain, restricted extraocular muscle movement, swelling of the eyelids, and conjunctiva, as well as disk edema. A case of a benign orbital fibrous histiocytoma is presented. The patient reported intermittent pain and occasional diplopia; severe edema of the right upper eyelid; and proptosis and inferior vertical displacement of the right globe were observed. Computed tomography revealed a well-defined mass that was subsequently surgically removed and histopathological results from the Armed-Forces Pathology Institute confirmed the diagnosis. Orbital fibrous histiocytomas are rare periocular tumors that can manifest multiple ocular signs and symptoms. Careful histologic examination is necessary for diagnosis, since these tumors have a wide range of morphology. Differential diagnoses include orbital masses with similar radiologic or histologic findings.


Integration of newly learned L2 words into the mental lexicon is modulated by vocabulary learning method.
The aim of the study was to investigate both L2 word integration and the effect of learning method on it. For this purpose, an L2 word-learning paradigm was designed with two learning methods: L2 words were paired with videos in the first one and their translation-equivalent L1 words in the second. To test L2 word integration, a lexical decision task associated with form priming was administered before and after the learning phase. The L2 words to be learned were used as primes. Forty-eight participants participated in the study. Before learning, a facilitation effect was obtained with pseudowords (not already learned L2 words) as primes and L1 words as targets. After learning, L2 words no longer facilitated L1 word recognition when learned with the video method, while they still had this effect when learned with the L1 words - L2 words method. In accordance with the prime lexicality effect (PLE), this absence of a facilitation effect indicates that L1 words and L2 words are involved in a lexical competition process common to the two languages. This result highlights swift lexicalisation and demonstrates the effect of learning method in lexicalisation.


Markers of intestinal inflammation, not bacterial burden, correlate with clinical outcomes in Clostridium difficile infection.
Clostridium difficile is a leading hospital-acquired infection. Many patients remain symptomatic for several days on appropriate antibiotic therapy. To assess the contribution of ongoing infection vs persistent inflammation, we examined the correlation between fecal cytokine levels, fecal C. difficile burden, and disease outcomes in C. difficile infection (CDI). We conducted a prospective cohort study in Barnes Jewish Hospital between June 2011 and May 2012 of hospitalized adults with CDI. We determined fecal interleukin 8 (IL-8) and lactoferrin protein concentrations by enzyme immunoassay. We used real-time polymerase chain reaction (PCR) to measure relative fecal IL-8 and CXCL-5 RNA transcript abundances, and quantitative PCR to enumerate C. difficile burden. Of 120 study subjects, 101 (84%) were started on metronidazole, and 33 of those (33%) were subsequently given vancomycin. Sixty-two (52%) patients had diarrhea persistent for 5 or more days after starting CDI therapy. Initial fecal CXCL-5 messenger RNA (mRNA), IL-8 mRNA, and IL-8 protein correlated with persistent diarrhea and use of vancomycin. Time to diarrhea resolution was longer in patients with elevated fecal cytokines at diagnosis. Fecal cytokines were more sensitive than clinical severity scores in identifying patients at risk of treatment failure. Clostridium difficile burden did not correlate with any measure of illness or outcome at any point, and decreased equally with metronidazole and vancomycin. Persistent diarrhea in CDI correlates with intestinal inflammation and not fecal pathogen burden. These findings suggest that modulation of host response, rather than adjustments to antimicrobial regimens, might be a more effective approach to patients with unremitting disease.


Hypoglutamatergic activity in the STOP knockout mouse: a potential model for chronic untreated schizophrenia.
In mice, the deletion of the STOP protein leads to hyperdopaminergia and major behavioral disorders that are alleviated by neuroleptics, representing a potential model of schizophrenia. The reduction of the glutamatergic synaptic vesicle pool in the hippocampus could reflect a disturbance in glutamatergic neurotransmission in this model. Here we examined potential disturbances in energy metabolism and interactions between neurons and glia in 15-week-old STOP KO, wild-type, and heterozygous mice. Animals received [1-(13)C]glucose and [1,2-(13)C]acetate, the preferential substrates of neurons and astrocytes, respectively. Extracts from the whole forebrain and midbrain were analyzed by HPLC, (13)C and (1)H NMR spectroscopy. Amounts and labeling of most metabolites were unchanged. However, glutamine concentration and amount of [4,5-(13)C]glutamine derived from [1,2-(13)C]acetate significantly decreased by 17% and 18%, respectively, in STOP KO compared with wild-type mice. The amount of [4-(13)C]glutamate was decreased in STOP KO and heterozygous compared with wild-type mice. gamma-Aminobutyric acid labeling was not influenced by the genotype. Because STOP-deficient mice have a lower synaptic vesicle density, less glutamate is released to the synaptic cleft, leading to decreased stimulation of the postsynaptic glutamate receptors, reflecting increased glutamine metabolism only in the vicinity of the postsynapse of STOP KO mice.


Liver biopsy for assessment of suspected drug-induced liver injury in metabolic dysfunction-associated steatohepatitis clinical trials: Expert consensus from the Liver Forum.
Causality assessment of suspected drug-induced liver injury (DILI) during metabolic dysfunction-associated steatohepatitis (MASH) clinical trials can be challenging, and liver biopsies are not routinely performed as part of this evaluation. While the field is moving away from liver biopsy as a diagnostic and prognostic tool, information not identified by non-invasive testing may be provided on histology. To address the appropriate utilisation of liver biopsy as part of DILI causality assessment in this setting. From 2020 to 2022, the Liver Forum convened a series of webinars on issues pertaining to liver biopsy during MASH trials. The Histology Working Group was formed to generate a series of consensus documents addressing these challenges. This manuscript focuses on liver biopsy as part of DILI causality assessment. Expert opinion, guidance and recommendations on the role of liver biopsy as part of causality assessment of suspected DILI occurring during clinical trials for a drug(s) being developed for MASH are provided. Lessons learned from prior MASH programs are reviewed and gaps identified. Although there are no pathognomonic features, histologic evaluation of suspected DILI during MASH clinical trials may alter patient management, define the pattern and severity of injury, detect findings that favour a diagnosis of DILI versus MASH progression, identify prognostic features, characterise the clinicopathological phenotype of DILI, and/or define lesions that influence decisions about trial discontinuation and further development of the drug.


Alcohol-related cognitive mechanisms underlying adolescent alcohol use and alcohol problems: Outcome expectancy, self-schema, and self-efficacy.
The objective of this study was to empirically test a theoretical model to determine the cognitive mechanisms that are associated with adolescent alcohol use and alcohol problems. We posited that alcohol outcome expectancies would affect alcohol-refusal self-efficacy through the drinker self-schema. We also posited that alcohol outcome expectancies and the drinker self-schema would affect alcohol use and problems through alcohol-refusal self-efficacy. A survey was administered to 225 adolescents in a public junior high school in Taiwan at two-time points, six months apart. Path analysis was used to determine the mechanisms underlying the alcohol-related cognitive constructs on the alcohol use and alcohol problems separately, controlling for appropriate alcohol-related personal and environmental factors. Indirect effects were estimated using the bootstrapping method. Higher positive alcohol outcome expectancies and lower negative alcohol outcome expectancies predicted higher drinker self-schema scores. Higher positive alcohol outcome expectancies and drinker self-schema scores predicted lower alcohol-refusal self-efficacy. Lower alcohol-refusal self-efficacy was associated with a history of drinking and alcohol problems in the past six months. Effects of alcohol outcome expectancies on alcohol use and alcohol problems were partially mediated through the drinker self-schema and alcohol-refusal self-efficacy. Findings support the proposed theoretical cognitive mechanisms underlying alcohol use and alcohol problems in a sample of Taiwanese adolescents. Given that alcohol-related cognitive constructs are modifiable, the findings also provide a foundation to suggest that interventions to reduce positive alcohol outcome expectations and prevent the formation of a drinker self-schema may facilitate alcohol-refusal self-efficacy and mitigate drinking behaviors in this adolescent population.


Gastric by-pass surgery in morbidly obese patients markedly decreases serum levels of vitamins A and C and iron in the peri-operative period.
We studied changes in serum levels of carotene, vitamins A, C, B12 and folate and iron in the immediate perioperative period after gastric by-pass surgery in nine morbidly obese patients and in six patients undergoing abdominal surgery. All parameters were measured preoperatively and 48 h postoperatively. Marked decreases occurred for vitamin A (2.04 +/- 0.45 mumol/l to 0.92 +/- 0.39 mumol/l, P less than 0.01), vitamin C (26 +/- 11.2 mumol/l to 4.9 +/- 4.0 mumol/l, P less than 0.01), iron (15 +/- 5.8 mumol/l to 4 +/- 2.7 mumol/l, P less than 0.01). A significant change also occurred for carotene (1.7 +/- 0.42 mumol/l to 1.25 +/- 0.48 mumol/l, P less than 0.05). No significant changes were seen for vitamins B12 and folate. Only changes in vitamins A and C were significantly greater in the morbidly obese patients compared to the control group of six patients. These changes likely represent redistribution of vitamins rather than enhanced urinary excretion.


A revised phylogeny of Antilopini (Bovidae, Artiodactyla) using combined mitochondrial and nuclear genes.
Antilopini (gazelles and their allies) are one of the most diverse but phylogenetically controversial groups of bovids. Here we provide a molecular phylogeny of this poorly understood taxon using combined analyses of mitochondrial (CYTB, COIII, 12S, 16S) and nuclear (KCAS, SPTBN1, PRKCI, MC1R, THYR) genes. We explore the influence of data partitioning and different analytical methods, including Bayesian inference, maximum likelihood and maximum parsimony, on the inferred relationships within Antilopini. We achieve increased resolution and support compared to previous analyses especially in the two most problematic parts of their tree. First, taxa commonly referred to as "gazelles" are recovered as paraphyletic, as the genus Gazella appears more closely related to the Indian blackbuck (Antilope cervicapra) than to the other two gazelle genera (Nanger and Eudorcas). Second, we recovered a strongly supported sister relationship between one of the dwarf antelopes (Ourebia) and the Antilopini subgroup Antilopina (Saiga, Gerenuk, Springbok, Blackbuck and gazelles). The assessment of the influence of taxon sampling, outgroup rooting, and data partitioning in Bayesian analyses helps explain the contradictory results of previous studies.


Herpesvirus type 1 serum antibodies and brain tumors in humans.
Sixty-five patients underwent craniotomy for brain tumors; of these 35 had glioblastoma multiforme (GM). The GM cases, as a group, showed significantly higher serum titers for herpesvirus type 1 (HSV-1) neutralizing antibodies (NT) than the non-glioblastoma cases. Both the GM group and the pituitary adenoma group had high levels of HSV-1 serum antibodies with the enzyme-linked immunosorbent assay (ELISA). These data, combined with other evidence, lead us to speculate that HSV-1 infections may be associated with certain brain tumors in humans. However, coincidental causes for these elevated HSV-1 titers must be ruled out.


VEGF-mediated angiogenesis and vascularization of a fumarate-crosslinked polycaprolactone (PCLF) scaffold.
Revascularization of natural and synthetic scaffolds is a critical part of the scaffold's incorporation and tissue ingrowth. Our goals were to create a biocompatible polymer scaffold with 3D-printing technology, capable of sustaining vascularization and tissue ingrowth. We synthesized biodegradable polycaprolactone fumarate (PCLF) scaffolds to allow tissue ingrowth via large interconnected pores. The scaffolds were prepared with Poly(lactic-co-glycolic acid)(PLGA) microspheres seeded with or without different growth factors including VEGF,FGF-2, and/or BMP-2. Scaffolds were implanted into the subcutaneous tissues of rats before undergoing histologic and microCT angiographic analysis. At harvest after 12 weeks, scaffolds had tissue infiltrating into their pores without signs of scar tissue formation, fibrous capsule formation, or immune responses against PCLF. Histology for M1/M2 macrophage phenotypes confirmed that there were no overt signs of immune responses. Both microCT angiography and histologic analysis demonstrated marked tissue and vessel ingrowth throughout the pores traversing the body of the scaffolds. Scaffolds seeded with microspheres containing VEGF or VEGF with either BMP-2 or FGF-2 had significantly higher vascular ingrowth and vessel penetration than controls. All VEGF-augmented scaffolds were positive for Factor-VIII and exhibited collagen tissue infiltration throughout the pores. Furthermore, scaffolds with VEGF and BMP-2 had high levels of mineral deposition throughout the scaffold that are attributable to BMP-2. PCLF polymer scaffold can be utilized as a framework for vascular ingrowth and regeneration of multiple types of tissues. This novel scaffold material has promise in tissue regeneration across all types of tissues from soft tissue to bone.


Estimation of genetic risk for Type 1 diabetes mellitus in newborns on dried blood spot.
The main contribution to genetic susceptibility for Type 1 Diabetes Mellitus (T1DM) is conferred by the Human Leukocyte Antigens (HLA). We evaluated the feasibility of large scale screening on Dried Blood Spot (DBS) to estimate the genetic risk for T1DM in newborns. Peripheral blood DBS samples from 256 newborns, were genotyped for HLA DRB1 and DQB1 alleles identification by a commercially available assay based on a dissociation enhancer lanthanide fluorescence system available in many newborn screening laboratories. Results were compared with those obtained in two wide multicentric studies on cord blood (DIABFIN and PREVEFIN). Genotyping on DBS revealed 6 subjects at high risk for T1DM, 99 at moderate risk for T1DM and the remaining at low risk for T1DM. We found 100% concordance between both techniques for HLA-DQB1 and DRB1 determination, confirming the feasibility of large scale screening on DBS. DBSs represent a resource for future studies about new genetics markers. This assay for estimate the genetic risk of T1DM on DBS showed an excellent sensitivity, specificity and accuracy compared with conventional techniques. Moreover, this assay resulted less expensive, and it could be easily performed on material already collected for newborn screening programs.