Divergent Evolution of the Transcriptional Network Controlled by Snf1-Interacting Protein Sip4 in Budding Yeasts.
Cellular responses to starvation are of ancient origin since nutrient limitation has always been a common challenge to the stability of living systems. Hence, signaling molecules involved in sensing or transducing information about limiting metabolites are highly conserved, whereas transcription factors and the genes they regulate have diverged. In eukaryotes the AMP-activated protein kinase (AMPK) functions as a central regulator of cellular energy homeostasis. The yeast AMPK ortholog SNF1 controls the transcriptional network that counteracts carbon starvation conditions by regulating a set of transcription factors. Among those Cat8 and Sip4 have overlapping DNA-binding specificity for so-called carbon source responsive elements and induce target genes upon SNF1 activation. To analyze the evolution of the Cat8-Sip4 controlled transcriptional network we have compared the response to carbon limitation of Saccharomyces cerevisiae to that of Kluyveromyces lactis. In high glucose, S. cerevisiae displays tumor cell-like aerobic fermentation and repression of respiration (Crabtree-positive) while K. lactis has a respiratory-fermentative life-style, respiration being regulated by oxygen availability (Crabtree-negative), which is typical for many yeasts and for differentiated higher cells. We demonstrate divergent evolution of the Cat8-Sip4 network and present evidence that a role of Sip4 in controlling anabolic metabolism has been lost in the Saccharomyces lineage. We find that in K. lactis, but not in S. cerevisiae, the Sip4 protein plays an essential role in C2 carbon assimilation including induction of the glyoxylate cycle and the carnitine shuttle genes. Induction of KlSIP4 gene expression by KlCat8 is essential under these growth conditions and a primary function of KlCat8. Both KlCat8 and KlSip4 are involved in the regulation of lactose metabolism in K. lactis. In chromatin-immunoprecipitation experiments we demonstrate binding of both, KlSip4 and KlCat8, to selected CSREs and provide evidence that KlSip4 counteracts KlCat8-mediated transcription activation by competing for binding to some but not all CSREs. The finding that the hierarchical relationship of these transcription factors differs between K. lactis and S. cerevisiae and that the sets of target genes have diverged contributes to explaining the phenotypic differences in metabolic life-style.


Failure of human cervical endplates: a cadaveric experimental model.
An in vitro biomechanical study using a servohydraulic testing machine on cadaveric endplates. To characterize the effects of bone mineral density, endplate geometry, and preparation technique on endplate failure load. The effects of endplate preparation methods on failure loads are only partly characterized in the literature. Endplate burring has been recommended to increase fusion rates. However, graft subsidence may complicate anterior reconstruction procedures. After radiographic screening, 21 cadaveric cervical spines underwent dual-energy x-ray absorptiometry scanning to quantify mineral content. Endplate geometry was calculated in 55 randomly selected endplates from the inferior C2 to the superior T1 levels. These vertebrae were embedded in polyester resin and randomly left intact, perforated, or burred. The cervical endplates were loaded at a rate of 0.2 mm/s on an Instron materials tester with an attached 9 mm diameter polycarbonate rod (an area of 64 mm2). A stepwise, univariate linear regression was used to compare the point of endplate failure with the vertebral level, endplate area, gender, age, bone mineral density, and preparation technique. Mean bone mineral density, as measured by dual-energy x-ray absorptiometry, was 0.713 g/cm2 (+/- 0.173 g/cm2). Mean endplate area was calculated at 323 mm2. A mean compressive force of 754 N (+/- 445 N) was required before endplate failure. Trends toward increasing compressive loads were noted with decreasing endplate area and increasing bone mineral density. Increasing age (P = 0.0203), caudal vertebral level (P < 0.0001), endplate burring (P = 0.0068), and female gender (P = 0.0452) were associated with significantly lower endplate fracture loads in compression. Bone quality was predictive of endplate compressive failure loads. Intact endplates failed at significantly higher loads than their perforated or burred counterparts.


Hermeneutic Constructivism: One ontology for authentic understanding.
Nursing and nurses rely upon qualitative research to understand the intricacies of the human condition. Acknowledging the subjective nature of reality and commonly founded in a constructivist epistemology, qualitative approaches offer opportunities for uncovering insights from the perspective of the individual participants, the insider's view, and the construction of representations that maintain an intimacy with the subject's realities. Debate continues, however, about what is needed for a qualitative construction to be considered an authentic understanding of a subject's realities. Authenticity in the context of qualitative research has been described as entailing consideration of a number of well-trodden dimensions: fairness, ontological, educative, catalytic and tactical. Taking these dimensional requirements as key, this paper argues that authenticity may not always be as well-developed through some of the standard practices in qualitative research as perhaps expected. In particular, qualitative understandings of authenticity stress that participants should not be merely reported on but instead should be dynamically involved in and changed by the constructions and interpretations of data developed throughout the research process. As this paper illustrates, such engagements appear problematic for qualitative research approaches that are beholden to designative commitments in the context of language and meaning-making and which tend to prioritise commonality and generality at the expense of individual authenticity. An alternative qualitative approach, Hermeneutic Constructivism, is proposed as better able to achieve the requirements of the dimensions of authenticity. As outlined, this approach is well-placed to present an understanding of human experience through a genuinely expressivist approach and transcends the stress upon the common or the general that can be pervasive and problematic.


Exploring the implicit interlayer regulatory mechanism between cells and tissue: stochastic mathematical analyses of the spontaneous ordering in beating synchronization.
The present study focused on beating synchronization, and tried to elucidate the interlayer regulatory mechanisms between the cells and clump in beating synchronization with using the stochastic simulations which realize the beating synchronizations in beating cells with low cell-cell conductance. Firstly, the fluctuation in interbeat intervals (IBIs) of beating cells encouraged the process of beating synchronization, which was identified as the stochastic resonance. Secondly, fluctuation in the synchronized IBIs of a clump decreased as the number of beating cells increased. The decrease in IBI fluctuation due to clump formation implied both a decline of the electrophysiological plasticity of each beating cell and an enhancement of the electrophysiological stability of the clump. These findings were identified as the community effects. Because IBI fluctuation and the community effect facilitated the beating stability of the cell and clump, these factors contributed to the spontaneous ordering in beating synchronization. Thirdly, the cellular layouts in clump affected the synchronized beating rhythms. The synchronized beating rhythm in clump was implicitly regulated by a complicated synergistic effect among IBI fluctuation of each beating cell, the community effect and the cellular layout. This finding was indispensable for leading an elucidation of mechanism of emergence. The stochastic simulations showed the necessity of considering the synergistic effect, to elucidate the interlayer regulatory mechanisms in biological system.


Breast cancer patients' narratives about positive and negative communication experiences.
Health staff-patient communication is increasingly considered an important issue in cancer research. However, questionnaires addressing satisfaction with communication limit the issues patients can raise, do not address the context of communication and often show a strong positive skew in responses. Thus, qualitative studies of communication are also needed. Fifteen breast cancer patients were interviewed 3 months after finishing adjuvant treatment. They were asked to tell a 10 minute narrative and recall five experiences from treatment. Themes were extracted using categories derived from previous research while at the same time being sensitive to new elaborations and categories. The participants reported both positive and negative communication-related experiences from a wide range of treatment situations. Two major themes emerged: Information giving as professional care-giving and meeting emotional needs. The analysis suggests that appropriate information giving may have several functions, such as re-establishing the patient's future and reducing worst-case fantasies. Meeting emotional needs was seldom reported as directly talking about negative emotions, but rather through a variety of health staff behaviours. Also, the analysis points to problems in expecting or even pressurizing patients to feel and display negative emotions. The results highlight that meeting medical and emotional needs of patients may be closely intertwined in concrete treatment situations.


A novel mechanism of tissue inhibitor of metalloproteinases-1 activation by interleukin-1 in primary human astrocytes.
Reactive astrogliosis is the gliotic response to brain injury with activated astrocytes and microglia being the major effector cells. These cells secrete inflammatory cytokines, proteinases, and proteinase inhibitors that influence extracellular matrix (ECM) remodeling. In astrocytes, the expression of tissue inhibitor of metalloproteinases-1 (TIMP-1) is up-regulated by interleukin-1 (IL-1), which is a major neuroinflammatory cytokine. We report that IL-1 activates TIMP-1 expression via both the IKK/NF-kappaB and MEK3/6/p38/ATF-2 pathways in astrocytes. The activation of the TIMP-1 gene can be blocked by using pharmacological inhibitors, including BAY11-7082 and SB202190, overexpression of the dominant-negative inhibitor of NF-kappaB (IkappaBalphaSR), or by the knock-down of p65 subunit of NF-kappaB. Binding of activated NF-kappaB (p50/p65 heterodimer) and ATF-2 (homodimer) to two novel regulatory elements located -2.7 and -2.2 kb upstream of the TIMP-1 transcription start site, respectively, is required for full IL-1-responsiveness. Mutational analysis of these regulatory elements and their weak activity when linked to the minimal tk promoter suggest that cooperative binding is required to activate transcription. In contrast to astrocytes, we observed that TIMP-1 is expressed at lower levels in gliomas and is not regulated by IL-1. We provide evidence that the lack of TIMP-1 activation in gliomas results from either dysfunctional IKK/NF-kappaB or MEK3/6/p38/ATF-2 activation by IL-1. In summary, we propose a novel mechanism of TIMP-1 regulation, which ensures an increased supply of the inhibitor after brain injury, and limits ECM degradation. This mechanism does not function in gliomas, and may in part explain the increased invasiveness of glioma cells.


Forensic pathology--de profundis.
This review of the literature discusses the evolution of the professional role of the forensic pathologist. This role is defined as that of an expert who mainly assists the legal process of investigating suspicious or unexpected deaths. The forensic pathologist is independent, bringing his expertise to various parts of the judicial process. His role in the development of forensic investigation and promotion of human rights is important.


Gammadelta lymphocyte response to porcine reproductive and respiratory syndrome virus.
Porcine reproductive and respiratory syndrome virus (PRRSV) continues to be one of the most important diseases facing swine industry today. Following PRRSV infection pigs develop both humoral and cell-mediated responses following PRRSV exposure; however, the relative importance in protection and clearance of the virus is not yet completely understood. Swine contain a large percentage of gammadelta T-lymphocytes in peripheral circulation capable of responding to various pathogens in both an innate and specific immune response. The objectives of this study were to determine whether gammadelta lymphocytes functionally respond to PRRSV upon initial exposure and re-exposure. Four month old PRRSV free gilts were intranasally inoculated with a field isolate MN-30100 then assessed at various time points post infection. On day 120, pigs were re-exposed with MN-30100 PRRSV strain and subsequently were bled on days 0, 7, and 14 post re-exposure. Lymphocyte subpopulations, antigen specific proliferation, and IFN-gamma production were evaluated throughout the study. Circulating gammadelta lymphocytes in PRRSV exposed animals expanded between days 14 to 70 (d14-d70, p = 0.016); following antigen stimulation, gammadelta lymphocyte proliferated by day 14 (d0-d14, p = 0.001) continuing through day 60. gammadelta lymphocytes produced IFN-gamma by day 14 pi continuing through day 50 (d0-d50, p = 0.004). Following re-exposure both gammadelta+ and CD4+ lymphocytes increased in IFN-gamma production. These results are not fully conclusive on the role of gammadelta lymphocytes against PRRSV; the data indicate that gammadelta lymphocytes specifically respond to PRRSV.


[A 13-year-old-girl with Pickwickian syndrome and Asperger syndrome].
We reported a 13-year-old girl with Pickwickian syndrome and Asperger syndrome. The chief complaint on admission was apnea attacks during sleep. She had severe obesity. Whole night polysomnography showed that the apnea attacks occurred during light and REM sleep, and that slow wave sleep and REM sleep volumes were decreased. These findings were the same as those on adult cases. Weight control was very difficult because of Asperger syndrome.


Opportunities and Challenges for Psychiatry in the Connectomic Era.
Most major psychiatric disorders arise from disturbances of anatomically distributed neural systems rather than isolated dysfunction of circumscribed brain regions. The past decade has witnessed rapid advances in our capacity to measure, map, and model neural connectivity in diverse species and at different resolution scales, from the level of individual neurons and synapses to large-scale systems spanning the entire brain. In this review, we consider how these techniques, when grounded in the theory and methods of network science, can contribute to a biological understanding of mental illness. We focus in particular on attempts to accurately map brain network disturbances in clinical populations and to model the mechanistic causes of these changes. This work suggests that pathology within highly connected hub regions is a consistent finding across a broad array of phenotypically diverse disorders, and that disparate changes in brain network organization can sometimes be explained by a surprisingly small and simple set of mechanisms.


Induction of prophage in Streptococcus lactis C2 by ultraviolet irradiation.
Streptococcus lactis C2 has been used extensively by many laboratories in studies on the metabolism of lactic streptococci. By using ultraviolet irradiation as the inducing agent, this organism was shown to release a small bacteriophage, indicating that it is a lysogenic strain. The induced phage had a head approximately 40 nm in diameter and a tail length and width of about 180 and 6 nm, respectively. A structure resembling a collar was observed. Attempts to isolate a sensitive indicator strain for the virus were unsuccessful.


[Metabolic engineering of (E)-β-farnesene synthase genes for aphid-resistant genetically modified plants].
Aphids are major agricultural pests that cause significant yield losses of crops each year. (E)-β-farnesene (EβF), as the main component of the aphid alarm pheromones, can interrupt aphid feeding and cause other conspecies in the vicinity to become agitated or disperse from their host plant. Furthermore, EβF can function as a kairomone in attracting aphid predators. EβF synthase genes, which encode enzymes that convert farnesyl diphosphate (FPP) to the acyclic sesquiterpene EβF, have been isolated and characterized from peppermint (Mentha × piperita and Mentha asiatica), Yuzu (Citrus junos), Douglas fir (Pseudotsuga menziesii), sweet wormwood (Artemisia annua) and chamomile (Matricaria recutita), respectively. Transgenic plant overexpressing EβF synthase genes has been one of the most efficient strategies for aphid management. In this review, the current statuses of transgenic plants engineered for aphid resistance were summarized. The plant-derived EβF synthase genes with their potential roles in aphid management via genetic-modified (GM) approaches were reviewed. The existing problem in GM plants with EβF synthase gene, such as low EβF emission was usually detected in the transgenic plant, was discussed and the development direction in this area was proposed.


Transrectal real-time tissue elastography targeted biopsy coupled with peak strain index improves the detection of clinically important prostate cancer.
The focus of the present study was to evaluate transrectal real-time tissue elastography (RTE)-targeted two-core biopsy coupled with peak strain index for the detection of prostate cancer (PCa) and to compare this method with 10-core systematic biopsy. A total of 141 patients were enrolled for evaluation. The diagnostic value of peak strain index was assessed using a receiver operating characteristic curve. The cancer detection rates of the two approaches and corresponding positive cores and Gleason score were compared. The cancer detection rate per core in the RTE-targeted biopsy (44%) was higher compared with that in systematic biopsy (30%). The peak strain index value of PCa was higher compared with that of the benign lesion. PCa was detected with the highest sensitivity (87.5%) and specificity (85.5%) using the threshold value of a peak strain index of ≥5.97 with an area under the curve value of 0.95. When the Gleason score was ≥7, RTE-targeted biopsy coupled with peak strain index detected 95.6% of PCa cases, but 84.4% were detected using systematic biopsy. Peak strain index as a quantitative parameter may improve the differentiation of PCa from benign lesions in the prostate peripheral zone. Transrectal RTE-targeted biopsy coupled with peak strain index may enhance the detection of clinically significant PCa, particularly when combined with systematic biopsy.


[Disaster nursing: one nurse's role and experience during the Fukushima Power Plant disaster following the Great East Japan Earthquake].
The Richter-scale 9.0 earthquake that struck Northeast Japan on March 11th, 2011 caused a tsunami that damaged the Fukushima No. 1 Power Plant and released enormous amounts of radiation into the environment. Many area residents were evacuated to several protected fallout shelters. Prior to the tsunami, Fukashima had around 505,760 residents over 65 years of age, comprising 24.9% of the city's pre-tsunami population of Fukushima (City of Fukushima, 2011). The high proportion of elderly contributed to difficulties encountered in evacuating and caring for Fukushima citizens in the immediate aftermath of the disaster. The first author participated in disaster relief efforts in two fallout shelters in Fukushima. This article was written to share her post-disaster care experience and learned knowledge with medical care professionals in Taiwan and other high earthquake risk areas. The article also offers guidelines on appropriate medical personnel response and behavior with regard to disaster response. We hope this experience-sharing offers positive suggestions for the future and facilitates improved disaster-care education in East Asia and enhanced international cooperation on disaster rescue.


Characterization of the 5'-flanking region of the human PTK6 gene.
PTK6 (also known as Brk) is a non-receptor protein tyrosine kinase, whose mRNA was expressed in the limited normal tissues such as colon and small intestine, and in breast carcinomas and breast cancer cell lines. The 813 bp region upstream from the translation initiation codon, which constitutes a functional promoter of the human PTK6 gene, was progressively deleted and fused to the luciferase reporter gene and transient expression of the resultant constructs was measured upon transfection into a breast carcinoma cell line, T-47D. Comparative analysis of luciferase activity revealed two major regions, -93 to -76 and -702 to -655, important for transcriptional regulation. The proximal -93 to -76 region was found to be essential for the function of the minimal promoter. By primer extension and PCR, it was shown that a PTK6 transcript started at the most 5' upstream is located around base -104. Therefore, the proximal -93 to -76 region is thought to function as a downstream cis-acting element. Luciferase analysis showed that the distal -702 to -655 region contained at least two cis-acting elements. Gel mobility shift assays with T-47D nuclear extract including competition analyses with consensus and mutant oligonucleotides and supershift analyses with NF-kappaB and Sp1 antibodies showed that NF-kappaB binds to the sequence from -706 to -688 and Sp1 binds to the sequence from -688 to -669. This study thus provides the first molecular insights into the transcriptional regulation of the human PTK6 gene.


Widely targeted metabolomics based on large-scale MS/MS data for elucidating metabolite accumulation patterns in plants.
Metabolomics is an 'omics' approach that aims to analyze all metabolites in a biological sample comprehensively. The detailed metabolite profiling of thousands of plant samples has great potential for directly elucidating plant metabolic processes. However, both a comprehensive analysis and a high throughput are difficult to achieve at the same time due to the wide diversity of metabolites in plants. Here, we have established a novel and practical metabolomics methodology for quantifying hundreds of targeted metabolites in a high-throughput manner. Multiple reaction monitoring (MRM) using tandem quadrupole mass spectrometry (TQMS), which monitors both the specific precursor ions and product ions of each metabolite, is a standard technique in targeted metabolomics, as it enables high sensitivity, reproducibility and a broad dynamic range. In this study, we optimized the MRM conditions for specific compounds by performing automated flow injection analyses with TQMS. Based on a total of 61,920 spectra for 860 authentic compounds, the MRM conditions of 497 compounds were successfully optimized. These were applied to high-throughput automated analysis of biological samples using TQMS coupled with ultra performance liquid chromatography (UPLC). By this analysis, approximately 100 metabolites were quantified in each of 14 plant accessions from Brassicaceae, Gramineae and Fabaceae. A hierarchical cluster analysis based on the metabolite accumulation patterns clearly showed differences among the plant families, and family-specific metabolites could be predicted using a batch-learning self-organizing map analysis. Thus, the automated widely targeted metabolomics approach established here should pave the way for large-scale metabolite profiling and comparative metabolomics.


The effect of trimipramine, cimetidine and atropine on gastric secretion.
A comparison was made of increasing doses of trimipramine (0.05-0.40 mg/kg/h), atropine (7-28 microgram/kg/h) and cimetidine (0.30-2.40 mg/kg/h) on the gastric secretion stimulated by 3 microgram/kg/h of histamine dihydrochloride as continuous infusion, each dose step lasting 30 minutes. In 9 healthy subjects it was found that trimipramine had no significant effect on the output of acid. The largest dose of atropine caused a reduction by 77%, and cimetidine a reduction of 89% during the last 15 minutes portions. The study suggests that the healing of peptic ulcer by trimipramine is not linked to the effect on the histamine-stimulated gastric acid secretion.


Preanalytical considerations in blood gas analysis.
Blood gas testing is a commonly ordered test in hospital settings, where the results almost always have the potential to dictate an immediate or urgent response. The preanalytical steps in testing, from choosing the correct tests to ensuring the specimen is introduced into the instrument correctly, must be perfectly coordinated to ensure that the patient receives appropriate and timely therapy in response to the analytical results. While many of the preanalytical steps in blood gas testing are common to all laboratory tests, such as accurate specimen labeling, some are unique to this testing because of the physicochemical properties of the analytes being measured. The common sources of preanalytical variation in blood gas testing are reviewed here.


Sudden cardiac arrest due to subtotal absence of left-sided pericardium--case report and review of the literature.
Congenital absence of the pericardium is a very rare cardiac malformation, usually diagnosed fortuitously on autopsy or surgery. Symptoms related to these abnormalities are usually benign, and fatalities reported in the literature are almost exclusively secondary to herniation of the heart through a partial defect. We present the unusual case of a 44-year-old woman admitted for sudden cardiac arrest. Initial evaluation suggested acute anterior myocardial infarction, but further investigation ruled out coronary heart disease. No arrhythmia could be initiated on electrophysiological study, and absence of most of the left pericardium was confirmed by cardiac magnetic resonance imaging. After the exclusion of common etiologies such as idiopathic ventricular fibrillation, transient great vessel torsion due to hypermobility of the heart with secondary malignant arrhythmia was considered to be the most likely mechanism for the sudden cardiac arrest. A review of the available literature on clinical presentation, diagnostic tools, and therapeutic options is also presented.


Expression of multidrug-resistance (MDR1) gene in normal epithelia and in invasive carcinomas of the uterine cervix.
Total RNA from 92 invasive cervical cancers was analyzed for the presence of multidrug-resistance (MDR1) gene (also known as PGY1) transcripts. A 4.5-kilobase MDR1 transcript band was detected in 40 (43%) of 92 invasive cervical carcinomas and in 15 (68%) of 22 normal cervices. MDR1 levels were low [mean, 2.5 arbitrary units (U)] except in one liver metastasis (50 U) treated with a drug regimen including vincristine. Of eight carcinomas treated by radiotherapy and/or chemotherapy, seven (88%) exhibited MDR1 transcripts as compared with 24 (35%) of 69 untreated carcinomas (Fisher's exact test; P = .01). In conclusion, our data suggest that the MDR1 gene plays a role in drug resistance of certain cervical cancers, but also that other mechanisms may be involved.


A Comparative Study of Contemporary Color Tongue Image Extraction Methods Based on HSI.
Tongue image with coating is of important clinical diagnostic meaning, but traditional tongue image extraction method is not competent for extraction of tongue image with thick coating. In this paper, a novel method is suggested, which applies multiobjective greedy rules and makes fusion of color and space information in order to extract tongue image accurately. A comparative study of several contemporary tongue image extraction methods is also made from the aspects of accuracy and efficiency. As the experimental results show, geodesic active contour is quite slow and not accurate, the other 3 methods achieve fairly good segmentation results except in the case of the tongue with thick coating, our method achieves ideal segmentation results whatever types of tongue images are, and efficiency of our method is acceptable for the application of quantitative check of tongue image.


Review of exertional rhabdomyolysis and a case in a rhesus monkey (Macaca mulatta).
This is the first confirmed report of exertional rhabdomyolysis in a non-human primate. The monkey was singly housed and presented with anorexia and reluctance to move. There was no external evidence of trauma. Clinicopathologic findings included mild azotemia, marked elevation in serum creatine phosphokinase (CPK), alanine aminotransferase, aspartate aminotransferase, and myoglobinuria. Two days post-incident, the peripheral skeletal muscle had marked multifocal myonecrosis and fibrillar disruption without an inflammatory reaction. Treatment included diuresis and pain relief, and urinary output was monitored. The monkey recovered over the next two weeks. The major significance of skeletal muscle damage is the potential of released myoglobin to cause acute renal failure in the presence of other co-factors such as hypovolemia, acidosis, or ischemia. CPK levels can be highly variable and are inconsistent with the degree of muscle damage; however, CPK is thought to be the most sensitive enzyme marker for muscle necrosis. Because of the potential life-threatening sequelae, exertional rhabdomyolysis should be included as a differential diagnosis when similar clinical and pathological signs are observed.


Prolapse Volume to Prolapse Height Ratio for Differentiating Barlow's Disease From Fibroelastic Deficiency.
As mitral valve (MV) repair for Barlow's disease remains surgically challenging, it is important to distinguish Barlow's disease from fibroelastic deficiency (FED) preoperatively. We hypothesized that the prolapse volume to prolapse height ratio (PV-PH ratio) may be useful to differentiate Barlow's disease and FED.Methods and Results:In 76 patients with MV prolapse who underwent presurgical transesophageal echocardiography, the 3D MV morphology was quantified: 19 patients were diagnosed with Barlow's disease and 57 with FED. The patients with Barlow's disease had greater prolapse volume and height than the patients with FED, as well as greater PV-PH ratio (0.61±0.35 vs. 0.17±0.10, P<0.001). Receiver-operating characteristic analysis revealed that with a cutoff value of 0.27, the PV-PH ratio differentiated Barlow's disease from FED with 84.2% sensitivity and 84.2% specificity. Net reclassification improvement showed that the differentiating ability of the PV-PH ratio was significantly superior to prolapse volume (1.30, P<0.001). After being adjusted by each of prolapse volume and height, annular area and shape, and the number of prolapsed segments, the PV-PH ratio had an independent association with Barlow's disease. The PV-PH ratio was able to differentiate Barlow's disease from FED with high accuracy. 3D quantification including this value should be performed before MV repair.


Identification of phenyl-pyridine-2-carboxylic acid derivatives as novel cell cycle inhibitors with increased selectivity for cancer cells.
Ro 41-4439, a phenyl-pyridine-2-carboxylic acid derivative, was identified by a cell-based screening approach that exploits the differences between normal and cancer cells in their sensitivity to cytotoxic agents. This compound showed low micromolar antiproliferative activity and cytotoxicity against a broad panel of human cancer cell lines in vitro, and over 10-fold selectivity to cancer cells when tested in parallel with a panel of proliferating normal human cells. Cytotoxicity of Ro 41-4439 is due to arrest of cell cycle progression in mitosis followed by induction of apoptosis. Four-week treatment of nude mice bearing established mammary tumor xenografts (MDA-MB-435) with well-tolerated doses of the compound showed 73% inhibition of tumor growth. Limited exploration of structure-activity relationships involving side chain length, and aryl and pyridine rings allowed for the identification of more potent analogs.


Formation of the calcarine sulcus: a potential marker to predict the progression in utero of isolated mild fetal ventriculomegaly.
Our previous study confirmed the negative association between the development of calcarine sulcus and the width of lateral ventricles. The purpose of current study was to evaluate the reliability of calcarine sulcus depth in the 2nd trimester to predict the prenatal enlargement of lateral ventricle in fetuses with isolated mild fetal ventriculomegaly (IMVM).This study used a retrospective cohort study design. A total of 97 pregnant women with IMVM diagnosed between 20 and 26 weeks' gestation returned for a 2nd examination at 30 to 32 weeks. Lateral ventricular size and calcarine sulcus depth were acquired from ultrasonography and magnetic resonance imaging (MRI) scans, respectively. Progression was defined as the process of developing from a lower group toward a higher (<10 mm, 10-12 mm, 13-15 mm, and ≥16 mm).Significant correlation was observed between calcarine sulcus depth and ventricular measurements at the 2nd scan (r = -0.71, P < .0001). Receiver-operating characteristic curves showed that calcarine sulcus depth (area under curve [AUC] = 0.83, 95% confidence interval [CI] = 0.74-0.92) had the best diagnostic performance in predicting the prenatal progression, as compared with lateral ventricle width (AUC = 0.69, 95%CI = 0.54-0.84) and gestational age (AUC = 0.70, 95%CI = 0.57-0.83) at the initial scan. The cutoff value for calcarine sulcus depth was 3.3 mm, with the corresponding sensitivity and specificity were 75.0% and 81.3%, respectively. Multivariate analyses showed that calcarine sulcus depth ≥3.3 mm (odds ratio = 0.09, 95%CI = 0.02-0.38, P = .001) was an independent predictor of the prenatal progression.For IMVM, calcarine sulcus depth might be a powerful marker to identify subjects at higher risk for worse prenatal progression.


Integrated spatial light receivers based on inverse design.
Photonic integrated spatial light receivers play a crucial role in free space optical (FSO) communication systems. In this paper, we propose a 4-channel and 6-channel spatial light receiver based on a silicon-on-insulator (SOI) using an inverse design method, respectively. The 4-channel receiver has a square receiving area of 4.4 µm × 4.4 µm, which enables receiving four Hermite-Gaussian modes (HG00, HG01, HG10, and HG02) and converting them into fundamental transverse electric (TE00) modes with insertion losses (ILs) within 1.6∼2.1 dB and mean cross talks (MCTs) less than -16 dB, at a wavelength of 1550 nm. The 3 dB bandwidths of the four HG modes range from 28 nm to 46 nm. Moreover, we explore the impact of fabrication errors, including under/over etching and oxide thickness errors, on the performance of the designed device. Simulation results show that the 4-channel receiver is robust against fabrication errors. The designed 6-channel receiver, featuring a regular hexagon receiving area, is capable of receiving six modes (HG00, HG01, HG10, HG02, HG20, and HG11) with ILs within 2.3∼4.1 dB and MCTs less than -15 dB, at a wavelength of 1550 nm. Additionally, the receiver offers a minimum optical bandwidth of 26 nm.


[Massive bleeding from the upper digestive tract in patients with pseudoaneurysm of splenic artery].
Pancreatic pseudocysts are common complication of both chronic and acute pancreatitis. Sanguination from damaged peripancreatic vessels into the lumen of pseudocyst results in pseudoaneurysm. The rupture of pancreatic pseudoaneurysm into the lumen of digestive tract causes massive bleeding witch source is often difficult to find during endoscopic examination. We present a case of patient with chronic alcohol pancreatitis, with pancreatic pseudocyst and of acute bleeding from upper digestive tract. In the endoscopy we found gastric ulcer with visible vessel. During hospitalization we observed increase the diameter of pseudocyst and circulation of it's liquid contence. Second-look endoscopy showed gastric fundic varices. Surgical operation revealed pseudoaneurysm of splenic artery inserting pressure on gastric wall.


The effect of curing intensity on mechanical properties of different bulk-fill composite resins.
The purpose of this study was to investigate the effects of two curing light intensities on the mechanical properties (Vickers microhardness, compressive strength, and diametral tensile strength) of bulk-fill resin-based composites (RBCs). Four commercially available bulk-fill RBCs (Tetric® N-Ceram, SonicFill™, Smart Dentin Replacement (SDR™) Posterior Flowable Material, and Filtek™ Posterior Restorative) were used in this study. A total of 72 cylindrical specimens of each RBC (n=288) were prepared and subjected to Vickers microhardness, compressive strength, and diametral tensile strength tests at high (1200 mW/cm2) and low (650 mW/cm2) curing light intensities (each n=12). Results were evaluated using independent and paired sample t-tests, one-way analysis of variance, and Tukey's post hoc test. All tests were performed at a significance level of P<0.05. The highest mean microhardness was observed for SonicFill (58.3 Vickers hardness number [VHN]) cured using high-intensity light. Although having the least mean microhardness values, a significant difference was observed between SDR cured using high-intensity light and that cured using low-intensity light (P<0.05). In the total sample, the highest mean compressive strength was obtained for SonicFill (262.6 MPa), followed by SDR (253.2 MPa), both cured using high-intensity light, and the least was measured for Tetric N-Ceram cured using low-intensity light (214.3 MPa). At high and low curing light intensities, diametral tensile strength for all RBCs except SonicFill was significant (P<0.001). A higher curing light intensity (1200 mW/cm2) had a positive influence on the compressive and tensile strength of the four bulk-fill RBCs and microhardness of two materials tested compared with lower curing light intensity (650 mW/cm2). SonicFill showed the greatest microhardness and compressive strength significantly for both curing light intensities and greater diametral tensile strength with high-intensity light, although not significant. SDR cured with high-intensity light showed the greatest diametral tensile strength among the four materials.


Cardiac Autonomic Neuropathy is Associated with Improved Diabetic Foot Ulcer Healing: A Single Centre Prospective Cohort Study.
People with diabetes and peripheral neuropathy (DPN) are at high risk of diabetic foot ulceration (DFU). The prevalence of cardiac autonomic neuropathy (CAN) in people with DFU is unknown and if CAN influences DFU healing is unclear. We investigated, in a prospective observational single-centre cohort study, if CAN predicts DFU healing in 47 (77% male) people with a DFU and DPN attending a university hospital foot clinic. CAN was diagnosed by 2 or more abnormal Ewing's tests. Baseline DFU severity was evaluated using the site, ischaemia, neuropathy, bacterial infection, area and depth (SINBAD) score. The primary outcome was defined as evidence of DFU healing on clinical examination. Median (interquartile) length of follow-up was 1150 (624-1331) days. The prevalence of CAN was 43%. Of the cohort, 70% had complete healing of their DFU. Participants with CAN had a shorter median (interquartile) duration time to heal compared to those without CAN [91 (44-164) days compared to 302 (135-413) (p=0.047)]. Minor/major amputation and mortality was similar in both groups. The presence of CAN increased DFU healing by two-fold [HR=2.05, 95% CI 1.01-4.16, p=0.046] in multivariable competing risk analyses. We demonstrate a high prevalence of CAN in a DFU cohort and that CAN is associated with improved DFU healing. The results of this study establish the scientific rationale for further studies to better understand the mechanisms between CAN and DFU outcomes.


[Standardization of the laboratory method for antithrombin activity measurement].
Measurement of antithrombin (AT) activity in plasma provides important information for the diagnosis of congential antithrombin deficiency and disseminated intravascular coagulation. When measured in the presence of heparin, plasma antithrombin activity includes heparin cofactor II activity as about 20-30% of the total activity. Antithrombin assay methods measuring plasma anti-F.Xa activity or those measured in the presence of 0.22 M NaCl exclude the influence of HC II activity. Dilution of the plasma sample is necessary to measure the heparin cofactor anti-thrombin activity of AT correctly. A laboratory method that precisely reflects plasma heparin cofactor anti-thrombin activity of AT should be established as the standard laboratory method.


Antimicrobial resistance among faecal enterococci from healthy individuals in Portugal.
Analysis of 247 faecal enterococcal isolates from 99 healthy Portuguese individuals during 2001 revealed the presence of enterococci resistant to vancomycin (5%) and highly resistant to streptomycin (52%), kanamycin (40%) or gentamicin (11%). Most isolates were also resistant to tetracycline, erythromycin, ciprofloxacin and quinupristin-dalfopristin. The vanA (two Tn1546 types), vanC1, erm(B), aac(6')-aph(2'')-Ia, aph(3')-IIIa, vat(E) and vat(D) genes were detected. Enterococcus faecalis and Enterococcus faecium isolates with high-level resistance to gentamicin were related to Portuguese poultry isolates described previously. E. faecium isolates that were highly resistant to vancomycin or gentamicin harboured different housekeeping purK alleles associated previously with different hosts.


Nucleolar organiser regions: new prognostic variable in breast carcinomas.
Nucleolar organiser regions (NORs), which are important for regulating protein synthesis, were identified in 20 breast carcinomas by means of a silver (Ag) staining technique. Infiltrating neoplasms with metastases in four or more axillary lymph nodes possessed, on average, a greater number of AgNORs per cell nucleus compared with neoplasms without nodal disease, or with one to three positive lymph nodes. The size, morphology, and distribution of AgNORs within the nucleus were also different in the two study groups. Overall, these findings suggest that breast carcinomas with multiple, irregular, and widely dispersed AgNORs tend to be of high grade malignancy.


A self-guided Internet-delivered intervention for adults with ADHD: A feasibility study.
Attention Deficit Hyperactivity Disorder (ADHD) is a prevalent neurodevelopmental disorder that tends to persist into adulthood. Many adults with ADHD seek non-pharmacological treatment in addition to pharmacological treatment. Still, there are few non-pharmacological treatment options available. The aim of the current study was to explore the feasibility of a self-guided Internet-delivered intervention for adults with ADHD. The study has an uncontrolled, within-group, pre-post design. Thirteen participants with an ADHD diagnosis were included and given access to the first three modules of a seven-module intervention. To explore the feasibility of the intervention, the adherence, credibility, and treatment satisfaction were examined. Preliminary efficacy of the intervention was examined through self-report measures of inattention, hyperactivity, depression, anxiety, stress, and quality of life. In terms of adherence, seven participants (54%) completed all three intervention modules (M = 1.85, SD = 1.3). The participants reported both good credibility and treatment satisfaction with the intervention. The participants also reported challenges related to usability and technical issues. Although the clinical outcomes must be interpreted with caution due to the study design and the small sample size, a statistically significant reduction in severity of inattention was reported by the participants following the intervention (p = .006, d = 1.57). The reduction was non-significant for hyperactivity (p = .326, d = 0.33). The participants who completed all three modules in the intervention (n = 7) also reported a significant decrease in stress (p = .042, d = 0.67) and a significant increase in quality of life (p = .016, d = 0.99). No significant changes were found on measures of anxiety and depression. The adherence to the intervention was relatively low, but the participants who completed the study reported good credibility and satisfaction with the intervention. These results indicate that there is a need to improve the intervention to make it more engaging before conducting a randomized-controlled trial investigating the clinical effects of the full seven-module intervention.


ctDNA analysis in the personalized clinical management of gastroesophageal adenocarcinoma: turning hope into reality.
Gastroesophageal adenocarcinoma (GEA) is a global health issue with a high fatality-to-case ratio and a 5-year overall survival that has only slightly improved. High-throughput molecular profiling has uncovered a profound complexity and heterogeneity in GEA biology, which limits considerably the treatment advances. Liquid biopsy with circulating tumor (ct)DNA analysis could elucidate GEA molecular heterogeneity and provide diagnostic, prognostic and predictive information to guide clinical decision-making. However, only a handful of studies have shown positive results for the application of ctDNA analysis in GEA clinical management. As a result, no comprehensive information is available to date on this continuously evolving topic. Here, we discuss the current state of knowledge, along with promises and challenges related to ctDNA analysis in GEA.


Distinct Regulations of HO-1 Gene Expression for Stress Response and Substrate Induction.
Heme oxygenase 1 (HO-1) is the key enzyme for heme catabolism and cytoprotection. Whereas HO-1 gene expression in response to various stresses has been investigated extensively, the precise mechanisms by which HO-1 gene expression is regulated by the HO-1 substrate heme remain elusive. To systematically examine whether stress-mediated induction and substrate-mediated induction of HO-1 utilize similar or distinct regulatory pathways, we developed an HO-1-DsRed-knock-in reporter mouse in which the HO-1 gene is floxed by loxP sites and the DsRed gene has been inserted. Myeloid lineage-specific recombination of the floxed locus led to fluorescence derived from expression of the HO-1-DsRed fusion protein in peritoneal macrophages. We also challenged general recombination of the locus and generated mice harboring heterozygous recombinant alleles, which enabled us to monitor HO-1-DsRed expression in the whole body in vivo and ex vivo. HO-1 inducers upregulated HO-1-DsRed expression in myeloid lineage cells isolated from the mice. Notably, analyses of peritoneal macrophages from HO-1-DsRed mice lacking NRF2, a major regulator of the oxidative/electrophilic stress response, led us to identify NRF2-dependent stress response-mediated HO-1 induction and NRF2-independent substrate-mediated HO-1 induction. Thus, the HO-1 gene is subjected to at least two distinct levels of regulation, and the available lines of evidence suggest that substrate induction in peritoneal macrophages is independent of CNC family-based regulation.


Malignant peripheral nerve sheath tumor of proximal third tibia.
A 16-year-old man had aswelling over the anterior aspect of the proximal third of the tibia for 1 year, which was peanut size initially and progressively increased to its present size of 10 cm × 8 cm. He underwent fine needle aspiration cytology (FNAC) twice during this period and reported aspindle cell sarcoma. Malignant peripheral nerve sheath tumor (MPNST) is a malignancy of the connective tissue surrounding the nerves. Previously, MPNST was also known as neurofibrosarcoma, malignant schwannoma, andneurogenic sarcoma. We are reporting this case for its rarity and peculiar mode of presentation. FNAC/core biopsy can be used as an effective tool to achievethe correct pathological diagnosis.


Durable antifog films from layer-by-layer molecularly blended hydrophilic polysaccharides.
Mechanically durable, long-lasting antifog coatings based on polysaccharides were developed using a layer-by-layer (LBL) assembly process. The unique properties of these coatings are a result of a molecular-level blending of the polysaccharides, with multilayers containing chitosan and carboxymethyl cellulose providing the best overall properties. The antifog properties resulted from a strong interaction between the polar and H-bonding elements of the assembled polymers and water molecules and the concomitant formation of thin films of water. Environmental scanning electron microscopy (ESEM) studies confirmed that fogging coatings are decorated with light scattering, micrometer-sized droplets of water whereas antifogging coatings remain droplet free. To improve the mechanical durability of the multilayer films on substrates, the surface was modified via self-assembly of epoxy-functionalized silane molecules. Cross-linking chemistry was then applied to improve the mechanical robustness of the LBL films on various surfaces. These films were characterized using several techniques: optical profilometery (PL), spectroscopic ellipsometry (EL), contact angle goniometry (CA), and atomic force microscopy (AFM). The antifog properties of the films were evaluated by several tests under different environmental conditions. This work demonstrates that the unique water-adsorbing properties of polysaccharides can be exploited to create permanent antifog properties, which may be useful for various applications.


Erectile dysfunction following Nd-YAG visual laser-assisted prostatectomy (VLAP).
We investigated if side fire Nd-YAG visual laser prostatectomy (VLAP) causes erectile dysfunction (ED) in patients who were sexually active prior to the procedure. The 36 study patients gave a detailed medical/sexual history: physical examination included bulbo-cavernous reflex (BCR) on rectal examination and routine blood tests. Lasing time, power of the applied laser beam (in watts), coagulation site and energy intensity were recorded. Patients with new onset ED during the one year study period underwent pudendal nerve conduction (PNC), color duplex Doppler ultrasonography, and NPT/RigiScan testing. In 6 out of 36 (16.7%) patients reporting significant post-operative ED, there was a tendency towards higher energy applied and longer lasing time but no correlation between prostate size or the site of energy application. Patients reported loss of night and/or morning erections (n = 5), retrograde ejaculations (n = 2), loss of ejaculate (n = 2), and decreased sensation of orgasm (n = 3). Three had abnormal PNC, duplex Doppler showed abnormal blood supply in four, and all six had abnormal NPT/RigiScan. We believe this is the first demonstration that VLAP may be associated with a high rate of ED and that the lasing time and intensity of applied laser energy may play a role in this outcome.


Effect of alpha-naphthalene acetic acid and thidiazuron on seedling of economic crops grown in endosulfan sulfate-spiked sand.
The effect of two plant growth regulators, alpha-naphthalene acetic acid (NAA) and thidiazuron (TDZ) on the growth of sweet corn (Zea mays), cowpea (Vigna sinensis) and cucumber (Cucurmis sativus) seedling planted in 1-100 mg kg(-1) of endosulfan sulfate spiked sand was investigated. Endosulfan sulfate had no apparent toxicity as seedlings of these crop plants grew normally in endosulfan sulfate spiked sand. Concentration of endosulfan sulfate in sand affected the response of seedling induction by NAA or TDZ. Induction of crop seeds by NAA or TDZ did not promote growth of sweet corn, cowpea and cucumber to an appreciable extent. Both plant regulators at concentration of 10 mg l(-1) seemed to exert adverse effect on crop seedling. TDZ decreased shoot length, root length and chlorophyll contents in leaves of sweet corn and cowpea growing in endosulfan sulfate spiked sand. In contrast, NAA was not toxic and promoted growth of sweet corn and cowpea seedling. However, cucumber was affected by NAA and TDZ more than other plants. TDZ significantly decreased biomass and root length of cucumber. Also, NAA significantly decreased cucumber root length and tended to increase cucumber root dried weight when grown in 100 mg kg(-1) of endosulfan sulfate spiked sand.


Key legal principles for hospitalists.
In a hospitalist system, when a patient leaves the hospital, he or she will return to a primary care provider (PCP) for follow-up and continuing care. The hand-off after discharge can compromise communication with the PCP. Physicians have a legal duty to provide follow-up care to patients with whom they have a relationship. The obligation to provide follow-up care endures even when the patient misses a scheduled appointment or does not adhere to the follow-up regimen. In general, the physician who began the care must fulfill that obligation. An essential component of follow-up care includes educating the patient about what symptoms require follow-up care and why it is important. The duty to provide adequate follow-up care is shared by the hospitalist and the PCP. Virtually no malpractice case law considers the obligations and practices of hospitalists. This article uses cases involving follow-up care for patients treated in an emergency department and general cases regarding liability for follow-up care to examine the potential legal obligations of both hospitalists and PCPs for follow-up care, including circumstances involving pending test results and incidental findings.


Assessing the magnitude of potential environmental impacts related to water and toxicity in the Peruvian hyper-arid coast: A case study for the cultivation of grapes for pisco production.
The environmental sustainability of the cultivation of grapes for the production of alcoholic beverages has been extensively analyzed in the literature from a Life Cycle Assessment perspective, although certain impact categories have been repeatedly neglected despite their importance, such as toxic emissions or the depletion of freshwater resources. Hence, the current study provides a detailed assessment of water footprint-related impact categories, including toxicity, for the cultivation of grapes for pisco production, an alcoholic beverage produced in coastal Peru in hyper-arid areas that suffer high levels of water scarcity. Characterization factors at a sub-watershed level were used to calculate water consumption impact assessment of grape production using the AWARE method. Site-specific toxic emissions were modelled using the PestLCI model, considering primary climate and soil data. The USEtox assessment method was then used to compute freshwater eco-toxicity with these data. Results demonstrate the high water footprint of irrigating vineyards in coastal Peru, especially considering the inefficient flooding irrigation process. In terms of water consumption, despite the high variability shown between sub-watersheds, the shift to other irrigation technologies must be analyzed with care due to the high competition for water existing in the area. Eutrophication potential showed particularly high values compared to the literature, whereas freshwater eco-toxicity impacts were relatively low due to the high volatilization of pesticides to air. Nevertheless, the lack of an adequate wastewater management system implies that the estimated potential toxic and eutrophying emissions may constitute a further environmental threat to water bodies.


Local function conservation in sequence and structure space.
We assess the variability of protein function in protein sequence and structure space. Various regions in this space exhibit considerable difference in the local conservation of molecular function. We analyze and capture local function conservation by means of logistic curves. Based on this analysis, we propose a method for predicting molecular function of a query protein with known structure but unknown function. The prediction method is rigorously assessed and compared with a previously published function predictor. Furthermore, we apply the method to 500 functionally unannotated PDB structures and discuss selected examples. The proposed approach provides a simple yet consistent statistical model for the complex relations between protein sequence, structure, and function. The GOdot method is available online (http://godot.bioinf.mpi-inf.mpg.de).


Variability in the quantification of abrasion on the Bruxcore device.
The rate of abrasion of dental surfaces during short periods of time is difficult to measure clinically, but one quantifiable method is the use of the Bruxcore bruxism monitoring device. The aim of this study was to estimate the interobserver and intraobserver variation in the Bruxcore system using different reading methods. Fifteen volunteers used individually fabricated Bruxcore devices during 4 consecutive nights, and this procedure was repeated after 6 weeks. The abraded areas of the 30 Bruxcore devices were measured by two observers on two occasions and with three methods: microscope without a reference scale; microscope with a reference scale; and a computer-aided system. Intraobserver variation was small (5%), but interobserver variation was statistically significant for all three methods. The computer-aided system was superior to the other two methods. The interaction between Bruxcore values and observers was statistically significant for the microscope methods but not for the computer method. This was a desired property, indicating stability of the computer-aided method over the range of Bruxcore values observed. Small measurement errors, independent of the size of the measurements, can be expected using a trained observer and a computer-aided method for reading the Bruxcore bruxism monitoring device.


Binding of Ca ions by Paramecium caudatum.
Binding of (45)Ca by live Paramecium caudatum was determined under various external ionic conditions. It was found that calcium uptake was separable into at least two components, a rapid and a slow one. The rapid component was influenced by the presence of certain other ions in a manner which agrees with the law of mass action. It appears that an ion exchange system may be involved in a binding equilibrium established between Paramecium, Ca(++), and certain other ions. K(+), Rb(+), and Ba(++) in the equilibrium medium are among those ions which inhibit calcium uptake. It is proposed that liberation of Ca(++) from binding sites on Paramecium by an exchange reaction with competing ions is the first step in the mechanism of ciliary reversal in the response to external application of these ions.


Risk factors for genital papillomavirus infection in populations at high and low risk for cervical cancer.
This study sought to determine risk factors for genital infection with papillomavirus (HPV) in Panamanian women 20-49 years old. Subjects were randomly selected from Herrera and Panama provinces (cervical cancer incidence 79 and 25/100,000, respectively). Participants were interviewed to determine sexual behavior. Cervicovaginal lavage specimens were obtained to test for HPV DNA by commercial dot blot hybridization. HPV-16/18 DNA was detected significantly more frequently (5%) in Panama than Herrera (2%) samples (P = .002). Clearly, infection with high-risk HPV types alone cannot account for the differences in cervical cancer incidence between the two populations. HPV-16/18 detection decreased with increasing years of sexual experience among all women in Panama and among women with multiple partners in Herrera. However, HPV-16/18 detection did not change with sexual experience among monogamous women in Herrera. Thus, the epidemiology of HPV is complex and reflects both virus- and population-specific factors.


Effects of date palm fruit extracts on skin mucosal immunity, immune related genes expression and growth performance of common carp (Cyprinus carpio) fry.
The aim of this study was to investigate the effects of date palm fruit extracts (DPFE) on skin mucosal immunity, immune related genes expression and growth performance of fry common carp (Cyprinus carpio). One hundred and twenty specimens (4.06 ± 0.13 g) were supplied and allocated into six aquaria; specimens in three aquaria were fed non-supplemented diet (control) while the fish in the other 3 aquaria were fed with DPFE at 200 ml kg(-1). At the end of feeding trial (8 weeks) skin mucus immune parameters (total immunoglobulins, lysozyme, protease and alkaline phosphatase activity) and immune related gene expression (tumor necrosis factor α [tnfa], lysozyme [ly] and interleukin-1-beta, [il1b]) in the head-kidney were studied. The results revealed that feeding carp fry with 200 ml kg(-1) DPFE remarkably elevated the three skin mucus immune parameters tested (P < 0.05). However, evaluation of immune related gene expression demonstrated that the expression of tnfa and il1b was considerably decreased (P < 0.05) in fish fed DPFE diet, while the expression of ly remained similar (P > 0.05) compared to control fish (fed control diet). Furthermore, growth performance parameters were significantly improved in fry fed DPFE (P < 0.05). More studies are needed to understand different aspects of DPFE administration in fry mucosal immunity.


Survey of nonconventional yeasts for lipid and hydrocarbon biotechnology.
Nonconventional yeasts have an untapped potential to expand biotechnology and enable process development necessary for a circular economy. They are especially convenient for the field of lipid and hydrocarbon biotechnology because they offer faster growth than plants and easier scalability than microalgae and exhibit increased tolerance relative to some bacteria. The ability of industrial organisms to import and metabolically transform lipids and hydrocarbons is crucial in such applications. Here, we assessed the ability of 14 yeasts to utilize 18 model lipids and hydrocarbons from six functional groups and three carbon chain lengths. The studied strains covered 12 genera from nine families. Nine nonconventional yeasts performed better than Saccharomyces cerevisiae, the most common industrial yeast. Rhodotorula toruloides, Candida maltosa, Scheffersomyces stipitis, and Yarrowia lipolytica were observed to grow significantly better and on more types of lipids and lipid molecules than other strains. They were all able to utilize mid- to long-chain fatty acids, fatty alcohols, alkanes, alkenes, and dicarboxylic acids, including 28 previously unreported substrates across the four yeasts. Interestingly, a phylogenetic analysis showed a short evolutionary distance between the R. toruloides, C. maltosa, and S. stipitis, even though R. toruloides is classified under a different phylum. This work provides valuable insight into the lipid substrate range of nonconventional yeasts that can inform species selection decisions and viability of lipid feedstocks.


Identity of "orbitozygomatic muscle".
Our exploration disclosed 17 (40%) stranded muscles of 42 hemifacial specimens of Korean cadavers. We name this distinct muscle "Orbitozygomatic Muscle (OZM)" that is either an aberrant orbicularis oculi muscle or a separate muscle. A triangular space bounded superiorly by the lowest edge of the orbicularis oculi muscle (OOM), laterally by Zygomaticus Minor, and inferiorly by OMZ gives way to neurovascular bundle and suborbicularis oculi fat pad (SOOF).


Development and validation of capillary electrophoresis method for tobramycin with precapillary derivatization and UV detection.
One of the major drawbacks in the analysis of aminoglycoside antibiotics is their lack of UV chromophore and/or fluorophore. Tobramycin, a representative member of this group, was examined in this study. To overcome the detection hurdle, a precapillary derivatization followed by capillary electrophoresis analysis with direct UV detection was investigated. A central composite design was applied to optimize the method and three parameters were selected in this study: buffer pH, temperature and % acetonitrile (ACN). Selectivity between tobramycin main component and its adjacent peaks as well as the peak efficiency and symmetry factors were established as responses. For each response, a model was obtained by a second-order mathematical expression. Successful results were obtained with a simple background electrolyte (BGE) containing 30 mM sodium tetraborate, pH 10.2, and ACN (75:25 v/v). Under these conditions, baseline separation of tobramycin from its adjacent kanamycin B and an unknown peak was achieved. A temperature of 20 degrees C and applied voltage of 28.0 kV were used. The method showed good validation data in terms of precision, limits of quantitation and detection, specificity and linearity and was found to be suitable for analysis of tobramycin bulk pharmaceutical samples.


CAR T-cell therapy for pancreatic cancer.
Chimeric antigen receptor (CAR) T-cell therapy utilizes genetic engineering to redirect a patient's own T cells to target cancer cells. The remarkable results in hematological malignancies prompted investigating this approach in solid tumors such as pancreatic cancer. The complex tumor microenvironment, stromal hindrance in limiting immune response, and expression of checkpoint blockade on T cells pose hurdles. Herein, we summarize the opportunities, challenges, and state of knowledge in targeting pancreatic cancer with CAR T-cell therapy.


Expression and molecular analysis of the Arabidopsis DXR gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase, the first committed enzyme of the 2-C-methyl-D-erythritol 4-phosphate pathway.
1-Deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) catalyzes the first committed step of the 2-C-methyl-D-erythritol 4-phosphate pathway for isoprenoid biosynthesis. In Arabidopsis, DXR is encoded by a single-copy gene. We have cloned a full-length cDNA corresponding to this gene. A comparative analysis of all plant DXR sequences known to date predicted an N-terminal transit peptide for plastids, with a conserved cleavage site, and a conserved proline-rich region at the N terminus of the mature protein, which is not present in the prokaryotic DXR homologs. We demonstrate that Arabidopsis DXR is targeted to plastids and localizes into chloroplasts of leaf cells. The presence of the proline-rich region in the mature Arabidopsis DXR was confirmed by detection with a specific antibody. A proof of the enzymatic function of this protein was obtained by complementation of an Escherichia coli mutant defective in DXR activity. The expression pattern of beta-glucuronidase, driven by the DXR promoter in Arabidopsis transgenic plants, together with the tissue distribution of DXR transcript and protein, revealed developmental and environmental regulation of the DXR gene. The expression pattern of the DXR gene parallels that of the Arabidopsis 1-deoxy-D-xylulose 5-phosphate synthase gene, but the former is slightly more restricted. These genes are expressed in most organs of the plant including roots, with higher levels in seedlings and inflorescences. The block of the 2-C-methyl-D-erythritol 4-phosphate pathway in Arabidopsis seedlings with fosmidomycin led to a rapid accumulation of DXR protein, whereas the 1-deoxy-D-xylulose 5-phosphate synthase protein level was not altered. Our results are consistent with the participation of the Arabidopsis DXR gene in the control of the 2-C-methyl-D-erythritol 4-phosphate pathway.


Identity and the ethics of gene therapy.
Some conditions detrimental to human well-being, such as sickle-cell anaemia, cystic fibrosis, muscular dystrophy, Lesch-Nyhan disease and various immunodeficiencies, are genetically determined. One potential means of preventing the development of such conditions is the manipulation of genetic material in the conceptus of an organism which would otherwise develop such conditions. Genetic manipulations could take the form either of excising and substituting genetic material, excising but not substituting genetic material, adding but not excising genetic material or reorganizing existing genetic material. To succeed, manipulation would have to change genetic structure so as to change its informational content. It might be thought, however, that all or some such manipulations would involve causing particular individuals to cease to exist and involve bringing into existence new, distinct individuals. Gene therapy could not, therefore, be a procedure which improved the circumstances of the particular individual to whom it is applied. It might be suggested that once the metaphysics of identity and the facts of gene therapy are understood, certain interesting conclusions concerning the ethics of gene therapy emerge. Some such conclusions have been discussed in this journal by Noam J. Zohar and Jeffrey P. Kahn. More, however, needs to be said about them since neither Zohar nor Kahn draws the correct conclusions. While both have pertinent things to say, neither has given a completely clear account of the metaphysics of gene therapy and so neither has completely traced out the implication of the metaphysics for the ethics of gene therapy. This paper attempts to remedy these defects.


Reverse phase protein microarrays and their utility in drug development.
The majority of human diseases, including cancer, are characterized by abnormal protein function. Proteins regulate virtually every cellular process and exhibit multiple kinds of post-translational modification that modulate expression levels and activation states, such as phosphorylation by protein kinases. Additionally proteins interact with each other in complex regulatory networks and signal transduction pathways modulated by feedback mechanisms. These pathways are disrupted in disease and altered by therapeutic drugs. Reverse phase protein microarray (RPMA) technology allows simultaneous measurement of numerous phosphorylated, glycosylated, cleaved, or total cellular proteins from complex mixtures in many samples at once. Therefore, RPMAs can provide a portrait of a cell's signaling pathways in diseased states, before and after treatment with drugs, and allows comparison of changes in drug-resistant and sensitive cells. Furthermore, the technology offers a means of connecting genomic abnormalities in cancer to targetable alterations in protein signaling pathways, even for genetic events that seem otherwise undruggable. Consequently, the RPMA platform has great utility in many steps of drug development including target identification, validation of a pharmaceutical agent's efficacy, understanding mechanisms of action, and discovery of biomarkers that predict or guide therapeutic response. RPMAs have become a powerful tool for drug development and are now being integrated into human clinical cancer trials, where they are being used to personalize therapy.


Effects of vitamin A and dexamethasone on capsule collagen metabolism in mouse mammary adenocarcinoma.
It was previously shown that the administration of dexamethasone (Dex) to mice bearing mammary adenocarcinoma caused the collagen content of the tumor capsule to be decreased by 50%, but collagenase and other neutral protease activities of the tumor were the same as in the controls. These events occurred with distinctly increased tumor invasion and metastasis. The present communication reports on the characterization of capsule collagen and the effects of agents [vitamin A (VA) and Dex] on capsule collagen metabolism and presents further evidence concerning the possible mechanisms by which the collagen content of the capsule was decreased in the Dex-treated hosts. The collagen extracted from capsules of untreated controls and mice (C3H/HeJ) treated with VA or Dex was primarily type I, as judged by the migration of protein bands in sodium dodecylsulfate-polyacrylamide gel electrophoresis and by patterns of elution peaks from an octadecyl C-18 column. The amino acid compositions of type I collagen of the capsule of treated and untreated controls were similar but not identical to those of mouse skin and guinea pig skin type I collagens. The specific activity of intracellular free [14C] proline, the extent of hydroxylation of [14C]proline residues of collagen, and the specific activity of [14C]hydroxyproline and proline in each case were similar in treated and in untreated controls. These results suggest that the observed 45% decrease in the conversion of [14C]proline into protein-bound [14C]hydroxyproline of the Dex-treated hosts apparently was due to decreased collagen synthesis. The data also suggest that the most critical effects of Dex on tumor invasion and metastasis appeared to occur at an early stage before full formation of the collagenous extracellular matrix.


Comprehensive Metabolite Profile Uncovers the Bioactive Components, Antioxidant and Antibacterial Activities in Wild Tree Peony Leaves.
Tree peonies (Paeonia Section Moutan)-including nine wild species, which belong to subsections Vaginatae and Delavayanae-are economically important plants with ornamental, nutritional, and medicinal applications. In this study, for the first time, we determined the bioactive components and antioxidant activities and antibacterial activities of the newly grown leaves of nine wild tree peony species (WTPS). A total of 276 bioactive components were identified through non-targeted metabolomics; more than 80% of the 276 metabolites identified are terpenoids and flavonoids. A total of 42 differential metabolites were quantitatively determined. The main differential metabolites were Paeoniflorin, Luteoloside, Hyperin, Apigenin-7-glucoside, Rhoifolin, and Cantharidin. Such a high terpenoid and flavonoid content of the leaf extracts renders them as species with strong antibacterial capacities, and most of the bacteria tested showed greater sensitivity derived from the members of subsection Vaginatae than those of subsection Delavayanae. All WTPS have significant antioxidant activity; this activity is attributed to high levels of the total phenolic content (TPC) and total flavonoid content (TFC), of which, among the nine WTPS, P. lutea has the strongest antioxidant capacity. Our results provided a theoretical basis for the in-deep application of tree peony leaves for food, medical, and pharmaceutical industries.


Mechanistic Insights Underlying Tolerance to Acetic Acid Stress in Vaginal Candida glabrata Clinical Isolates.
During colonization of the vaginal tract Candida glabrata cells are challenged with the presence of acetic acid at a low pH, specially when dysbiosis occurs. To avoid exclusion from this niche C. glabrata cells are expected to evolve efficient adaptive responses to cope with this stress; however, these responses remain largely uncharacterized, especially in vaginal strains. In this work a cohort of 18 vaginal strains and 2 laboratory strains (CBS138 and KUE100) were phenotyped for their tolerance against inhibitory concentrations of acetic acid at pH 4. Despite some heterogeneity has been observed among the vaginal strains tested, in general these strains were considerably more tolerant to acetic acid than the laboratory strains. To tackle the mechanistic insights behind this differential level of tolerance observed, a set of vaginal strains differently tolerant to acetic acid (VG281∼VG49 < VG99 < VG216) and the highly susceptible laboratory strain KUE100 were selected for further studies. When suddenly challenged with acetic acid the more tolerant vaginal strains exhibited a higher activity of the plasma membrane proton pump CgPma1 and a reduced internal accumulation of the acid, these being two essential features to maximize tolerance. Based on the higher level of resistance exhibited by the vaginal strains against the action of a β-1,3-glucanase, it is hypothesized that the reduced internal accumulation of acetic acid inside these strains may originate from them having a different cell wall structure resulting in a reduced porosity to undissociated acetic acid molecules. Both the vaginal and the two laboratory strains were found to consume acetic acid in the presence of glucose indicating that metabolization of the acid is used by C. glabrata species as a detoxification mechanism. The results gathered in this study advance the current knowledge on the mechanisms underlying the increased competitiveness of C. glabrata in the vaginal tract, a knowledge that can be used to guide more suitable strategies to treat infections caused by this pathogenic yeast.


Histamine stimulates phosphorylation of adherens junction proteins and alters their link to vimentin.
Histamine increases microvascular permeability by creating small transitory (100-400 nm) gaps between adjacent endothelial cells at sites of vascular endothelial (VE)-cadherin-based adhesion. We examined the effects of histamine on the proteins within the VE-cadherin-based adherens junction in primary human umbilical vein endothelial cells. VE-cadherin is linked not only by beta- and alpha-catenin to cortical actin but also by gamma-catenin to the intermediate filament vimentin. In mature human umbilical vein cultures, the VE-cadherin immunoprecipitate contained equivalent amounts of alpha- and beta-catenin, 130% as much beta- as gamma-catenin, and 50% as much actin as vimentin. Within 60 s, histamine decreased the fraction of VE-cadherin in the insoluble portion of the cell lysate by 35 +/- 1.5%. At the same time, histamine decreased the amount of vimentin that immunoprecipitated with VE-cadherin by 50 +/- 6%. Histamine did not affect the amount of actin or the amount of alpha-, beta-, or gamma-catenin that immunoprecipitated with VE-cadherin. Within 60 s, histamine simulated a doubling in the phosphorylation of VE-cadherin and beta- and gamma-catenin. The VE-cadherin immunoprecipitate contained kinase activity that phosphorylated VE-cadherin and gamma-catenin in vitro.


Classical Hodgkin lymphoma occurring in clusters of nodal marginal zone B-lymphocytes in association with progressive transformation of germinal center. A case report.
We present a case of a classical Hodgkin lymphoma occurring in clusters of marginal zone B-lymphocytes (MZBLs). Most lymphoid follicles possessed hyperplastic germinal centers, while a portion of the follicles exhibited a progressive transformation of the germinal center (PTGC). Clusters of MZBLs showed a perifollicular distribution. The classic Reed-Sternberg cells were found in clusters of MZBLs. A portion of the Reed-Sternberg cells were CD15+, CD20+, CD30+, CD79a+, fascin+, vimentin+, EMA-, and bcl-2-. Some Reed-Sternberg cells were surrounded by CD3+ CD45RO+ CD57-rosettes. In situ hybridization studies demonstrated strong expression of EBER in classic Reed-Sternberg cells and their variants. The overall morphological, immunohistological, and EBV findings confirmed that the present case is a classical Hodgkin lymphoma. The MZBLs were CD20+, CD79a+, sIgM+/-, sIgD-, CD5-, CD21-, CD43-, CD45RO-, and Bcl-2-. Some MZBLs had polytypic intracytoplasmic immunoglobulin. Problems arising in the differential diagnosis between lymphocyte-predominant Hodgkin lymphoma and PTGC have been described. An occasional association between MZBLs clusters and PTGC has been reported previously. This case suggests that classical Hodgkin lymphoma should be added to the differential diagnosis of PTGC.


[Determination of curcumin in urine by capillary electrophoresis].
The major component of the plant curcuma longa (a widely cultivated tropical plant in Asia and Central America) is curcumin. Curcumin has been reported to have very strong anti-inflammatory, anti-carcinogenic, anti-oxidant, antiallergic, anti-bacterial, and anti-tumor activities. Little is known about the absorption, distribution, and metabolism of curcumin in human beings. The first step in in vivo physiological and pharmacokinetic studies is to develop a method to measure curcumin in body fluid. A rapid capillary electrophoretic (CE) method with diode array detection was established for the determination of curcumin in human urine. It could be rapidly determined within 2.5 min. The optimized experimental conditions were as follows: 15 mmol/L Na2B4O7 as buffer, applied voltage 20 kV, temperature 25 degrees C and detection wavelength 262 nm. The method has been validated and shows good performance with respect to selectivity, reproducibility and limit of detection. Curcumin had good linearity in the range of 10 - 300 mg/L, and the recoveries of curcumin added in urine were more than 96.3% with relative standard deviations (RSDs) less than 2.3%. The method is sensitive, fast and accurate and can be used to determine curcumin in urine.


A factor analytic study of the Italian National Institute of Health Quality of Life - Core Evaluation Form (ISSQoL-CEF).
The Italian National Institute of Health Quality of Life - Core Evaluation Form (ISSQoL-CEF) is a specific questionnaire measuring health-related quality of life for human immunodeficiency virus-infected people in the era of highly active antiretroviral therapy. The main goal of this study was to examine the construct validity of this questionnaire by confirmation of its hypothesized dimensional structure. Baseline quality of life data from four clinical studies were collected and a confirmatory factor analysis of the ISSQoL-CEF items was carried out. Both first-order and second-order factor models were tested: Model 1 with nine correlated first-order factors; Model 2 with three correlated second-order factors (Physical, Mental, and Social Health); Model 3 with two correlated second-order factors (Physical and Mental/Social Health); Model 4 with only one second-order factor (General Health). A total of 261 patients were surveyed. Model 1 had a good fit to the data. Model 2 had an acceptable fit to the data and it was the best of all hierarchical models. However, Model 2 fitted the data worse than Model 1. The findings of in this study, consistent with the results of previous study, pointed out the construct validity of the ISSQoL-CEF.


Overexpression of osteoactivin protects skeletal muscle from severe degeneration caused by long-term denervation in mice.
We have previously shown that osteoactivin, a type I membrane glycoprotein expressed in myofibers, upregulated expression of matrix metalloprotease (MMP)-3 and MMP-9 in fibroblasts infiltrated denervated skeletal muscle in mice. To address whether osteoactivin-mediated increase in MMPs in skeletal muscle is useful for regeneration of denervated skeletal muscle, we subjected osteoactivin-transgenic mice to long-term denervation for 70 or 90 days. Long-term denervation caused severe degeneration of myofibers and fibrosis in skeletal muscle of wild-type mice. However, overexpression of osteoactivin protected skeletal muscle from such changes. Infiltration of fibroblast-like cells and collagen deposition were sustained at low levels after long-term denervation in skeletal muscle of osteoactivin-transgenic mice. This cytoprotective effect of osteoactivin was supported by the expression of regeneration/degeneration-associated genes in the gastrocnemius muscle during denervation. Denervation significantly upregulated the expression of anti-fibrotic genes, such as glypican-1 and decorin-1, in the gastrocnemius muscle of osteoactivin-transgenic mice, compared with wild-type mice. In contrast, overexpression of osteoactivin caused a significant reduction in denervation-induced expression of elongation factor 1A-1, an indicator for the persistence of degenerated cells. Our results suggest that an osteoactivin-mediated increase in MMPs in skeletal muscle might be useful for protecting injured muscle from fibrosis, leading to full regeneration after denervation.


IgE and FcepsilonRI regulation.
The high-affinity immunoglobulin (Ig)E receptor, FcepsilonRI, regulates the action of mast cells and basophils and therefore, regulates the expression of atopic disease. There have been several recent observations that demonstrate new behaviors for this receptor. The control of FcepsilonRI expression, control of cell function by FcepsilonRI, and expression of FcepsilonRI on other cell types are important new areas of understanding currently being explored.


Dual TCR T Cells: Identity Crisis or Multitaskers?
Dual TCR T cells are a common and natural product of TCR gene rearrangement and thymocyte development. As much as one third of the T cell population may have the capability to express two different TCR specificities on the cell surface. This discovery provoked a reconsideration of the classic model of thymic selection. Many potential roles for dual TCR T cells have since been hypothesized, including posing an autoimmune hazard, dominating alloreactive T cell responses, inducing allergy, and expanding the TCR repertoire to improve protective immunity. Yet, since the initial wave of publications following the discovery of dual TCR T cells, research in the area has slowed. In this study, we aim to provide a brief but comprehensive history of dual TCR T cell research, re-evaluate past observations in the context of current knowledge of the immune system, and identify key issues for future study.


Video-assisted anal fistula treatment.
Fistula in ano is a common disease seen in the surgical outpatient department. Many procedures are advocated for the treatment of fistula in ano. However, none of the procedures is considered the gold standard. The latest addition to the list of treatment options is video-assisted anal fistula treatment (VAAFT). It is a minimally invasive, sphincter-saving procedure with low morbidity. The aim of our study was to compare the results with a premier study done previously. The procedure involves diagnostic fistuloscopy and visualization of the internal opening, followed by fulguration of the fistulous tract and closure of the internal opening with a stapling device or suture ligation. The video equipment (Karl Storz, Tuttlingen, Germany) was connected to an illuminating source. The study was conducted from July 2010 to March 2014. Eighty-two patients with fistula in ano were operated on with VAAFT and were followed up according to the study protocol. The recurrence rate was 15.85%, with recurrences developing in 13 cases. Postoperative pain and discomfort were minimal. VAAFT is a minimally invasive procedure performed under direct visualization. It enables visualization of the internal opening and secondary branches or abscess cavities. It is a sphincter-saving procedure and offers many advantages to patients. Our initial results with the procedure are quite encouraging.


[A case of amyotrophic lateral sclerosis presenting as motor aphasia in its early stage].
A 76-year-old right-handed woman complained of speech disturbance and difficulity of singing was admitted to our hospital. Examination showed motor aphasia and mild cognitive impairment. After she was discharged, dementia and weakness of the extremities had rapidly progressed. She was readmitted eight month after the first visit, when she was almost abulic, her skeletal and bulbar muscles were remarkably atrophic, and hyperreflexia of the extremities were seen. Electromyographcal study showed neurogenic pattern. These findings suggest amyotrophic lateral sclerosis (ALS) with dementia. Pathological findings were atrophy at the anterior horn of the spinal cord. The brain was diffusely atrophic. The extent of degenerative change was not lateralized. This case is a discriminative type of ALS with dementia, that its first symptom is motor aphasia.


Autosomal Dominantly Inherited GREB1L Variants in Individuals with Profound Sensorineural Hearing Impairment.
Congenital hearing impairment is a sensory disorder that is genetically highly heterogeneous. By performing exome sequencing in two families with congenital nonsyndromic profound sensorineural hearing loss (SNHL), we identified autosomal dominantly inherited missense variants [p.(Asn283Ser); p.(Thr116Ile)] in GREB1L, a neural crest regulatory molecule. The p.(Thr116Ile) variant was also associated with bilateral cochlear aplasia and cochlear nerve aplasia upon temporal bone imaging, an ultra-rare phenotype previously seen in patients with de novo GREB1L variants. An important role of GREB1L in normal ear development has also been demonstrated by greb1l-/- zebrafish, which show an abnormal sensory epithelia innervation. Last, we performed a review of all disease-associated variation described in GREB1L, as it has also been implicated in renal, bladder and genital malformations. We show that the spectrum of features associated with GREB1L is broad, variable and with a high level of reduced penetrance, which is typically characteristic of neurocristopathies. So far, seven GREB1L variants (14%) have been associated with ear-related abnormalities. In conclusion, these results show that autosomal dominantly inherited variants in GREB1L cause profound SNHL. Furthermore, we provide an overview of the phenotypic spectrum associated with GREB1L variants and strengthen the evidence of the involvement of GREB1L in human hearing.


The role of the NF-κB, SAPK/JNK, and TLR4 signalling pathways in the responses of RAW 264.7 cells to extremely low-intensity microwaves.
To investigate the role of the toll-like receptor 4 (TLR4), nuclear factor κB (NF-κB), and stress activated protein kinases/Jun N-terminal kinase (SAPK/JNK) signalling pathways in the responses of RAW 264.7 macrophages to low-intensity microwaves (MW). Three inhibitors of TLR4, SAPK/JNK, and NF-κB signalling, namely CLI-095, SP600125, and IKK Inhibitor XII, respectively, were added to cultured RAW 264.7 macrophages before MW treatment. MW exposure resulted in stimulation of RAW 264.7 cell activity manifested by increases in cytokine production and the stimulation of cell signalling. The blocking of a key kinase of the NF-κB pathway by IKK Inhibitor XII resulted in decreased MW-induced TLR4 expression and increased SAPK/JNK and NF-κB phosphorylation in irradiated cells. In addition, IKK Inhibitor XII significantly decreased tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin 1α (IL-1α), interleukin 6 (IL-6), and interleukin 10 (IL-10) production in both exposed and unexposed RAW 264.7 macrophages. Inhibitor SP6000125 did not prevent an MW effect on signal proteins with the exception of decreased SAPK/JNK phosphorylation in RAW 264.7 cells. Cytokine production was markedly decreased in MW-exposed cells cultured with SP6000125. The inhibitor of TLR4, CLI-095, did not affect signal proteins and cytokine production changes in MW-exposed cells. The results suggest that low-intensity MW promotes macrophage activity via mechanisms involving cellular signalling, particularly the NF-κB pathway.


The paralytic shellfish toxin, saxitoxin, enters the cytoplasm and induces apoptosis of oyster immune cells through a caspase-dependent pathway.
Exposure of the toxin-producing dinoflagellate Alexandrium catenella (A. catenella) was previously demonstrated to cause apoptosis of hemocytes in the oyster species Crassostrea gigas. In this work, a coumarin-labeled saxitoxin appeared to spread throughout the cytoplasm of the hemocytes. PSTs, including saxitoxin, were also shown to be directly responsible for inducing apoptosis in hemocytes, a process dependent on caspase activation and independent of reactive oxygen species (ROS) production. A series of in vitro labelling and microscopy experiments revealed that STX and analogs there of induced nuclear condensation, phosphatidylserine exposure, membrane permeability, and DNA fragmentation of hemocytes. Unlike in vertebrates, gonyautoxin-5 (GTX5), which is present in high concentrations in A. catenella, was found to be more toxic than saxitoxin (STX) to oyster immune cells. Altogether, results show that PSTs produced by toxic dinoflagellates enter the cytoplasm and induce apoptosis of oyster immune cells through a caspase-dependent pathway. Because of the central role of hemocytes in mollusc immune defense, PST-induced death of hemocytes could negatively affect resistance of bivalve molluscs to microbial infection.


Pasteurella multocida ecthyma complicated by necrotizing fasciitis.
Necrotizing fasciitis is a serious infection of the skin and soft tissues. Pasteurella multocida is rarely reported to cause necrotizing fasciitis and is associated with high mortality. We describe a female patient with a past medical history of diabetes mellitus and myeloproliferative disorder presenting with bullae and erythema of the right forearm secondary to P. multocida infection after possible cat bite. Despite adequate antibiotic coverage she developed necrotizing fasciitis diagnosed clinically and on diagnostic imaging. Patient was taken to the operating room emergently and underwent irrigation and debridement with subsequent split-skin graft. She recovered well after the surgeries and was discharge on intravenous antibiotics. At clinic follow-up, her wounds were healing well without any significant new symptoms.


Constitutive non-inducible expression of the Arabidopsis thaliana Nia 2 gene in two nitrate reductase mutants of Nicotiana plumbaginifolia.
We have isolated a haploid cell line of N. plumbaginifolia, hNP 588, that is constitutive and not inducible for nitrate reductase. Nitrate reductase mutants were isolated from hNP 588 protoplasts upon UV irradiation. Two of these nitrate reductase-deficient cell lines, nia 3 and nia 25, neither of which contained any detectable nitrate reductase activity, were selected for complementation studies. A cloned Arabidopsis thaliana nitrate reductase gene Nia 2 was introduced into each of the two mutants resulting in 56 independent kanamycin-resistant cell lines. Thirty of the 56 kanamycin-resistant cell lines were able to grow on nitrate as the sole nitrogen source. Eight of these were further analyzed for nitrate reductase enzyme activity and nitrate reductase mRNA production. All eight lines had detectable nitrate reductase activity ranging from 7% to 150% of wild-type hNP 588 callus. The enzyme activity levels were not influenced by the nitrogen source in the medium. The eight lines examined expressed a constitutive, non-inducible 3.2 kb mRNA species that was not present in untransformed controls.


Ultraviolet-durable superhydrophobic zinc oxide-coated mesh films for surface and underwater-oil capture and transportation.
In this paper, we report a simple and an inexpensive method for fabricating superhydrophobic/superoleophilic mesh films from microstructured ZnO coatings. The microstructured ZnO coatings, which do not contain any fluorinated compounds, maintain their superhydrophobicity and superoleophilicity after ultraviolet irradiation and display environmental stability. Furthermore, those microstructured ZnO-coated mesh films exhibit good selectivity (even underwater) and excellent recyclability, making them promising candidates for many potential applications, including liquid-liquid separation, water treatment, and liquid transportation.


Anti-Angiogenics: Their Value in Colorectal Cancer Therapy.
Angiogenesis is a hallmark of cancer and is regulated by a balance of pro- and anti-angiogenic factors; among them, the vascular endothelial growth factor (VEGF) is the key angiogenic factor. VEGF plays an important role in colorectal cancer (CRC) biology, and its inhibition by using bevacizumab, an anti-VEGF antibody, proved for the first time to be effective and became indispensable for the treatment of metastatic CRC (mCRC). Several large phase III studies showed also relevant responses and tolerability of other anti-angiogenic drugs such as ramucirumab, aflibercept, and regorafenib, and led to the approval of these therapeutics. Nevertheless, the efficacy of anti-angiogenic therapies is rather limited and the high expectations raised by preclinical studies were not fulfilled in the clinics. Furthermore, to date, no predictive biomarkers for anti-angiogenic agents could be identified and validated. Thus, new mechanisms of action are discussed, such as tumor vasculature normalization to improve the accessibility of tumor tissue by drugs or to promote tumor infiltration by host immune cells. Cellular and molecular studies will be necessary to characterize the dynamic changes of the tumor microenvironment and the vascular architecture in individual patients in order to predict responsiveness to anti-angiogenic therapies. In this review, we tried to highlight the standard of care of using anti-angiogenics in mCRC patients and to provide an outlook on potential new substances and strategies.


The role of the tracheae and musculature during pathfinding of Drosophila embryonic sensory axons.
Developing sensory axons were studied in Drosophila embryos which carried a mutation in the trachealess and/or the twist gene. In these embryos, the tracheae and/or somatic muscles, which represent part of the substrate on which sensory axons normally grow, are absent. The results demonstrate that in each of these three mutant backgrounds, the majority of sensory nerves form normally. This indicates that neither the tracheae nor the somatic musculature is absolutely required for pathfinding of the embryonic sensory axons. On the other hand, the incidence of misrouted axons is significantly increased, most strongly in the trh, twi double mutant. Furthermore, axonal elongation is considerably slowed down, and sensory neurons which fail to send out an axon are frequent. We take our results to indicate that peripheral axons in the Drosophila embryo may be guided by multiple cues which, acting together, ensure the high fidelity of axonal pathways observed in normal development. The removal of one of these cues by itself does not necessarily lead to the total disability of axons to reach their target, although it enhances the frequency of error in pathfinding.