The Success of Treatment Free Remission in Chronic Myeloid Leukaemia in Clinical Practice: A Single-Centre Retrospective Experience from South Africa.
Chronic myeloid leukaemia (CML) management has evolved from a disease once considered to be incurable just over 2 decades ago to that of one of a "functional cure" as defined by the sustained molecular response on stopping tyrosine kinase inhibitor(TKI) therapy. The next goal of CML management has been treatment-free remission (TFR). The past 4 years have seen much international data on TFR attempts in CML in clinical practice. However, Africa as a continent has lagged behind the rest of the world, in keeping up with the latest trends in CML management, and so this study aims to address this gap by assessing the outcome of TFR in CML in a single centre in South Africa (SA). We conducted a retrospective cohort study in 12 CML patients in the chronic phase to assess the success of TKI discontinuation. The patients were treated in King Edward VIII Hospital (KEH), a tertiary, academic hospital in KwaZulu-Natal, South Africa, and the study period was from June 2020 to May 2022. Patients included had to have been on TKI therapy for a minimum of 5 years and achieved a deep molecular response (DMR) for a minimum period of 3 years. The overall TFR cohort showed a success rate of 75% at a median follow-up of 12 months. All patients who failed TFR, defined as a loss of major molecular remission (MMR), failed within 6 months of stopping TKI therapy. All patients who failed TFR regained DMR after retreatment with TKI, with no disease progression reported. The only factor influencing the success of TFR was the total period of TKI therapy. Despite our study having a small cohort of patients, this study demonstrated that TFR in CML is an attainable goal, even in a resource-limited setting.


"Influential" Intraoperative Educators and Variability of Teaching Styles.
Academic surgeons manage their role as intraoperative educators in a variety of ways. Such variability is neither idiosyncratic nor is there a single best approach. This study sought to explore the practices of surgeons deemed influential by their residents, allowing insight into a variety of potentially effective practices. Constructivist grounded theory guided data collection and analysis. Data sources included surveys from senior surgical residents (PGY3-6) and recent graduates from an academic hospital in Canada (36% response rate), intraoperative observations of teaching interactions, and semi-structured interviews with observed surgeons. Rigour was supported by data triangulation, constant comparison, and collection to theoretical sufficiency. We developed a framework grouping effective teaching into three overlapping approaches: exacting, empowering, and fostering. The approaches differ based on the level of independence granted and the degree of expectation placed on individual residents. Each demonstrates different strategies for balancing the multiple supervisory roles and patient care obligations faced by academic surgeons. We also identified strategies that could be used across approaches to enhance learning. For surgical educators seeking to improve upon the quality of the intraoperative supervision they provide, frameworks such as this may serve as models of effective supervision. Enhancing surgeons' knowledge of proven strategies, combined with reflecting on how they teach and how they balance responsibilities to patients and trainees, may allow them to broaden their educational practice.


Introduction of a nitroxide group on position 2 of 9-phenoxyacridine: easy access to spin labelled DNA-binding conjugates.
In the search for spin labelled intercalators of general use to construct DNA-binding conjugates, 6-chloro-2-[(1-oxyl-2,2,5,5-tetramethyl- pyrrolin-3-yl)methyloxy]-9-phenoxy-acridine 5, has been prepared. This key-intermediate reacts with amines to give the corresponding labelled 9-amino substituted acridines. Comparative EPR and fluorescence measurements show that the label causes only little modification of the binding properties of acridine.


Magnifying endoscopy, stereoscopic microscopy, and the microvascular architecture of superficial esophageal carcinoma.
In this study we clarify the microvascular architecture of superficial esophageal carcinoma as observed by ultra-high magnification endoscopy and stereoscopic microscopy with Microfil injection. We observed two surgically resected specimens of superficial esophageal cancer under stereoscopic microscopy with Microfil injection. In addition, in the histological investigation, we measured the caliber of the vessels at the surface of the tumor. We carried out ultra-high magnification before treatment in 82 patients with superficial esophageal neoplasms. We classified the depth of tumor penetration of superficial esophageal carcinoma into four categories: m1 to m3 (mucosal cancer) and sm (submucosal cancer). By observing the normal esophageal mucosa under a stereoscopic microscope and an ultra-high magnification endoscope, we were able to visualize the intrapapillary capillary loops (IPCL). In cancer lesions, we observed characteristic changes in the superficial microvascular architecture according to the depth of tumor invasion. In m1 invasion, there was dilatation of the IPCL; in m2 invasion, there was dilatation and elongation of the IPCL; in m3, there was a mixed appearance of the IPCL and tumor vessels; and in sm invasion, complete replacement by tumor vessels. On the basis of the above criteria, ultra-high magnification endoscopic observation before treatment showed a rate of agreement between histological depth of invasion and magnified appearance of 60/72 cases (83.3 %) for which satisfactory pictures were obtained. The histological investigation showed the caliber of the IPCL of the m1 cancer lesions (12.9 +/- 3.9 microm) to be significantly greater than that of the normal esophageal mucosa (6.9 +/- 1.5 microm) (P < 0.0001). Observation of the microvascular architecture of superficial esophageal carcinoma is useful in the diagnosis of the depth of invasion.


Transfusion and surgery.
In this survey of transfusion in surgery, we have attempted to provide the surgeon with an understanding of the problems associated with homologous transfusion and a practical knowledge of treatment strategies and alternatives designed to reduce homologous blood exposure. Such a review cannot be encyclopedic. Our hope is that it will serve the reader as a stimulus to examine his or her transfusion practices and as a guide for future self-learning.


Neuroanatomical development in the absence of PKC phosphorylation of the myristoylated alanine-rich C-kinase substrate (MARCKS) protein.
The myristoylated alanine-rich C-kinase substrate protein (MARCKS) is a widely expressed target of protein kinase C (PKC) phosphorylation. Disruption of Marcks in mice leads to a number of developmental defects within the central nervous system that are completely prevented by expression of an epitope-tagged wild-type human MARCKS transgene. In the present study, we investigated whether PKC phosphorylation of MARCKS is necessary for normal central nervous system development and postnatal survival. Expression at approximately twice normal levels of a mutant MARCKS protein in which the four PKC phosphorylatable serines were replaced by asparagines did not allow postnatal survival of Marcks(-/-) pups. Nonetheless, the rescued animals exhibited none of the characteristic anatomical defects seen in the brains and retinas of knockout mice, suggesting that PKC phosphorylation of MARCKS is not required for normal central nervous system development. Expression studies showed that transgene expression was limited to the central nervous system, which has implications for the lack of postnatal survival as well as for the pathogenesis of the neuronal ectopia characteristic of MARCKS deficiency. A novel aspect of the MARCKS-deficient phenotype was also noted, absence of the pontine nuclei; this was also largely reversed in Marcks(-/-) animals expressing the mutant transgene. These data raise the possibility of a role for MARCKS in the netrin-regulated process of pontine nuclei formation.


An inner-city school-based program to promote early awareness of risk factors for Sudden Infant Death Syndrome.
Adolescent, nonwhite women with less than high school education have infants at higher risk for Sudden Infant Death Syndrome (SIDS) but face barriers to risk reduction education. We implemented a novel school-based health education program (grades 4 to 12) and found an association between exposure and awareness of risk factors.


The periodic table and other wallcharts in the teaching of chemistry in St Andrews, 1884-1919.
The teaching of the chemistry of the elements at the University of St Andrews by Professor Thomas Purdie FRS is examined with reference to selections from a collection of recently discovered historic wallcharts and extracts from his detailed hand-written lecture notes. Together these reveal a comprehensive and exciting programme of lectures incorporating numerous practical demonstrations which were continually updated to reflect the latest state of knowledge in what was a rapidly changing field. This article is part of the theme issue 'Mendeleev and the periodic table'.


Engineering controllable oxygen vacancy defects in iron hydroxide oxide immobilized on reduced graphene oxide for boosting visible light-driven photo-Fenton-like oxidation.
Visible light-driven photo-Fenton-like technology is a promising advanced oxidation process for water remediation, while the construction of effective synergetic system remains a great challenge. Herein, iron hydroxide oxide (α-FeOOH) with controllable oxygen vacancy defects were engineered on reduced graphene oxide (rGO) nanosheets (named as OVs-FeOOH/rGO) through an in-situ redox method for boosting visible light-driven photo-Fenton-like oxidation. By adjusting the pH environment to modulate the redox reaction kinetics between graphene oxide (GO) and ferrous salt precursors, the oxygen vacancy concentration in α-FeOOH could be precisely controlled. With optimized oxygen vacancy defects obtained at pH 5, the OVs-FeOOH/rGO displayed superior photo-Fenton-like performance for Rhodamine B degradation (99% within 40 mins, rate constant of 0.2278 mg-1 L min-1) with low H2O2 dosage (5 mM), standing out among the reported photo-Fenton-like catalysts. The catalyst also showed excellent reusability, general applicability, and tolerance ability of realistic environmental conditions, which demonstrates great potential for practical applications. The results reveal that moderate oxygen vacancy defects can not only strengthen absorption of visible light and organic pollutants, but also promote the charge transfer to simultaneously accelerate the photogenerated electron-hole separation and Fe(III)/Fe(II) Fenton cycle, leading to the remarkable photo-Fenton-like oxidation performance. This work sheds light on the controllable synthesis and mechanism of oxygen vacancy defects to develop efficient photo-Fenton-like catalysts for wastewater treatment.


Setting research priorities to improve cost-effectiveness estimations of primary prophylaxis with clotting factor for people with severe haemophilia.
Primary prophylaxis with clotting factor is the clinical treatment of choice for people with severe haemophilia, as evidence suggests it can prevent the onset and progression of joint- and muscle-related problems caused by bleeding episodes. However, the major limitation of this approach is that it requires considerably more clotting factor compared with treating on-demand. Thus, there is a need to establish its cost-effectiveness. The aim of this paper is to review the published evidence on the cost-effectiveness of prophylaxis and to highlight areas for future research that would decrease the uncertainty around these findings.


Lactococcus lactis subsp. lactis JCM 5805 activates natural killer cells via dendritic cells.
Lactococcus lactis subsp. lactis JCM 5805 (JCM5805) has been shown to stimulate plasmacytoid dendritic cells (pDC). Here, we investigated the effect of JCM5805 on NK cells. In vitro studies suggested that JCM5805 activated natural killer (NK) cells via dendritic cells including pDC. Furthermore, the oral administration of JCM5805 enhanced the cytotoxic activity of NK cells.


Diagonal band of Broca modulates the cardiac component of the baroreflex in unanesthetized rats.
The diagonal band of Broca (DBB) is involved in cardiovascular control in rats. In the present study, we report the effect of acute and reversible neurotransmission inhibition in the DBB by bilateral microinjection of the nonselective neurotransmission blocker CoCl(2) (1mM, 100 nL) on the cardiac baroreflex response in unanesthetized rats. Local DBB neurotransmission inhibition did not affect baseline values of either blood pressure or heart rate, suggesting no tonic DBB influence on cardiovascular system activity. However, CoCl(2) microinjections enhanced both the reflex bradycardia associated with blood pressure increases caused by i.v. infusion of phenylephrine and tachycardiac response evoked by blood pressure decreases caused by i.v. infusion of sodium nitroprusside. An increase in baroreflex gain was also observed. Baroreflex returned to control values 60 min after CoCl(2) microinjections, confirming its reversible effect. In conclusion, our data suggest that synapses within DBB have a tonic inhibitory influence on both the cardiac parasympathetic and sympathetic components of the baroreflex.


Minimally invasive treatment of retrocardiac tension pneumomediastinum in an extremely low birth weight infant.
A 23.4-week premature and extremely low birth weight neonate was transferred to the Neonatal Intensive Care Unit for management of respiratory failure and retrocardiac pneumomediastinum, suspected to be the result of a low tracheal injury during intubation. Initial conservative management failed and chest radiographs demonstrated worsening retrocardiac pneumomediastinum. Due to the patient's extreme low birth weight and location of the pneumomediastinum, surgery was deemed to be very high risk and potentially fatal. We report the successful definitive percutaneous management of retrocardiac tension pneumomediastinum with a minimally invasive bedside method using an intercostal paraspinal approach under the guidance of ultrasound and plain radiography.


Combined distal tibial rotational osteotomy and proximal growth plate modulation for treatment of infantile Blount's disease.
Infantile Blount's disease is a condition that causes genu varum and internal tibial torsion. Treatment options include observation, orthotics, corrective osteotomy, elevation of the medial tibial plateau, resection of a physeal bar, lateral hemi-epiphysiodesis, and guided growth of the proximal tibial physis. Each of these treatment options has its disadvantages. Treating the coronal deformity alone (genu varum) will result in persistence of the internal tibial torsion (the axial deformity). In this report, we describe the combination of lateral growth modulation and distal tibial external rotation osteotomy to correct all the elements of the disease. This has not been described before for treatment of Blount's disease. Both coronal and axial deformities were corrected in this patient. We propose this combination (rather than the lateral growth modulation alone) as the method of treatment for early stages of Blount's disease as it corrects both elements of the disease and in the same time avoids the complications of proximal tibial osteotomy.


Science based guidance for the assessment of endocrine disrupting properties of chemicals.
The European legislation on plant protection products (Regulation (EC) No. 1107/2009) and biocides (Directive 98/8/EC), as well as the regulation concerning chemicals (Regulation (EC) No. 1907/2006 'REACH') only support the marketing and use of chemical products on the basis that they do not induce endocrine disruption in humans or non-target species. However, there is currently no agreed guidance on how to identify and evaluate endocrine activity and disruption. Consequently, an ECETOC task force was formed to provide scientific criteria that may be used within the context of these three legislative documents. Specific scientific criteria for the determination of endocrine disrupting properties that integrate information from both regulatory (eco)toxicity studies and mechanistic/screening studies are proposed. These criteria combine the nature of the adverse effects detected in studies which give concern for endocrine toxicity with an understanding of the mode of action of toxicity so that adverse effects can be explained scientifically. The criteria developed are presented in the form of flow charts for assessing relevant effects for both humans and wildlife species. In addition, since not all chemicals with endocrine disrupting properties are of equal hazard, assessment of potency is also proposed to discriminate chemicals of high concern from those of lower concern. The guidance presented in this paper includes refinements made to an initial proposal following discussion of the criteria at a workshop of invited regulatory, academic and industry scientists.


Transient deficiency of peripheral blood accessory cells in supporting T cell mitogenesis in patients suffering from chronic idiopathic thrombocytopenic purpura after intravenous gammaglobulin treatment.
Mitogenic response of blood lymphocytes to phytohemagglutinin (PHA) and to OKT3 monoclonal antibodies was investigated in 7 patients suffering from chronic idiopathic thrombocytopenic purpura (ITP) before, during and after high-dose intravenous (i.v.) immunogammaglobulin (IgG) infusion. The platelet count rose above the pre-treatment values during infusion therapy in all patients but one. Five out of seven patients presented elevated platelet-associated IgG (PA-IgG) levels at the time of the first infusion; four of these showed an increase in platelet count and a transient reduction or normalization of PA-IgG after IgG infusion. Five out of seven patients showed an impairment of T lymphocyte mitogenic response to PHA and OKT3 before therapy. All patients responded to IgG therapy with a transient deficiency of FcR mediated monocytes (Mo) in supporting T cell mitogenesis induced by both mitogens during and after IgG infusion. This reduced cooperative capability of Mo disappeared at various times after the end of therapy (range 3-12 days). The transient alteration of Mo function, possibly due to a modification in the surface number or in the affinity of Fc-receptors, can explain in part, the increase in platelet count during and after IgSRK infusion.


Functional assay of cytotoxic lymphocytes involved in antibody-mediated cytotoxicity in normal and rheumatoid subjects.
Antibody-mediated lymphocyte-induced cytotoxicity (K cell activity) was measured in a system consisting of Chang human liver cells, rabbit anti-Chang antiserum, and Triosil-Ficoll purified human lymphocytes. It was concluded that one K cell probably inactivates or kills one target cell, and that the number of active K cells is similar in the peripheral blood of normal and rheumatoid subjects and is about 0-5%.


Baseline IL-6 is a biomarker for unfavourable tuberculosis treatment outcomes: a multisite discovery and validation study.
Biomarkers of unfavourable tuberculosis (TB) treatment outcomes are needed to accelerate new drug and regimen development. Whether plasma cytokine levels can predict unfavourable TB treatment outcomes is unclear. We identified and internally validated the association between 20 a priori selected plasma inflammatory markers and unfavourable treatment outcomes of failure, recurrence and all-cause mortality among adults with drug-sensitive pulmonary TB in India. We externally validated these findings in two independent cohorts of predominantly diabetic and HIV co-infected TB patients in India and South Africa, respectively. Pre-treatment interferon-γ, interleukin (IL)-13 and IL-6 were associated with treatment failure in the discovery analysis. Internal validation confirmed higher pre-treatment IL-6 concentrations among failure cases compared with controls. External validation among predominantly diabetic TB patients found an association between pre-treatment IL-6 concentrations and subsequent recurrence and death. Similarly, external validation among predominantly HIV co-infected TB patients found an association between pre-treatment IL-6 concentrations and subsequent treatment failure and death. In a pooled analysis of 363 TB cases from the Indian and South African validation cohorts, high pre-treatment IL-6 concentrations were associated with higher risk of failure (adjusted OR (aOR) 2.16, 95% CI 1.08-4.33; p=0.02), recurrence (aOR 5.36, 95% CI 2.48-11.57; p<0.001) and death (aOR 4.62, 95% CI 1.95-10.95; p<0.001). Adding baseline IL-6 to a risk prediction model comprised of low body mass index, high smear grade and cavitation improved model performance by 15% (C-statistic 0.66 versus 0.76; p=0.02). Pre-treatment IL-6 is a biomarker for unfavourable TB treatment outcomes. Future studies should identify optimal IL-6 concentrations for point-of-care risk prediction.


Plasmonic Coupling on an Optical Microfiber Surface: Enabling Single-Molecule and Noninvasive Dopamine Detection.
Optical fibers can be effective biosensors when employed in early-stage diagnostic point-of-care devices as they can avoid interference from molecules with similar redox potentials. Nevertheless, their sensitivity needs to be improved for real-world applications, especially for small-molecule detection. This work demonstrates an optical microfiber biosensor for dopamine (DA) detection based on the DA-binding-induced aptamer conformational transitions that occur at plasmonic coupling sites on a double-amplified nanointerface. The sensor exhibits ultrahigh sensitivity when detecting DA molecules at the single-molecule level; additionally, this work provides an approach for overcoming optical device sensitivity limits, further extending optical fiber single-molecule detection to a small molecule range (e.g., DA and metal ions). The selective energy enhancement and signal amplification at the binding sites effectively avoid nonspecific amplification of the whole fiber surface which may lead to false-positive results. The sensor can detect single-molecule DA signals in body-fluids. It can detect the released extracellular DA levels and monitor the DA oxidation process. An appropriate aptamer replacement allows the sensor to be used for the detection of other target small molecules and ions at the single-molecule level. This technology offers alternative opportunities for developing noninvasive early-stage diagnostic point-of-care devices and flexible single-molecule detection techniques in theoretical research.


Oral epithelial cell segmentation from fluorescent multichannel cytology images using deep learning.
Cytology is a proven, minimally-invasive cancer screening and surveillance strategy. Given the high incidence of oral cancer globally, there is a need to develop a point-of-care, automated, cytology-based screening tool. Oral cytology image analysis has multiple challenges such as, presence of debris, blood cells, artefacts, and clustered cells, which necessitate a skilled expertise for single-cell detection of atypical cells for diagnosis. The main objective of this study is to develop a semantic segmentation model for Single Epithelial Cell (SEC) separation from fluorescent, multichannel, microscopic oral cytology images and classify the segmented images. We have used multi-channel, fluorescent, microscopic images (number of images; n = 2730), which were stained differentially for cytoplasm and nucleus. The cytoplasmic and cell membrane markers used in the study were Mackia Amurensis Agglutinin (MAA; n: 2364) and Sambucus Nigra Agglutinin-1 (SNA-1; n: 366) with a nuclear stain DAPI. The cytology images were labelled for SECs, cluster of cells, artefacts, and blood cells. In this study, we used encoder-decoder models based on the well-established U-Net architecture, modified U-Net and ResNet-34 for multi-class segmentation. The experiments were performed with different class combinations of data to reduce imbalance. The derived MAA dataset (n: 14,706) of SEC, cluster, and artefacts/blood cells were used for developing a classification model. InceptionV3 model and a new custom Convolutional-Neural-Network (CNN) model (Artefact-Net) were trained to classify SNA-1 marker stained segmented images (n:6101). For segmentation models, Intersection Over Union (IoU) and F1 score were used as the evaluation matrices, while the classification models were evaluated using the conventional classification metrics like precision, recall and F1-Score. The U-Net and the modified U-Net models gave the best IoU overall (0.73-0.76) as well as for SEC segmentation (079). The images segmented using the modified U-Net model were classified by Artefact-Net and Inception V3 model with F1 scores of 0.96 and 0.95 respectively. The Artefact-Net, when compared to InceptionV3, provided a better precision and F1 score in classifying clusters (Precision: 0.91 vs 0.80; F1: 0.91 vs 0.86). This study establishes a pipeline for SEC segmentation with the segmented component containing only single cells. The pipline will enable automated, cytology-based early detection with reduced bias.


A comparison of the asthmatic response to methacholine and exercise.
The airway responses to methacholine and to exercise challenges were compared in 45 young adults with asthma. The spirometric response to five minutes of treadmill exercise was first documented. On a separate day methacholine dose-response relationships were determined. All asthmatics had an abnormal response to methacholine, and 36 had an abnormal response to exercise. Methacholine sensitivity and exercise-induced asthma were significantly related (r = 0.69, p less than 0.001), but the relationship was nonlinear; the increased response to exercise related to the logarithm of the methacholine response. Between asthmatics with generally unreactive airways, small variation in methacholine sensitivity was associated with large variations in the severity of exercise-induced asthma; between more responsive asthmatics, there was a smaller effect. It is suggested that exercise-induced asthma is dependent on two factors: a stimulus generated during exercise and a response from abnormal bronchi. The bronchial response may be a limiting factor in asthmatics with less responsive airways.


Combination of copeptin, placental growth factor and total annexin V microparticles for prediction of preeclampsia at 10-14 weeks of gestation.
Preeclampsia (PE) remains to be an enigmatic puzzle for clinicians and researchers perplexing them for decades. As delivery remains only choice of treatment, early prediction of PE will offer timely therapeutic invention and hence extensive research efforts have been put in identification of biomarkers which will facilitate early prediction of PE. Serum levels of CPP, PlGF and plasma total annexin V MPs were assessed in women who subsequently developed PE (n = 33), IUGR (n = 81) and normal pregnancy outcome (n = 112) at 10-14 weeks of gestation. Comparison of biomarker levels between patients and control group was done using Mann Whitney test. Receiver operating curve (ROC) analysis and binary logistic regression analysis were used to evaluate predictive utility of combination of CPP, PlGF and total annexin V MPs for prediction of PE. Women who subsequently developed PE showed significantly elevated levels of total annexin V MPs [2766.04 (2086.88-3794) versus 1090.74 (631.91-2197.16)] and CPP [440.98 (365.12-488.92) versus 217.8 (171.13-308.98)] compared to controls. Serum PlGF levels were significantly reduced in women with PE 17.68 (12.66-22.32) compared to controls 105.22 (35.02-255.1). Using logistic regression analysis, the combination of CPP, PlGF and total annexin V MPs gave high predictive value with AUC of 0.970, 93.1% sensitivity, 90.7% specificity, 77.50% Positive predictive value, 98.10% Negative predictive value, 11.69 Positive likelihood ratio and 0.07 Negative likelihood ratio for PE prediction at 10-14 weeks. The combination of serum markers and plasma microparticles can be used for 10-14 weeks prediction and discrimination of PE from other pregnancy complications.


Continuous deep sedation for patients nearing death in the Netherlands: descriptive study.
To study the practice of continuous deep sedation in 2005 in the Netherlands and compare it with findings from 2001. Questionnaire study about random samples of deaths reported to a central death registry in 2005 and 2001. Nationwide physician study in the Netherlands. Reporting physicians received a questionnaire about the medical decisions that preceded the patient's death; 78% (n=6860) responded in 2005 and 74% (n=5617) in 2001. Characteristics of continuous deep sedation (attending physician, types of patients, drugs used, duration, estimated effect on shortening life, palliative consultation). Requests for euthanasia. The use of continuous deep sedation increased from 5.6% (95% confidence interval 5.0% to 6.2%) of deaths in 2001 to 7.1% (6.5% to 7.6%) in 2005, mostly in patients treated by general practitioners and in those with cancer (in 2005, 47% of sedated patients had cancer v 33% in 2001). In 83% of cases sedation was induced by benzodiazepines, and in 94% patients were sedated for periods of less than one week until death. Nine per cent of those who received continuous deep sedation had previously requested euthanasia but their requests were not granted. Nine per cent of the physicians had consulted a palliative expert. The increased use of continuous deep sedation for patients nearing death in the Netherlands and the limited use of palliative consultation suggests that this practice is increasingly considered as part of regular medical practice.


Enhancement of S-adenosylmethionine production by deleting thrB gene and overexpressing SAM2 gene in Bacillus amyloliquefaciens.
To improve the S-adenosylmethionine (SAM) production in methionine-free medium, effects of deleting genes of SAM decarboxylase (speD) and homoserine kinase (thrB) on SAM titers were investigated, and the SAM synthetase gene (SAM2) was also overexpressed. In B. amyloliquefaciens HSAM2, deleting speD to block the SAM utilization pathway significantly reduced the SAM titer. After knockout of thrB to block the branched pathway, the resulted mutant HSAM4 produced 143.93 mg/L SAM, increasing by 42% than HSAM2. Further plasmid-based expression of SAM2 improved the SAM titer to 226.92 mg/L, and final optimization of key fermentation parameters resulted in the maximum SAM titer of 412.01 mg/L in flasks batch fermentation. Deleting thrB and overexpressing SAM2 gene were efficient for enhanced SAM production in B. amyloliquefaciens. The maximum SAM titer in flasks batch fermentation was much higher than that of previous reports.


High-dose methotrexate with leucovorin rescue: effectiveness in relapsed hairy cell leukemia.
Six patients with relapsed hairy cell leukemia after splenectomy were treated with high-dose methotrexate and leucovorin rescue. Five patients had objective responses as determined by improved blood counts. In two of them, the response has continued for more than 14 and 44 months, respectively. Intensive treatment with high-dose methotrexate and leucovorin rescue was well tolerated and no significant myelosuppression was seen.


Prevotella marseillensis sp. nov., a new bacterium isolated from a patient with recurrent Clostridium difficile infection.
Prevotella marseillensis strain Marseille-P8229T (= CSURP8229) is a new species isolated from a patient with recurrent Clostridium difficile infection. It is an anaerobic, non-motile, non-spore-forming Gram-negative coccobacillus isolated from the stool of patient with recurrent Clostridium difficile infection in Marseille. We present herein its phenotypic description together with MALDI-TOF mass spectrometry analysis and genome sequencing and comparison. The genome of P. marseillensis is 4.1607 Mbp long with 45.80 mol% of G+C content, and it contains 3078 protein-coding genes.


The sinister story of a gauche deliverer and his corrie-fisted tribesmen: Ehud and the left-handed artillery.
Bias against left-handers is well-documented and seen in the etymology of "left" and "right" in most languages. The subject of this study, Ehud, lived between the exodus of the Hebrew slaves from Egypt and the establishment of the Israelite kingdom (c1200-1000 BC), at the transition between the Late Bronze Age and Iron Age 1. His left-handedness was crucial to his deliverance of the proto-nation from tyranny, recorded in Judges in the Hebrew Bible. The description of Ehud as left-handed ('iṭṭēr yaḏ-yεmînô) is used once more in the Hebrew Bible, also in Judges, to describe the artillery of Ehud's tribe. The words apparently mean "bound/restricted in the right hand", sometimes interpreted as "ambidextrous". This is unlikely: ambidexterity is uncommon. The artillery used the sling with either hand, but Ehud used his left (śεm'ōl) hand to draw his sword. śεm'ōl, used throughout the Hebrew Bible, means left, without bias or derogatory implications. We suggest that 'iṭṭēr yaḏ-yεmînô was a right-handed bias towards left-handed people, but Ehud's left-handed victory was recognized as significant. Significant enough that (a) the language changed and the biased description was dropped in favour of simple description, and (b) the army changed with the development of left-handed slingers (artillery).


Phosphodiesterase inhibitor KMUP-3 displays cardioprotection via protein kinase G and increases cardiac output via G-protein-coupled receptor agonist activity and Ca(2+) sensitization.
KMUP-3 (7-{2-[4-(4-nitrobenzene) piperazinyl]ethyl}-1, 3-dimethylxanthine) displays cardioprotection and increases cardiac output, and is suggested to increase cardiac performance and improve myocardial infarction. To determine whether KMUP-3 improves outcomes in hypoperfused myocardium by inducing Ca(2+) sensitization to oppose protein kinase (PK)G-mediated Ca(2+) blockade, we measured left ventricular systolic blood pressure, maximal rates of pressure development, mean arterial pressure and heart rate in rats, and measured contractility and expression of PKs/RhoA/Rho kinase (ROCK)II in beating guinea pig left atria. Hemodynamic changes induced by KMUP-3 (0.5-3.0 mg/kg, intravenously) were inhibited by Y27632 [(R)-(+)-trans-4-1-aminoethyl)-N-(4-Pyridyl) cyclohexane carboxamide] and ketanserin (1 mg/kg, intravenously). In electrically stimulated left guinea pig atria, positive inotropy induced by KMUP-3 (0.1-100μM) was inhibited by the endothelial NO synthase (eNOS) inhibitors N-nitro-l-arginine methyl ester (L-NAME) and 7-nitroindazole, cyclic AMP antagonist SQ22536 [9-(terahydro-2-furanyl)-9H-purin-6-amine], soluble guanylyl cyclase (sGC) antagonist ODQ (1H-[1,2,4] oxadiazolo[4,3-a] quinoxalin-1-one), RhoA inhibitor C3 exoenzyme, β-blocker propranolol, 5-hydroxytryptamine 2A antagonist ketanserin, ROCK inhibitor Y27632 and KMUP-1 (7-{2-[4-(2-chlorobenzene) piperazinyl]ethyl}-1, 3-dimethylxanthine) at 10μM. Western blotting assays indicated that KMUP-3 (0.1-10μM) increased PKA, RhoA/ROCKII, and PKC translocation and CIP-17 (an endogenous 17-kDa inhibitory protein) activation. In spontaneous right atria, KMUP-3 induced negative chronotropy that was blunted by 7-nitroindazole and atropine. In neonatal myocytes, L-NAME inhibited KMUP-3-induced eNOS phosphorylation and RhoA/ROCK activation. In H9c2 cells, Y-27632 (50μM) and PKG antagonist KT5823 [2,3,9,10,11,12-hexahydro-10R- methoxy-2,9-dimethyl-1-oxo-9S,12R-epoxy-1H-diindolo(1,2,3-fg:3',2',1'-kl) pyrrolo(3,4-i)(1,6)benzodiazocine-10-carboxylic acid, methyl ester] (3μM) reversed KMUP-3 (1-100μM)-induced Ca(2+)-entry blockade. GPCR agonist activity of KMUP-3 appeared opposed to KMUP-1, and increased cardiac output via Ca(2+) sensitization, and displayed cardioprotection via cyclic GMP/PKG-mediated myocardial preconditioning in animal studies.


THE EFFECT OF A RHYTHMIC GYMNASTICS-BASED POWER-FLEXIBILITY PROGRAM ON THE LOWER LIMB FLEXIBILITY AND POWER OF CONTEMPORARY DANCERS.
There is an increasing cross-fertilization between the movements in contemporary dance and aesthetic sports such as rhythmic gymnastics. In such sports, supplementary training is commonly used to enable athletes to achieve all of their training goals, however dancers typically do not engage in cross training from other disciplines. The aim of this research was to test the effectiveness of a rhythmic gymnastics-based supplementary training program on lower limb flexibility and power of contemporary dancers. Experimental study using a between-groups design. Training Intervention: Prahran Rhythmic Gymnastics Specialist Centre, Testing: Deakin University. Eleven female contemporary dancers were randomly assigned to either a control or intervention group. The intervention group (n = 6) participated in an eight-week rhythmic gymnastics-based supplementary training program comprising two one-hour sessions per week in addition to their usual training. The control group (n = 5) continued their usual dance training. Pre and post measures of range of motion (ROM) and power were taken via a dance-specific kick test using an isokinetic dynamometer and a customized grand jeté test in a 3D-motion laboratory. Significant increases in right and left leg grand jeté ROM, right leg peak kicking torque and left leg grand jeté height were recorded in the intervention compared to the control group. The results of the study suggest that rhythmic gymnastics training could provide a viable means for contemporary dancers to target active ROM and power of explosive dance movements. Future research should include subjective evaluations of dance performers to confirm that training adaptions are transferred to improvements in performance quality. III.


18-Substituted progesterone derivatives as inhibitors of aldosterone biosynthesis.
The synthesis of new progesterone derivatives substituted at the 18 methyl group is described. These compounds are designed as 18-monooxygenase, cytochrome P-450-dependent potential kcat inhibitors. Preliminary results on the in vitro biological investigation of these modified progesterones are presented.


The association between low frequency of having breakfast and dyslipidemia in South Korean men and women.
This study investigated the association between frequency of having breakfast and dyslipidemia in South Korean adults aged 30 or over. This study, including 10,874 participants, was based on the Seventh Korea National Health and Nutrition Examination Survey conducted between 2013 and 2016. Using multiple logistic regression analysis, we examined the associations between frequency of having breakfast (and other covariates) and dyslipidemia in men and women. Dyslipidemia was more common among male participants with lower frequency of having breakfast in a week and was significantly high in "0 times a week" group (OR = 1.42, 95% CI, 1.13-1.78) compared with 5-7 times/week. This trend was observed in those aged 40-59 years, white collar workers, those living with their spouse, and overweight or obese participants among males and in females aged 40-49 and ≥ 60 years. Statistically significant tendency of having dyslipidemia was observed with decreasing frequency of having breakfast in total cholesterol, low-density lipoprotein cholesterol (LDL-C), and triglycerides in males (LDL-C: p for trend < 0.0001, TG: p for trend = 0.0004), but not in females. Frequency of having breakfast was associated with reduced dyslipidemia rate. The risk of dyslipidemia with a low frequency of breakfast was particularly observed in males aged 40-59, white collar workers, those living with a spouse, and postmenopausal females aged ≥ 60.


Is depression associated with microvascular disease in patients with type 2 diabetes?
We hypothesize that late-life depression is a manifestation of microvascular disease in patients with type 2 diabetes. We conducted a clinic-based cross-sectional study, comparing retinal vascular caliber, a marker of microvascular disease, in participants with type 2 diabetes with major depression (n=34), without depression (n=27) and healthy non-diabetic controls (n=38). Retinal vascular caliber was measured from digital retinal photographs using a validated computer-assisted method. After adjusting for age and gender, there was a trend of increasing retinal arteriolar caliber from healthy controls (132.6 microm), to diabetic patients without depression (139.2 microm), and diabetic patients with major depression (145.3 microm, P=0.008). The trend in retinal arteriolar caliber remains significant after adjusting for duration of diabetes, but not after further adjusting for vascular risk factors. Our findings suggest that there is variation in the retinal vascular caliber between type 2 diabetic patients with and without major depression and non-diabetic controls. This variation was largely related to poorer diabetes control and a higher frequency of vascular risk factors in diabetic patients, particularly those with depression. Studies with larger sample size may provide further insights into this association.


ROS/NF-κB Signaling Pathway-Mediated Transcriptional Activation of TRIM37 Promotes HBV-Associated Hepatic Fibrosis.
Hepatic fibrosis is an inflammatory response that leads to liver cirrhosis in the most advanced condition. Liver cirrhosis is a leading cause of deaths associated with liver diseases; hence, understanding the underlying mechanisms of hepatic fibrosis is critical to develop effective therapies. Tripartite motif (TRIM) family proteins have been shown to be involved in liver fibrosis; however, the exact role of several TRIM proteins in this process remained unexplored. In this study, we investigated the role of TRIM37 in hepatitis B virus (HBV)-associated hepatic fibrosis. We analyzed TRIM37 expression in hepatic fibrosis patients and performed functional and mechanistic studies in tissue culture and mouse models to identify the role of TRIM37 in hepatic fibrosis. We found an increased expression of TRIM37 in hepatic fibrosis patients. Mechanistically, we showed that TRIM37 physically interacts with SMAD7 and promotes ubiquitination-mediated degradation of SMAD7, and that SMAD7 is a key mediator of TRM37-induced hepatic fibrosis. Furthermore, we showed nuclear factor κB (NF-κB) activation mediated by reactive oxygen species (ROS) is necessary for the transcriptional induction of TRIM37 during HBV infection. Our study shows TRIM37 as an important promoter of HBV-associated hepatic fibrosis.


Repeated stereotactic radiosurgery (SRS) using a non-coplanar mono-isocenter (HyperArc™) technique versus upfront whole-brain radiotherapy (WBRT): a matched-pair analysis.
Stereotactic radiosurgery (SRS) is an effective treatment option for multiple brain metastases (BMs). Modern mono-isocentric techniques allow the delivery of multiple stereotactic courses, in the event of intracranial failure. Nevertheless, limited data on effectiveness and toxicity have been reported in comparison to WBRT. Aim of this retrospective matched-pair analysis was to compare patients affected by limited BMs treated with multiple SRS courses using a mono-isocentric, non-coplanar technique (HyperArc™, Varian Medical System) to upfront WBRT. One hundred and two patients accounting for 677 BMs were treated with HyperArc™. In case of further intracranial progression, 44 treatment courses of 201 metastases in 19 patients, were treated by subsequent HyperArc™ courses. This population was matched with 38 patients treated with WBRT. The median BMs number was 4 (range 2-10) for HyperArc™ and 5 (range 2-10) for WBRT. Overall survival (OS) and toxicity were evaluated. The median follow-up was 9 months (range 3-40 months). The median OS was not reached (range 5-22 months) for HyperArc™ patients and 8 months (range 3-40 months) for WBRT patients, while the 1-year OS was 77% and 34.6% for HyperArc™ and WBRT, respectively (p = 0.001; HR 4.77, 95% CI 1.62-14.00). There was one case of radionecrosis. HyperArc™ is an effective and safe technique for the treatment of multiple BMs. In selected cases of intracranial oligorecurrence, further subsequent courses can be safely delivered with the same technical approach. Moreover, in patients with a limited number of BMs, SRS showed an improved survival outcome when compared to WBRT.


Unilateral BEST1-Associated Retinopathy.
To describe a series of patients with molecularly confirmed mutation in BEST1 causing Best disease but with unilateral clinical manifestation. Retrospective observational case series. Moorfields Eye Hospital and Great Ormond Street Hospital, London (United Kingdom). Five patients (10 eyes) with uniocular manifestation of BEST1 mutation causing Best disease were ascertained retrospectively from the clinical and genetic databases. Patients had full ophthalmologic examination, color fundus photography, fundus autofluorescence imaging, spectral-domain optical coherence tomography, and detailed electrophysiological assessment. Genetic testing was performed. All cases had a clinical appearance typical of and consistent with Best disease at various stages, except that the presentation was unilateral. The reduced electrooculogram light rise was bilateral and in the context of normal electroretinograms therefore indicates generalized dysfunction at the level of the retinal pigment epithelium. Mutation in BEST1 has variable penetrance and expressivity, and can be uniocular. The clinical and electrophysiological features described assist targeted mutational screening and alert to the potential diagnosis even when there is an atypical unilateral presentation.


Impact of interventricular delay on repolarization parameters in patients with biventricular pacing.
A 48-year-old man who was previously a responder to cardiac resynchronization therapy (CRT) presented with worsening of heart failure and reduction in the percentage biventricular (BiV) pacing. Device interrogation revealed T-wave oversensing that was eliminated by optimizing the interventricular (VV) delay. Compared to simultaneous BiV pacing, both sequential left ventricle-right ventricle (LV-RV) and univentricular LV pacing shortened the QTc, T peak-end intervals and reduced the T wave amplitude. Modification of VV delays can result in changes in wavefront of activation and thereby affect ventricular repolarization patterns.


Ocular characteristics associated with the location of focal lamina cribrosa defects in open-angle glaucoma patients.
PurposeTo investigate the clinical characteristics according to the location of focal lamina cribrosa (LC) defects and its associated ocular features.Patients and methodsA total of 139 open-angle glaucoma patients underwent Spectralis optical coherence tomography (OCT) with enhanced depth imaging. Alterations in the contour of the LC were investigated to find focal LC defects. The location of the visible LC defect from the neural canal wall (far-peripheral and mid-peripheral) and clock-hour position (superotemporal, temporal and inferotemporal) were classified. Disc ovality ratio and disc-foveal angle were measured from disc and retinal nerve fiber layer (RNFL) photographs. The disc tilt degree was measured using a Heidelberg Retina Tomograph (HRT) III system. The en face OCT image of the disc scans was registered to the disc and RNFL photographs, to determine whether the focal LC defects corresponded spatially to the glaucomatous damage location.ResultsEyes with far-peripheral LC defects were significantly myopic and had a higher disc ovality ratio. The disc tilt degree obtained by HRT revealed significant temporal disc tilt in eyes with temporal LC defects (P<0.001). Eyes with inferotemporal LC defects had a significantly larger disc-foveal angle (P=0.027). The inferotemporal LC defects corresponded to the location of glaucomatous damage in 81.6%; however, only 46.2% of eyes with a superotemporal LC defect and 3.2% of eyes with a temporal LC defect corresponded spatially with the glaucomatous damage location.ConclusionsThe clinical characteristics and association with glaucomatous damage location were different according to the location of focal LC defect.


Addition of either gastric lipase or cholesterol esterase to improve both β-cryptoxanthin ester hydrolysis and micellarization during in vitro digestion of fruit pulps.
Using the INFOGEST in vitro digestion protocol adapted to carotenoids, the impact of additional rabbit gastric lipase (RGL) on the hydrolysis extent of β-cryptoxanthin esters was evaluated for the first time, and compared with the addition of porcine cholesterol esterase (CEL). Both the modifications increased the hydrolysis of (all-E)-β-cryptoxanthin esters from mandarin and peach pulps, although the outcomes were different. Addition of RGL consistently increased the average hydrolysis extent from 55.2% to 59.5% in mandarin pulp and from 22.7% to 48.8% in peach pulp (p < 0.05). The addition of CEL produced lower hydrolysis extents, i.e., 58.5% in mandarin (not statistically significant) and 28.4% in peach (p < 0.05), compared to those obtained with RGL. The hydrolysis extent positively correlated with the carotenoid ester concentration in both matrices. Bioaccessibility values were higher in mandarin pulp (range 32-34%) compared to those in peach pulp (range 16-21%), and were associated with the hydrolysis extent of the carotenoid esters during digestion. Addition of RGL and CEL produced no significant (p < 0.05) effect on the overall carotenoid bioaccessibility values of mandarin, while positively affected those in peach. Altogether these results corroborate that the hydrolysis extent of xanthophyll esters limits bioaccessibility. Additionally, hydrophobicity of the carotenoid inversely correlates with micellarization, as free (all-E)-xanthophylls micellarized in a higher extent compared to (all-E)-β-carotene and xanthophyll esters. The new information of our results is that the addition of rabbit gastric lipase substantially contributes to the hydrolysis of β-cryptoxanthin esters from fruit pulps, and consequently, to increase carotenoid bioaccessibility, being even more effective than CEL.


Serum levels of VEGF and TNF-alpha and their association with C-reactive protein in patients with endometriosis.
To compare serum levels of vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-alpha), and C-reactive protein (CRP) throughout the menstrual cycle in women with and without endometriosis and to quantify the association between these serum markers. Twenty five endometriosis patients (ASRM stage III-IV) and 13 controls were followed through one menstrual cycle. VEGF, TNF-alpha and CRP concentrations were measured with enzyme-linked immunosorbent assay in each of the four cycle phases (early and late proliferative and early and late secretory). Non-parametric methods and Pearson's correlation coefficients were used for statistical analysis. Increased serum levels of VEGF in secretory phases and of TNF-alpha in all cycle phases were observed. A positive correlation between CRP and VEGF was found in endometriosis patients but not in controls. Our results suggest that excessive systemic angiogenic activity is a feature of patients with endometriosis.


MicroRNA-155 is a predictive marker for survival in patients with clear cell renal cell carcinoma.
To investigate the clinical significance of micro-ribonucleic acid-155 in clear cell renal cell carcinoma, in particular focusing on the association of expression levels of micro-ribonucleic acid-155 with clinicopathological factors, cancer-specific survival and therapeutic outcomes in clear cell renal cell carcinoma patients. Quantitative reverse transcription polymerase chain reaction of micro-ribonucleic acid-155 was carried out on 137 clear cell renal cell carcinoma cases, containing 77 matched pairs of clear cell renal cell carcinoma and normal adjacent kidney tissues from the same patients. Significant overexpression of micro-ribonucleic acid-155 was found in clear cell renal cell carcinoma compared with normal kidney tissue. Expression of micro-ribonucleic acid-155 was not associated with prognosis in all stage groups. However, in 43 patients with stage III and IV clear cell renal cell carcinoma, low expression levels of micro-ribonucleic acid-155 correlated with a poor prognosis. Regarding cancer-free survival of 26 patients with stage III and IV clear cell renal cell carcinoma who received curative resection and cancer-specific survival of 31 patients who received postoperative therapy with interferon-α after radical nephrectomy, low expression levels of micro-ribonucleic acid-155 correlated with poor clinical outcomes in these two groups. Low expression of micro-ribonucleic acid-155 represents a valuable marker of poor clinical outcomes in patients with stage III and IV clear cell renal cell carcinoma.


Executive Summary of the Ovarian Cancer Evidence Review Conference.
The Centers for Disease Control and Prevention awarded funding to the American College of Obstetricians and Gynecologists to develop educational materials for clinicians on gynecologic cancers. The American College of Obstetricians and Gynecologists convened a panel of experts in evidence review from the Society for Academic Specialists in General Obstetrics and Gynecology and content experts from the Society of Gynecologic Oncology to review relevant literature, best practices, and existing practice guidelines as a first step toward developing evidence-based educational materials for women's health care clinicians about ovarian cancer. Panel members conducted structured literature reviews, which were then reviewed by other panel members and discussed at a virtual meeting of stakeholder professional and patient advocacy organizations in February 2022. This article is the executive summary of the relevant literature and existing recommendations to guide clinicians in the prevention, early diagnosis, and special considerations of ovarian cancer. Substantive knowledge gaps are noted and summarized to provide guidance for future research.


Two-stage evaluation of the impact of a nursing information system in Taiwan.
This study evaluated the post-implementation impact of a nursing information system and identified issues related to the technology adoption process. Given the high level of investment necessary to implement information systems, evaluation has become vital to ensure successful adoption and use. Improved understanding of implementation difficulties/barriers and factors leading to them can serve as a platform for the development of strategies and education programs for users. The study design was a two-stage data comparison analysis of a nursing information system focusing on computerized nursing care plan use. Data were collected from nurses by questionnaire during the first month (December 2004) and 1 year after system installation (December 2005). While nurses gave the information technology a higher evaluation score at the second stage than at the early stage, the overall ratings were slightly negative at both stages. Age and pressure to use the system were critical factors at both stages, whereas computer skills and perceived time using the system were vital at the beginning stage of implementation. Issues of concern at both stages were system functionality, efficiency, usability, and user support. Implementation of an information system requires consideration of issues involving hardware and software, staff training, organizational policy, and workflow changes.


Argyrosis of the lacrimal sac.
Silver deposition in the wall of the lacrimal sac after prolonged application of mild silver protein (Argyrol) eye drops is documented at the light and electron microscopical level. Silver granules were found in the extracellular matrix predominantly on elastic fibres and within cells forming conglomerates in secondary lysosomes. Most of the silver-containing cells showed a prominent rough endoplasmic reticulum, suggesting their fibroblastic origin. Investigation by energy-dispersive analysis of X-rays (EDAX) indicated the precipitation of silver as selenide.


Measurement of gain characteristics of semiconductor lasers by amplified spontaneous emissions from dual facets.
In this letter, we describe a novel gain measurement approach for semiconductor edge-emitting lasers, with which TE and TM gain spectra can be easily obtained by collecting the amplified spontaneous emissions at dual facets of the device. An unstrained and continuously-operated GaAs/AlGaAs single quantum well laser strip is used to illustrate this method. The measured gain spectra are compared with theoretical gain curves to analyze the gain polarization characteristics and the relevant subband structure in the valence band of the well using the measured gain spectra.


An evaluation of a computer based education program for the diagnosis and management of dementia in primary care. An international study of the transcultural adaptations necessary for European dissemination.
The aim of this study is to make an inventory of the changes that are needed to make an interactive computer based training program (ICBT) with a specific educational content, acceptable to professional communities with different linguistic,cultural and health care backgrounds in different European countries. Existing educational software, written in two languages was reviewed by GPs and primary care professionals in three different countries. Reviewers worked through the program using a structured critical reading grid. A 'simple' translation of the program is not sufficient. Minor changes are needed to take account of linguistic differences and medical semantics. Major changes are needed in respect of the existing clinical guidelines in every country related to differences in the existing health care systems. ICTB programs cannot easily be used in different countries and cultures. The development of a structured educational program needs collaboration between educationalists, domain experts, information technology advisers and software engineers. Simple validation of the content by local expert groups will not guarantee the program's exportability. It is essential to involve different national expert groups at every phase of the development process in order to disseminate it in other countries.


Quality assurance in cardiovascular CT: a practical guide.
Cardiovascular computed tomography (CCT) is a cutting-edge imaging technique providing important, non-invasive, diagnostic information. Concerns exist regarding radiation exposure to patient populations, but achieving optimal image quality at the lowest doses can be challenging. This guide provides practical advice about how quality can be assured in any CCT unit or radiology department. Illustrated by real-world vignettes and data analysis from our own experience, we highlight a multidisciplinary team approach to each stage of the patient journey, the effectiveness of regular dose audit overseen by a CT optimisation group, and the importance of underused systolic scanning techniques, in order to drive significant dose reduction without loss of image quality or clinical confidence.


Structural characterization of a P-selectin and EGFR dual-targeting fucoidan from Sargassum fusiforme.
In this study, we obtained fucoidans SFP, SHP, STP, and FVP from Sargassum fusiforme, Sargassum horneri, Sargassumthunbergii, and Fucus vesiculosus, respectively. Chitosan/fucoidan nanoparticles (Cs/F NPs) were prepared using the fucoidans mentioned above. SFP NPs and SHP NPs showed strong binding abilities to P-selectin and epithelial growth factor receptor (EGFR). Given the yields from the alga, SFP was first selected to explore the structural characteristics of the P-selectin and EGFR dual-targeting fucoidan. SFP had an estimated molecular weight of 739 kDa and was mainly composed of galactose (26.57%, mol%) and fucose (66.81%), with minor amounts of mannose (2.54%), glucosamine (0.42%), and glucose (3.66%). Galactose and fucose accounted for thevast majority. Further investigation, including methylation analysis, one- and two-dimensional nuclear magnetic resonance, and mass spectroscopy, was performed to reveal the fine structure of SFP. The results indicated that SFP mainly consisted of → 3)-α-l-Fucp-(1→, →4)-α-l-Fucp-(1→, →3,4)-α-l-Fucp-(1→, →3)-β-d-Galp-(1→, and minor → 6)-β-d-Galp-(1→, partially sulfated at the C-4 of → 3)-α-l-Fucp-(1→, C-3 of → 4)-α-l-Fucp-(1→, C-3 of → 6)-β-d-Galp-(1→, and C-6 of → 3)-β-d-Galp-(1 → . Sulfated fuco- and galactofuco-segments formed the branches.


Progesterone utilizes the PI3K-AKT pathway in human spermatozoa to regulate motility and hyperactivation but not acrosome reaction.
Progesterone is a physiologic regulator of sperm hyperactivation and acrosome reaction and it does so by activating a range of kinases present in the spermatozoa. In the present study, the involvement of the AKT- phosphatidylinositol 3-kinase (PI3K) signaling pathway in mediating progesterone response in human spermatozoa was investigated. In capacitated spermatozoa, progesterone transiently and concentration dependently lead to phosphorylation of AKT at both Thr 308 and Ser 473 in the tail region. This phosphorylation was inhibited by the PI3K inhibitor wortmannin, suggesting that progesterone leads to activation of PI3K-AKT pathway. The activation of AKT in response to progesterone is calcium dependent and the CatSper channel inhibitor mibefradil significantly reduced progesterone mediated AKT phosphorylation. Preincubation of spermatozoa with wortmannin inhibited the progesterone mediated increase in tyrosine phosphorylation and also attenuated the increase in number of motile, progressively motile and hyperactive spermatozoa but not the number of acrosome reacted spermatozoa. These observations imply that progesterone via CatSper activates the PI3K-AKT pathway required for motility and hyperactivation but not for acrosome reaction.


Management recommendations for knee osteoarthritis: how usable are they?
Despite the availability of practice guidelines for the management of knee osteoarthritis, inadequacies in practices of clinicians and patients have been found, leading to suboptimal outcomes. Literature has shown that simply disseminating management recommendations does not lead to adherence. Research suggests that barriers to use should be identified and addressed to improve adherence. The objective of this study was to identify barriers to use of conservative management recommendations for knee osteoarthritis by patients, general practitioners and physiotherapists. Following systematic reviews of evidence and guidelines, 12 key management recommendations were elaborated on four themes: medication, exercise, self-management and occupation. Focus groups were separately done with patients with knee osteoarthritis, general practitioners and physiotherapists to assess barriers to the use of recommendations. Patients and general practitioners appeared generally fatalistic with regards to knee osteoarthritis, with physiotherapists being more positive regarding long-term improvement of knee osteoarthritis. For medication, discrepancies were found between recommendations and views of clinicians. Both patients and general practitioners appeared ambivalent towards exercise and activity, recognizing its usefulness but identifying it at the same time as a cause of knee osteoarthritis. Patients and general practitioners appeared to consider weight loss particularly difficult. Barriers specific to each knee osteoarthritis management recommendation and stakeholder group were identified. Recommendations to address these barriers were elaborated. Results of this study can be used to develop implementation strategies to overcome identified barriers, with the goal of facilitating the use of guideline recommendations and improving outcomes.


Calcium pump kinetics determined in single erythrocyte ghosts by microphotolysis and confocal imaging.
The activity of the plasma membrane calcium pump was measured in single cells. Human red blood cell ghosts were loaded with a fluorescent calcium indicator and either caged calcium and ATP (protocol A) or caged ATP and calcium (protocol B). In a suitably modified laser scanning microscope either calcium or ATP were released by a short UV light pulse. The time-dependent fluorescence intensity of the calcium indicator was then followed in single ghosts by repetitive confocal imaging. The fluorescence intensity was converted into calcium concentration, which in turn was used to derive the kinetic parameters of the calcium pump, the Michaelis-Menten constant Km, and the maximal transport rate vmax. Km and vmax values derived in this manner were 24 +/- 14 microM and 1.0 +/- 0.6 microM/(ghost s) for protocol A, and 4 +/- 3 microM and 1.0 +/- 0.6 microM/(ghost s) for protocol B, respectively. The difference between A and B is presumably caused by calmodulin, which is inactive in the experiments with protocol A. The possibilities to extend the new method to living nucleus-containing cells transiently transfected with mutants of the plasma membrane calcium pump are discussed.


Coxsackie B3 virus can replicate in cultured human foetal heart cells and is inhibited by interferon.
Coxsackie B viruses (types 1 to 5) are the most frequent reported cause of acute viral myocarditis. To study the pathogenesis of the disease at the cellular level, we simulated an infectious situation by infecting cultured human foetal heart cells with Coxsackie B3 (CB3) virus. Successful replication of this virus could be demonstrated by the presence of virus particles inside cultivated foetal myocytes together with high titres of progeny virus of 10(8) plaque-forming units (PFU) per millilitre culture medium. Within 9 h of infection networks of myocytes lost their ability to contract spontaneously followed by disintegration and replacement by overgrowing fibroblasts which survived the infection. These cells produced CB3 virus continuously over several months, indicating carrier state infection of human myocardial fibroblasts. Human fibroblasts interferon (IFN-beta) was found to act as a potent inhibitor of the replication of this virus. Virus yields could be reduced from 1.2 x 1.8 x 10(5) PFU/ml culture medium when human heart cells were incubated with IFN-beta 20 h prior to challenge with a high input multiplicity of 50 PFU of CB3 virus per cell, demonstrating the major protective role of IFN-beta in CB3 viral infection. It thus appear that IFN-beta might become useful as an antiviral agent in the treatment of Coxsackie myocarditis.


A coupled peroxidatic oxidation technique for the histochemical localization of monoamine oxidase A and B and benzylamine oxidase.
A coupled peroxidatic oxidation technique is presented which employs benzylamine and tyramine as substrates and clorgyline, deprenyl, phenelzine and pargyline as specific inhibitors. Using this technique with frozen sections of human term placenta and rat liver, the histochemical localization of monocamine oxidase A and B and bnezylamine oxidase has been demonstrated.


Antimicrobial activity and biocompatibility of polyurethane and silicone catheters containing low concentrations of silver: a new perspective in prevention of polymer-associated foreign-body-infections.
In modern medicine, infection is one of the most serious complications of implanted plastic devices. The host is not able to overcome this special type of opportunistic infection despite having a normal immune response and a low virulence of most of the bacteria involved. Antimicrobial therapy alone generally cannot cure the infection and the removal of catheters often remains the only choice of therapy. Bacterial adhesion to the polymer surface of the catheter, be it luminal or external, is an important step in the pathogenesis of catheter-associated infections. In this report, we describe new approaches to the prevention of infections by impregnation of polyurethane and silicone with silver by two different methods. The antimicrobial activity of these silver-impregnated catheters is more than 10 fold higher for coagulase-negative staphylococci (CNS) compared to catheters without silver. Similar results are obtained with other microbial organisms like Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Candida albicans. The new polymers show no cytotoxic or thrombogenic side effects in vitro.


Spontaneous immunomodulation and regulation of angiogenesis and osteogenesis by Sr/Cu-borosilicate glass (BSG) bone cement to repair critical bone defects.
Injectable bone biomaterials like bone cement should be designed and fabricated with certain biological criteria, which include: 1) recruitment and polarization of the macrophages from M1 (pro-inflammatory) to M2 (anti-inflammatory) phenotype, 2) enhance vascularization, and 3) activate osteogenic differentiation of bone marrow-derived stem cells to promote bone healing. So far, no injectable biomaterials could spontaneously regulate the entire bone healing process that involves inflammation, angiogenesis, and osteogenesis. Therefore, in this study, we designed bone cement comprised of strontium and copper-incorporated borosilicate glass (Sr/Cu-BSG) in the liquid phase of chitosan to modulate bone healing. In vitro studies showed that the controlled release of Sr and Cu ions up-regulated anti-inflammatory genes(IL-1Ra and TGF-β1) while down-regulating pro-inflammatory genes(IL-1β and IL-6) in macrophages at 3 days. Sr and Cu ions also increased the expressions of angiogenic genes (VEGF and bFGF) in HUVECs at 5 days and osteogenic genes (Runx-2, OCN, and OPN) in hBMSCs at 7, 14, and 21 days. 5Sr3Cu-BSG bone cement exhibited the best anti-inflammatory, angiogenic, and osteogenic properties among the bone cement groups with different Sr and Cu ratios. Short-term and long-term implantation of Sr/Cu-BSGs in femoral condylar bone defects of rats and rabbits confirmed the in vitro results, where the degradation rate of Sr/Cu-BSG matched the bone healing rate. Similar to in vitro, the 5Sr3Cu-BSG group also showed the highest bone formation in vivo. Excellent physical and chemical properties, along with its bone repairing ability, make the Sr/Cu-BSG bone cement a good candidate biomaterial for treating bone defects.


Interactions of ganglioside GM1 with human and fetal calf sera. Formation of ganglioside-serum albumin complexes.
The interactions of ganglioside GM1 with human and fetal calf sera were studied, the following main results being obtained: (a) GM1, upon incubation with both sera gave origin to two GM1-protein complexes, which also occurred after interaction of GM1 with the albumin fractions prepared from the same sera. Instead no complex formation occurred using the albumin-free fractions. Therefore GM1 appeared to specifically bind serum albumin and to form GM1-albumin complexes. (b) GM1 binding to serum albumin started at ganglioside concentrations surely micellar (above 10(-6) M), was time and concentration dependent, and resulted in a relevant degree of GM1 complexation (up to 80% of total GM1 in human serum and up to 18% in fetal calf serum). (c) the binding kinetics appeared, in both serum and the correspondent albumin fraction, to be biphasic: in the first phase, occurring till about 2 . 10(-4) M GM1, the ratio between bound and total GM1 increased linearly with increasing GM1 concentration; in the second phase, occurring above 2 . 10(-4) M, the ratio remained practically constant. After these findings it should be expected that GM1, when present in serum containing systems, forms complexes with albumin. This should be approximately considered when studying the effects of exogeneous GM1 in in vivo and in vitro (tissue cultures) systems.


Fungi in Rice Field Open Irrigation System: Ecological Implications and Biosecurity.
Italy is the leading producer of rice in Europe, but this crop is increasingly threatened by many factors such as pathogens' resistance, pollution and climate change. To date, few works keep in consideration the ecological role that the open irrigation system can play in the dispersion of important opportunistic species, and if it is affected by agricultural management and environmental seasonal changing. This work carried out the mycological characterization of a rice field irrigation system located in Vistarino (Pavia, Lombardy, Italy). Three main sections of an irrigation system (canal, ditch and paddy) were sampled during the summer 2018 (irrigation season of the rice crop). Water samples processing underlined how the irrigation system is rich of fungal diversity (59 species isolated). In order of abundance, the canal samples are characterized by the dominance of Aspergillus, Cladosporium, Fusarium and Trichoderma genera, while the ditch samples by Alternaria, Cladosporium, Fusarium, and Penicillium genera, and the paddy samples by Alternaria, Cladosporium, Fusarium and Trichoderma genera. Results showed that the three environments are mycologically independent of each other: fungi do not exploit the irrigation system for their dispersion in paddy. Probably fungi prefer others dispersion systems such as air dispersion. This means that an open irrigation system is not to be considered as a continue system with free circulation of fungi. Indeed, each sector of the system appears characterized by a typical funga, which undergoes variations during the sampled season due to agricultural management and environmental conditions.


Interaction between a picornavirus and a wild population ofDrosophila melanogaster.
Drosophila C virus (DCV) has a considerable impact on ovarian morphogenesis inDrosophila melanogaster host populations. This virus also affects the developmental time and the fresh weight of infected females. In order to investigate the hypothesis that DCV may play a role in the dynamics ofDrosophila populations, the fertility and embryonic and larvo-pupal death rates of a host population and that of five DCV-free populations were determined. A comparison of two populations, one of them DCV-free, the other infected, suggested that the fertility of the DCV-infected flies was higher than that of uninfected flies, despite a greater larvo-pupal death rate. Fertility of the infected flies was greater among the infected population than for the DCV-free populations. The DCV-free populations originated from five different localities. The virus clearly does have an impact on the biotic potential of its host population. This paper reports for the first time a positive interaction between a viral population and a host population as it increases certain parameters of host population dynamics.


Synthesis and anticonvulsant activity evaluation of 8-alkoxy-5-(4H-1,2,4-triazol-4-yl)quinoline derivatives.
Two series of 8-alkoxy-5-(4H-1,2,4-triazol-4-yl)quinolines and 8-alkoxy-5-(2H-1,2,4-triazol-3-one-4-yl)quinolines were synthesized. The anticonvulsant activity of these compounds was evaluated with maximal electroshock seizure test and rotarod test. Among the synthesized compounds, 8-octoxy-5-(4H-1,2,4-triazol-4-yl)quinoline (4g) was the most active compound with ED(50) of 8.80 mg/kg, TD(50) of 176.03 mg/kg and protective index of 20.0. Its neurotoxicity was lower than all other synthesized compounds and also markedly lower than that of the reference drug carbamazepine. In addition, the potency of compound 4g against seizures induced by pentylenetetrazole, 3-mercaptopropionic acid, and bicuculline suggested its broad spectrum activity, and the mechanisms of action including inhibition of voltage-gated ion channels and modulation of GABAergic activity might involve in its anticonvulsant activity.


Studies of nutrients and their relationship to cancer in the multiethnic population of Hawaii.
Because of the many ethnic cultures in Hawaii, there is great dietary variation in the population, even within individual ethnic groups. Epidemiologic studies in this setting are providing useful information on the role of macro- and micronutrients in the risk for certain cancers. The results of several recent and ongoing case-control studies are described, especially with regard to the influence of dietary fat, vitamin A, and vitamin C. The degree of consistency of the findings with other reports in the literature is discussed.


Trypanosoma cruzi: metabolic labeling of trypomastigote sialoglycolipids.
Trypomastigote forms (infective) of Trypanosoma cruzi incorporate (3H)-palmitic acid and D-(3H)-galactose into glycolipids. Palmitic acid-labeled acidic glycolipids were partially hydrolyzed with neuraminidase. The labeling of these compounds when the intact cell surface was labeled with galactose oxidase plus NaB3H4 indicates the membrane location of the sialoglycolipids.


A new, pellet-forming fungal strain: its isolation, molecular identification, and performance for simultaneous sludge-solids reduction, flocculation, and dewatering.
Filamentous and nonfilamentous microorganisms can cause bulking and foaming in wastewater sludge settling and dewatering. In this research, sludge degradation and bioflocculation was studied using pellet-forming filamentous fungi isolated from municipal wastewater sludge. To understand the role of filamentous fungi in sludge settling and dewatering, the isolated fungi was inoculated with both spores and pellets (beads) into sterilized and nonsterilized sludge having different suspended-solids concentrations. Biofloc formation, sludge settling, sludge degradation, change in pH of fungal-grown medium, zeta potential, and microscopic analysis of bioflocs were performed. The suspended-solids concentration was found to decrease over 5 d of incubation because of use and biodegradation by fungal biomass. The isolated fungal strain was well adapted to forming biofloc and to interacting with natural microbial flora and exhibited low capillary-suction time for sludge dewatering.


In vivo and in vitro studies exploring the pharmacokinetic interaction between bosentan, a dual endothelin receptor antagonist, and glyburide.
In a clinical trial with patients with chronic heart failure, a higher incidence of elevated levels of liver transaminases was observed during concomitant treatment with bosentan, a dual endothelin receptor antagonist, and glyburide (INN, glibenclamide), a sulfonylurea-type antidiabetic drug, than with treatment with bosentan alone. This study was conducted to investigate a possible pharmacokinetic interaction between bosentan and glyburide. In a randomized, 2-way crossover study, 12 healthy volunteers received treatments A and B. Treatment A consisted of 125 mg bosentan twice a day for 10 days plus concomitant 2.5 mg glyburide twice a day on days 6 to 10. Treatment B consisted of 2.5 mg glyburide twice a day for 10 days plus concomitant 125 mg bosentan twice a day on days 6 to 10. Plasma concentrations of bosentan and its metabolites and of glyburide were measured on days 5 and 10 of treatment A and treatment B, respectively. Bosentan reduced the area under the concentration-versus-time curve of glyburide approximately 40% (P <.05). Glyburide decreased the area under the concentration-versus-time curve of bosentan and its metabolites 20% to 30% (P <.05). Results of in vitro experiments showed that glyburide is metabolized by cytochrome P450 (CYP) 2C9, 2C19, and 3A4. No interaction was observed on the level of serum protein binding. The plasma levels of both bosentan and glyburide were reduced after concomitant administration. This finding is consistent with a CYP3A4-inducing potential of both drugs. The observed pharmacodynamic interaction between bosentan and glyburide in patients with chronic heart failure cannot be explained by a pharmacokinetic interaction.


Toxicity of cadmium oxide instilled into the rat lung. I. Metabolism of cadmium oxide in the lung and its effects on essential elements.
The metabolism of cadmium oxide (CdO, insoluble form) and cadmium chloride (CdCl2, soluble form) instilled intratracheally into the rat lung was investigated. CdO might be solubilized rapidly in the lung and consequently pulmonary clearance rate of CdO was not so different from that of CdCl2. At a dose of 5 micrograms Cd/rat about 20% of the dose was translocated to the liver within 12 h, whereas gradual and consistent accumulation of Cd was observed in the kidney up to 7 days. Both pulmonary clearance and translocation of Cd to the liver were accelerated with the dose of instilled CdO, however, Cd accumulated in the kidney was proportional to the dose. Lung weight was increased by the instillation of CdO. Lung essential elements such as S, P, Mg, Zn and Mn were not affected in the inflammatory-reparative proliferative process, but Cu content of unit lung weight was slightly decreased.


Comparative Analysis of Different Isolated Oleaginous Mucoromycota Fungi for Their γ-Linolenic Acid and Carotenoid Production.
γ-Linolenic acid (GLA) and carotenoids have attracted much interest due to their nutraceutical and pharmaceutical importance. Mucoromycota, typical oleaginous filamentous fungi, are known for their production of valuable essential fatty acids and carotenoids. In the present study, 81 fungal strains were isolated from different Egyptian localities, out of which 11 Mucoromycota were selected for further GLA and carotenoid investigation. Comparative analysis of total lipids by GC of selected isolates showed that GLA content was the highest in Rhizomucor pusillus AUMC 11616.A, Mucor circinelloides AUMC 6696.A, and M. hiemalis AUMC 6031 that represented 0.213, 0.211, and 0.20% of CDW, respectively. Carotenoid analysis of selected isolates by spectrophotometer demonstrated that the highest yield of total carotenoids (640 μg/g) was exhibited by M. hiemalis AUMC 6031 and M. hiemalis AUMC 6695, and these isolates were found to have a similar carotenoid profile with, β-carotene (65%), zeaxanthin (34%), astaxanthin, and canthaxanthin (5%) of total carotenoids. The total fatty acids of all tested isolates showed moderate antimicrobial activity against Staphylococcus aureus and Salmonella Typhi, and Penicillium chrysogenum. To the best of our knowledge, this is the first report on the highest yield of total lipid accumulation (51.74% CDW) by a new oleaginous fungal isolate R. pusillus AUMC 11616.A. A new scope for a further study on this strain will be established to optimize and improve its total lipids with high GLA production. So, R. pusillus AUMC 11616.A might be a potential candidate for industrial application.


Ultrastructural and biochemical analysis of "undifferentiated" pulmonary carcinomas.
Seven cases of "undifferentiated" pulmonary carcinoma were studied ultrastructurally; five were of the typical oat cell variety and the remaining two consisted of larger cells. In three of the former and both of the latter cases neurosecretory-like granules were demonstrated. Biochemical analysis of tumor tissue extracts revealed 5-hydroxy-3-indoleacetic acid, vanilylmandelic acid, and catecholamine activity in all instances. No hormonal syndrome or metabolic abnormality was detected in any of the patients. The concomitant morphologic demonstration of neurosecretory-like granules and the presence of 5-hydroxy-3-indoleacetic acid, vanilylmandelic acid, and catecholamines in neoplastic tissue would provide further evidence that these tumors may indeed arise from bronchial endocrine cells and could therefore be classified within the group of neuroendocrine carcinomas. Also it seems apparent that these neuroendocrine bronchial carcinomas may include tumors consisting of cells somewhat larger than the typical oat cell. The observation of 5-hydroxy-3-indoleacetic acid, vanilylmandelic acid, and catecholamine activity in two oat cell carcinomas in which neurosecretory granules could not be demonstrated poses an interesting problem whose solution may only derive from further studies.


Comparison of mode estimation methods and application in molecular clock analysis.
Distributions of time estimates in molecular clock studies are sometimes skewed or contain outliers. In those cases, the mode is a better estimator of the overall time of divergence than the mean or median. However, different methods are available for estimating the mode. We compared these methods in simulations to determine their strengths and weaknesses and further assessed their performance when applied to real data sets from a molecular clock study. We found that the half-range mode and robust parametric mode methods have a lower bias than other mode methods under a diversity of conditions. However, the half-range mode suffers from a relatively high variance and the robust parametric mode is more susceptible to bias by outliers. We determined that bootstrapping reduces the variance of both mode estimators. Application of the different methods to real data sets yielded results that were concordant with the simulations. Because the half-range mode is a simple and fast method, and produced less bias overall in our simulations, we recommend the bootstrapped version of it as a general-purpose mode estimator and suggest a bootstrap method for obtaining the standard error and 95% confidence interval of the mode.


SPECT imaging of herpes simplex virus type1 thymidine kinase gene expression by [(123)I]FIAU(1).
Introduction of suicide genes, such as herpes simplex virus type1 thymidine kinase (HSV1-tk), in tumor cells has provided a useful method for tumor gene therapy. Several L-nucleosides, such as Lamivudine (3TC) and Clevudine (L-FMAU), have been successfully tested as high-potency antiviral agents. To investigate the potential differences between D- and L-isomers of nucleosides, [(125/123)I]-2'-fluoro-2'-deoxy-1beta-D/L-arabino-furanosy-5-iodo-uracil (D/L-FIAU) have been synthesized and evaluated as potential SPECT agents for imaging HSV1-tk gene expression. [(125/123)I]D- and L-FIAU were prepared by iododestannylation of the respective tin precursors with (125/123)I-sodium iodide. In vitro cell uptake studies were performed by incubation of [(125)I]D- and L-FIAU in RG2 cells expressing HSV1-tk (RG2TK+). In vivo studies including biodistribution and SPECT were performed in RG2TK+ and RG2TK- tumor-bearing nude mice using [(123)I]D- and L-FIAU. Cell uptake and biodistribution studies indicated that [(125/123)I]L-FIAU did not show any high accumulation (sensitivity) or uptake ratios (selectivity) in HSV1-TK-positive (RG2TK+) tumors as compared to control tumors. In contrast, [(125/123)I]D-FIAU displayed both sensitivity and selectivity to RG2TK+ tumors. The selective in vivo accumulation of [(123)I]D-FIAU increased with time and the tumor uptake ratios (RG2TK+/RG2TK-) for 2, 4, and 24 hours averaged 6.2, 22.7, and 58.8, respectively. High-resolution SPECT of four nude tumor-bearing mice demonstrated a very high uptake of [(123)I]D-FIAU in the RG2TK+ tumor, while no significant tracer accumulation was observed in the RG2TK- tumor and other organs. The data suggest that only the D-isomer of [(123)I]FIAU is useful for imaging HSV1-tk gene expression in mice by high-resolution SPECT imaging.


Myosin light chain kinase is expressed in neurons and glia: immunoblotting and immunocytochemical studies.
The contractile protein myosin is thought to subserve motility-related functions in a wide range of eukaryotic non-muscle cells including both neurons and glia. To determine if the Ca2+/calmodulin-dependent enzyme, myosin light chain kinase (MLCK) is involved in the regulation of neural myosin we investigated the presence and localization of MLCK in a variety of neural tissues by immunoblotting and immunocytochemistry. A specific immunoreactive protein (M(r) = 146,000) was detected in blotted homogenates from many regions of rat brain and from primary cultures of either astrocytes or cerebellar granule cells grown in the absence of other cell types. At the light microscopic level, MLCK-immunoreactivity was evident in many regions of rat brain, as well as in the cultured astrocytes and cerebellar granule cells. MLCK-immunoreactivity was observed to be largely cytosolic in astrocytes but with a proportion associated with the cytoskeleton. In the cerebellar granule cells immunoreactivity was present in neuronal processes as well as somata. The detection of MLCK in neural cells suggests that MLCK-catalyzed myosin phosphorylation may couple changes in intracellular calcium concentrations to motility-related functions of neurons and glia.


Nutritional lipid-induced oxidative stress leads to mitochondrial dysfunction followed by necrotic death in FaO hepatocytes.
Mitochondrial dysfunction and hepatocyte cell death have been reported in fatty liver and non-alcoholic steatohepatitis. Our aim in this study was to evaluate whether direct exposure of hepatocytes to extracellular fat could facilitate such deleterious effects. FaO hepatic cells treated with fat was used as an in vitro model for steatosis. FaO hepatocytes were exposed to 0.1% triacylglycerols using commercially available lipid emulsion (LE) for various periods and studied for production of reactive oxygen species (ROS), mitochondrial function, and cell death parameters. To study the type of cell death, high-mobility group box chromosomal protein 1cellular levels, DNA fragmentation, and caspase activity were evaluated. Cells incubated with LE for 6 h exhibited a marked increase in the production of intracellular ROS. Using treatments with peroxisome proliferator-activated receptor activators, mitochondrial electron-transfer chain inhibitor, and different sources of LE that did or did not contain medium-chain triacylglycerols, the mitochondria were found to be the source of ROS. LE treatment resulted in phosphorylation of adenosine monophosphate-activated protein kinase, accompanied by a decrease in adenosine triphosphate levels. Changes in intracellular ROS and energy levels were followed by cell death. FaO hepatocytes showed a significant reduction in high-mobility group box chromosomal protein-1 and little DNA fragmentation. Incubation with LE for 24 h did not change caspase-3 activity, indicating that hepatocyte death was necrotic. The antioxidant N-acetylcysteine was able to attenuate the changes in intracellular energy levels and ROS levels and to prevent cell death after exposure to LE. These results suggest that exposure of FaO cells to LE leads to an increase in mitochondrial ROS production and a decrease in cellular energy levels followed by necrotic cell death.


Gene expression patterns in juvenile American alligators (Alligator mississippiensis) exposed to environmental contaminants.
Reproductive and developmental abnormalities have been reported in the American alligator (Alligator mississippiensis) population from Lake Apopka, FL, that is chronically exposed to a complex mixture of environmental contaminants. To begin to understand the molecular mechanisms that could lead to the observed abnormalities of the reproductive and endocrine system, we quantified concentrations of the steroid hormones testosterone (T) and estradiol-17beta (E(2)) and expression of steroid hormone receptors and genes relating to steroidogenesis in gonadal tissue from juvenile alligators from three lakes in Florida using enzyme immunoassay and quantitative real-time polymerase chain reaction. Alterations of ESR2 (estrogen receptor beta) and SF1 (steroidogenic factor 1) mRNA expression in male gonadal tissue, without an observed difference in plasma concentrations of T, from the different lakes, begin to provide insight into potential mechanisms underlying the alterations of the reproductive system previously observed. Likewise, alterations in P450 aromatase and DAX1 (dosage-sensitive sex reversal, adrenal hypoplasia congenita critical region on the X chromosome, gene 1) mRNA expression, with elevated plasma E(2) concentrations in females, provide leads to the potential mechanisms modifying folliculogenesis and ovarian development. The investigation of these genes also helps clarify normal endocrine and reproductive system function in the American alligator.


Subtype Selective γ-Aminobutyric Acid Type A Receptor (GABAAR) Modulators Acting at the Benzodiazepine Binding Site: An Update.
γ-Aminobutyric acid (GABA) is the major inhibitory neurotransmitter within the central nervous system (CNS) with fast, transsynaptic, and modulatory extrasynaptic effects being mediated by the ionotropic GABA type A receptors (GABAARs). These receptors are of particular interest because they are the molecular target of a number of pharmacological agents, of which the benzodiazepines (BZDs), such as diazepam, are the best described. The anxiolytic, sedating, and myorelaxant effects of BZDs are mediated by separate populations of GABAARs containing either α1, α2, α3, or α5 subunits and the molecular dissection of the pharmacology of BZDs indicates that subtype-selective GABAAR modulators will have novel pharmacological profiles. This is best exemplified by α2/α3-GABAAR positive allosteric modulators (PAMs) and α5-GABAAR negative allosteric modulators (NAMs), which were originally developed as nonsedating anxiolytics and cognition enhancers, respectively. This review aims to summarize the current state of the field of subtype-selective GABAAR modulators acting via the BZD binding site and their potential clinical indications.


Relative effectiveness of powered and manual toothbrushes in elderly patients with implant-supported mandibular overdentures.
The aim of this study was to compare the clinical effectiveness of a powered toothbrush (Braun Oral-B Plaque Remover 3-D) and a manual soft toothbrush (Oral-B Squish-grip brush) for the control of supragingival plaque and soft tissue inflammation around implants supporting mandibular overdentures. The study sample involved 40 edentulous subjects, aged 55-80 years, having 2 unsplinted mandibular implants supporting a complete removable overdenture opposed by a maxillary complete denture. In this single-blinded, randomised, cross-over clinical trial, two 6-week experimental phases were separated by a 2-week wash-out period. 2 weeks prior to each experimental phase (pre-entry visits), implant abutments were polished to remove all plaque and a standardised instruction in the use of the toothbrush was given. Modified plaque and bleeding indices were recorded at the start and end of each experimental period. Mean index scores at each phase were analysed using paired t-test, and the mean number of sites showing a change in plaque or mucositis were compared using the Mann-Whitney U-test. Combined data from 2 different implant systems were considered after controlling for implant type. Only minor changes in plaque and bleeding scores were observed following the two test periods. There were no statistically significant differences between the manual and powered toothbrushes. Manual and powered brushes were found to be of comparable efficacy with regard to improvement in peri-implant bleeding and plaque indices.


Retrograde peri-implantitis associated with residual cysts: 3 Case reports.
The authors describe 3 cases of retrograde peri-implantitis associated with residual cysts and their successful surgical and regenerative management. Implants were originally placed with both immediate and delayed protocols. Three patients sought treatment at the authors' specialist private practices with the chief symptoms of pain and swelling in association with implant-retained restorations. Clinical, radiographic, and histopathologic findings supported the diagnoses of infected residual cysts. The lesions were enucleated in toto, the implant surfaces were decontaminated, and bone regeneration procedures were carried out. Long-term recall visits, ranging from 3 through 10 years, confirmed successful retention of all implants treated accordingly. The authors describe an unusual clinical manifestation and emphasize the role of local procedures during implant placement in areas compromised with infection, cysts, or both and the importance of long-term recall visits.


Changes in UK ophthalmology surgical training: analysis of cumulative surgical experience 2009-2015.
To investigate changes in the patterns of cumulative surgical experience for ophthalmologists in the UK following the introduction of a new national training scheme. Retrospective review of all surgical training records submitted to the UK Royal College of Ophthalmologists by trainees for the award of Certificate of Completion of Training (CCT) for the period 2009-2015. Secondary level care, UK. 539 trainees achieving CCT over the 7-year study period. Higher specialist training or ophthalmology specialist training. Number of CCT awards by years and procedures performed for cataract surgery, strabismus, corneal grafts, vitreoretinal (VR) procedures, oculoplastics and glaucoma. Cataract surgical experience showed little change with median number performed/performed supervised (P/PS) 592, IQR: 472-738; mean: 631. Similarly, the median number of strabismus (P/PS 34), corneal grafts (assisted, 9) and VR procedures (assisted, 34) appeared constant. There was a trend towards increasing surgical numbers for oculoplastics (median 116) and glaucoma (57). Overall case numbers for ophthalmic specialist training (OST) trainees (7-year training programme) were higher than higher surgical training (HST) trainees (4.5-year programme) with the exception of squint (P/PS), corneal grafts (P/PS) and VR cases (P/PS). Overall case numbers reported at time of CCT application appear stable or with a marginal trend towards increasing case numbers. HST (4.5-year programme) case numbers do not include those performed before entry to HST, and although case numbers tended to be higher for OST trainees (7-year programme) compared with HST trainees, they were not proportionately so.


Ba2+ selectively inhibits receptor-mediated contraction of the esophageal muscularis mucosae.
The aim of the present study was to examine the effect of Ba2+ on acetylcholine- and KCl-induced contractions of the guinea-pig esophageal muscularis mucosae. When the muscularis mucosae was pretreated with nicardipine (1 microM), Ba2+ (0.1-30 mM) markedly inhibited the acetylcholine (3 microM)-induced tone, and at 10-30 mM the tone returned to its basal level. In contrast, Ba2+ (0.1-30 mM) slightly increased the KCl (60 mM)-induced tone. Moreover, the Ba2+ (30 mM)-increased KCl tone was completely inhibited by treatment with nicardipine (0.3-1 microM). In conclusion, Ba2+ both selectively inhibits receptor-mediated contraction of the muscularis mucosae and itself permeates through voltage-dependent Ca2+ channels.