Soyacystatin, a novel cysteine proteinase inhibitor in soybean, is distinct in protein structure and gene organization from other cystatins of animal and plant origin.
Cystatins, cysteine proteinase inhibitors, deserve note because of their regulatory and protective functions in plant tissues. We isolated both genomic DNA and cDNA clones from soybean that encode a cystatin consisting of 245 amino acid residues (soyacystatin). It is, while basically similar in sequence to known cystatins that are generally in the range of 12-15 kDa, characterized by having extremely large extension sequences in both its amino and carboxyl termini. The genomic DNA encoding soyacystatin is also unique in that it consists of four exons with three introns in its coding regions. The mRNA for soyacystatin is distinctly expressed in soybean seeds 2 weeks after flowering. Soyacystatin purified from mature soybean seeds had a molecular mass of about 26 kDa on SDS/PAGE which suggests that it contains the extension sequences. Papain-inhibition experiments demonstrate that this endogenous soyacystatin has almost the same inhibitory activity as that of its deletion mutant (102 amino acid residues) recombinantly produced by truncation of the amino and carboxyl terminal extensions, indicating that the occurrence of the extensions does not affect the cystatin activity. Immunohistochemical experiments reveal that soyacystatin is expressed nearly uniformly in the cotyledons. These results also suggest the possible occurrence of a cysteine proteinase as the target enzyme of soyacystatin.


Long-term follow-up of coronary angioplasty in patients with diabetes compared with nondiabetics.
Some reports have indicated that in patients with diabetes mellitus and multivessel disease, coronary artery bypass surgery is preferred over percutaneous coronary intervention (PCI). We retrospectively compared outcome PCI in diabetic and nondiabetic patients. Ninety-seven diabetics and 971 nondiabetics were included and randomised before PCI to aspirin alone or aspirin plus coumadin. Fifty diabetics and 481 nondiabetics underwent follow-up angiography. The primary endpoint comprised all-cause mortality, myocardial infarction or targetvessel revascularisation. Baseline characteristics were similar between the groups except for significantly more males and smokers among the nondiabetics. The diabetics had significantly more previous strokes, more left anterior descending coronary artery disease as well as more restenotic lesions and multivessel disease. At 30 days, the primary endpoint had occurred in five diabetics (5.2%) and 47 nondiabetics (4.9%), (p=0.8) and at one year in 17 (17.5%) and in 165 (17.1%), respectively (p=0.9). Event-free survival remained comparable during long-term follow-up (four years). Multivariate analysis showed no differences for the occurrence of any event (p=0.9, 95% CI 0.6-1.7). At six months, the minimal luminal diameter was significantly smaller in the diabetics (1.55±0.76 mm vs. 1.78±0.66 mm, p=0.01). Diabetics also had more restenosis (41% vs. 23%, p=0.003). Despite angiographical differences at six months between the diabetics and nondiabetics, both short-term and long-term clinical follow-up appeared to be similar.


Effect of Asbestos Exposure on the Frequency of EGFR Mutations and ALK/ROS1 Rearrangements in Patients With Lung Adenocarcinoma: A Multicentric Study.
The aim of this study is to investigate the effect of asbestos exposure on cancer-driver mutations. Between January 2014 and September 2018, epidermal growth factor receptor (EGFR), anaplastic lymphoma receptor tyrosine kinase (ALK), and c-ros oncogene 1 receptor tyrosine kinase gene (ROS1) alterations, demographic characteristics, asbestos exposure, and asbestos-related radiological findings of 1904 patients with lung adenocarcinoma were recorded. The frequencies of EGFR mutations, ALK, and ROS1 rearrangements were 14.5%, 3.7%, and 0.9%, respectively. The rates of EGFR mutations and ALK rearrangements were more frequent in asbestos exposed non-smokers (48.7% and 9%, respectively). EGFR mutation rate was correlated to female gender and not-smoking, ALK rearrangement rate was correlated to younger age, not-smoking, and a history of asbestos exposure. The higher rate of ALK rearrangements in asbestos-exposed lung adenocarcinoma cases shows that asbestos exposure may most likely cause genetic alterations that drive pulmonary adenocarcinogenesis.


Use of a machine learning model to predict retention in care in an urban HIV clinic.
Identifying barriers to retention in care (RIC) is critical to ending the HIV epidemic in the United States. Therefore, we developed a machine learning model (MLM) to identify predictive factors for RIC in an urban HIV clinic. Our MLM yielded a positive predictive value of 84%, higher than previously reported MLMs. We found that MLM can be used to develop interventional strategies to enhance RIC in HIV care.


Molecular cloning and functional characterization of murine cDNA encoding transcription factor NFATc.
Transcription factors of the NFAT (nuclear factor of activated T cells) family play important roles in immune and inflammatory responses by regulating the expression of genes encoding cytokines and immunoregulatory proteins. Here we describe cloning and characterization of full-length cDNA encoding murine (m) NFATc which predicts that the protein has all the conserved structural motifs of NFAT family members, including the rel homology domain, the NFAT homology domain and the nuclear translocation signals. mNFATc complexed with AP-1 bound specifically to the murine IL-2 NFAT recognition sequence and activated transcription from the co-transfected IL-2 promoter in COS-7 cells. Northern blot analysis showed that the cDNA probe hybridized with a 4.5 kb transcript which is highly inducible in murine T cells. By Northern and in situ hybridization, mNFATc transcript was detected from the early stage of development. In the mouse embryo, mNFATc transcript was strongly expressed in thymus, lung and submandibular gland and weakly in skeletal muscle and heart suggesting that mNFATc may have a role both in embryogenesis and in mature T cells.


Management of bone-invasive, hyperostotic sphenoid wing meningiomas.
The hyperostosis frequently associated with sphenoid wing meningiomas is actual invasion of bone by the tumor. The intracranial portion of the tumor is usually thin with en plaque spread, and the tumor tends to invade the orbit through the superior orbital fissure. The authors reviewed the records of 67 patients with sphenoid wing meningiomas who underwent surgery at the University of Arkansas for Medical Sciences between 1994 and 2004. In all 67 cases, the surgery was performed by the senior author. Seventeen of the patients had the distinguishing characteristics of hyperostotic sphenoid wing meningiomas-extensive bone invasion, en plaque dural involvement, and a minimal intracranial mass with minimal orbital involvement. In all patients, hyperostosis was determined on the basis of preoperative neuroimaging. Histopathological evaluation of bone specimens was performed in 14 cases. Estrogen and progesterone receptor expression and Ki 67 labeling were evaluated in all specimens. Chromosome analysis was performed in all tumors resected since 2001 (seven cases). Particular attention was paid to removing all involved bone and dura mater. Total removal was achieved in 14 cases (82.3%), with only one recurrence (7.1%) over a mean follow-up period of 36 months (range 5-72 months). Radical resection was followed by cranioorbital reconstruction to prevent enophthalmos and to obtain good cosmetic results. No deaths or serious complications occurred in association with surgery. Proptosis was corrected in all cases and visual acuity improved in seven (70%) of 10 cases. Revision of the orbital reconstruction was required because of postoperative enophthalmos (two cases) or restricted postoperative ocular movement (one case). Sphenoid wing meningiomas frequently invade bone, although such invasion does not represent malignancy. These lesions are generally histologically benign. Total removal with a prospect for cure and visual preservation should be the goal of treatment. This requires extensive drilling of the invaded bone and extensive excision of the involved dura. When the optic canal is involved, it should be decompressed. Extensive bone resection should be followed by cranioorbital reconstruction for good cosmesis and to prevent enophthalmos.


Bone status and adipokine levels in children on vegetarian and omnivorous diets.
Measurements of bone mineral density (BMD) reflect bone status but not the dynamics of bone turnover. Biochemical markers, which show global skeletal activity, were validated for the assessment of bone formation and resorption processes. Adipokines also play a significant role in the regulation of bone metabolism. To assess body composition, bone mineral density, bone turnover markers and adipokine levels in relation to vegetarian and omnivorous diets. The study included 53 vegetarian and 53 omnivorous prepubertal healthy children matched for age and sex (median age 7.0 years). Body composition and BMD were assessed by dual-energy X-ray absorptiometry. 25-hydroxyvitamin D and parathormone levels were measured by chemiluminescence method. Serum carboxy-terminal propeptide of type I collagen (CICP), total osteocalcin (OC) and its forms carboxylated (c-OC) and undercarboxylated (uc-OC), C-terminal cross-linking telopeptide of collagen type I (CTX), leptin and adiponectin levels were determined using immunoenzymatic assays. Both groups of children were comparable in terms of body composition, except for the percentage of fat mass, which was lower (19.24 vs. 21.77%, p = 0.018) in vegetarians. Mean values of total BMD z-score and lumbar spine BMD z-score were lower (-0.583 vs. -0.194, p = 0.009 and -0.877 vs. -0.496, p = 0.019, respectively) in vegetarians compared with omnivores. Serum leptin level was about 2-fold lower (1.39 vs. 2.94 ng/mL, p < 0.001) in vegetarians, however, adiponectin concentration was similar in both groups. Vegetarians had similar concentration of 25-hydroxyvitamin D, but higher parathormone (40.8 vs. 32.1 pg/mL, p = 0.015) and CTX (1.94 vs. 1.76 ng/mL, p = 0.077) levels than omnivores. Total osteocalcin and CICP concentrations were comparable in both groups, however, c-OC/uc-OC ratio was higher (1.43 vs. 1.04 ng/mL, p < 0.05) in vegetarians. We found positive correlation between c-OC and nutritional parameters adjusted for total energy intake (plant protein, phosphorus, magnesium and fiber intakes) in vegetarian children. Prepubertal children on a vegetarian diet had significantly lower total and lumbar spine BMD z-scores, but absolute values of bone mineral density did not differ. BMD z-scores did not correlate with bone metabolism markers and nutritional variables, but were positively associated with anthropometric parameters. Lower leptin levels in vegetarian children reflect lower body fat. Longitudinal studies are necessary to evaluate the impact of the observed association on bone health at adulthood.


Comparison of two assessments of real-world data and real-world evidence for regulatory decision-making.
Real-world data (RWD) and real-world evidence (RWE) are increasingly used to support regulatory decision-making, but regulatory agencies and stakeholders may apply different definitions for RWD and use different criteria to determine when analysis of such data are considered RWE in decisions on drug approvals. To explore this issue, we reviewed two prominent publications that operationalized the definitions of RWD and RWE when describing the use of RWE in drug approvals by the United States Food & Drug Administration (FDA) and the European Medicines Agency (EMA). Both publications considered non-interventional (observational) studies, RWD as a comparator arm for a single-arm trial, product-related literature reviews, and RWD to support clinical trial implementation (e.g., to identify potential participants) as generating RWE. In contrast, inconsistencies were identified regarding types of data sources and study designs that were considered as not generating RWE. For example, a lack of agreement existed regarding whether RWE is generated when RWD describe therapeutic contexts or are used in phase I/II interventional trials, open-label extension studies, or pharmacovigilance activities. These discrepancies highlight opportunities to develop a consistent understanding of the role of RWE in regulatory decision-making for drug approvals among regulatory agencies and stakeholders.


Safety and efficacy of a novel drug elores (ceftriaxone+sulbactam+disodium edetate) in the management of multi-drug resistant bacterial infections in tertiary care centers: a post-marketing surveillance study.
In India, Elores (CSE-1034: ceftriaxone+sulbactam+disodium edetate) was approved as a broad spectrum antibiotic in year 2011 and is used for management of Extended Spectrum Beta Lactamases/Metallo Beta lactamases infections in tertiary care centers. The objective of this study was to investigate the efficacy of this drug in patients with Extended Spectrum Beta Lactamases/Metallo Beta lactamases infections and identify the incidence of adverse events in real clinical settings. This Post Marketing Surveillance study was conducted at 17 centers across India and included 2500 patients of all age groups suffering from various bacterial infections and treated with Elores (CSE1034). Information regarding demographic, clinical and microbiological parameters, dosage and treatment duration, efficacy and adverse events (AEs) associated with the treatment were recorded. A total of 2500 patients were included in the study and efficacy was evaluated in 2487 patients. In total, 409 AEs were reported in 211 (8.4%) patients. The major AEs reported were vomiting (3.0%), pain at injection site (2.5%), nausea (2.3%), redness at site (1.96%), thrombophlebitis (1.4%). Of total reported AEs, 40 (5.3%) AEs were reported in pediatric, 310 (20.6%) in adult, and 59 (23.6%) in geriatric group. No AE belonging to grade IV or V was reported in any patient. In terms of efficacy, 1977 (79.4%) patients were cured, 501 (20.1%) patients showed clinical improvement and 5 (0.2%) patients were complete failure. The treatment duration varied from 5 to 7 days in different patients depending on the infection type. In this post-marketing surveillance study, CSE-1034 was found to be an effective and safe option against Pip tazo and meropenem in management of patients with multi-drug resistant (MDR) bacterial infections under routine ward settings.


Selective trapping of single mammalian breast cancer cells by insulator-based dielectrophoresis.
The trapping or immobilization of individual cells at specific locations in microfluidic platforms is essential for single cell studies, especially those requiring cell stimulation and downstream analysis of cellular content. Selectivity for individual cell types is required when mixtures of cells are analyzed in heterogeneous and complex matrices, such as the selection of metastatic cells within blood samples. Here, we demonstrate a microfluidic device based on direct current (DC) insulator-based dielectrophoresis (iDEP) for selective trapping of single MCF-7 breast cancer cells from mixtures with both mammalian peripheral blood mononuclear cells (PBMC) as well MDA-MB-231 as a second breast cancer cell type. The microfluidic device has a teardrop iDEP design optimized for the selective capture of single cells based on their differential DEP behavior under DC conditions. Numerical simulations adapted to experimental device geometries and buffer conditions predicted the trapping condition in which the dielectrophoretic force overcomes electrokinetic forces for MCF-7 cells, whereas PBMCs were not trapped. Experimentally, selective trapping of viable MCF-7 cells in mixtures with PBMCs was demonstrated in good agreement with simulations. A similar approach was also executed to demonstrate the selective trapping of MCF-7 cells in a mixture with MDA-MB-231 cells, indicating the selectivity of the device for weakly invasive and highly invasive breast cancer cells. The DEP studies were complemented with cell viability tests indicating acceptable cell viability over the course of an iDEP trapping experiment.


Detailed Phytochemical Evaluation of a Locally Endemic Species (Campanula baskilensis) by LC-MS/MS and Its In-Depth Antioxidant and Enzyme Inhibitory Activities.
The importance of bioactive plant species in the scientific world is increasing day by day. The relationship between health and traditional-modern life, promotes the creation of new value-added natural products. This is the first research to conduct a bioactivity and chemical composition analysis of Campanula baskilensis species, which belongs to the medicinally important genus Campanula L (Campanulaceae). The aim of the current study is to quantitatively investigate the phytochemical contents of aerial and root parts of different C. baskilensis extracts (ethanol, methanol, and water) by LC-MS/MS and to evaluate their total phenolic and flavonoid contents, antioxidant and enzyme inhibitory activities. Remarkably, LC-MS/MS results revealed that, high amounts of quinic acid (53.6 mg/g aerial-MeOH extract), fumaric acid (6.3 mg/g aerial-H2 O extract, 2.5 mg/g root-H2 O extract), protocatechuic acid (11.4 mg/g aerial-H2 O extract), vanillic acid (1.4 mg/g aerial-EtOH extract), quercetin-3-O-rutinoside (rutin) (2.3 mg/g aerial-EtOH extract), hesperetin 7-rutinoside (hesperidin) (2.0 mg/g aerial-EtOH extract), kaempferol-3-O-rutinoside (nicotiflorin) (5.5 mg/g aerial-EtOH extract) were detected in the extracts of the species. Considering the bioactivity tests performed on C. baskilensis extracts, aerial-H2 O extract showed significant activity in all antioxidant assays. However, ethanol extracts of root and aerial parts exhibited the highest activities in all enzyme inhibitory tests.


Home care nurses' experiences with and perceptions of elder self-neglect.
Self-neglect is a poorly understood phenomenon evident to home healthcare nurses who describe older adults who self-neglect as disheveled, unkempt individuals living in cluttered, filthy homes. In spite of the concerns nurses report about these individuals and their situations, individuals who self-neglect give no indication there is any reason for concern for their welfare, and in fact some refuse intervention. The purpose of this study was to determine how home healthcare nurses perceive elder self-neglect, their experiences with this phenomenon, and to explore the steps nurses take when self-neglect is suspected.


Have historical climate changes affected Gentoo penguin (Pygoscelis papua) populations in Antarctica?
The West Antarctic Peninsula (WAP) has been suffering an increase in its atmospheric temperature during the last 50 years, mainly associated with global warming. This increment of temperature trend associated with changes in sea-ice dynamics has an impact on organisms, affecting their phenology, physiology and distribution range. For instance, rapid demographic changes in Pygoscelis penguins have been reported over the last 50 years in WAP, resulting in population expansion of sub-Antarctic Gentoo penguin (P. papua) and retreat of Antarctic Adelie penguin (P. adeliae). Current global warming has been mainly associated with human activities; however these climate trends are framed in a historical context of climate changes, particularly during the Pleistocene, characterized by an alternation between glacial and interglacial periods. During the last maximal glacial (LGM∼21,000 BP) the ice sheet cover reached its maximum extension on the West Antarctic Peninsula (WAP), causing local extinction of Antarctic taxa, migration to lower latitudes and/or survival in glacial refugia. We studied the HRVI of mtDNA and the nuclear intron βfibint7 of 150 individuals of the WAP to understand the demographic history and population structure of P. papua. We found high genetic diversity, reduced population genetic structure and a signature of population expansion estimated around 13,000 BP, much before the first paleocolony fossil records (∼1,100 BP). Our results suggest that the species may have survived in peri-Antarctic refugia such as South Georgia and North Sandwich islands and recolonized the Antarctic Peninsula and South Shetland Islands after the ice sheet retreat.


Pediatric Myxopapillary Ependymomas: A Clinicopathologic Evaluation.
Myxopapillary ependymomas (MPEs) have an indolent clinical course, corresponding to World Health Organization Grade I. A total of 13 pediatric MPEs have been reported in the literature with "anaplastic features," including elevated proliferative activity (≥5 mitoses/10 high-power fields), necrosis, and microvascular proliferation. No consensus exists regarding the prognostic significance of such features. A retrospective clinicopathologic review of pediatric MPEs diagnosed between 1996 and 2018 at Mayo Clinic was performed. Totally, 8 pediatric MPEs (6 male; age: 7.52 to 16.88 y) were identified. Totally, 3 had disseminated disease at presentation. All patients underwent surgical resection (7 gross total; 1 subtotal). Totally, 5 cases harbored ≥5 mitoses/10 high-power fields (range: 5 to 9), 3 of which showed necrosis (2 with disseminated disease). Postsurgery, 2 patients received radiation; one with disseminated disease and another with increased mitotic activity/necrosis; neither has recurred (follow-up: 1.18 and 3.19 y). In all, 2 patients with disseminated disease, elevated mitotic activity, and necrosis had new metastatic disease/progression of nonresected metastatic foci (2.6 and 26.8 mo), received radiation therapy, and remain progression free (3.01 and 9.34 y). All patients are alive (median follow-up 1.31 y, range: 0.66 to 11.75). Among pediatric MPEs, the concurrent presence of elevated mitotic activity and necrosis may be associated with an aggressive clinical course, warranting closer surveillance and consideration of adjuvant therapies.


Cats and owners interact more with each other after a longer duration of separation.
Little is known about the cat's (Felis silvestris catus) need for human contact, although it is generally believed that cats are more independent pets than e.g. dogs. In this study, we investigated the effect of time left alone at home on cat behaviour (e.g. social and distress-related) before, during and after separation from their owner. Fourteen privately owned cats (single-housed) were each subjected to two treatments: the cat was left alone in their home environment for 30 min (T0.5) and for 4 h (T4). There were no differences between treatments in the behaviour of the cat (or owner) before owner departure, nor during the first 5 min of separation. During separation, cats were lying down resting proportionally less (T = 22.5, P = 0.02) in T0.5 (0.27±0.1 (mean±SE)) compared to in T4 (0.58±0.08), probably due to a similar duration of higher activity early in the separation phase in both treatments. Comparisons of the time interval (min 20-25) in both treatments indicated no differences across treatments, which supports such an explanation. Towards the end of the separation phase (the last two 5-min intervals of separation in both treatments), no differences were observed in the cats' behaviour, indicating that cats were unaffected by separation length. At reunion however, cats purred more (T = 10.5, P = 0.03) and stretched their body more (T = 17, P = 0.04) after a longer duration of separation (T4:0.05±0.02; 0.03±0.01; T0.5: 0.01±0.007; 0.008±0.003). Also, owners initiated more verbal contact (T = 33.5, P = 0.04) after 4 h (0.18±0.05) compared to after 30 min (0.12±0.03). There was no evidence of any correlations between the level of purring or body stretching by the cat and verbal contact by the owner implying that the behavioural expressions seen in the cats are independent of the owner's behaviour. Hence, it seemed as cats coped well with being left alone, but they were affected by the time they were left alone, since they expressed differences in behaviour when the owner returned home. The increased level of social contact initiated by the cats after a longer duration of separation indicates a rebound of contact-seeking behaviour, implying that the owner is an important part of the cat's social environment.


Relationship between electromyographic activity of the vastus lateralis while standing and the extent of bilateral simulated knee-flexion contractures.
The effect of simulated bilateral knee-flexion contractures (KFC) on the electromyographic (EMG) activity of the vastus lateralis was studied by testing 10 normal subjects using surface EMG to test the hypothesis that the activity of the knee extensors would increase as a function of the severity of the contracture. The root mean square of the EMG activity was determined from four 4-s samples taken at 30-s intervals, during 2 min of standing in each of five positions of simulated KFC (0 degree, 10 degrees, 20 degrees, 30 degrees and 40 degrees). A randomly balanced order of conditions was used. KFC were simulated in each subject by means of an adjustable line from the subject's waist to the sole of each foot. An analysis of variance was used to contrast EMG activity, and a significant difference was found between each of the positions (P less than 0.05). The mean (+/- 1 SD) EMG activity, expressed as a percentage of the maximum voluntary contraction, was 0.3% (+/- 0.2) at 0 degree, 7.6% (+/- 5.6) at 10 degrees, 10.9% (+/- 7.6) at 20 degrees, 16.6% (+/- 12.4) at 30 degrees and 24.0% (+/- 14.0) at 40 degrees. A linear relationship was found (r2 = 0.986), expressed by the equation y = 0.62 + 0.56 x, where y represents EMG activity and x represents the extent of simulated KFC (P = 0.0007). The results provide insight into the increased knee extensor activity necessary to stand with KFC and underline the importance of treating this common disorder.


Development of a machine learning-based model for predicting risk of early postoperative recurrence of hepatocellular carcinoma.
Surgical resection is the primary treatment for hepatocellular carcinoma (HCC). However, studies indicate that nearly 70% of patients experience HCC recurrence within five years following hepatectomy. The earlier the recurrence, the worse the prognosis. Current studies on postoperative recurrence primarily rely on postoperative pathology and patient clinical data, which are lagging. Hence, developing a new pre-operative prediction model for postoperative recurrence is crucial for guiding individualized treatment of HCC patients and enhancing their prognosis. To identify key variables in pre-operative clinical and imaging data using machine learning algorithms to construct multiple risk prediction models for early postoperative recurrence of HCC. The demographic and clinical data of 371 HCC patients were collected for this retrospective study. These data were randomly divided into training and test sets at a ratio of 8:2. The training set was analyzed, and key feature variables with predictive value for early HCC recurrence were selected to construct six different machine learning prediction models. Each model was evaluated, and the best-performing model was selected for interpreting the importance of each variable. Finally, an online calculator based on the model was generated for daily clinical practice. Following machine learning analysis, eight key feature variables (age, intratumoral arteries, alpha-fetoprotein, pre-operative blood glucose, number of tumors, glucose-to-lymphocyte ratio, liver cirrhosis, and pre-operative platelets) were selected to construct six different prediction models. The XGBoost model outperformed other models, with the area under the receiver operating characteristic curve in the training, validation, and test datasets being 0.993 (95% confidence interval: 0.982-1.000), 0.734 (0.601-0.867), and 0.706 (0.585-0.827), respectively. Calibration curve and decision curve analysis indicated that the XGBoost model also had good predictive performance and clinical application value. The XGBoost model exhibits superior performance and is a reliable tool for predicting early postoperative HCC recurrence. This model may guide surgical strategies and postoperative individualized medicine.


A comprehensive review of microbial contamination in the indoor environment: sources, sampling, health risks, and mitigation strategies.
The quality of the indoor environment significantly impacts human health and productivity, especially given the amount of time individuals spend indoors globally. While chemical pollutants have been a focus of indoor air quality research, microbial contaminants also have a significant bearing on indoor air quality. This review provides a comprehensive overview of microbial contamination in built environments, covering sources, sampling strategies, and analysis methods. Microbial contamination has various origins, including human occupants, pets, and the outdoor environment. Sampling strategies for indoor microbial contamination include air, surface, and dust sampling, and various analysis methods are used to assess microbial diversity and complexity in indoor environments. The review also discusses the health risks associated with microbial contaminants, including bacteria, fungi, and viruses, and their products in indoor air, highlighting the need for evidence-based studies that can relate to specific health conditions. The importance of indoor air quality is emphasized from the perspective of the COVID-19 pandemic. A section of the review highlights the knowledge gap related to microbiological burden in indoor environments in developing countries, using India as a representative example. Finally, potential mitigation strategies to improve microbiological indoor air quality are briefly reviewed.


Three-dimensional vectorial holography based on machine learning inverse design.
The three-dimensional (3D) vectorial nature of electromagnetic waves of light has not only played a fundamental role in science but also driven disruptive applications in optical display, microscopy, and manipulation. However, conventional optical holography can address only the amplitude and phase information of an optical beam, leaving the 3D vectorial feature of light completely inaccessible. We demonstrate 3D vectorial holography where an arbitrary 3D vectorial field distribution on a wavefront can be precisely reconstructed using the machine learning inverse design based on multilayer perceptron artificial neural networks. This 3D vectorial holography allows the lensless reconstruction of a 3D vectorial holographic image with an ultrawide viewing angle of 94° and a high diffraction efficiency of 78%, necessary for floating displays. The results provide an artificial intelligence-enabled holographic paradigm for harnessing the vectorial nature of light, enabling new machine learning strategies for holographic 3D vectorial fields multiplexing in display and encryption.


Designing research studies on psychosocial interventions in autism.
To address methodological challenges in research on psychosocial interventions for autism spectrum disorder (ASD), a model was developed for systematically validating and disseminating interventions in a sequence of steps. First, initial efficacy studies are conducted to establish interventions as promising. Next, promising interventions are assembled into a manual, which undergoes pilot-testing. Then, randomized clinical trials test efficacy under controlled conditions. Finally, effectiveness studies evaluate outcomes in community settings. Guidelines for research designs at each step are presented. Based on the model, current priorities in ASD research include (a) preparation for efficacy and effectiveness trials by developing manuals for interventions that have shown promise and (b) initial efficacy studies on interventions for core features of ASD such as social reciprocity.


Flow cytometric detection of membrane potential changes in murine lymphocytes induced by concanavalin A.
The effect of the mitogenic lectin concanavalin A on the membrane potential of murine lymphocytes was investigated by observing the fluorescence of cells stained with carbocyanine and oxonol dyes. We describe a rapid and reliable method for detecting lectin-induced membrane potential changes in individual cells by flow cytometric analysis of oxonol fluorescence. By 10 min after addition of lectin to suspensions of isolated cells from lymph node, 7-15% of the cells have responded by releasing oxonol dye, indicating a membrane hyperpolarization. The dose onset of this response is similar to that for mitogenesis, which was assessed by measuring [3H]thymidine incorporation. The effect is abolished by alpha-methyl mannoside (100mM), which prevents concanavalin A from binding to the cells, but not by fucose (100mM). When cells are treated with lectin in medium from which Ca2+ has been omitted or to which quinine (0.5mM) has been added, a membrane depolarization is observed. Since these are conditions under which activation of plasma membrane Ca2+-dependent K+ channels is prevented, these findings support the view that the early hyperpolarization of these cells is brought about by an increase in intracellular free [Ca2+].


[Effect of rapid maxillary expansion on respiratory problems: 10-year retrospective study].
Rhinomanometric studies have demonstrated that the maxillary fast expantion (ERM) reduces the nasal resistance of the air conduct and thus achieving the normal limits, in must cases. The hypotesis is that breathing will change from buccal to nasal, concluding that the nasal defense mechanisms (filtration, humidification and warming of inhaled air) will reduce some respiratory pathologies.


7-aa peptide mimic from HVR1 of HCV protects hepatic injury in rats by reduced expression of key pro-inflammatory factors.
Hepatitis C virus (HCV) infection related hepatitis remains one of the most serious health problems world-wide. During investigation of hypervariable region 1 (HVR1) of HCV, a seven amino acids mimic peptide (GQTYTSG, named 7-P) was identified within the N-terminal of HVR1 that could protect porcine serum (PS)-induced hepatic injury in rats. 7-P could not only decrease the serum levels of aminotransferases (ALT, AST), alkaline phosphatase (AKP), and billirubin, but were also effective in reversing histopathological evidence of PS-induced rat chronic liver injury. The protective mechanism was investigated in detail using Affymetrix microarrays analysis. The bioinformatic data was confirmed by western blotting. Further studies revealed that IL-1beta, tumor necrosis factor receptor superfamily 14 (TNFRSF14), intercellular adhesion molecule 2 (ICAM2), sterol regulatory element-binding protein 1 (SREBP 1), Prohibitin (PHB), and inhibitor of DNA binding 2 (Id2) were the key factors for 7-P-induced hepatoprotective effect. Together these data suggest that 7-P mimic from HVR1 of HCV protects hepatic injury by reducing the expression of key pro-inflammatory factors. 7-P might have potential therapeutic application for hepatitis in human beings.


Pharmacological properties of ACh receptors on the heart of the marine bivalve Meretrix lusoria.
The cholinergic receptors on the heart of the marine bivalve Meretrix lusoria were used as a model to investigate their pharmacological properties in bivalves. By recording method, we measured the contractile force (CF) and the heart rate (HR) of the heart to examine the characteristics of cholinergic receptors of a marine bivalve. The known ACh receptor agonists or antagonists in vertebrate nervous system were then introduced into the heart by perfusion, to monitor their effects upon heart activities. Of the six tested agonists, two mAChR agonists (OXA-22, bethanechol) and two nAChR agonists (methylcarbachol, nicotine) mimicked the response induced by ACh. Among the seven tested antagonists, three mAChR antagonists (trihexyphenidy1, atropine and scopolamine) and three nAChR antagonists (TEA, curare and hexamethonium bromide) showed the antagonistic effects on the inhibition induced by ACh. However, there was one mAChR agonist, metoclopramide, that behaved like an ACh antagonist. Both APE (mAChR agonist) and PTMA (nAChR antagonist) showed no significant effects. This differences suggest that the pharmacological characteristics of ACh receptors on the heart of bivalve are different from those in vertebrates. It may be a novel type of ACh receptors.


The renewal of photoreceptor cell outer segments.
The utilization of methionine-(3)H by retinal photoreceptor cells has been studied by radioautographic technique in the rat, mouse, and frog. In all three species, the labeled amino acid is concentrated initially in the inner segment of the cell. Within 24 hr, the radioactive material is displaced to the base of the outer segment, where it accumulates as a distinct reaction band. The reaction band then gradually moves along the outer segment and ultimately disappears at the apex of the cell, which is in contact with the retinal pigment epithelium. These findings are interpreted to indicate that the photoreceptor cell outer segment is continually renewed, by the repeated lamellar apposition of material (membranous discs) at the base of the outer segment, in conjunction with a balanced removal of material at its apex. The outer segment renewal rate is accelerated in frogs when ambient temperature is raised, and is elevated in both frogs and rats when the intensity of retinal illumination is increased.


Estrogen and progesterone treatment mimicking pregnancy for protection from breast cancer.
Early age at full-term pregnancy lowers the risk of breast cancer in women; lactation seems to be of marginal importance and aborted pregnancy is not associated with reduced risk. Although early full-term pregnancy provides protection against breast cancer, first full-term pregnancy in older women appears to increase the risk. The protective effect of pregnancy has also been observed in rats and mice; in these animals, lactation has an additive effect and interrupted pregnancy provides partial but significant protection. Pregnancy at a young age (< or = 3 months) is highly effective, but pregnancy in older animals (> or = 4 months) is less effective. Parity-induced protection against mammary cancer in rodents can be reproduced by short-term treatment (approximately equivalent to gestational period of rodent or shorter) with the pregnancy hormones, estrogen and progesterone. Administration of pregnancy hormones to nulliparous women may be a useful strategy for protection against breast cancer. However, estrogen and progesterone are thought to play major roles in promotion of the proliferation of breast epithelial cells. Thus, the duration of such treatment and the age at which it is administered are essential factors that require further study. Experimental data suggest that short-term treatment of older rats (aged 6 months) with estrogen and progesterone accelerates mammary carcinogenesis and that long-term (>20 weeks) treatment abolishes the cancer-suppressing effect or even accelerates mammary carcinogenesis. Thus, the available evidence suggests that age and duration of estrogen and progesterone treatment are particularly important factors for protection from breast cancer.


Comprehensive sequence and expression profile analysis of PEX11 gene family in rice.
PEX11 gene family has been shown to be involved in peroxisome biogenesis but very little is known about this gene family in rice. Here we show that five putative PEX11 genes (OsPEX11-1-5) present in rice genome and each contain three conserved motifs. The PEX11 sequences from rice and other species can be classified into three major groups. Among the five rice PEX11 genes, OsPEX11-2 and -3 are most likely duplicated. Expression profile and RT-PCR analysis suggested that the members of PEX11 family in rice had differential expression patterns: OsPEX11-1 and OsPEX11-4 had higher expression levels in leaf tissues than in the other tissues, OsPEX11-2 was detected only in germinated seeds, OsPEX11-3 was expressed predominantly in endosperm and germinated seeds, and OsPEX11-5 was expressed in all the tissues investigated. We also observed that the rice PEX11 genes had differential expression patterns under different abiotic stresses. OsPEX11-1 and OsPEX11-4 were induced by abscisic acid (ABA), hydrogen peroxide (H2O2), salt and low nitrogen stress conditions. OsPEX11-3 was responsive to ABA and H2O2 treatments, and OsPEX11-5 was responsive to ABA, H2O2, and salt treatments. However, OsPEX11-2 had no response to any of the stresses. Our results suggest that the rice PEX11 genes have diversification not only in sequences but also in expression patterns under normal and various stress conditions.


High temporal resolution and streak-free four-dimensional cone-beam computed tomography.
Cone-beam computed tomography (CBCT) has been clinically used to verify patient position and to localize the target of treatment in image-guided radiation therapy (IGRT). However, when the chest and the upper abdomen are scanned, respiratory-induced motion blurring limits the utility of CBCT. In order to mitigate this blurring, respiratory-gated CBCT, i.e. 4D CBCT, was introduced. In 4D CBCT, the cone-beam projection data sets acquired during a gantry rotation are sorted into several respiratory phases. In these gated reconstructions, the number of projections for each respiratory phase is significantly reduced. Consequently, undersampling streaking artifacts are present in the reconstructed images, and the image contrast resolution is also significantly compromised. In this paper, we present a new method to simultaneously achieve both high temporal resolution ( approximately 100 ms) and streaking artifact-free image volumes in 4D CBCT. The enabling technique is a newly proposed image reconstruction method, i.e. prior image constrained compressed sensing (PICCS), which enables accurate image reconstruction using vastly undersampled cone-beam projections and a fully sampled prior image. Using PICCS, a streak-free image can be reconstructed from 10-20 cone-beam projections while the signal-to-noise ratio is determined by a denoising feature of the selected objective function and by the prior image, which is reconstructed using all of the acquired cone-beam projections. This feature of PICCS breaks the connection between the temporal resolution and streaking artifacts' level in 4D CBCT. Numerical simulations and experimental phantom studies have been conducted to validate the method.


A survival-adjusted quantal-response test for analysis of tumor incidence rates in animal carcinogenicity studies.
In rodent cancer bioassays, groups of animals are exposed to different doses of a chemical of interest and followed for tumor occurrence. The resulting tumor rates are commonly analyzed using a survival-adjusted Cochran-Armitage (CA) trend test. The CA trend test has reasonable power when the tumor-response curve is linear in dose, but it may be underpowered for a nonlinear response. An alternative survival-adjusted test procedure based on isotonic regression methodology has previously been proposed. Although this alternative procedure performs well when the tumor response is nonlinear in dose, it has less power than the CA trend test when the response is linear in dose. Here, we introduce a new survival-adjusted test procedure that makes use of both the CA trend test and the isotonic regression-based trend test. Using a broad range of experimental conditions typical of National Toxicology Program (NTP) bioassays, we conducted extensive computer simulations to compare the false-positive error rate and power of the proposed procedure with the survival-adjusted CA trend test. The new procedure competes well with the survival-adjusted CA trend test when observed tumor rates are linear in dose and performs substantially better when observed tumor rates are nonlinear in dose. Further, the proposed trend test almost always has a smaller false-positive rate than does the survival-adjusted CA trend test. We also developed an order-restricted inference-based procedure for performing multiple pairwise comparisons between each of the dose groups and the control group. The trend test and the multiple pairwise comparisons test are demonstrated using an example from a study conducted by the NTP.


Metformin and the ATM DNA damage response (DDR): accelerating the onset of stress-induced senescence to boost protection against cancer.
By activating the ataxia telangiectasia mutated (ATM)-mediated DNA Damage Response (DDR), the AMPK agonist metformin might sensitize cells against further damage, thus mimicking the precancerous stimulus that induces an intrinsic barrier against carcinogenesis. Herein, we present the new hypothesis that metformin might function as a tissue sweeper of pre-malignant cells before they gain stem cell/tumor initiating properties. Because enhanced glycolysis (the Warburg effect) plays a causal role in the gain of stem-like properties of tumor-initiating cells by protecting them from the pro-senescent effects of mitochondrial respiration-induced oxidative stress, metformin's ability to disrupt the glycolytic metabotype may generate a cellular phenotype that is metabolically protected against immortalization. The bioenergetic crisis imposed by metformin, which may involve enhanced mitochondrial biogenesis and oxidative stress, can lower the threshold for cellular senescence by pre-activating an ATM-dependent pseudo-DDR. This allows an accelerated onset of cellular senescence in response to additional oncogenic stresses. By pushing cancer cells to use oxidative phosphorylation instead of glycolysis, metformin can rescue cell surface major histocompatibility complex class I (MHC-I) expression that is downregulated by oncogenic transformation, a crucial adaptation of tumor cells to avoid the adaptive immune response by cytotoxic T-lymphocytes (CTLs). Aside from restoration of tumor immunosurveillance at the cell-autonomous level, metformin can activate a senescence-associated secretory phenotype (SASP) to reinforce senescence growth arrest, which might trigger an immune-mediated clearance of the senescent cells in a non-cell-autonomous manner. By diminishing the probability of escape from the senescence anti-tumor barrier, the net effect of metformin should be a significant decrease in the accumulation of dysfunctional, pre-malignant cells in tissues, including those with the ability to initiate tumors. As life-long or late-life removal of senescent cells has been shown to prevent or delay the onset or progression of age-related disorders, the tissue sweeper function of metformin may inhibit the malignant/metastatic progression of pre-malignant/senescent tumor cells and increase the human lifespan.


[How to develop molecular epidemiology in conventional epidemiology survey on eye diseases in China].
Molecular epidemiology is a new branch of eye epidemiology that combines theories and methods in both epidemiology and molecular biology. The definition of molecular epidemiology is to use biological and, in particular, genetic markers (nucleic acid, protein) as measures for detecting the propensity of a disease developing, or as an indicator of a disease or an exposure in the studies of disease distribution. Molecular epidemiology has the same objectives as conventional ophthalmic epidemiology in a defined population. The main designs used in ophthalmic molecular epidemiology include descriptive study, case control study, and nested case control study. Currently, molecular experimental techniques mainly include single nucleotide polymorphism, ELISA, protein and mRNA array, microRNA and the study of epigenetic markers. Gene susceptibility biomarker is one of the most commonly used molecular markers. The findings of ophthalmic molecular epidemiology studies can be used to design personalized therapy. Undoubtedly, ophthalmic molecular epidemiology will evolve and develop in the new era for the prevention and control of complex eye diseases such as age-related macular degeneration, cataract, glaucoma and diabetic retinopathy.


[Generic drug policy implementation in Brazil].
A generic drug policy has been implemented in Brazil since 1999. Several political and administrative stages transpired between enactment of the legislation and the actual marketing and consumption of these drugs. This article describes the policy implementation process and examines the country's generic drug legislation, approved from 1999 to 2002. To contextualize these measures, the study compares articles published by two national periodicals and interviews with a government representative involved in drafting the legislation and a representative from the pharmaceutical industry. Generic drugs quickly gained considerable space in the Brazilian pharmaceutical market. Ongoing adaptation of the legislation, media support, and the government's involvement in spreading the policy were key success factors. The population's access to medicines did not increase significantly, but people can now purchase medicines at more affordable prices and with quality assurance and interchangeability.


Reducing effect of artichoke extract on heterocyclic aromatic amine formation in beef and chicken breast meat.
The aim of this study was to investigate the inhibitory effect of different levels of artichoke extract (0, 0.5, and 1.0%) on the formation of heterocyclic aromatic amines (HAAs) in beef and chicken breast meat cooked by either pan-frying or oven-roasting. All meat samples were cooked at three different temperatures (150, 200, and 250°C) and the levels of 12 HAAs (IQ, IQx, MeIQ, MeIQx, 4,8-DiMeIQx, 7,8-DiMeIQx, PhIP, harman, norharman, AαC, MeAαC, and Trp-P-2) were assessed. The total HAA content in beef and chicken breast ranged from not detectable to 49.26ng/g, and not detectable to 83.06ng/g, respectively. The inhibitory effects of 0.5 and 1.0% artichoke extracts on total HAAs levels were found to be 6-46% and 25-98% in beef, and 5-97% and 14-95% in chicken breast, respectively. The present study showed that artichoke extracts could mitigate HAA formation especially in oven-roasted beef and chicken breast meat.


Stress-induced β cell early senescence confers protection against type 1 diabetes.
During the progression of type 1 diabetes (T1D), β cells are exposed to significant stress and, therefore, require adaptive responses to survive. The adaptive mechanisms that can preserve β cell function and survival in the face of autoimmunity remain unclear. Here, we show that the deletion of the unfolded protein response (UPR) genes Atf6α or Ire1α in β cells of non-obese diabetic (NOD) mice prior to insulitis generates a p21-driven early senescence phenotype and alters the β cell secretome that significantly enhances the leukemia inhibitory factor-mediated recruitment of M2 macrophages to islets. Consequently, M2 macrophages promote anti-inflammatory responses and immune surveillance that cause the resolution of islet inflammation, the removal of terminally senesced β cells, the reduction of β cell apoptosis, and protection against T1D. We further demonstrate that the p21-mediated early senescence signature is conserved in the residual β cells of T1D patients. Our findings reveal a previously unrecognized link between β cell UPR and senescence that, if leveraged, may represent a novel preventive strategy for T1D.


Perceived stress, stressors, and coping strategies among nursing students in the Middle East and North Africa: an overview of systematic reviews.
In nursing students, high stress levels can lead to burnout, anxiety, and depression. Our objective is to characterize the epidemiology of perceived stress, stressors, and coping strategies among nursing students in the Middle East and North Africa region. We conducted an overview of systematic reviews. We systematically searched PubMed, Embase, PsycInfo, and grey literature sources between January 2008 and June 2020 with no language restrictions. We included any systematic review reporting measurable stress-related outcomes including stress prevalence, stressors, and stress coping strategies in nursing students residing in any of the 20 Middle East and North Africa countries. We also included additional primary studies identified through a hand search of the reference lists of relevant primary studies and systematic reviews. Seven systematic reviews and 42 primary studies with data from Bahrain, Egypt, Iraq, Jordan, Oman, Pakistan, Palestine, Saudi Arabia, and Sudan were identified. Most studies included nursing students undergoing clinical training. The prevalence range of low, moderate, and high perceived stress among nursing students was 0.8-65%, 5.9-84.5%, and 6.7-99.2%, respectively. Differences related to gender, training period, or the type of tool used to measure stress remain unclear given the wide variability in the reported prevalence measures across all stress levels. Common clinical training stressors were assignments, workload, and patient care. Academic training-related stressors included lack of break/leisure time, low grades, exams, and course load. Nursing students utilized problem focused (dealing with the problem), emotion focused (regulating the emotion), and dysfunctional (venting the emotions) stress coping mechanisms to alleviate their stress. Available data does not allow the exploration of links between stress levels, stressors, and coping strategies. Limited country-specific prevalence data prevents comparability between countries. Reducing the number or intensity of stressors through curriculum revision and improving students' coping response could contribute to the reduction of stress levels among students. Mentorship, counseling, and an environment conducive to clinical training are essential to minimize perceived stress, enhance learning, and productivity, and prevent burnout among nursing students.


Familial pseudohyperkalemia in blood donors: a novel mutation with implications for transfusion practice.
Familial pseudohyperkalemia (FP) is a dominantly inherited condition in which red blood cells (RBCs) have an increased cold-induced permeability to monovalent cations. Potassium leaks into the supernatant of all stored blood with time, but FP RBCs leak potassium more rapidly. We investigated two unrelated blood donors whose RBC donations demonstrated unexpectedly high potassium after 5 and 6 days' storage. We matched the observed pattern of RBC cation leak to a previously recognized family with FP (FP-Cardiff) and investigated the likely cause with targeted DNA analysis. Cation leakage from the donor RBCs and from standard donor units was measured. DNA analysis of donors and family members with FP-Cardiff was performed. Allele frequencies were obtained from human variation databases. Both implicated donors were found to have increased cold-induced potassium leak identical in pattern to affected members of the family with FP-Cardiff. We found a heterozygous substitution Arg723Gln in the ATP-binding cassette, Subfamily B, Member 6 protein that segregated with FP in the Cardiff family and was also present in both blood donors. Arg723Gln is listed in human variation databases with an allele frequency of approximately 1:1000. We describe a novel FP mutation that may affect 1:500 European blood donors and causes rapid loss of potassium from stored RBCs. This finding has implications for neonates and infants receiving large-volume RBC transfusions. Genomic screening of donors could be used to identify donors with this mutation and potentially improve the quality and safety of donor units.


Use of the proportion of patients dying on an End of Life Pathway as a quality marker: considerations for interpretation.
The Department of Health as part of its End of Life Care Strategy has developed a set of markers as a quality of care proxy for adults at the end of life. 'The number/proportion of patients dying with the Liverpool Care Pathway (or equivalent) in place' is suggested as a quality metric for all care providers. A retrospective audit of uptake of use of the Liverpool Care Pathway (LCP) in an NHS hospital in the North of England showed that 39% of all patients who died had been placed on the LCP. Overall 58% of patients who died were judged to meet the criteria for LCP use. This represented 81% of patients dying with cancer as a primary cause compared to 51% of patients dying with non-cancer. This difference was statistically very significant. In the Trust under study, 67% of dying patients who fulfilled the LCP criteria were placed on the pathway. The results of this study suggest that a simple percentage of deaths on the pathway is an unsophisticated statistic which needs to be interpreted with care. In particular it will be influenced by the proportion of people dying with cancer or non-cancer. This should be considered particularly when presenting the results to the public and to healthcare stakeholders or when making comparisons between provider organizations.


The nonskeletal consequences of osteoporotic fractures. Psychologic and social outcomes.
The prevalence of osteoporosis is rising as the population of the United States and other developed countries ages. These increasing numbers of people have motivated pharmaceutical companies to develop and market several antiresorptive medications that can slow down the bone loss associated with osteoporosis. Although these are not cures for this disease, they are an important first step in a vital ongoing public health effort to prevent osteoporosis in the future and to manage osteoporosis now. We cannot expect to remediate the problems caused by this disease if we attend only to its skeletal implications. Like any other chronic disease, osteoporosis has significant psychologic and social consequences. From anxiety and depression to social withdrawal and isolation, if these problems are left unresolved, they can have a significant negative impact not only on health issues but also on overall quality of life. No quick fixes exist for the numerous ways in which osteoporosis can transform an autonomous person into a dependent and hopeless patient. In part, responsibility for helping this patient rests with the medical community. Referrals to appropriate providers can improve a patient's physical and emotional well-being. Physician specialists can help the patient manage comorbid conditions. Physical and occupational therapists can teach exercises, home safety, and safe movement. Social workers can provide a framework for coping that enables individuals to improve their interpersonal interactions and minimize stress in their lives. Nutritionists, pharmacists, nurses, and other health care professionals can make major contributions to the quality of life of people with osteoporosis and should be encouraged to do so. Unfortunately, managed care has set policies that deprive patients with osteoporosis of the kinds of care that would be most useful to them. As we have advocated for the last 15 years, a multidisciplinary approach offers patients the most positive overall way to manage osteoporosis. Therefore, new alternatives need to be examined, alternatives that provide both low-cost and high-quality care. In the long run, patients who practice self-management, that is, those who take responsibility for their own calcium and vitamin D intake, are compliant with medications, exercise, and practice home safety, and who have a healthy outlook, can control their osteoporosis. The most effective intervention for the future may be to teach individuals how to use self-management strategies so that they can take charge of their osteoporosis and positively influence their quality of life.


On the margins of health economics: a response to 'resolving NICE'S nasty dilemma'.
In a 2011 article published in this journal, Baker et al. set out to resolve a nasty dilemma for NICE by reconciling two approaches for determining whether adopting a new intervention would increase total health gains produced from available resources and hence increase system efficiency. In this response we show how the proposed reconciliation, as well as the two approaches on which it is based, fail to inform decision makers about the efficiency of a new intervention. We show how this arises from the misuse of incremental costs and effects of between-intervention comparisons as measures of changes in costs and effects associated with marginal adjustments to the scale of an intervention. Ironically, incremental data represent the choices faced by decision makers and we illustrate a method for determining unambiguously whether a new intervention represents an improvement in efficiency.


How to reduce mortality in ST-elevation myocardial infarction patients treated with primary percutaneous coronary interventions: cut the bleeding.
In-hospital mortality for ST-elevation myocardial infarction (STEMI) has declined thanks to a greater use of primary percutaneous coronary interventions (PCI) associated with more effective antiplatelet and anticoagulant drugs. In this regard, bivalirudin has been shown to decrease total and cardiac mortality as compared to unfractionated heparin (UFH). The primary purpose of this analysis is to evaluate the hypothesis that the reduction of in-hospital bleeding is the most plausible explanation for the improved survival of STEMI patients treated with bivalirudin during primary PCI. The secondary objective is to reconsider the prognostic significance of the radial access alone or in association with bivalirudin on the basis of the published data. We have done a comprehensive evaluation of the main and related publications of the HORIZONS-AMI trial in addition to an extensive research by Medline of randomized trials evaluating the prognostic impact of radial access as compared with the femoral one in primary PCI. In the HORIZONS-AMI trial bivalirudin resulted in significantly lower rates of the 30 day primary endpoint (defined as major adverse ischemic outcomes plus major bleeding) over UFH plus GPI, largely due to the significantly lower rate of the protocol-defined major bleeding. All-cause and cardiac mortality were also reduced in the bivalirudin arm at 3 year follow-up. Recent studies have also shown that the use of the radial instead of the femoral approach for primary PCI is associated with reduced bleeding as well as reduced mortality. Our research suggests that decreasing bleeding by either a pharmacologic strategy (use of bivalirudin) or a technical approach (the transradial access) improves survival in STEMI patients undergoing primary PCI. The validity of this hypothesis should be confirmed by specific randomized trials.


Unraveling sub-site specificities of peptidic serine protease inhibitors by substitutional and structural analysis.
The interaction of peptidic inhibitors with serine proteases is investigated using peptide spot synthesis on cellulose sheets. This method permits a highly parallel analysis of subsite specificities and an optimization of peptidic inhibitors towards higher affinity and specificity for a given target protease. Information from crystal structures of corresponding protease/inhibitor complexes may help to explain the observed binding pattern.


Shiga toxin-associated hemolytic-uremic syndrome: combined cytotoxic effects of Shiga toxin, interleukin-1 beta, and tumor necrosis factor alpha on human vascular endothelial cells in vitro.
This study explores the relationship between Shiga toxin-producing Shigella or Escherichia coli strains and the development of vascular complications in humans following bacillary dysentery. We propose that endotoxin-elicited interleukin-1 or tumor necrosis factor alpha (TNF) may combine with Shiga toxin to facilitate vascular damage characteristic of hemolytic-uremic syndrome. This study examines the cytotoxic effects of Shiga toxin, interleukin-1, and TNF on cultured human umbilical vein endothelial cells (HUVEC). Both Shiga toxin and TNF were cytotoxic to HUVEC, although HUVEC obtained from individual umbilical cords differed in their sensitivities to these agents. With Shiga toxin-sensitive HUVEC, combinations of TNF with Shiga toxin resulted in a synergistic cytotoxic effect. In contrast, interleukin-1 was not cytotoxic to HUVEC, nor did it enhance cell death in combination with Shiga toxin. The synergistic cytotoxic response of HUVEC to Shiga toxin and TNF was dose and time dependent for both agents and could be neutralized by monoclonal antibodies directed against either Shiga toxin or TNF. This synergistic response was delayed, being maximal on day 2. Preincubation (24 h) of HUVEC with TNF sensitized the cells to Shiga toxin. TNF alone had no effect on HUVEC protein synthesis but enhanced the inhibitory activity of Shiga toxin. These results are consistent with a role for Shiga toxin in the development of hemolytic-uremic syndrome at the level of the vascular endothelium in humans.


The femR315 gene from Staphylococcus aureus, the interruption of which results in reduced methicillin resistance, encodes a phosphoglucosamine mutase.
The femR315 gene was recently identified by Tn551 insertional mutagenesis as one of the new auxiliary genes, the alteration of which resulted in a drastically reduced methicillin resistance of the Staphylococcus aureus strain COL. femR315 (also known as femD) theoretically encoded a protein of 451 amino acids showing significant amino acid sequence homology with phosphoglucomutases and similar enzymes catalyzing the isomerization of hexoses and hexosamine phosphates (S. Wu, H. de Lencastre, A. Sali, and A. Tomasz, Microb. Drug Resist. 2:277-286, 1996). We describe here the overproduction and purification of the FemR315 protein as well as its identification as the phosphoglucosamine mutase which catalyzes the formation of glucosamine-1-phosphate from glucosamine-6-phosphate, the first step in the reaction sequence leading to the essential peptidoglycan precursor UDP-N-acetylglucosamine. On the basis of these findings, we propose to change the names femR315 and femD to the functionally more appropriate name glmM.


Experimental allergic encephalomyelitis in agammaglobulinemic chickens.
White Leghorn chickens were subjected to bursectomy and total body irradiation at hatching, thus rendering them agammaglobulinemic. At 4 weeks of age, these chickens and control chickens received lyophilized bovine spinal cord emulsified with complete Freund's adjuvant. Agammaglobulinemic chickens developed the lesions of allergic encephalomyelitis with the same frequency and intensity as the controls. Our findings support the contention that antibodies are not necessary for the development of experimental allergic encephalomyelitis.


Evasion from NK cell immunity by MHC class I chain-related molecules expressing colon adenocarcinoma.
Evasion of host immune responses is well documented for viruses and may also occur during tumor immunosurveillance. The mechanisms involve alterations in MHC class I expression, Ag processing and presentation, chemokine and cytokine production, and lymphocyte receptor expression. Epithelial tumors overexpress MHC class I chain-related (MIC) molecules, which are ligands for the activating receptor NKG2D on NK and T cells. We report that NK cells from patients with colorectal cancer lack expression of activating NKG2D and chemokine CXCR1 receptors, both of which are internalized. Serum levels of soluble MIC (sMIC) are elevated and are responsible for down-modulation of NKG2D and CXCR1. In contrast, high serum levels of CXC ligands, IL-8, and epithelial-neutrophil-activating peptide (ENA-78) do not down-modulate CXCR1. In vitro, internalization of NKG2D and CXCR1 occurs within 4 and 24 h, respectively, of incubating normal NK cells with sMIC-containing serum. Furthermore, natural cytotoxicity receptor NKp44 and chemokine receptor CCR7 are also down-modulated in IL-2-activated NK cells cocultured in MIC-containing serum-an effect secondary to the down-modulation of NKG2D and not directly caused by physical association with sMIC. The patients' NK cells up-regulate expression of NKG2D, NKp44, CXCR1, and CCR7 when cultured in normal serum or anti-MIC Ab-treated autologous serum. NKG2D(+) but not NKG2D(-) NK cells are tumoricidal in vitro, and in vivo they selectively traffic to the xenografted carcinoma, form immunological synapse with tumor cells, and significantly retard tumor growth in the SCID mice. These results suggest that circulating sMIC in the cancer patients deactivates NK immunity by down-modulating important activating and chemokine receptors.


Anaphylactic and anaphylactoid reactions associated with lepirudin in patients with heparin-induced thrombocytopenia.
Lepirudin (Refludan) is a hirudin derivative. It is a direct thrombin inhibitor obtained by recombinant technology from the medicinal leech and is approved for treatment of heparin-induced thrombocytopenia complicated by thrombosis. Because 3 cases of fatal anaphylaxis possibly associated with use of lepirudin have been reported, we initiated an investigation of putative lepirudin-associated anaphylaxis. Aided by the manufacturer (Schering AG, Berlin, Germany), we used the lepirudin study databases to identify all patients in whom possible anaphylaxis/severe allergy was recorded from 1994 to September 2002. The 26 possible cases identified were reviewed independently by 2 investigators. After excluding patients with mild skin reactions, reactions likely caused by concomitant medications, poorly documented cases, and reactions that did not correspond temporally with lepirudin use, there remained 9 patients judged to have had severe anaphylaxis in close temporal association with lepirudin. All reactions occurred within minutes of intravenous lepirudin administration, with 4 fatal outcomes (3 acute cardiorespiratory arrests, 1 hypotension-induced myocardial infarction). In these 4 cases, a previous uneventful treatment course with lepirudin was identified (1 to 12 weeks earlier). We recorded high-titer IgG-anti-lepirudin antibodies in an additional patient with anaphylaxis. Because lepirudin has been used in approximately 35 000 patients, the risk of anaphylaxis is approximately 0.015% (5 of 32 500) on first exposure and 0.16% (4 of 2500) in reexposed patients (7.5% estimated reexposures). Lepirudin can cause fatal anaphylaxis, particularly in patients who are treated within 3 months of a previous exposure. The overall risk/benefit assessment of lepirudin as a treatment for heparin-induced thrombocytopenia remains favorable.


Reprogrammed quiescent B cells provide an effective cellular therapy against chronic experimental autoimmune encephalomyelitis.
Activated B cells can regulate immunity and have been envisaged as a potential cell-based therapy for treating autoimmune diseases. However, activated human B cells can also propagate immune responses, and the effects resulting from their infusion into patients cannot be predicted. This led us to consider resting B cells, which in contrast are poorly immunogenic, as an alternative cellular platform for the suppression of unwanted immunity. Here, we report that resting B cells can be directly engineered with lentiviral vectors to express antigens in a remarkably simple, rapid, and effective way. Notably, this neither required nor induced activation of the B cells. With this approach we were able to produce reprogrammed resting B cells that inhibited antigen-specific CD4(+) T cells, CD8(+) T cells, and B cells upon adoptive transfer in mice. Furthermore, resting B cells engineered to ectopically express myelin oligodendrocyte glycoprotein antigen protected recipient mice from severe disability and demyelination in EAE, and even induced complete remission from disease in mice lacking functional natural Tregs, which otherwise developed chronic paralysis. In conclusion, our study introduces reprogrammed quiescent B cells as a novel tool for suppressing undesirable immunity.


Evaluation of antioxidant properties of dermocosmetic creams by direct electrochemical measurements.
Cyclic voltammetry and linear sweep voltammetry were preliminarily used in order to evaluate the global antioxidant properties of dermocosmetic creams. Experiments were performed by introducing electrodes directly into the creams without any pretreatment of the samples. Current-potential curves showed significant anodic current depending on the antioxidant-containing cream studied. In comparison, little amperometric response was recorded with an antioxidant-free cream base. Aqueous solutions of the corresponding antioxidants showed analogous anodic waves and similar peak potentials. A correlation between the global anodic peak and the presence of the antioxidant species in the cream was made with eleven skin creams, attesting to the reliability of the method. Among the tested electrode materials, platinum gave the best results in terms of electrochemical kinetics and measurement precision (current peak standard deviation less than 5%). Exposure of a depilatory cream to oxidizing agents (e.g., hydrogen peroxide, air, or light) caused a decrease in peak current as expected. This methodology enabled us to evaluate the evolution of the total antioxidant capacity under oxidative stress and gives encouragement to further development of a voltammetric method to quantify cream antioxidant power.


The reemerging and outbreak of genotypes 4 and 5 of Japanese encephalitis virus.
The Japanese encephalitis virus (JEV) is classified into five distinct genotypes, with genotypes 1 and 3 historically showing higher activity. These genotypes are the primary agents of viral encephalitis in the Asian continent. Genotypes 4 and 5 have remained silent in low-latitude tropical regions since their discovery. From 2009, the hidden genotype 5 suddenly emerged simultaneously in mosquitoes from the Tibetan region of China and those from South Korea in East Asia. The detection of genotype 5 of JEV in these mosquitoes was associated with cases of viral encephalitis in the local population. Similarly, in 2022, the long-silent genotype 4 of JEV emerged in Australia, resulting in a local outbreak of viral encephalitis that primarily affected adults and caused fatalities. The emergence and outbreaks of genotypes 4 and 5 of JEV present new challenges for the prevention and control of Japanese encephalitis (JE). This study not only analyzes the recent emergence of these new genotypes but also discusses their implications in the development of JE vaccines and laboratory tests for newly emerging JEV infections.


Loss of Cdc13 causes genome instability by a deficiency in replication-dependent telomere capping.
In budding yeast, Cdc13, Stn1, and Ten1 form the telomere-binding heterotrimer CST complex. Here we investigate the role of Cdc13/CST in maintaining genome stability by using a Chr VII disome system that can generate recombinants, chromosome loss, and enigmatic unstable chromosomes. In cells expressing a temperature sensitive CDC13 allele, cdc13F684S, unstable chromosomes frequently arise from problems in or near a telomere. We found that, when Cdc13 is defective, passage through S phase causes Exo1-dependent ssDNA and unstable chromosomes that are then the source for additional chromosome instability events (e.g. recombinants, chromosome truncations, dicentrics, and/or chromosome loss). We observed that genome instability arises from a defect in Cdc13's function during DNA replication, not Cdc13's putative post-replication telomere capping function. The molecular nature of the initial unstable chromosomes formed by a Cdc13-defect involves ssDNA and does not involve homologous recombination nor non-homologous end joining; we speculate the original unstable chromosome may be a one-ended double strand break. This system defines a link between Cdc13's function during DNA replication and genome stability in the form of unstable chromosomes, that then progress to form other chromosome changes.


Reappraising the role of albumin for resuscitation.
Fluid resuscitation is a common intervention in acute medical practice. The optimum fluid for resuscitation remains hotly debated and it is likely to vary from one clinical situation to another. Human albumin solutions have been available since the 1940s, but their use varies greatly around the world. This review examines the current evidence for and against the use of albumin as a resuscitation fluid. Fluid resuscitation with albumin has been compared to resuscitation with saline in large high-quality trials in adult ICU patients and in African children. Within overall equivalent effects, albumin may offer a slight mortality benefit in adult ICU patients with severe sepsis whilst increasing mortality in patients with traumatic brain injury. There are no recent high-quality trials comparing albumin to synthetic colloid solutions. In African children with febrile illness and compensated shock, the effects of bolus resuscitation with albumin and saline are similar, but both increase mortality compared to treatment that avoids fluid boluses. Fluid resuscitation with albumin is well tolerated and produces similar results to resuscitation with saline. Albumin should be avoided in patients with traumatic brain injury; possible benefits in adults with severe sepsis remain to be confirmed.


Observation Versus Embolization in Patients with Blunt Splenic Injury After Trauma: A Propensity Score Analysis.
Non-operative management (NOM) is the standard of care in hemodynamically stable patients with blunt splenic injury after trauma. Splenic artery embolization (SAE) is reported to increase observation success rate. Studies demonstrating improved splenic salvage rates with SAE primarily compared SAE with historical controls. The aim of this study was to investigate whether SAE improves success rate compared to observation alone in contemporaneous patients with blunt splenic injury. We included adult patients with blunt splenic injury admitted to five Level 1 Trauma Centers between January 2009 and December 2012 and selected for NOM. Successful treatment was defined as splenic salvage and no splenic re-intervention. We calculated propensity scores, expressing the probability of undergoing SAE, using multivariable logistic regression and created five strata based on the quintiles of the propensity score distribution. A weighted relative risk (RR) was calculated across strata to express the chances of success with SAE. Two hundred and six patients were included in the study. Treatment was successful in 180 patients: 134/146 (92 %) patients treated with observation and 48/57 (84 %) patients treated with SAE. The weighted RR for success with SAE was 1.17 (0.94-1.45); for complications, the weighted RR was 0.71 (0.41-1.22). The mean number of transfused blood products was 4.4 (SD 9.9) in the observation group versus 9.1 (SD 17.2) in the SAE group. After correction for confounders with propensity score stratification technique, there was no significant difference between embolization and observation alone with regard to successful treatment in patients with blunt splenic injury after trauma.


Identification, diagnosis, treatment, and in-hospital outcomes of acute pulmonary embolism: Results from a single integrated health system.
Although the high-risk acute pulmonary embolism (PE) population has been described, little is known about the contemporary inpatient experience and practice patterns of the PE population as a whole. All patients with a diagnosis of acute PE from January 1, 2016, to June 30, 2017 within our academic, multihospital health system were retrospectively identified using International Classification of Diseases, 10th Revision, codes, and data were manually abstracted by 2 clinical investigators. Descriptive analyses were performed according to clinical risk stratification categories from the European Society of Cardiology. Of 829 total patients, 372 (44.8%) patients had intermediate or high-risk PE. Mean age was 62.1 years old, and 42.1% of patients had a history of malignancy. One hundred fifty-three (18.5%) patients had an acute PE during a hospitalization for another indication. A total of 6.0% underwent invasive PE therapies, 26.1% required intensive care unit admission, and 9.0% experienced in-hospital death or hospice discharge. In a subgroup description, patients who developed acute PE during a hospitalization for another indication had a higher incidence of incomplete risk stratification and a higher mortality (9.8%) than the primary cohort. Mortality was attributed to PE in 48.4% of cases. This contemporary description of acute PE managed at a single large, multihospital academic health system highlights substantial health care utilization and high mortality despite the available of advanced therapeutics. Additional work is needed to standardize care for the heterogeneous PE population to ensure appropriate allocation of resources and improved outcomes for all PE patients.


Monoclonal antibodies against murine IFN-gamma abrogate in vivo tumor immunity against RSV-induced murine sarcomas.
Previous work has shown that immunization of syngeneic mice with v-src-induced sarcomas gives rise to specific protection against a lethal dose of v-src-transformed fibroblasts. This immune response is mediated by Lyt-1+, Lyt-2,3+ T lymphocytes, with no apparent involvement of cytotoxic T cells, as shown in Winn-type assays. Immune cells mediating tumor rejection in this system have now been further characterized, and it was found that L3T4+ T lymphocytes alone provided full protection against v-src-induced sarcomas. Moreover, the role of interferon-gamma (IFN-gamma) in the tumor rejection was analyzed. A monoclonal antibody directed against this lymphokine was able to reverse the protective effect displayed by immune T lymphocytes, by eliciting highly effective T suppressor cells. It was thus concluded that T cells with L3T4 surface marker are the main thing responsible for the adoptive immunity in this tumor system, and the activity of these cells is positively modulated by lymphokines such as IFN-gamma.


Activation of human endothelial cells by mobilized porcine leukocytes in vitro: implications for mixed chimerism in xenotransplantation.
The induction of immunologic tolerance to pig antigens in primates may facilitate the development of successful clinical xenotransplantation protocols. The infusion of mobilized porcine peripheral blood leukocytes (PBPC, consisting of approximately 2% peripheral blood progenitor cells) into preconditioned baboons, intended to induce mixed hematopoietic cell chimerism, however, results in a severe thrombotic microangiopathy (TM) that includes vascular injury, microvascular thrombosis, and pronounced thrombocytopenia. Because the mechanisms responsible for TM are unclear, we have explored the effects of PBPC on human umbilical vein endothelial cell (HUVEC) activation. Confluent HUVEC monolayers were established in 96-well cell culture clusters. PBPC were mobilized from miniature swine with porcine interleukin 3 (pIL-3), porcine stem cell factor (pSCF), and human granulocyte-colony stimulating factor (hG-CSF) and were collected by leukapheresis. PBPC were added to HUVEC (0-1x10(7) PBPC/well) for 3- to 24-hr periods and, with cell-based ELISA techniques, surface levels of E-selectin, vascular cell adhesion molecule 1 (VCAM-1), and intercellular adhesion molecule 1 (ICAM-1) were measured. In some cases, peripheral blood leukocytes (PBL) were collected from pigs that did not receive pIL-3, pSCF, or hG-CSF and were added to HUVEC. PBPC were also sorted into subsets of CD2- cells, CD2+ cells, and cellular debris, each of which were added separately to HUVEC. Transwell permeable membrane inserts were placed over HUVEC to prevent direct cell-cell contact with PBPC in some instances. PBPC from different pigs (n=6) induced an increase in the expression of E-selectin, VCAM-1, and ICAM-1 to levels 5, 4, and 2 times greater than baseline, respectively. ICAM-1 expression reached maximum levels after the addition of 6x10(5) PBPC/well. Expression of E-selectin and VCAM-1 increased further with the addition of greater numbers of PBPC, reaching maximum levels after the addition of 1x10(7) PBPC/well. PBPC-induced up-regulation of E-selectin, VCAM-1, and ICAM-1 had a maximum effect after approximately 6 hr, 12 hr, and 6 to 9 hr, respectively (n=3). The effects of fresh and frozen PBPC on HUVEC were similar (n=2). Compared to PBPC, PBL induced higher levels of E-selectin, VCAM-1, and ICAM-1 on HUVEC (n=2). The addition of CD2- cells to HUVEC induced an increase in E-selectin and VCAM-1 to levels 4 times greater than baseline, whereas the addition of CD2+ cells or debris did not elicit a substantial effect (n=2). Transwell permeable membranes prevented PBPC-induced up-regulation of E-selectin, VCAM-1, and ICAM-1 on HUVEC (n=2), suggesting that the mechanism of activation requires direct cell-cell contact. Porcine PBPC activate HUVEC, as suggested by an increase in surface E-selectin, VCAM-1, and ICAM-1 levels, and have a maximum effect after 9 hr. Freezing of PBPC does not affect PBPC-induced activation of HUVEC. PBL induce greater activation of HUVEC than do PBPC. CD2- cells are primarily responsible for PBPC-induced activation of HUVEC and direct cell-cell contact is required. Removal of CD2- cells before the administration of PBPC or the use of agents that interrupt PBPC-endothelial cell interactions may prevent or treat TM in baboons.


[Creation of maize lines inducing maternal haploids with high frequency].
The new inducers of maternal haploids have been created. The results on their inducing capacity in crossing with MK01u and A619 lines and MK01u x A619 hybrid are presented. Pollination with pollen of the new inducers allowed to obtain in average 6.5% of haploids among the three maternal genotypes. The ZMS and KMS initial lines induced 2.3% and 2.0% of haploids respectively. The frequency of haploids was studied in different parts of ears. It was found that in the higher part of an ear where the egg cells are the youngest during flowering the frequency of haploids was almost three times higher than in the lower part of an ear and more than twice higher than in the middle part of an ear. It was assumed that early pollination may increase frequency of obtaining haploids.


Performance of the Black Soldier Fly (Diptera: Stratiomyidae) on Vegetable Residue-Based Diets Formulated Based on Protein and Carbohydrate Contents.
The black soldier fly [Hermetia illucens L. (Diptera: Stratiomyidae)] can be sustainably reared on organic waste streams and thereby provide a novel animal protein source for animal feed. Black soldier fly's performance and body composition depend to a large extent on the nutrient composition of the waste stream. Black soldier fly larvae were fed on four diets consisting of vegetable by-products from the food industry. All four diets contained dried distiller's grains with solubles plus one or two other main ingredients; for diet 1 the additional ingredient was grape pulp; diet 2: potato peels; diet 3: bean seeds; and diet 4: cabbage leaves plus old bread. The diets were formulated based on an optimal summed protein and carbohydrate (P+C) concentration resulting from previous experiments and a 1:2 protein:carbohydrate (P:C) ratio. We quantified both larval and adult performance. Diet 4 was the best performing vegetable waste-based diet. In a follow-up experiment, we re-formulated diet 4 to have P:C ratios of 1:2 or 1:3 and 40 or 47% dry matter of P+C. In the first experiment, although there were differences in larval performance among the diets, all diets supported a higher larval performance than reported previously, with diet 4 having 47% P+C being the best performing diet. We infer that not only total macronutrient content but also nutritional quality of proteins and carbohydrates affected performance. In the second experiment, the two vegetable residue-based larval diets resulted in similar larval and adult performance. High dietary protein resulted in increases in larval crude fat content.


[Functional visual explorations of Bardet-Biedl syndrome. A study of three cases].
Laurence-Moon syndrome, which is very rare, and Bardet-Biedl syndrome, which is more frequent are now well-recognized as two distinct entities in pediatric neurology. Bardet-Biedl syndrome includes a number of common clinical signs it shares with Laurence-Moon syndrome but also with other syndromes, particularly Alströme syndrome. These signs are retinitis pigmentosa, mental retardation, obesity, and hypogonadism. Ophthalmological and electrophysiological examinations are essential for confirmation and correct diagnosis of Bardet-Biedl syndrome. We present three case histories. Our third case illustrates the possibility of below normal yet discernable electroretinogram amplitudes which do not infirm the diagnosis of Bardet-Biedl syndrome.


Prognostic significance of peripheral blood CD8highCD57+ lymphocytes in bladder carcinoma patients after intravesical IL-2.
The objective of this study was to evaluate the recurrence-preventing effect of intravesical instillations of interleukin-2 (IL-2) in patients with non-muscle-invasive bladder carcinoma. In addition, this study aimed to determine the significance of immune parameters for recurrence-free interval. Twenty-six patients with non-muscle-invasive bladder carcinoma were treated with intravesical instillations of IL-2 (Proleukin®, Novartis, formerly Chiron) in doses of 9 × 10(6) IU on 5 consecutive days, beginning on the second day after transurethral resection (TUR) of tumours. CD8(high)CD57(+) lymphocytes in peripheral blood were determined before TUR and compared with the recurrence-free interval after treatment. The multivariate analysis showed that CD8(high)CD57(+) lymphocytes had a prognostic significance in combination with number of bladder tumours, prior recurrence rate and age of patients. Peripheral blood CD8(high)CD57(+) lymphocytes have prognostic significance for recurrence-free survival in patients with non-muscle-invasive bladder carcinoma after TUR and intravesical IL-2.


Foxc1 controls the growth of the murine frontal bone rudiment by direct regulation of a Bmp response threshold of Msx2.
The mammalian skull vault consists of several intricately patterned bones that grow in close coordination. The growth of these bones depends on the precise regulation of the migration and differentiation of osteogenic cells from undifferentiated precursor cells located above the eye. Here, we demonstrate a role for Foxc1 in modulating the influence of Bmp signaling on the expression of Msx2 and the specification of these cells. Inactivation of Foxc1 results in a dramatic reduction in skull vault growth and causes an expansion of Msx2 expression and Bmp signaling into the area occupied by undifferentiated precursor cells. Foxc1 interacts directly with a Bmp responsive element in an enhancer upstream of Msx2, and acts to reduce the occupancy of P-Smad1/5/8. We propose that Foxc1 sets a threshold for the Bmp-dependent activation of Msx2, thus controlling the differentiation of osteogenic precursor cells and the rate and pattern of calvarial bone development.


Hammock effect and complete cusp prolapse: Rare mechanisms of Melody valve failure demonstrated by intracardiac echocardiography.
Transcatheter pulmonary valve replacement using Melody valve (Medtronic, Minneapolis MN) has significantly increased in the recent decades. Melody valve failures, although rare, can be problematic and require re-intervention. Through intracardiac echocardiography, we present two patients who each had a rare etiology for dysfunction of their Melody valve. Hammock effect, wherein the valve does not oppose the stent and complete cusp failure causing severe regurgitation have not been previously described as causes of Melody valve failure in the absence of endocarditis. Awareness and knowledge of these mechanisms is pivotal in the management of this patient population.


Intraperitoneal administration of daptomycin in recurrent peritonitis with suspected biofilm.
Forms of peritonitis are the most problematic infections in patients undergoing peritoneal dialysis since they can jeopardise the technique. Current treatment includes administering vancomycin, cephalosporins and aminoglycosides empirically until the cause of the infection is known. However, the current situation with regard to emerging bacterial resistances makes it necessary to include new drugs in the therapeutic array for complicated forms of peritonitis that may become recurrent and compromise dialyser efficacy. Daptomycin is a lipopeptide antibiotic used to treat gram-positive bacterial infections. It has not yet been approved for treatment of infections of this type, but it is starting to be used in this area due to being highly effective against methicillin-resistant bacteria with intermediate sensitivity to vancomycin, particularly when the bacteria are associated with biofilm formation.


Proline: an essential intermediate in arginine degradation in Saccharomyces cerevisiae.
Results of studies on proline-nonutilizing (Put-) mutants of the yeast Saccharomyces cerevisiae indicate that proline is an essential intermediate in the degradation of arginine. Put- mutants excreted proline when grown on arginine or ornithine as the sole nitrogen source. Yeast cells contained a single enzyme, delta 1-pyrroline-5-carboxylate (P5C) dehydrogenase, which is essential for the complete degradation of both proline and arginine. The sole inducer of this enzyme was found to be proline. P5C dehydrogenase converted P5C to glutamate, but only when the P5C was derived directly from proline. When the P5C was derived from ornithine, it was first converted to proline by the enzyme P5C reductase. Proline was then converted back to P5C and finally to glutamate by the Put enzymes proline oxidase and P5C dehydrogenase.


Transcortical sensory aphasia with relatively spared spontaneous speech and naming.
Patients with transcortical sensory aphasia have relatively preserved repetition ability but have poor comprehension and naming ability. Their spontaneous speech contains paraphasic errors and lacks content. We describe a patient with a left parietal lesion who had poor comprehension but who was able to repeat. However, unlike previously reported cases of transcortical aphasia, the patient had relatively normal naming and spontaneous speech. We believe that this distinct and previously unreported form of transcortical aphasia is induced by a one-way dissociation between phonemic and semantic processors.


[Hepatitis C--standard therapy].
Infection with the hepatitis C virus leads to chronic hepatitis in the majority of patients. Diagnosis is based on the presence of anti-HCV antibodies and confirmed by positive HCV RNA. The natural course of the disease is slow. Cirrhosis is found in a minority of patients two decades post infection. Nevertheless, cirrhosis is much more frequently observed than in patients with hepatitis B infection. Treatment of choice is interferon-alpha. Today combination with ribavirin is recommended for most patients. In combination therapy the sustained response rate six months after stop of treatment is about 40% in naive patients with respect to virus elimination. In patients treated with high doses of interferon-a for one year the sustained response rate is comparable. The response rate is higher in patients with HCV infections of non-1 genotype and in patients with lower virus titers, e.g. less than 2 Mill. genome equivalents per ml. Interferon-a treatment also leads to an improvement of liver histology. Necro-inflammatory scores are reduced. It has also an antifibrogenic effect. Progression of fibrosis is reduced. The antiproliferative effect of interferon-a leads a lower rate of hepatocellular carcinomas, which has been demonstrated in several retrospective studies. In patients with Child A cirrhosis the time till decompensation is delayed. Because of the slow progression, the relatively low response rate and the adverse events of interferon-a and ribavirin treatment should be instituted on an individual base depending on host factors such as age, co-morbidity and stage of liver disease.


Outcome of patients with Fanconi anemia developing myelodysplasia and acute leukemia who received allogeneic hematopoietic stem cell transplantation: A retrospective analysis on behalf of EBMT group.
Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is curative for bone marrow failure in patients with Fanconi anemia (FA), but the presence of a malignant transformation is associated with a poor prognosis and the management of these patients is still challenging. We analyzed outcome of 74 FA patients with a diagnosis of myelodysplastic syndrome (n = 35), acute leukemia (n = 35) or with cytogenetic abnormalities (n = 4), who underwent allo-HSCT from 1999 to 2016 in EBMT network. Type of diagnosis, pre-HSCT cytoreductive therapies and related toxicities, disease status pre-HSCT, donor type, and conditioning regimen were considered as main variables potentially influencing outcome. The 5-year OS and EFS were 42% (30-53%) and 39% (27-51%), respectively. Patients transplanted in CR showed better OS compared with those transplanted in presence of an active malignant disease (OS:71%[48-95] vs 37% [24-50],P = .04), while none of the other variables considered had an impact. Twenty-two patients received pre-HSCT cytoreduction and 9/22 showed a grade 3-4 toxicity, without any lethal event or negative influence on survival after HSCT(OS:toxicity pre-HSCT 48% [20-75%] vs no-toxicity 51% [25-78%],P = .98). The cumulative incidence of day-100 grade II-IV a-GvHD and of 5-year c-GvHD were 38% (26-50%) and 40% (28-52%). Non-relapse-related mortality and incidence of relapse at 5-years were 40% (29-52%) and 21% (11-30%) respectively, without any significant impact of the tested variables. Causes of death were transplant-related events in most patients (34 out of the 42 deaths, 81%). This analysis confirms the poor outcome of transformed FA patients and identifies the importance of achieving CR pre-HSCT, suggesting that, in a newly diagnosed transformed FA patient, a cytoreductive approach pre-HSCT should be considered if a donor have been secured.