The Health Systems Funding Platform: Is this where we thought we were going?
In March 2009, the Task Force for Innovative International Financing for Health Systems recommended "a health systems funding platform for the Global Fund, GAVI Alliance, the World Bank and others to coordinate, mobilize, streamline and channel the flow of existing and new international resources to support national health strategies." Momentum to establish the Health Systems Funding Platform was swift, with the World Bank convening a Technical Workshop on Health Systems Strengthening (HSS), and serial meetings organized to progress the agenda. Despite its potential significance, there has been little comment in peer-reviewed literature, though some disquiet in the international development community around the scope of the Platform and the capacity of the partners, which appears disproportionate to the available information. This case study uses documentary analysis, participant observation and 24 in-depth interviews to examine the processes of development and key issues raised by the Platform. The findings show a fluid and volatile process, with debate over whether ongoing engagement in HSS by Global Fund and GAVI represents a dilution of organizational focus, risking ongoing support, or a paradigm shift that facilitates the achievement of targeted objectives, builds systems capacity, and will attract additional resources. Uncertainty in the development of the Platform reflects the flexibility of the recently formed global health initiatives, and the instability of donor commitments, particularly in the current financial climate. But implicit in the conflict is tension between key global stakeholders over defining and ownership of the health systems agenda. The tensions appear to have been resolved through a focus on national planning, applying International Health Partnership principles, though the global financial crisis and key personnel changes may yet alter outcomes. Despite its dynamic evolution, the Platform may offer an incremental path towards increasing integration around health systems, that has not been previously possible.


Treatment Resistance: A Time-Based Approach for Early Identification in First Episode Psychosis.
Although approximately 1/3 of individuals with schizophrenia are Treatment Resistant (TR), identifying these subjects prospectively remains challenging. The Treatment Response and Resistance in Psychosis working group defines <20% improvement as an indicator of TR, though its utility in First Episode Schizophrenia (FES) remains unknown. In a prospective cohort of FES (n = 129) followed up for 5 years, we evaluated two improvement thresholds for 'probable TR'; <20% and <50% based on positive, negative, and total symptoms. We ascertained (1) the ecological validity (i.e., the ability to identify an expected subgroup of 1/3rd of patients); (2) the predictive validity (i.e., ability to predict poor global functioning) and (3) the clinical utility (association with clozapine use at the 5th year). Using the criteria of a total symptom reduction of <50% or negative symptom reduction of <20% resulted in 'probable TR' rates of 37% and 33%, respectively. Using <20% positive or total symptoms criteria resulted in very low rates, indicating minimal utility in FES. <50% total symptom criterion best predicted the global functioning over 5 years. Clozapine use was only predicted by positive symptom criterion. Prospective characterization of TRS is possible at 6 months after FES through a time-based approach using a 50% threshold for symptom change in treatment-adherent patients.


Neuronal intranuclear inclusion disease: two cases of dopa-responsive juvenile parkinsonism with drug-induced dyskinesia.
There are very few conditions that present with dopa-responsive juvenile parkinsonism. We present two such children with neuronal intranuclear inclusion disease (NIID) who had an initial good levodopa response that was soon complicated by disabling dopa-induced dyskinesia. One child was diagnosed by rectal biopsy in life, and the other diagnosis was confirmed at postmortem. In this patient, dopamine transporter imaging showed severely decreased binding of the radiotracer in the striatum on both sides. Bilateral subthalamic deep brain stimulation in this patient produced initial improvement, but this was not sustained. Both patients died within 10 years of symptom onset. As well as levodopa responsiveness with rapid onset of dyskinesia, clues to the diagnosis of NIID in patients presenting with parkinsonism include the presence of gaze-evoked nystagmus, early onset dysarthria and dysphagia and oculogyric crises. Differential diagnosis of clinical symptoms and neuropathological findings are discussed including the approach to rectal biopsy for early diagnosis.


A hybrid IMRT/VMAT technique for the treatment of nasopharyngeal cancer.
Hybrid IMRT/VMAT technique which combined intensity modulated radiotherapy (IMRT) and volumetric modulated arc therapy (VMAT) was developed for the treatment of nasopharyngeal cancer (NPC). Two-full-arc VMAT (2ARC-VMAT), 9-field IMRT (9F-IMRT), and Hybrid IMRT/VMAT plans for NPC were compared in terms of the dosimetric quality, sparing of organs at risk (OARs), and delivery efficiency. The Hybrid IMRT/VMAT technique can improve the target dose homogeneity and conformity compared with 9F-IMRT and 2ARC-VMAT. It can reduce the dose delivered to the TMJ, mandible, temporal lobe, and unspecified tissue with fewer MUs compared with 9F-IMRT and dose delivered to parotids, brainstem, and spinal cord compared with 2ARC-VMAT technique. The mean delivery time of Hybrid plans was shorter than that of 9F-IMRT plans (408 s versus 812 s; P=0.00) and longer than that of 2ARC-VMAT plans (408 s versus 179 s; P=0.00). Hybrid IMRT/VMAT technique could be a viable radiotherapy technique with better plan quality.


Intracerebroventricular administration of bromocriptine ameliorates the insulin-resistant/glucose-intolerant state in hamsters.
Bromocriptine, a potent dopamine D2 receptor agonist, suppresses lipogenesis and improves glucose intolerance and insulin resistance. Recent evidence suggests that bromocriptine may produce these effects by altering central nervous system (CNS) regulation of metabolism. To determine whether or not the CNS plays a critical role in these bromocriptine-mediated effects on peripheral metabolism, we compared the metabolic responses to bromocriptine when administered peripherally versus centrally in naturally obese and glucose intolerant Syrian hamsters. Male hamsters (BW 194 +/- 5 g) were treated with bromocriptine or vehicle either intraperitoneally (i.p., 800 microgram/animal) or intracerebroventricularly (i.c.v., 1 microgram/animal) daily at 1 h after light onset for 14 days while held on 14-hour daily photoperiods. Glucose tolerance tests (GTTs, 3 g glucose/kg BW) were conducted after treatment. Compared to control animals, bromocriptine i.p. significantly reduced weight gain (11.7 vs. -2.4 g) and the areas under the glucose and insulin GTT curves by 29 and 48%, respectively. Similarly, compared with vehicle-treated controls, bromocriptine i.c.v. at 1 microgram/animal substantially reduced weight gain (8.7 vs. -6.3 g), the areas under the glucose and insulin GTT curves by 31 and 44% respectively, and the basal plasma insulin concentration by 41% (p < 0.05). Furthermore, both treatments significantly improved insulin-mediated suppression of hepatic glucose production during a hyperinsulinemic-euglycemic clamp. Thus, daily administration of bromocriptine at a very low dose i.c.v. replicates the metabolic effects of bromocriptine administered i.p. at a much higher dose. This finding demonstrates for the first time that the CNS is a critical target of bromocriptine's metabolic effects.


Application of methylene blue -vitamin C -N-acetyl cysteine for treatment of critically ill COVID-19 patients, report of a phase-I clinical trial.
COVID-19 is a global catastrophic event that causes severe acute respiratory syndrome. The mechanism of the disease remains unclear, and hypoxia is one of the main complications. There is no currently approved protocol for treatment. The microbial threat as induced by COVID-19 causes the activation of macrophages to produce a huge amount of inflammatory molecules and nitric oxide (NO). Activation of macrophages population into a pro-inflammatory phenotype induces a self-reinforcing cycle. Oxidative stress and NO contribute to this cycle, establishing a cascade inflammatory state that can kill the patient. Interrupting this vicious cycle by a simple remedy may save critical patients' lives. Nitrite, nitrate (the metabolites of NO), methemoglobin, and prooxidant-antioxidant-balance levels were measured in 25 ICU COVID-19 patients and 25 healthy individuals. As the last therapeutic option, five patients were administered methylene blue-vitamin C-N-acetyl Cysteine (MCN). Nitrite, nitrate, methemoglobin, and oxidative stress were significantly increased in patients in comparison to healthy individuals. Four of the five patients responded well to treatment. In conclusion, NO, methemoglobin and oxidative stress may play a central role in the pathogenesis of critical COVID-19 disease. MCN treatment seems to increase the survival rate of these patients. Considering the vicious cycle of macrophage activation leading to deadly NO, oxidative stress, and cytokine cascade syndrome; the therapeutic effect of MCN seems to be reasonable. Accordingly, a wider clinical trial has been designed. It should be noted that the protocol is using the low-cost drugs which the FDA approved for other diseases. TRIAL REGISTRATION NUMBER: NCT04370288.


A physiological model of granulopoiesis to predict clinical drug induced neutropenia from in vitro bone marrow studies: with application to a cell cycle inhibitor.
Neutropenia is one of the most common dose-limiting toxocities associated with anticancer drug therapy. The ability to predict the probability and severity of neutropenia based on in vitro studies of drugs in early drug development will aid in advancing safe and efficacious compounds to human testing. Toward this end, a physiological model of granulopoiesis and its regulation is presented that includes the bone marrow progenitor cell cycle, allowing for a mechanistic representation of the action of relevant anticancer drugs based on in vitro studies. Model development used data from previously reported tracer kinetic studies of granulocyte disposition in healthy humans to characterize the dynamics of neutrophil margination in the presence of endogenous granulocyte-colony stimulating factor (G-CSF). In addition, previously published data from healthy volunteers following pegfilgrastim and filgrastim were used to quantify the regulatory effects of support G-CSF therapies on granulopoiesis. The model was evaluated for the cell cycle inhibitor palbociclib, using an in vitro system of human bone marrow mononuclear cells to quantify the action of palbociclib on proliferating progenitor cells, including its inhibitory effect on G1 to S phase transition. The in vitro results were incorporated into the physiological model of granulopoiesis and used to predict the time course of absolute neutrophil count (ANC) and the incidence of neutropenia observed in three previously reported clinical trials of palbociclib. The model was able to predict grade 3 and 4 neutropenia due to palbociclib treatment with 86% accuracy based on in vitro data.


Differences in protein structural regions that impact functional specificity in GT2 family β-glucan synthases.
Most cell wall and secreted β-glucans are synthesised by the CAZy Glycosyltransferase 2 family (www.cazy.org), with different members catalysing the formation of (1,4)-β-, (1,3)-β-, or both (1,4)- and (1,3)-β-glucosidic linkages. Given the distinct physicochemical properties of each of the resultant β-glucans (cellulose, curdlan, and mixed linkage glucan, respectively) are crucial to their biological and biotechnological functions, there is a desire to understand the molecular evolution of synthesis and how linkage specificity is determined. With structural studies hamstrung by the instability of these proteins to solubilisation, we have utilised in silico techniques and the crystal structure for a bacterial cellulose synthase to further understand how these enzymes have evolved distinct functions. Sequence and phylogenetic analyses were performed to determine amino acid conservation, both family-wide and within each sub-family. Further structural analysis centred on comparison of a bacterial curdlan synthase homology model with the bacterial cellulose synthase crystal structure, with molecular dynamics simulations performed with their respective β-glucan products bound in the trans-membrane channel. Key residues that differentially interact with the different β-glucan chains and have sub-family-specific conservation were found to reside at the entrance of the trans-membrane channel. The linkage-specific catalytic activity of these enzymes and hence the type of β-glucan chain built is thus likely determined by the different interactions between the proteins and the first few glucose residues in the channel, which in turn dictates the position of the acceptor glucose. The sequence-function relationships for the bacterial β-glucan synthases pave the way for extending this understanding to other kingdoms, such as plants.


Doxorubicin loaded dual pH- and thermo-responsive magnetic nanocarrier for combined magnetic hyperthermia and targeted controlled drug delivery applications.
Magnetic nanocarriers have attracted increasing attention for multimodal cancer therapy due to the possibility to deliver heat and drugs locally. The present study reports the development of magnetic nanocomposites (MNCs) made of an iron oxide core and a pH- and thermo-responsive polymer shell, that can be used as both hyperthermic agent and drug carrier. The conjugation of anticancer drug doxorubicin (DOX) to the pH- and thermo-responsive MNCs via acid-cleavable imine linker provides advanced features for the targeted delivery of DOX molecules via the combination of magnetic targeting, and dual pH- and thermo-responsive behaviour which offers spatial and temporal control over the release of DOX. The iron oxide cores exhibit a superparamagnetic behaviour with a saturation magnetization around 70 emu g(-1). The MNCs contained 8.1 wt% of polymer and exhibit good heating properties in an alternating magnetic field. The drug release experiments confirmed that only a small amount of DOX was released at room temperature and physiological pH, while the highest drug release of 85.2% was obtained after 48 h at acidic tumour pH under hyperthermia conditions (50 °C). The drug release kinetic followed Korsmeyer-Peppas model and displayed Fickian diffusion mechanism. From the results obtained it can be concluded that this smart magnetic nanocarrier is promising for applications in multi-modal cancer therapy, to target and efficiently deliver heat and drug specifically to the tumour.


Recombinant bovine/human parainfluenza virus type 3 (B/HPIV3) expressing the respiratory syncytial virus (RSV) G and F proteins can be used to achieve simultaneous mucosal immunization against RSV and HPIV3.
Recombinant bovine/human parainfluenza virus type 3 (rB/HPIV3), a recombinant bovine PIV3 (rBPIV3) in which the F and HN genes were replaced with their HPIV3 counterparts, was used to express the major protective antigens of respiratory syncytial virus (RSV) in order to create a bivalent mucosal vaccine against RSV and HPIV3. The attenuation of rB/HPIV3 is provided by the host range restriction of the BPIV3 backbone in primates. RSV G and F open reading frames (ORFs) were placed under the control of PIV3 transcription signals and inserted individually into the rB/HPIV3 genome in the promoter-proximal position preceding the nucleocapsid protein gene. The recombinant PIV3 expressing the RSV G ORF (rB/HPIV3-G1) was not restricted in its replication in vitro, whereas the virus expressing the RSV F ORF (rB/HPIV3-F1) was eightfold restricted compared to its rB/HPIV3 parent. Both viruses replicated efficiently in the respiratory tract of hamsters, and each induced RSV serum antibody titers similar to those induced by RSV infection and anti-HPIV3 titers similar to those induced by HPIV3 infection. Immunization of hamsters with rB/HPIV3-G1, rB/HPIV3-F1, or a combination of both viruses resulted in a high level of resistance to challenge with RSV or HPIV3 28 days later. These results describe a vaccine strategy that obviates the technical challenges associated with a live attenuated RSV vaccine, providing, against the two leading viral agents of pediatric respiratory tract disease, a bivalent vaccine whose attenuation phenotype is based on the extensive host range sequence differences of BPIV3.


Immobilization-free sequence-specific electrochemical detection of DNA using ferrocene-labeled peptide nucleic acid.
An electrochemical method for sequence-specific detection of DNA without solid-phase probe immobilization is reported. This detection scheme starts with a solution-phase hybridization of ferrocene-labeled peptide nucleic acid (Fc-PNA) and its complementary DNA (cDNA) sequence, followed by the electrochemical transduction of Fc-PNA-DNA hybrid on indium tin oxide (ITO)-based substrates. On the bare ITO electrode, the negatively charged Fc-PNA-DNA hybrid exhibits a much reduced electrochemical signal than that of the neutral-charge Fc-PNA. This is attributed to the electrostatic repulsion between the negatively charged ITO surface and the negatively charged DNA, hindering the access of Fc-PNA-DNA to the electrode. On the contrary, when the transduction measurement is done on the ITO electrode coated with a positively charged poly(allylamine hydrochloride) (PAH) layer, the electrostatic attraction between the (+) PAH surface and the (-) Fc-PNA-DNA hybrid leads to a much higher electrochemical signal than that of the Fc-PNA. The measured electrochemical signal is proportional to the amount of cDNA present. In terms of detection sensitivity, the PAH-modified ITO platform was found to be more sensitive (with a detection limit of 40 fmol) than the bare ITO counterpart (with a detection limit of 500 fmol). At elevated temperatures, this method was able to distinguish fully matched target DNA from DNA with partial mismatches. Unpurified PCR amplicons were detected using a similar format with a detection limit down to 4.17 amol. This detection method holds great promise for single-base mismatch detection as well as electrochemistry-based detection of post-PCR products.


Effect of hydroalcoholic extract of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability and activity of microcosm biofilm and on enamel demineralization.
The aim of this study was to assess the effect of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves hydroalcoholic extracts on viability and metabolism of a microcosm biofilm and on enamel demineralization prevention. Microcosm biofilm was produced on bovine enamel using inoculum from pooled human saliva mixed with McBain saliva, under 0.2% sucrose exposure, for 14 days. The biofilm was daily-treated with the extracts for 1 min. At the end, it was analyzed with respect to viability by fluorescence, CFU counting and extracellular polysaccharides (phenol-sulphuric acid colorimetric assay) and lactic acid (enzymatic assay) production. The demineralization was measured by TMR. The data were compared using ANOVA or Kruskal-Wallis (p<0.05). M. urundeuva All. at 100, 10 and 0.1 μg/mL and Q. grandiflora Mart. at 100 and 0.1 μg/mL reduced biofilm viability similarly to positive control (chlorhexidine) and significantly more than the negative-vehicle control (35% ethanol). M. urundeuva at 1000, 100 and 0.1 μg/mL were able to reduce both lactobacilli and mutans streptococci CFU counting, while Q. grandiflora (1000 and 1.0 μg/mL) significantly reduced mutans streptococci CFU counting. On the other hand, the natural extracts were unable to significantly reduce extracellular polysaccharides and lactic acid productions neither the development of enamel carious lesions. The extracts showed antimicrobial properties on microcosm biofilm, however, they had no effect on biofilm metabolism and caries protection.


Virulence of Streptococcus mutans: a sensitive method for evaluating cariogenicity in young gnotobiotic rats.
Gnotobiotic rats infected with Streptococcus mutans 6715 at 19 days of age and fed a purified diet (305) containing 5% sucrose developed extensive caries lesions on all molar surfaces within 16 days (35 days of age). Approximately twice as many lesions developed when infected rats were maintained until 45 days of age, whereas noninfected rats did not develop caries when fed diet 305. Gnotobiotic rats infected with S. mutans 6715 and fed a purified diet containing no sucrose (300) until day 25 and subsequently fed diet 305 for 10 days developed lesions similar to rats fed diet 305 for 16 days. Furthermore, rats infected with S. mutans 6715 and fed diet 300 until 45 days of age developed approximately one-half the smooth surface lesions as infected rats fed diet 305 for the same length of time. The level of caries on buccal and proximal molar surfaces in 45-day-old gnotobiotic rats varied when animals were infected with S. mutans AHT, BHT, NCTC 10449, 6715, or LM-7. Animals infected with S. mutans AHT showed more severe lesions on the buccal surfaces than those observed in animals infected with the other strains of S. mutans tested, whereas S. mutans 6715 caused significantly more caries on proximal surfaces. On the other hand, rats infected with S. mutans LM-7 exhibited the lowest level of caries on all molar surfaces of the five strains of S. mutans tested.


Aberrant protamine content in sperm and consequential implications for infertility treatment.
Human sperm express two types of protamine: protamine 1 (P1) and the family of protamine 2 (P2) proteins, with P1 and P2 normally existing in a ratio of approximately 1:1. Both the elevation and reduction of this ratio have been linked with male infertility suggesting that abnormalities in protamine expression, processing and replacement may be responsible for effects on semen parameters observed in infertile males affected by deficient protamination, along with abnormalities in associated regulatory processes. Abnormal protamination may result in insufficient condensation in the sperm nucleus, thus rendering paternal DNA susceptible to damage, which could have detrimental consequences upon embryogenesis. Consequently, it is imperative that Assisted Reproductive Technologies (ARTs) endeavour to utilise sperm devoid of protamine abnormalities, especially because retained histones are present in imprinted gene clusters. Emerging evidence indicates that abnormalities in protamine content may influence epigenetic signals transmitted via paternal DNA. Indeed, an increase in rare imprinting disorders has been observed in children conceived via in vitro fertilisation (IVF). This review examines the links between male infertility, abnormal protamine expression and replacement, the implications of abnormal sperm DNA packaging on fertility treatments and the potential iatrogenic effects of ART procedures on sperm function.


Psychology's Replication Crisis and the Grant Culture: Righting the Ship.
The past several years have been a time for soul searching in psychology, as we have gradually come to grips with the reality that some of our cherished findings are less robust than we had assumed. Nevertheless, the replication crisis highlights the operation of psychological science at its best, as it reflects our growing humility. At the same time, institutional variables, especially the growing emphasis on external funding as an expectation or de facto requirement for faculty tenure and promotion, pose largely unappreciated hazards for psychological science, including (a) incentives for engaging in questionable research practices, (b) a single-minded focus on programmatic research, (c) intellectual hyperspecialization, (d) disincentives for conducting direct replications, (e) stifling of creativity and intellectual risk taking, (f) researchers promising more than they can deliver, and (g) diminished time for thinking deeply. Preregistration should assist with (a), but will do little about (b) through (g). Psychology is beginning to right the ship, but it will need to confront the increasingly deleterious impact of the grant culture on scientific inquiry.


Pay-it-forward to increase uptake among 15-18-year-old adolescent girls compared with user-paid vaccination: The pilot results of a two-arm randomized controlled trial in China.
China has low human papillomavirus(HPV) vaccination rate due to lack of public funding and mistrust in domestic vaccines. This pilot study evaluated the feasibility and preliminary effectiveness of an innovative pay-it-forward strategy, which has a participant receive a subsidized vaccine and offers her an opportunity to donate to support other girls, in improving HPV vaccine uptake among 15-18-year-old adolescent girls. A two-arm randomized controlled pilot trial was performed in one vaccination clinic in Western China. Adolescent girls (via caregivers) were invited to participate the study via online dissemination of the pilot study. Eligible ones were then randomly allocated using a sealed envelope method into standard-of-care or pay-it-forward arm in a 1:1 ratio. Pay-it-forward participants received hand-written postcard messages, a subsidized vaccine, and an opportunity to donate and (or) write postcards for future recipient girls. Standard-of-care participants self-paid for vaccines. The primary outcome was the first-dose HPV vaccine uptake, which was calculated using multivariable logistic regression, presented as crude/adjusted odds ratios (cORs/aORs) and 95% con dence intervals (CIs). Standard scales were used to assess the feasibility of the program. A total of 100 participants (50 in each arm) were recruited from Jan 4, 2022 to Feb 18, 2022. The HPV vaccine uptake rate was 98% (49/50) in the pay-it-forward arm and 82% (41/50) in the standard-of-care arm (cOR = 10.76, 95% CI: 1.31-88.47, P = 0.027; aOR = 12.12, 95% CI: 1.37-107.29, P = 0.025). The completion rate of full schedule of HPV vaccination in the two arms was 100% (49/49) and 95.1% (39/41), respectively. Of all 49 vaccinated girls in the pay-it-forward arm, 38 (77.6%) donated to support future participants, and the total donation accounted for 33.3% of prepaid subsidization. Among caregivers in the pay-it-forward arm, 97.6% (41/42) believed that this strategy was feasible. The pilot showed feasibility and preliminary effectiveness of a pay-it-forward strategy to increase HPV vaccination rate. The high uptake rate in the standard-of-care arm is likely caused by the selection bias associated with the online dissemination approach and secured vaccine supply through the program. Further adaption of the intervention package and a population-based recruitment pathway are needed to better reflect local contexts and enhance the generalizability of the subsequent formal trial. Chinese Clinical Trial Registry (ChiCTR), ChiCTR2200055542. Retrospectively registered on 11 January 2022, https://www.chictr.org.cn/showproj.html?proj=139738.


Ligand-receptor interactions in live cells by fluorescence correlation spectroscopy.
Receptor binding studies most often require the use of radioactively labeled ligands. In certain cases, the numbers of receptors are few per cell and no specific binding is detected because of a high background. Specific interactions between certain ligands (e.g. peptides, hormones, natural products) and their receptors are, therefore, often overlooked by the conventional binding technique. Fluorescence correlation spectroscopy (FCS) allows detection of the interaction of ligands with receptors in their native environment in live cells in a tiny confocal volume element (0.2 fl) at single-molecule detection sensitivity. This technique permits the identification of receptors which were not possible before to detect by isotope labeling. The beauty of the FCS technique is that there is no need for separating an unbound ligand from a bound one to calculate the receptor bound and free ligand fractions. This review will show FCS as a sensitive and a rapid technique to study ligand-receptor interaction in live cells and will demonstrate that the FCS-analysis of ligand-receptor interactions in live cells fulfils all the criteria of a ligand binding to its receptor i.e. it is able to provide information on the affinity and specificity of a ligand, binding constant, association and dissociation rate constants as well as the number and mobility of receptors carrying a fluorescently labeled ligand. This review is of pharmaceutical significance since it will provide insights on how FCS can be used as a rapid technique for studying ligand-receptor interactions in cell cultures, which is one step forward towards a high throughput drug screening in cell cultures.


Simvastatin attenuates TGF-β1-induced epithelial-mesenchymal transition in human alveolar epithelial cells.
Transforming growth factor-β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) of alveolar epithelial cells (AEC) may contribute to idiopathic pulmonary fibrosis (IPF). TGF-β1-induced EMT in A549 cells (a human AEC cell line) resulted in the adoption of mesenchymal responses that were predominantly mediated via the TGF-β1-Smad2/3 signaling pathway. Simvastatin (Sim), a 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase inhibitor, has been previously reported to inhibit EMT in human proximal tubular epithelial cells and porcine lens epithelial cells and to suppress Smad2/3 phosphorylation in animal models. However, whether Sim can attenuate TGF-β1-induced EMT in A549 cells and its underlying mechanisms remains unknown. Cells were incubated with TGF-β1 in the presence or absence of Sim. The epithelial marker E-cadherin (E-Cad) and the mesenchymal markers, α-smooth muscle actin (α-SMA), vimentin (Vi) and fibronectin (FN), were detected using western blotting analyses and immunofluorescence. Phosphorylated Smad2 and Smad3 levels and connective tissue growth factor (CTGF) were analyzed using western blotting. In addition, a cell migration assay was performed. Moreover, the levels of matrix metalloproteinase (MMP)-2 and -9 in the culture medium were examined using ELISA. Sim significantly attenuated the TGF-β1-induced decrease in E-Cad levels and elevated the levels of α-SMA, Vi and FN via the suppression of Smad2 and Smad3 phosphorylation. Furthermore, Sim inhibited the mesenchymal-like responses in A549 cells, including cell migration, CTGF expression and secretion of MMP-2 and -9. However, Sim failed to reverse the cell morphologial changes induced by TGF-β1 in A549 cells. Sim attenuated TGF-β1-induced EMT in A549 cells and might be a promising therapeutic agent for treating IPF.


89Zr-Labeled Anti-PD-L1 Antibody Fragment for Evaluating In Vivo PD-L1 Levels in Melanoma Mouse Model.
The rise of programmed death-1 (PD-1)/PD-L1 immune checkpoint inhibitor therapy has been one of the most promising developments in melanoma research. However, not all the melanoma patients respond to such immune checkpoint blockade. There is a great need of biomarkers for appropriate melanoma patient selection and therapeutic efficacy monitoring. The objective of this study is to develop a novel radiolabeled anti-PD-L1 antibody fragment, as an imaging biomarker, for evaluating the in vivo PD-L1 levels in melanoma. The Df-conjugated F(ab')2 fragment of the anti-mouse PD-L1 antibody was successfully synthesized and radiolabeled with 89Zr. Both Df-F(ab')2 and 89Zr-Df-F(ab')2 maintained the nano-molar murine PD-L1 targeting specificity and affinity. 89Zr-Df-F(ab')2 showed less uptake in normal liver tissue in mice compared with its full antibody counterpart 89Zr-Df-anti-PD-L1. Positron emission tomography (PET)/computed tomography images clearly showed that 89Zr-Df-F(ab')2 possessed superior pharmacokinetics and imaging contrast over the radiolabeled full antibody, with much earlier and higher tumor uptake (5.5 times more at 2 h post injection) and much lower liver background (51% reduction at 2 h post injection). The specific and high murine PD-L1-targeting uptake at tumor foci coupled with fast clearance of 89Zr-Df-F(ab')2 highlighted its potential for in vivo PET imaging of murine PD-L1 levels and future development of radiolabeled anti-human PD-L1 fragment for potential application in melanoma patients.


Endovascular therapy of acute vertebrobasilar occlusion: early treatment onset as the most important factor.
In view of the poor prognosis for patients with acute intracranial vertebrobasilar occlusion (VBO), factors were sought that predict survival and good neurologic outcome after acute endovascular treatment by means of local intra-arterial fibrinolysis (LIF) and percutaneous transluminal angioplasty (PTA). LIF was performed in 83 patients with angiographically established acute VBO. A significant residual stenosis after LIF was treated by additional PTA in 8 patients. The types of occlusion were classified as either embolic occlusion (EO) or atherothrombotic occlusion (AO). Outcome was evaluated after 3 months by the Barthel Index (BI) as favorable (BI >90), unfavorable (BI <90) or death and compared for each of 3 diagnostic or treatment variables: recanalization success, occlusion type and time to treatment. Four fibrinolytic treatment modes [urokinase, low-dose and high-dose recombinant tissue-type plasminogen activator (rt-PA), rt-PA + Lys-plasminogen] were also analyzed. The outcome was favorable in 19 patients (23%), unfavorable in 14 (17%) and 50 died (60%). Recanalization was successful in 54 patients (66%). The neurologic outcome was better in recanalized than in nonrecanalized patients (favorable outcome: 30 vs. 10%, mortality: 54 vs. 72%; p = 0.118). The neurologic outcome was better in EO than in AO (favorable outcome: 31 vs. 17%, mortality: 47 vs. 70%, p = 0.112). Under combined treatment by LIF and PTA in 8 patients with AO, 4 survived, 3 with a favorable outcome (38%). Early treatment onset (<or=6 h) led to a significantly better neurologic outcome than delayed treatment onset (>6 h; favorable outcome: 36 vs. 7%, mortality: 52 vs. 70%, p = 0.005). Although no statistically significant differences were found between the types of fibrinolytic agents, treatment with rt-PA and Lys-plasminogen tended toward better results. Early treatment onset proved to be the most important factor for successful endovascular therapy in acute VBO, whereas recanalization and presence of an embolic occlusion also tended toward better results. Additional PTA may be a promising therapy in cases of significant residual stenosis after LIF.


Demography of vascular Behcet's disease with different gender and age: an investigation with 166 Chinese patients.
The Clinical features of vascular Behcet's disease (BD) are not well understood because there are few studies. Our study aimed to investigate characteristics of vascular BD in both genders in different age groups. We enrolled 923 patients with BD who presented to our hospital with adequate medical histories and proper vascular screening exams. The raw incidence rate of vascular BD was 17.98% (166/923). The ratio of vascular BD in male to female patients was 1.868 (p = 0.0004, 95% confidence interval (CI): 1.317 to 2.625). There was a tendency towards higher ESR and CRP in vascular BD patients than in mucocutaneous, but the difference was not significant. The most susceptible affected vessels were cerebral (29.6% in males, 59.4% in females) and lower limb vessels (31.2% in males, and 17.2% in females). The incidence of vascular involvement in younger (< 50 years old) and older (≥ 50 years old) patients were similar, with ratios of 16.58% (122/736) and 23.53% (44/187) respectively. However, in females, younger patients were less likely to have vascular involvement than were older patients (11.43% vs. 20% p = 0.0328, OR: 0.5161, 95% CI: 0.2874 to 0.912). Aneurysm or pseudoaneurysm was diagnosed in 1.84% (17/923) patients, mostly in male patients (p < 0.05, OR: 3.221, 95% CI: 1.097 to 9.112). Twenty vascular BD patients were followed up, and the age at BD diagnosis was 33.23 ± 11.56 year. This did not differ statistically with their age at vascular involvement (36.15 ± 9.52 years). Ages of vascular BD patients did not differ significantly from those of mucocutaneous BD patients (n = 143) in both males and females. Vascular BD, including lethal types of aneurysm is more likely to occur in male patients. The female patients has a similar incidence rate with the males in their postmenopausal age. There was no evidence of progression course from mucocutaneous BD to vascular involvement.


Inhibition of fibronectin-activated migration of microvascular endothelial cells by interleukin-1alpha, tumour necrosis factor alpha and interferon gamma.
The effect of interferon gamma (IFN) and the inflammatory cytokines tumour necrosis factor alpha (TNF) and interleukin 1alpha (IL-1) on micro- and macrovascular endothelial cell (EC) proliferation and migration was analysed. Whereas both micro- and macrovascular EC were growth-inhibited in response to the aforementioned cytokines, only microvascular EC were sensitive to TNF, IL-1 and IFN as inhibitors of fibronectin-activated cell migration. In addition, because microvascular EC play a crucial role in angiogenesis, and the formation of new capillaries depends upon the presence of angiogenic polypeptides, we evaluated the synthesis of fibroblast growth factor (FGF) type 1 and 2, Vascular Endothelial Growth Factor (VEGF) and Hepatocyte Growth Factor (HGF) in our system. Both micro- and macrovascular EC produce large amounts of FGF-2, which is mainly localized in the nucleus, and almost undetectable levels of FGF-1. In addition, the two cell types synthesize notable levels of VEGF and no HGF. Whether these findings are relevant to the different in vivo functions of EC residing different districts remains the focus of additional studies.


Timing of voiding cystourethrogram after urinary tract infection.
Of the 363 Thai children upon whom a voiding cystouretrogram was performed, a vesicoureteral reflux was detected in 22.8% (17.1%-28.5%) of those for whom it was performed within 7 days (n = 215) of a urinary tract infection diagnosis and in 24.3% (17.4%-31.2%) of those for whom it was performed 7 days (n = 148) after diagnosis. There was no statistically significant difference in reflux prevalence between these two groups.


Outcome and growth of infants with severe chronic renal failure.
We aimed to assess the outcome and growth of infants with severe chronic renal failure (CRF). One hundred and one children presented between January 1, 1986, and December 12, 1998, with a glomerular filtration rate (GFR) of <20 mL/min/1.73 m2. The median (range) age at presentation was 0.3 (0 to 1.5) years, and follow-up was 7.6 (1.5 to 13) years. One- and five-year survival rates were 87 and 78%, respectively. The growth of the 81 children who survived over two years was evaluated. Eighty-one percent were enterally fed from age 0.7 (0 to 4.5) years for 1.9 (0.1 to 6.8) years. Forty-six percent had a gastrostomy, and 22% a Nissen fundoplication. Twenty-five were managed conservatively. Twenty were transplanted without dialysis at age 4 (1.7 to 8.5) years, and 36 were dialyzed at age 1.1 (0 to 9.8) before transplantation at age 2. 4 (1.3 to 10) years. The mean (SD) height standard deviation score increased from -2.16 (1.34) at 6 months (N = 63) to -1.97 (1.37) at 1 year (N = 75), -1.79 (1.29) at 2 years (N = 75), -1.33 (1.29) at 3 years (N = 68, P = 0.0006), -1.27 (1.04) at 5 years (N = 47, P = 0.0001), and -0.85 (0.82) at 10 years (N = 18, P = 0.001). The body mass index was in the normal range in the majority of patients. Mortality in infants with CRF occurs mainly in the first year of life. With early enteral feeding, the mean height standard deviation score is within the normal range from one year of age.


[Reliability and Validity of the Behavioral Check List for Preschool Children to Measure Attention Deficit Hyperactivity Behaviors].
Nowadays, attention deficit hyperactivity (ADH) problems are observed commonly among school-age children. However, questionnaires specific to ADH behaviors among preschool children are very few. The aim of this study was to investigate the reliability and validity of the 25-item Behavioral Check List (BCL), which was developed from interviews of parents with children who were diagnosed as having Attention-deficit/hyperactivity disorder (ADHD) and measures ADH behaviors in preschool age. We recruited 22 teachers from 10 nurseries/kindergartens in Miyagi Prefecture, Japan. A total of 138 preschool children were assessed using the BCL. To investigate inter-rater reliability, two teachers from each facility assess seven to twenty children in their class, and intraclass correlation coefficients (ICCs) were calculated. The teachers additionally answered questions in the 1/5-5 Caregiver-Teacher Report Form (C-TRF) to investigate the criterion validity of the BCL. To investigate structural validity, exploratory factor analysis with promax rotation and confirmatory factor analysis were performed. The internal consistency reliability of the BCL was good (α = 0.92) and correlation analyses also confirmed its excellent criterion validity. Although exploratory factor analysis for the BCL yielded a five-factor model that consisted of a factor structure different from that of the original one, the results were similar to the original six factors. The ICCs of the BCL were 0.38-0.99 and it was not high enough for inter-rater reliability in some facilities. However, there is a possibility to improve it by giving raters adequate explanations when using BCL. The present study showed acceptable levels of reliability and validity of the BCL among Japanese preschool children.


In situ lipid-loading activates peripheral dendritic cell subsets characterized by cellular ROS accumulation but compromises their capacity to prime naïve T cells.
Dendritic cells (DCs), professional antigen-presenting cells, play an important role in pathologies by controlling adaptive immune responses. However, their adaptation to and functionality in hypercholesterolemia, a driving factor in disease onset and progression of atherosclerosis remains to be established. In this study, we addressed the immediate impact of high fat diet-induced hypercholesterolemia in low-density lipoprotein receptor deficient (Ldlr-/-) mice on separate DC subsets, their compartmentalization and functionality. While hypercholesterolemia induced a significant rise in bone marrow myeloid and dendritic cell progenitor (MDP) frequency and proliferation rate after high fat diet feeding, it did not affect DC subset numbers in lymphoid tissue. Hypercholesterolemia led to almost immediate and persistent augmentation in granularity of conventional DCs (cDCs), in particular cDC2, reflecting progressive lipid accumulation by these subsets. Plasmacytoid DCs were only marginally and transiently affected. Lipid loading increased co-stimulatory molecule expression and ROS accumulation by cDC2. Despite this hyperactivation, lipid-laden cDC2 displayed a profoundly reduced capacity to stimulate naïve CD4+ T cells. Our data provide evidence that in hypercholesterolemic conditions, peripheral cDC2 subsets engulf lipids in situ, leading to a more activated status characterized by cellular ROS accumulation while, paradoxically, compromising their T cell priming ability. These findings will have repercussions not only for lipid driven cardiometabolic disorders like atherosclerosis, but also for adaptive immune responses to pathogens and/or endogenous (neo) antigens under conditions of hyperlipidemia.


The nature of naming errors in primary progressive aphasia versus acute post-stroke aphasia.
To compare the distribution of error types across subgroups of primary progressive aphasia and poststroke aphasia in different vascular locations. We analyzed naming errors in 49 individuals with acute left hemisphere ischemic stroke and 55 individuals with three variants of primary progressive aphasia. Location of atrophy or ischemic stroke was characterized using MRI. We found that distribution of error types was very similar across all subgroups, irrespective of the site or etiology of the lesion. The only significant difference across groups was the percentage of circumlocutions (F(7, 96) = 3.02, p = .005). Circumlocution errors were highest among logopenic variant PPA (24%) and semantic variant PPA (24%). Semantic coordinate errors were common in all groups, probably because they can arise from disruption of different cognitive processes underlying naming and, therefore, from different locations of brain damage. Semantic errors are common among all types of primary progressive aphasia and poststroke aphasia, and the type of error depends in part on the location of damage.


Rapid 3D phenotyping of cardiovascular development in mouse embryos by micro-CT with iodine staining.
Microcomputed tomography (micro-CT) has been used extensively in research to generate high-resolution 3D images of calcified tissues in small animals nondestructively. It has been especially useful for the characterization of skeletal mutations but limited in its utility for the analysis of soft tissue such as the cardiovascular system. Visualization of the cardiovascular system has been largely restricted to structures that can be filled with radiopaque intravascular contrast agents in adult animals. Recent ex vivo studies using osmium tetroxide, iodinated contrast agents, inorganic iodine, and phosphotungstic acid have demonstrated the ability to stain soft tissues differentially, allowing for high intertissue contrast in micro-CT images. In the present study, we demonstrate the application of this technology for visualization of cardiovascular structures in developing mouse embryos using Lugol solution (aqueous potassium iodide plus iodine). We show the optimization of this method to obtain ex vivo micro-CT images of embryonic and neonatal mice with excellent soft-tissue contrast. We demonstrate the utility of this method to visualize key structures during cardiovascular development at various stages of embryogenesis. Our method benefits from the ease of sample preparation, low toxicity, and low cost. Furthermore, we show how multiple cardiac defects can be demonstrated by micro-CT in a single specimen with a known genetic lesion. Indeed, a previously undescribed cardiac venous abnormality is revealed in a PlexinD1 mutant mouse. Micro-CT of iodine-stained tissue is a valuable technique for the characterization of cardiovascular development and defects in mouse models of congenital heart disease.


[Fibromyalgia (generalized tendomyopathy): defect of a program of movements and their realization].
It was established that fulfillment of any motor action implies a design of the program of action in CNS and its realization by peripheral elements of locomotor system. A program includes an acceptor of the result of action and efferent synthesis on a central level (parietal-premotor area, pallidostriatal system, lymbico-reticular complex, ex cetera). A very motor act is realized by the activity of a segmental apparatus of the spinal cord and skeletal muscles. Acceptor of the result of action provides a control of the coincidence between afferent parameters of the result of an action and its real characteristics by means of the reverse feed-back (P.K. Anokhin, 1975). Incoordination of its parameters is a condition for the existence of a physiologic functional system for the construction of the necessary movement, while a coincidence of their indices resulted in the disintegration of this system. Appearance of myofascicular hypertonus in this system distorts a real parameters of the movement because of a deficit and imbalance of the proprioception in a segmental apparatus and central formations. A proprioceptive desafferentation promotes disinhibition of the neurons and formation of the generators of pathologically increased irritation with positive feed-back on rubro-segmental (A), pallido-thalamic (B), strio-piramidal (C) and parietal-premotor (D) levels of the construction of the movement according to N.A. Bernshtein (1966). Its appearance on the B and C levels prevents destruction of the physiologic system of organization after realization of its activity and promotes its reformation into the pathologic one. It manifested clinically in appearance of a pathologic dynamic stereotype. Additional integration of lymbico-reticular complex, and high centers of autonomic supplement into this system promotes a formation of the stable neurotic, depressive reactions and autonomic disorders. Such reactions are transitory and have adaptive character at normal program of construction and fulfillment of the motion, while at pathologic one they have desadaptive character. Clinically they are known as multiple psychoautonomic syndromes combined with muscular, fascial and ligamental pains of different location.


lncRNA MALAT1 modulates oxaliplatin resistance of gastric cancer via sponging miR-22-3p.
Various regulatory mechanisms have been demonstrated to be associated with cancer progression. ncRNA and mRNA play important roles in gastric cancer (GC) cell growth and drug resistance, respectively. However, the regulatory network of ncRNA and mRNA in GC oxaliplatin (OXA) resistance has not been fully clarified. The expression of miR-22-3p, MALAT1, and zinc finger protein 91 (ZFP91) was detected in tissues and cells using quantitative real-time PCR. The protein level of ZFP91 was measured by Western blot analysis. Luciferase reporter, pull-down, and RNA immunoprecipitation assays were used to determine the relationship between MALAT1, miR-22-3p, and ZFP91. MTT assay was applied to measure cell survival and proliferation. Cell apoptosis was detected using flow cytometry. Tumor xenograft assay was used to detect the function of miR-22-3p in vivo. In this study, we found that MALAT1 and ZFP91 expression was upregulated while the expression of miR-22-3p was downregulated in GC/OXA tissues and cells. Additionally, miR-22-3p was a target miRNA of MALAT1 and ZFP91 was a target mRNA of miR-22-3p. Functional studies showed that the knockdown of MALAT1 or overexpression of miR-22-3p inhibited GC/OXA cell survival, proliferation, and drug resistance as well as induced apoptosis, which could be reversed by the inhibition of miR-22-3p or overexpression of ZFP91. We observed a new regulatory network for MALAT1 in drug resistance of GC. MALAT1 modulates ZFP91 to promote GC cells OXA resistance via sponging miR-22-3p.


Impact of COVID-19 Pandemic on Adolescents and Young Adults Living With Type 2 Diabetes.
The aim of this study was to assess the impacts of the COVID-19 pandemic on adolescents and young adults living with type 2 diabetes (T2D) involved in the national Improving Renal Complications in Adolescents with T2D through REsearch (iCARE) study. The Environmental influences on Child Health Outcomes (ECHO) COVID-19 Questionnaire developed by the National Institutes of Health ECHO COVID-19 Task Force was administered to participants (n=85) from the iCARE study between June 2020 and October 2020. Children 12 years old (via parent report) and adolescents and young adults ≥13 years old (via self-report) participated. The questionnaire assessed the impact of the pandemic on health-care appointments, lifestyle, internet use, social connections and mental health. Participants were 17.0±3.1 (range, 12 to 27) years of age and predominantly female (61.3%). During the pandemic, 69.4% were able to attend their health-care appointments by telephone or virtual platforms, 31.7% ate more, 45.1% slept more and 29.3% spent less time on physical activities. There was an increase in internet use for both educational (42.0%) and noneducational purposes (54.9%). Participants felt less socially connected (64.6%). Participants also felt sometimes (59.2%), often (19.7%) and very often (6.7%) satisfied with their lives. Our study revealed that the COVID-19 pandemic has had various impacts on the daily lives of adolescents and young adults living with T2D. Future research should include longitudinal studies of the health burden of the COVID-19 pandemic on this population, with a more in-depth evaluation of mental health outcomes and clinical outcomes.


Interpersonal counselling in general practice.
Interpersonal counselling (IPC) derives from interpersonal psycho-therapy (IPT) but is briefer in the number and duration of sessions and is particularly suited to the primary care setting. While depression and other psychological symptoms are not necessarily 'caused' by interpersonal problems, they do occur in a social and interpersonal context. Problem areas commonly associated with the onset of depression are unresolved grief, interpersonal disputes, role transition and interpersonal deficits such as social isolation. This article discusses IPC and how it can be used in the general practice setting. The structure of IPC is of a brief treatment of six sessions, each with an explicit focus: assessment, education about the interaction between interpersonal relationships and psychological symptoms, identifying current stress areas and helping the patient deal with these more positively and termination of the IPC relationship. Interpersonal counselling can be utilised general practice to reduce psychological symptoms, restore morale, improve self esteem and the quality of the patient's social adjustment and interpersonal relationships.


Four-dimensional transcatheter intraarterial perfusion MR imaging for monitoring chemoembolization of hepatocellular carcinoma: preliminary results.
Angiographic endpoints for chemoembolization of hepatocellular carcinoma (HCC) are subjective, and optimal endpoints remain unknown. Transcatheter intraarterial perfusion (TRIP) magnetic resonance (MR) imaging, when performed in a combined MR/interventional radiology (MR-IR) suite, offers an objective method to quantify intraprocedural tumor perfusion changes, but was previously limited to two spatial dimensions. This study prospectively tested the hypothesis that a new volumetric acquisition over time, four-dimensional TRIP MR imaging, can measure HCC perfusion changes during chemoembolization. Seven men (mean age, 53 years; range, 42-65 y) with eight tumors (mean size, 2.5 x 2.4 cm(2); diameter range, 1.5-5.2 cm) underwent chemoembolization in an MR-IR suite between February and December 2007, with intraprocedural tumor perfusion reductions monitored with four-dimensional TRIP MR imaging. Microcatheter chemoembolization was performed with a 1:1 mixture of chemotherapy agent and emulsifying contrast agent, followed by the administration of gelatin microspheres. Pre- and post-chemoembolization time-intensity curves were generated for each tumor. Semiquantitative measures of tumor perfusion, including area under the curve (AUC), peak signal intensity (SI), time to peak SI, and maximum upslope (MUS), were calculated, and mean differences before and after chemoembolization were compared with paired t tests. Four-dimensional TRIP MR imaging-monitored chemoembolization was successful in all cases. Calculated AUCs before and after chemoembolization (439 vs 221, P = .004, 50% reduction), peak SI (32 vs 19, P = .012, 41% reduction), and MUS (11 vs 3, P = .028, 73% reduction) showed significant reductions after chemoembolization. Time to peak SI did not significantly change (23 sec vs 36 sec, P = .235, 57% increase). Four-dimensional TRIP MR imaging can successfully measure semiquantitative changes in HCC perfusion during MR-IR-monitored chemoembolization. Future studies may correlate changes in these objective functional parameters with tumor response.


[Cytomegalovirus infection in Omsk region].
Epidemiologic studies of prevalence of cytomegalovirus infection (CMVI) in Omsk region during a period of 15 years showed 1.6-fold increase of population seropositivity to CMV with rate of increase +2.94. In recent years antibodies to CMV were detected in 90.3 - 94.8% of cases. Diagnostics of innate and acquired CMVI was improved. Feasibility of detection of infectious process activity during complex use of new laboratory methods (IgA detection, assessment of IgG avidity, IgG to individual antigens in immunoblotting as well as detection of DNA and "early phase" viral ptoteins) was demonstrated. High rates of infection in population determine the necessity of revision of strategy for examination of children and adults, including screening of pregnant woman, on this infection.


Biochemical rationale and myocardial tissue data on the effective therapy of cardiomyopathy with coenzyme Q10.
The tissue levels of coenzyme Q10 (CoQ10) in endomyocardial biopsy samples and blood from 43 patients with cardiomyopathy were determined by steps of extraction, purification, and HPLC. The biopsy samples were obtained from the patients after a routine heart catheterization. Six patients were of class I, 18 of class II, 11 of class III, and 8 of class IV (classified according to guidelines of the New York Heart Association). True control biopsies of healthy hearts are not available for ethical reasons, but the data of the four classes by severity of disease may be justifiably compared. Patients of class IV had lower (P less than 0.01) levels of CoQ10 than those of class I. Patients of classes III and IV had a lower (P less than 0.0001) level than those of classes I and II. Biopsy samples were obtained from five patients after treatment with CoQ10 for 2-8 months. The increases of CoQ10 levels ranged from 20% to 85%; the mean value was higher (P less than 0.02) than before treatment. Blood deficiencies also increase with severity of disease, but not as markedly as for the biopsies. These data reveal a myocardial deficiency of CoQ10, which is higher with increasing severity of disease and is reduced by therapy. This biochemistry correlates with the effective treatment of cardiomyopathy with CoQ10.


Sludge conditioning by using alumina nanoparticles and polyelectrolyte.
The combined use of nanoparticle and polyelectrolyte as retention aids in pulp and paper industry tend to give better flocculation (retention) and drainage (dewatering) than conventional polyelectrolyte flocculation. The combined use of alumina nanoparticle and polyelectrolyte in conditioning waste activated sludge was investigated in the current study. Alumina with three different sizes (139.5, 241.7, and 326.4 nm) was utilized in combination with a cationic polyelectrolyte (T3052) of molecular weight of 1.1 x 10(7) and charge density of 2.1 meq/g. It was found from capillary suction time (CST) and specific resistance to filtration (SRF) measurement that sludge conditioned with alumina/polyelectrolyte showed a better dewaterability than polyelectrolyte alone. In addition, it was found that the better dewaterability was obtained as alumina became smaller. Floc size and fractal dimension of flocs were examined as well. The mechanism of enhanced dewaterability was proposed that alumina nanoparticles became adsorbed onto sludge and positive patches were formed. The electrostatic repulsion made the polyelectrolyte more stretched, and resulted in more effective flocculation and bridging as polyelectrolyte was added. Dosing sequence also affected the conditioning effectiveness. However, the combined use of silica nanoparticles and a cationic polelectrolyte, KP-201C, or alumina nanoparticles and an anionic polyelectrolyte, AP410, did not result in improved dewaterability of waste activated sludge. Possible explanation was discussed. Preliminary results show that some in-depth work is needed regarding the introduction of nanoparticles in sludge conditioning.


Serious doubts over "Eggs forever?".
A recent commentary in this journal by Byskov et al. (2005) claims that, despite published results from numerous independent lines of investigation from our laboratory and others, there does not "exist any evidence for neo-folliculogenesis in the adult mammalian ovary." While we agree with Byskov et al. that our work represents a radical departure from the age-old dogma that mammalian females permanently lose the capacity for oocyte and follicle production during the perinatal period, careful examination of all of the available data leaves no doubt that adult female mammals retain the capacity for oogenesis and folliculogenesis. These findings do not change the fact that exhaustion of the oocyte pool occurs with advancing chronological age--a process responsible for driving the menopause in women--but rather question the basic mechanism underlying age-related ovarian failure. In this regard, studies of aging male mice have demonstrated that testicular atrophy is associated with a dramatic decline in the number, activity and quality of germline stem cells that maintain spermatogenesis during adulthood (Zhang et al., 2006). Therefore, to the contrary of the opinion of Byskov et al. that such a process would be "considered exceptional among stem cells," it is certainly reasonable to hypothesize that a similar deterioration of female germline stem cell function underlies the decline in oocyte quality and the onset of ovarian failure in aging females. Further, while we accept that a departure from conventional thinking can take years to gain widespread acceptance, we feel this resistance to change should not be construed as the sole means to voice opinions about the validity of our data or the maturity of our principal conclusion.


Long term effect (more than five years) of intrathecal baclofen on impairment, disability, and quality of life in patients with severe spasticity of spinal origin.
To evaluate long term change in impairment, disability, and health related functional status in patients with severe spasticity who received intrathecal baclofen. A long term (more than five years) observational longitudinal follow up study assessing 21 patients who received intrathecal baclofen given by programmable pump. Patients had chronic disabling spasticity which did not respond to oral antispasmolytic agents. Clinical efficacy was assessed by the Ashworth scale and spasm score; disability by the expanded disability status scale (EDSS), ambulation index (AI), and incapacity status scale (ISS); and health related quality of life by the sickness impact profile (SIP) and the Hopkins symptom checklist (HSCL). Compared with pretreatment values, there was a significant improvement in clinical efficacy (Ashworth scale and spasm score, p<0.05) but a small but significant worsening of disability (EDSS, AI, and ISS, p<0.05). Comparing pretreatment with 26 weeks after pump implantation, a worsening was observed in disability (EDSS and ISS, p<0.05) and perceived health status (SIP, psychosocial dimension, p<0.05). Long term administration of intrathecal baclofen delivered by an implanted programmable pump resulted in improved clinical efficacy but not in improvement in disability or perceived health status.


Sessile serrated adenomas: demographic, endoscopic and pathological characteristics.
To study the demographic and endoscopic characteristics of patients with sessile serrated adenoma (SSA) in a single center. Patients with SSA were identified by review of the pathology database of Mayo Clinic Arizona from 2005 to 2007. A retrospective chart review was performed to extract data on demographics, polyp characteristics, presence of synchronous adenomatous polyps or cancer, polypectomy methods, and related complications. One hundred and seventy-one (2.9%) of all patients undergoing colonoscopy had a total of 226 SSAs. The mean (SE) size of the SSAs was 8.1 (0.4) mm; 42% of SSAs were < or = 5 mm, and 69% were < or = 9 mm. Fifty-one per cent of SSAs were located in the cecum or ascending colon. Approximately half of the patients had synchronous polyps of other histological types, including hyperplastic and adenomatous polyps. Synchronous adenocarcinoma was present in seven (4%) cases. Ninety-seven percent of polyps were removed by colonoscopy. Among patients with colon polyps, 2.9% were found to have SSAs. Most of the SSAs were located in the right side and were safely managed by colonoscopy.


The genus Amegilla (Hymenoptera, Apidae, Anthophorini) in Australia: a revision of the subgenus Asaropoda.
The species in the subgenus Amegilla (Asaropoda) are revised. Species delineation was decided based on diagnostic morphological characters as well as an incomplete phylogeny based on mitochondrial cytochrome oxidase 1 sequence data. Strong support was obtained for separating the Australian species of Amegilla into the three subgenera previously proposed on the basis of morphology. The subgenus Asaropoda was found to comprise 21 species, including ten new species: A. albiclypeata Leijs, sp. nov., A. aurantia Leijs, sp. nov., A. batleyi Leijs, sp. nov., A. crenata Leijs, sp. nov., A. griseocincta Leijs, sp. nov., A. incognita Leijs, sp. nov., A. nitidiventris Leijs, sp. nov., A. scoparia Leijs, sp. nov., A. xylocopoides Leijs, sp. nov., and A. youngi Leijs, sp. nov. The subspecies A. preissi frogatti is raised to species level, and 16 new synonymies are proposed. Keys to the species of both sexes and descriptions or redescriptions are provided. Distribution maps, data on flower visitation and phenology are given.


Early microbiological response to linezolid vs vancomycin in ventilator-associated pneumonia due to methicillin-resistant Staphylococcus aureus.
Methicillin-resistant Staphylococcus aureus (MRSA) is a common cause of ventilator-associated pneumonia (VAP). This prospective, open-label, multicenter clinical trial compared the early microbiological efficacy of linezolid (LZD) therapy with that of vancomycin (VAN) therapy in patients with MRSA VAP. A total of 149 patients with suspected MRSA VAP were randomized to receive either LZD, 600 mg, or VAN, 1 g every 12 h. Patients with baseline bronchoscopic BAL (BBAL) fluid quantitative culture findings that were positive for MRSA (>or= 10(4) cfu/mL) comprised the study population. The primary outcome was microbiological response (<or= 10(2) cfu/mL) in a second BBAL performed 72 to 96 h following the start of treatment. Thirty LZD-treated patients and 20 VAN-treated patients had microbiologically confirmed MRSA at baseline; 23 and 19 patients, respectively, underwent repeat BBAL. While a greater number of LZD-treated patients than VAN-treated patients achieved a microbiological cure (56.5% vs 47.4%, respectively; p = 0.757; 95% confidence interval, -21.1 to 39.4), this difference was not statistically significant. Nonstatistically significant differences were also seen for LZD-treated patients vs VAN-treated patients in terms of clinical cure (66.7% vs 52.9%, respectively), survival rate (86.7% vs 70.0%, respectively), and the mean duration of ventilation (10.4 vs 14.3 d, respectively), hospitalization (18.8 vs 20.1 d, respectively), ICU stay (12.2 vs 16.2 d, respectively), and time spent alive and not receiving mechanical ventilation (15.5 vs 11.1 d, respectively). Three patients who had been extubated prior to undergoing repeat BBAL had been randomized to receive LZD therapy. Early microbiological cure rates were not statistically significantly higher with LZD therapy than with VAN therapy despite trends in all secondary clinical outcomes favoring LZD therapy. These results suggest that any beneficial effect of LZD therapy may be due to factors other than increased bacterial clearance. Clinicaltrials.gov Identifier: NCT00572559.


The innovative use of qualitative and mixed methods research to advance improvements along the HIV prevention and care continua.
Despite enormous advances in prevention and care modalities, HIV continues to burden populations around the globe and is largely driven by social and behavioral processes. Mixed methods and qualitative research endeavors are best suited to uncovering and making sense of these dynamics, producing unique and actionable findings to alleviate the burden of HIV. We reviewed the global literature published on PubMed from 2020 to 2021 to identify studies that produced new insights into the social and behavioral dynamics that drive the HIV epidemic, focusing on mixed methods or purely qualitative study designs. Mixed methods and qualitative studies have revealed important nuances in the social and behavioral dynamics associated with the HIV prevention and care continua, from preexposure prophylaxis uptake and adherence to engagement in HIV care and treatment, and have important implications for attaining goals for controlling the epidemic. Articles reviewed contribute to advancing our understanding of complex social dynamics, structural level factors such as healthcare systems and policy, as well as the research endeavor itself and the need to diversify and sustain research to truly represent the perspectives of those most impacted by HIV. Numerous studies represent the unique ability of qualitative and mixed methods research to expand our understanding of and empathy for individuals living with and affected by HIV, offering new insights to help alleviate the burden of HIV.


Short-term macroinvertebrate recruitment and sediment accumulation: a novel field chamber approach.
Stream-deposited sediment is one of the major stressors affecting stream biota. Several methods exist to quantify stream sediment embeddedness, but they are relatively qualitative and operationally defined. The authors developed a short-term in situ embeddedness chamber method to measure aquatic insect recruitment and associated sediment accumulation in a more quantitative, better replicated manner. With sediment accumulation and aquatic insect recruitment as endpoints, three exposure periods were evaluated (4, 7, and 14 d) on a low-order stream (Honey Creek, New Carlisle, Ohio, USA) and a medium-order stream (Stillwater River, Covington, Ohio, USA). Chamber results show significant positive correlations between newly deposited fine sediment and insect recruitment. Embeddedness was also measured using the more conventional techniques of the Burns method and the U.S. Geological Survey National Water Quality Assessment Program method. This in situ chamber method allows for increased experimental options for assessing the stress of embeddedness and siltation on benthic communities and may prove useful for investigating the resilience of benthic communities after disturbances.


Functional genomics of drug-induced ion homeostasis identifies a novel regulatory crosstalk of iron and zinc regulons in yeast.
Pyrrolidine dithiocarbamate (PDTC), a known inhibitor of NFκB activation, has antioxidative as well as antiviral activities. PDTC is effective against several virus families, indicating that its antiviral mechanism targets host rather than viral functions. To investigate its mode of action, we used baker's yeast as a simple eukaryotic model system and two types of genome-wide analysis. First, expression profiling using whole-genome DNA microarrays identifies more than 200 genes differentially regulated upon PDTC exposure. Interestingly, the Aft1-dependent iron regulon is a main target of PDTC, indicating a lack of iron availability. Moreover, the PDTC-caused zinc influx triggers a strong regulatory effect on zinc transporters due to the cytoplasmic zinc excess. Second, phenotypic screening the EUROSCARF collection for PDTC hypersensitivity identifies numerous mutants implicated in vacuolar maintenance, acidification as well as in transport, mitochondrial organization, and translation. Notably, the screening data indicate significant overlaps of PDTC-sensitive genes and those mediating zinc tolerance. Hence, we show that PDTC induces cytoplasmic zinc excess, eliciting vacuolar detoxification, which in turn, disturbs iron homeostasis and activates the iron-dependent regulator Aft1. Our work reveals a complex crosstalk in yeast ion homeostasis and the underlying regulatory networks.


The local impact of a coal-fired power plant on inorganic mercury and methyl-mercury distribution in rice (Oryza sativa L.).
Emission from coal-fired power plants is one of the major anthropogenic sources of mercury (Hg) in the environment, because emitted Hg can be quickly deposited nearby the source, attention is paid to the effects of coal-burning facilities on levels of toxic methyl-mercury (MeHg) in biota near such sources. Since rice is an agricultural crop that can bio-accumulate MeHg, the potential effects of a large Hg-emitting coal-fired power plant in Hunan Province, China on both inorganic Hg (Hg(II)) and MeHg distributions in rice was investigated. Relatively high MeHg (up to 3.8 μg kg-1) and Hg(II) (up to 22 μg kg-1) concentrations were observed in rice samples collected adjacent to the plant, suggesting a potential impact of Hg emission from the coal fired power plant on the accumulation of Hg in rice in the area. Concentrations of MeHg in rice were positively correlated with soil MeHg, soil S, and gaseous elemental Hg (GEM) in ambient air. Soil MeHg was the most important factor controlling MeHg concentrations in rice. The methylation of Hg in soils may be controlled by factors such as the chemical speciation of inorganic Hg, soil S, and ambient GEM.


Photoaffinity ligands in the study of cytochrome p450 active site structure.
While photoaffinity ligands have been widely used to probe the structures of many receptors and nucleic acid binding proteins, their effective use in the study of cytochrome p450 structure is less established. Nevertheless, significant advances in this field have been made since the technique was first applied to p450cam in 1979. In several cases, especially studies involving p450s of the 1A and 2B families, peptides covalently modified with photoaffinity ligands have been isolated and characterized. Some of these peptides were predicted by molecular modeling to line substrate binding regions of the enzymes. Other data obtained from such studies were more difficult to reconcile with theory. This review addresses the status of photoaffinity labeling as a tool for studying cytochrome p450 structure. In addition, potential future directions in this field are discussed, including the development of heme-directed agents and validation of their effectiveness as photoaffinity ligands using sperm whale myoglobin as a test protein. The potential for hydroxyaromatic compounds to serve as photoactivated probes of active site nucleophiles is also discussed. This class of compounds and its derivatives has long been known in the fields of photochemistry and photophysics to be precursors of reactive radicals and quinone methides that are likely to serve as effective active site probes of the p450s.


Antibody response in a family with COVID-19.
Robust serological assays for SARS-CoV-2 are essential for determining prior infection and the suitability of donated convalescent plasma for plasma therapy. We compared two in-house and three commercial serological assays in a family cohort with SARS-CoV-2-infected members. Three individuals in a family of five developed COVID-19 confirmed by PCR, following a trip abroad. Three to four weeks after the onset of symptoms, three commercial ELISAs and an in-house immunofluorescence test demonstrated antibodies to SARS-CoV-2. An in-house neutralisation test also demonstrated neutralising antibodies. The in-house assays and one commercial assay gave congruent results, which were also consistent with the initial PCR and/or clinical evaluation, indicating prior infection in three of the five family members. The other commercial assays indicated possible infection in a fourth family member, but this result was likely due to cross-reactivity. The neutralising antibodies suggest complete or partial immunity against reinfection.


St John's Wort (Hypericum perforatum L.) photomedicine: hypericin-photodynamic therapy induces metastatic melanoma cell death.
Hypericin, an extract from St John's Wort (Hypericum perforatum L.), is a promising photosensitizer in the context of clinical photodynamic therapy due to its excellent photosensitizing properties and tumoritropic characteristics. Hypericin-PDT induced cytotoxicity elicits tumor cell death by various mechanisms including apoptosis, necrosis and autophagy-related cell death. However, limited reports on the efficacy of this photomedicine for the treatment of melanoma have been published. Melanoma is a highly aggressive tumor due to its metastasizing potential and resistance to conventional cancer therapies. The aim of this study was to investigate the response mechanisms of melanoma cells to hypericin-PDT in an in vitro tissue culture model. Hypericin was taken up by all melanoma cells and partially co-localized to the endoplasmic reticulum, mitochondria, lysosomes and melanosomes, but not the nucleus. Light activation of hypericin induced a rapid, extensive modification of the tubular mitochondrial network into a beaded appearance, loss of structural details of the endoplasmic reticulum and concomitant loss of hypericin co-localization. Surprisingly the opposite was found for lysosomal-related organelles, suggesting that the melanoma cells may be using these intracellular organelles for hypericin-PDT resistance. In line with this speculation we found an increase in cellular granularity, suggesting an increase in pigmentation levels in response to hypericin-PDT. Pigmentation in melanoma is related to a melanocyte-specific organelle, the melanosome, which has recently been implicated in drug trapping, chemotherapy and hypericin-PDT resistance. However, hypericin-PDT was effective in killing both unpigmented (A375 and 501mel) and pigmented (UCT Mel-1) melanoma cells by specific mechanisms involving the externalization of phosphatidylserines, cell shrinkage and loss of cell membrane integrity. In addition, this treatment resulted in extrinsic (A375) and intrinsic (UCT Mel-1) caspase-dependent apoptotic modes of cell death, as well as a caspase-independent apoptotic mode that did not involve apoptosis-inducing factor (501 mel). Further research is needed to shed more light on these mechanisms.


Engraftment and Repopulation Potential of Late Gestation Fetal Rat Hepatocytes.
The limited availability of donor organs has led to a search for alternatives to liver transplantation to restore liver function and bridge patients to transplantation. We have shown that the proliferation of late gestation (embryonic day 19) fetal rat hepatocytes is mitogen-independent and that mechanisms regulating mRNA translation, cell cycle progression, and gene expression differ from those of adult rat hepatocytes. In the present study, we investigated whether E19 fetal hepatocytes can engraft and repopulate an injured adult liver. Fetal hepatocytes were isolated using a monoclonal antibody against a hepatic surface protein, leucine amino peptidase (LAP). LAP+ and LAP- fractions were analyzed by immunofluorescence and microarray. Immunopurified E19 liver cells from DPPIV+ rats were transplanted via splenic injection into partial hepatectomized DPPIV- rats that had been pretreated with mitomycin C. More than a third of LAP+ fetal hepatocytes expressed ductal markers. Transcriptomic analysis revealed that these dual-expressing cells represent a population of less well-differentiated hepatocytes. Upon transplantation, LAP+ late gestation fetal hepatocytes formed hepatic, endothelial, and ductal colonies within 1 month. By 10 months, colonies derived from LAP+ cells increased so that up to 35% of the liver was repopulated by donor-derived cells. Late gestation fetal hepatocytes, despite being far along in the differentiation process, possess the capacity for extensive liver repopulation. This is likely related to the unexpected presence of a significant proportion of hepatocyte marker-positive cells maintaining a less well-differentiated phenotype.


Modulation of experimental colon tumorigenesis by types and amounts of dietary fatty acids.
Epidemiological studies and laboratory animal model assays suggest that a high intake of dietary fat promotes colorectal cancer. Several in vivo and in vitro studies support the hypothesis that omega-6 fatty acids promote colon tumorigenesis, whereas omega-3 fatty acids lack promoting activity. Fat intake in the United States traditionally includes high amounts (30% of total caloric intake) of saturated fat rather than omega-6 fatty acids. Therefore, the present study was designed to compare the modulatory effects of a high-fat diet containing mixed lipids (HFML), a diet rich in saturated fatty acids (the average American diet), a diet with fish oil (HFFO) that is rich in omega-3 fatty acids, and a low-fat corn oil diet (LFCO) on the formation of chemically induced colonic aberrant crypt foci (ACF) and tumors, cyclooxygenase (COX)-2 activity, and apoptosis during experimental colon carcinogenesis. At 5 weeks of age, groups of male F344 rats were fed a 5% corn oil diet (LFCO). At 7 weeks of age, rats intended for carcinogen treatment received s.c. injections of azoxymethane at a dose level of 15 mg/kg of body weight once weekly for 2 weeks. Beginning 1 day after the carcinogen treatment, groups of rats were then maintained on experimental diets containing 20% HFML or 20% HFFO. Rats were killed at 8, 23, or 38 weeks after azoxymethane treatment. Colonic ACF and tumors were evaluated histopathologically, and apoptosis was evaluated by the terminal deoxynucleotidyl transferase-mediated nick end labeling method. Colonic mucosae and tumor samples harvested at week 38 were analyzed for COX-2 synthetic activity and expression. The rats fed the HFML diet showed significantly increased total colonic ACF (P < 0.001-0.0001) with a multiplicity of > or = 4 aberrant crypts/focus (P < 0.0001) compared with the effects of the HFFO or LFCO diets at week 8, 23, and 38. Interestingly, there was a 2- to 3-fold increase (> or = 4) in multicrypt foci in rats given the HFML diet as compared with such foci in rats fed the HFFO or LFCO diets. By week 23, the HFML diet had significantly increased the incidence of colonic tumors (30-60%) and their multiplicity (100-141%) when compared with the effects of the LFCO or HFFO diets. At week 38, the HFML diet had induced 100% colon tumor incidence and a 4-fold multiplicity of adenocarcinomas compared with the LFCO and HFFO diets. At weeks 23 and 38, a significantly lower percentage of apoptotic colonic epithelial cells were observed in the tumors of animals fed the HFML diet as compared with those fed the HFFO diet. The HFML diet caused significantly increased levels of COX-2 activity in colon tumors (P < 0.05-0.01), and these tumors had enhanced levels of COX-2 expression as compared with those in assays with LFCO or HFFO diets. These observations demonstrate for the first time that HFML diets containing high levels of saturated fatty acids (such as those in Western diets) promote colon carcinogenesis. Although the mechanisms involved in colon tumor promotion by a HFML diet are not fully known, our results indicate that the modulation of eicosanoid production via the influence on COX activity and the suppression of apoptosis may play a key role in HFML diet-induced colon tumorigenesis.


Pancreaticobiliary endoscopic ultrasound in England 2007 to 2016: Changing practice and outcomes.
Background and study aims Population-level data on the outcomes of pancreaticobiliary endoscopic ultrasound (PB-EUS) are limited. We examined national PB-EUS and fine-needle aspiration (FNA) activity, its relation to pancreatic cancer therapy, associated mortality and adverse events. Patients and methods Adults undergoing PB-EUS in England from 2007-2016 were identified in Hospital Episode Statistics. A pancreatic cancer cohort diagnosed within 6 months of PB-EUS were studied separately. Multivariable logistic regression models examined associations with 30-day mortality and therapies for pancreatic cancer. Results 79,269 PB-EUS in 68,908 subjects were identified. Annual numbers increased from 2,874 (28 % FNA) to 12,752 (35 % FNA) from 2007 to 2016. 8,840 subjects (13 %) were diagnosed with pancreatic cancer. Sedation related adverse events were coded in 0.5 % and emergency admission with acute pancreatitis in 0.2 % within 48 hours of PB-EUS. 1.5 % of subjects died within 30 days of PB-EUS. Factors associated with 30-day mortality included increasing age (odds ratio 1.03 [95 % CI 1.03-1.04]); male sex (1.38 [1.24-1.56]); increasing comorbidity (1.49 [1.27-1.74]); EUS-FNA (2.26 [1.98-2.57]); pancreatic cancer (1.39 [1.19-1.62]); increasing deprivation (least deprived quintile 0.76 [0.62-0.93]) and lower provider PB-EUS volume (2.83 [2.15-3.73]). Factors associated with surgical resection in the pancreatic cancer cohort included lower provider PB-EUS volume (0.44 [0.26-0.74]) and the least deprived subjects (1.33 [1.12-1.57]). 33 % of pancreatic cancer subjects who underwent EUS, did not subsequently receive active cancer treatment. Conclusions Lower provider PB-EUS volume was associated with higher 30-day mortality and reduced rates of both pancreatic cancer surgery and chemotherapy. These results suggest potential issues with case selection in lower-volume EUS providers.


Dynamic reconfiguration of security policies in wireless sensor networks.
Providing security and privacy to wireless sensor nodes (WSNs) is very challenging, due to the heterogeneity of sensor nodes and their limited capabilities in terms of energy, processing power and memory. The applications for these systems run in a myriad of sensors with different low-level programming abstractions, limited capabilities and different routing protocols. This means that applications for WSNs need mechanisms for self-adaptation and for self-protection based on the dynamic adaptation of the algorithms used to provide security. Dynamic software product lines (DSPLs) allow managing both variability and dynamic software adaptation, so they can be considered a key technology in successfully developing self-protected WSN applications. In this paper, we propose a self-protection solution for WSNs based on the combination of the INTER-TRUST security framework (a solution for the dynamic negotiation and deployment of security policies) and the FamiWare middleware (a DSPL approach to automatically configure and reconfigure instances of a middleware for WSNs).We evaluate our approach using a case study from the intelligent transportation system domain.


ESHRE PGD consortium data collection VII: cycles from January to December 2004 with pregnancy follow-up to October 2005.
The seventh report of the ESHRE PGD Consortium is presented documenting cycles collected for the calendar year 2004 and follow-up of the pregnancies and babies born subsequent to these cycles up to October 2005. Since the beginning of the data collections, there has been a steady increase in the number of cycles, pregnancies and babies reported. For data collection VII, 45 centres have participated, reporting on 3358 cycles to oocyte retrieval (OR), 679 pregnancies and 528 babies born. Five hundred and fifty nine OR were reported for chromosomal abnormalities, 113 OR for sexing for X-linked diseases, 520 OR for monogenic diseases, 2087 OR for PGS, and 79 OR for social sexing. Data VII is compared with the cumulative data for data collections I-VI.


A Simple Approach for Predicting the Spin State of Homoleptic Fe(II) Tris-diimine Complexes.
We propose a simple method for predicting the spin state of homoleptic complexes of the Fe(II) d6 ion with chelating diimine ligands. The approach is based on the analysis of a single metric parameter within a free (noncoordinated) ligand: the interatomic separation between the N-donor metal-binding sites. An extensive analysis of existing complexes allows the determination of critical N···N distances that dictate the regions of stability for the high-spin and low-spin complexes, as well as the intermediate range in which the magnetic bistability (spin crossover) can be observed. The prediction has been tested on several complexes that demonstrate the validity of our method.


Transneuronal transport of herpes simplex virus from the cervical vagus to brain neurons with axonal inputs to central vagal sensory nuclei in the rat.
The recent introduction of live viruses as intra-axonal tracing agents has raised questions concerning which central neurons are transneuronally labelled after application of the virus to peripheral organs or peripheral nerves. Since the central connections of the vagus nerve have been well described using conventional neuronal tracing agents, we chose to inject Herpes Simplex Virus Type 1 into the cervical vagus of the rat. After survival times of up to 3 days the rat brains were processed immunohistochemically using a polyclonal antiserum against herpes simplex virus. Two days after injection of the virus we observed viral antigen in the area postrema and in the nucleus tractus solitarius and the dorsal motor nucleus of the vagus (dorsal vagal complex), principally ipsilaterally. At this survival time the viral antigen in the dorsal vagal complex was largely confined to glial cells. After 3 days the viral antigen was localized both in glia and in nerve cells within the dorsal vagal complex and in brain regions previously demonstrated, using conventional tracing procedures, to contain neurons with axonal projections to the dorsal vagal complex. This was true for medullary, pontine, midbrain and hypothalamic regions and for telencephalic regions including the amygdala, the bed nucleus of the stria terminalis, and the insular and medial frontal cortices. Many of the nerve cells containing viral antigen were displayed in a Golgi-like manner, with excellent visualization of the dendritic tree. Axonal processes, in contrast, were not visualized. We used co-localization studies to confirm previous findings concerning monoamine neurotransmitter-related antigens present in medullary and pontine neurons projecting to the dorsal vagal complex. After 3 days there were many Herpes Simplex Virus Type 1-containing glial cells along the intra-medullary course of the vagal rootlets. However, no viral antigen was found in brain regions containing neurons whose axons pass through the region of glial cell-labelled rootlets. Glial cells containing viral antigen were particularly numerous in brain regions known to receive an input from neurons in the area postrema and the dorsal vagal complex. Taken together with our observation concerning the early appearance of viral antigen within glial cells in the dorsal vagal complex, this suggests that when the virus reaches the axon terminal portion it is transferred to nearby glial cells and possibly enters central neurons by way of these structures.


Role of transthoracic Doppler echocardiography in patients with a proximal left coronary artery lesion that cannot be diagnosed by computed tomography angiography.
The diagnosis of lesions with severe calcium or in-stent stenosis using coronary computed tomography angiography (CCTA) is still difficult. The aim of the present study was to evaluate the accuracy of transthoracic Doppler echocardiography (TTDE) in patients with suspected angina pectoris, who had a proximal left coronary artery (LCA) site that could not be evaluated by CCTA. Fifty-eight patients were evaluated. The proximal LCA was defined as the left main coronary artery and proximal left anterior descending coronary artery. All patients underwent TTDE and had coronary angiography performed as a reference method. We measured the proximal left coronary flow velocity (CFV) by both color and pulse Doppler methods. Proximal coronary flow was detected in 45 (78%) of 58 patients. CFVs measured by both methods were significantly greater in the group with severe stenosis (percent diameter stenosis >70%) than in the groups with moderate stenosis (percent diameter stenosis 40% to 70%) or without stenosis (color Doppler: 148 ± 42 cm/s, 89 ± 40 cm/s, and 41 ± 22 cm/s, respectively, p <0.05; pulse Doppler: 143 ± 61 cm/s, 82 ± 33 cm/s, and 39 ± 17 cm/s, respectively, p <0.05). Receiver operating characteristic curve analysis showed that the optimal CFV cut-off values obtained by color and pulse Doppler to diagnose severe stenosis were 92 cm/s (sensitivity, 100%; specificity, 90%) and 81 cm/s (sensitivity, 100%; specificity, 85%), respectively. In conclusion, TTDE could diagnose proximal LCA stenosis with good accuracy in patients who could not be evaluated by CCTA.


Evolution of male pheromones in moths: reproductive isolation through sexual selection?
Central to our understanding of the species concept is knowledge of the nature and evolution of reproductive isolating mechanisms. The once widely accepted model of Dobzhansky, which holds that isolation evolves through selection against hybrids of differentially adapted populations, is now largely rejected. This rejection is due to both theoretical difficulties and a paucity of examples of the predicted pattern of reproductive character displacement. From a survey of five families of Lepidoptera, entailing more than 800 species, evidence is given that male courtship pheromones have evolved within the context of sexual isolation as an adaptive response to mating mistakes between differentially adapted populations; however, distinct from the natural selection model of Dobzhansky, this report suggests the mechanism for change to be sexual selection.


The evolution of neuropeptide signalling: insights from echinoderms.
Neuropeptides are evolutionarily ancient mediators of neuronal signalling that regulate a wide range of physiological processes and behaviours in animals. Neuropeptide signalling has been investigated extensively in vertebrates and protostomian invertebrates, which include the ecdysozoans Drosophila melanogaster (Phylum Arthropoda) and Caenorhabditis elegans (Phylum Nematoda). However, until recently, an understanding of evolutionary relationships between neuropeptide signalling systems in vertebrates and protostomes has been impaired by a lack of genome/transcriptome sequence data from non-ecdysozoan invertebrates. The echinoderms-a deuterostomian phylum that includes sea urchins, sea cucumbers and starfish-have been particularly important in providing new insights into neuropeptide evolution. Sequencing of the genome of the sea urchin Strongylocentrotus purpuratus (Class Echinoidea) enabled discovery of (i) the first invertebrate thyrotropin-releasing hormone-type precursor, (ii) the first deuterostomian pedal peptide/orcokinin-type precursors and (iii) NG peptides-the 'missing link' between neuropeptide S in tetrapod vertebrates and crustacean cardioactive peptide in protostomes. More recently, sequencing of the neural transcriptome of the starfish Asterias rubens (Class Asteroidea) enabled identification of 40 neuropeptide precursors, including the first kisspeptin and melanin-concentrating hormone-type precursors to be identified outside of the chordates. Furthermore, the characterization of a corazonin-type neuropeptide signalling system in A. rubens has provided important new insights into the evolution of gonadotropin-releasing hormone-related neuropeptides. Looking forward, the discovery of multiple neuropeptide signalling systems in echinoderms provides opportunities to investigate how these systems are used to regulate physiological and behavioural processes in the unique context of a decentralized, pentaradial bauplan.


Synthesis of phosphorylated recognition marker in lysosomal enzymes is located in the cis part of Golgi apparatus.
Rat liver membranes were subjected to centrifugation in a sucrose density gradient in which the Golgi apparatus was separated into several subfractions. Two enzymes involved in the synthesis of the phosphorylated recognition marker in lysosomal enzymes, UDP-N-acetylglucosamine:lysosomal enzyme precursor N-acetylglucosamine-1-phosphotransferase and alpha-N-acetylglucosaminyl phosphodiesterase fractionated with alpha-1,2-mannosidase, a marker enzyme of cis Golgi membranes and differently from galactosyltransferase, a marker enzyme of trans Golgi membranes.


Anticipation of racism and sexism: Factors related to setting career goals for urban youth of color.
This study examined the effects of anticipated racial and gender discrimination on variables related to the development of career goals. For a sample of 195 urban youth of color we used path analysis to test a career decision self-efficacy and goal-setting model informed by the social cognitive career theory self-management model of career exploration and decision-making behavior (Lent & Brown, 2013). Specifically, we examined how students' anticipation of racial and gender discrimination in the workplace were related to the process variables of career decision self-efficacy and outcome expectations, and how self-efficacy and outcome expectations were related to setting clear, viable goals. The results indicate different pathways for boys and girls. For boys, an anticipated hostile racial employment climate was negatively related to career decision self-efficacy, with career decision self-efficacy and vocational outcome expectations positively related to goals. For girls, an anticipated hostile racial employment climate was negatively related to vocational outcome expectations, with vocational outcome expectations positively related to career goals. Analyses showed that for boys, career decision-making self-efficacy fully mediated the effects of racial discrimination on vocational outcome expectations, and vocational outcome expectations partially mediated the effects of career decision-making efficacy on goals. While career decision making was related to vocational outcome expectations, there were no indirect effects for girls. For this sample, anticipated employment-related gender discrimination was not significantly related to career decision self-efficacy or outcome expectations among either boys or girls. Implications for training, practice, and research are discussed. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Should hepatitis B surface antigen positive mothers breast feed?
Breast fed infants may be at greater risk of mother to infant hepatitis B virus infection compared with formula fed infants. We studied 85 infants born to 84 hepatitis B surface antigen positive mothers (only two of whom were hepatitis B e positive), and who had received immunisation against hepatitis B virus. Our results indicate that breast feeding does not increase the risk of developing hepatitis B virus infection in infants born to these mothers if immunisation is carried out.