Inference of site frequency spectra from high-throughput sequence data: quantification of selection on nonsynonymous and synonymous sites in humans.
Sequencing errors and random sampling of nucleotide types among sequencing reads at heterozygous sites present challenges for accurate, unbiased inference of single-nucleotide polymorphism genotypes from high-throughput sequence data. Here, we develop a maximum-likelihood approach to estimate the frequency distribution of the number of alleles in a sample of individuals (the site frequency spectrum), using high-throughput sequence data. Our method assumes binomial sampling of nucleotide types in heterozygotes and random sequencing error. By simulations, we show that close to unbiased estimates of the site frequency spectrum can be obtained if the error rate per base read does not exceed the population nucleotide diversity. We also show that these estimates are reasonably robust if errors are nonrandom. We then apply the method to infer site frequency spectra for zerofold degenerate, fourfold degenerate, and intronic sites of protein-coding genes using the low coverage human sequence data produced by the 1000 Genomes Project phase-one pilot. By fitting a model to the inferred site frequency spectra that estimates parameters of the distribution of fitness effects of new mutations, we find evidence for significant natural selection operating on fourfold sites. We also find that a model with variable effects of mutations at synonymous sites fits the data significantly better than a model with equal mutational effects. Under the variable effects model, we infer that 11% of synonymous mutations are subject to strong purifying selection.


A novel f -divergence based generative adversarial imputation method for scRNA-seq data analysis.
Comprehensive analysis of single-cell RNA sequencing (scRNA-seq) data can enhance our understanding of cellular diversity and aid in the development of personalized therapies for individuals. The abundance of missing values, known as dropouts, makes the analysis of scRNA-seq data a challenging task. Most traditional methods made assumptions about specific distributions for missing values, which limit their capability to capture the intricacy of high-dimensional scRNA-seq data. Moreover, the imputation performance of traditional methods decreases with higher missing rates. We propose a novel f -divergence based generative adversarial imputation method, called sc- f GAIN, for the scRNA-seq data imputation. Our studies identify four f -divergence functions, namely cross-entropy, Kullback-Leibler (KL), reverse KL, and Jensen-Shannon, that can be effectively integrated with the generative adversarial imputation network to generate imputed values without any assumptions, and mathematically prove that the distribution of imputed data using sc- f GAIN algorithm is same as the distribution of original data. Real scRNA-seq data analysis has shown that, compared to many traditional methods, the imputed values generated by sc- f GAIN algorithm have a smaller root-mean-square error, and it is robust to varying missing rates, moreover, it can reduce imputation bias. The flexibility offered by the f -divergence allows the sc- f GAIN method to accommodate various types of data, making it a more universal approach for imputing missing values of scRNA-seq data.


Treatment of military-related post-traumatic stress disorder: challenges, innovations, and the way forward.
Post-traumatic stress disorder (PTSD) is one of the common mental disorders in military and veteran populations. Considerable research and clinical opinion has been focused on understanding the relationship between PTSD and military service and the implications for prevention, treatment, and management. This paper examines factors associated with the development of PTSD in this population, considers issues relating to engagement in treatment, and discusses the empirical support for best practice evidence-based treatment. The paper goes on to explore the challenges in those areas, with particular reference to treatment engagement and barriers to care, as well as treatment non-response. The final section addresses innovative solutions to these challenges through improvements in agreed terminology and definitions, strategies to increase engagement, early identification approaches, understanding predictors of treatment outcome, and innovations in treatment. Treatment innovations include enhancing existing treatments, emerging non-trauma-focused interventions, novel pharmacotherapy, personalized medicine approaches, advancing functional outcomes, family intervention and support, and attention to physical health.


Comorbidity: implications for the importance of primary care in 'case' management.
Although comorbidity is very common in the population, little is known about the types of health service that are used by people with comorbid conditions. Data from claims on the nonelderly were classified by diagnosis and extent of comorbidity, using a case-mix measure known as the Johns Hopkins Adjusted Clinical Groups, to study variation in extent of comorbidity and resource utilization. Visits of patients (adults and children) with 11 conditions were classified as to whether they were to primary care physicians or to other specialists, and whether they involved the chosen condition or other conditions. Comorbidity varied within each diagnosis; resource use depended on the degree of comorbidity rather than the diagnosis. When stratified by degree of comorbidity, the number of visits for comorbid conditions exceeded the number of visits for the index condition in almost all comorbidity groups and for visits to both primary care physicians and to specialists. The number of visits to primary care physicians for both the index condition and for comorbid conditions almost invariably exceeded the number of visits to specialists. These patterns differed only for uncommon conditions in which specialists played a greater role in the care of the condition, but not for comorbid conditions. In view of the high degree of comorbidity, even in a nonelderly population, single-disease management does not appear promising as a strategy to care for patients. In contrast, the burden is on primary care physicians to provide the majority of care, not only for the target condition but for other conditions. Thus, management in the context of ongoing primary care and oriented more toward patients' overall health care needs appears to be a more promising strategy than care oriented to individual diseases. New paradigms of care that acknowledge actual patterns of comorbidities as well as the need for close coordination between generalists and specialists require support.


Association of p73 and MDM2 polymorphisms with the risk of epithelial ovarian cancer in Chinese women.
This study was to investigate the association of p73 G4C14-to-A4T14 and Murine Double Minute2 (MDM2) 309T/G, Del1518+/- single nucleotide polymorphisms with the risk of epithelial ovarian cancer (EOC) in Chinese. This hospital-based case-control study included 257 ovarian cancer patients and 257 healthy women who were matched for age. p73 and MDM2 genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism. There were no significant differences in allele frequencies and genotype distributions of the p73 G4C14-to-A4T14 polymorphism between cases and control women (P = 0.55 and 0.20, respectively). The frequencies of the G allele of the MDM2 309T/G polymorphism were significantly lower in ovarian cancer cases (46.7%) than those in healthy controls (54.7%), there was a statistical difference between the 2 groups (P = 0.01). Compared with the T/T genotype, the G allelotype (T/G+G/G genotype) significantly decreased the risk of developing EOC (odds ratio, 0.65; 95% confidence interval, 0.44-0.97). Although MDM2 Del1518+/- genotypes and allele frequencies did not differ between the case and the control groups (P = 0.68 and P = 0.45), Del1518 +/+ genotype tended to increase the risk of mucinous ovarian cancer or earlier ovarian cancer by stratification analysis according to histological subtypes or clinical stage. Besides, there was a significant interaction between p73 G4C14-to-A4T14 and MDM2 309T/G polymorphisms by the likelihood ratio test (P = 0.03; odds ratio, 0.89; 95% confidence interval, 0.80-0.99). The MDM2 SNP309G allele significantly decreased the risk of EOC and might be a potentially protective factor for EOC development in Chinese women.


Mechanisms of Nickel Toxicity in the Highly Sensitive Embryos of the Sea Urchin Evechinus chloroticus, and the Modifying Effects of Natural Organic Matter.
A 96 h toxicity test showed that the embryos of the New Zealand sea urchin (Evechinus chloroticus) are the most sensitive of all studied marine species to waterborne nickel (Ni), with the EC50 for the development of fully formed pluteus larvae found to be 14 μg L(-1). Failure to develop a standard larval shape suggested skeletal impairment. Whole body ions (Na, Mg) increased with Ni exposure and calcium influx was depressed. The effects of natural organic matter (NOM) on Ni accumulation and toxicity were also examined in three different seawater sources (nearshore, offshore, and near the outlet of a "brown water" stream). At low dissolved organic carbon (DOC) concentrations the brown water NOM was protective against Ni toxicity, however at higher DOC concentrations it exacerbated developmental toxicity in the presence of Ni. These results show that sea urchin development is highly sensitive to Ni via a mechanism that involves ionoregulatory disturbance, and that Ni toxicity is influenced by environmental factors such as NOM. These data will be critical for the development of water quality guidelines for Ni in the marine environment.


Intraocular pressure and outflow facility are unchanged following acute and chronic intracameral chondroitinase ABC and hyaluronidase in monkeys.
We determined the effect of chronic and acute loss of glycosaminoglycans from the aqueous outflow pathway on intraocular pressure (IOP) and outflow facility in the subhuman primate eye. For the study of the chronic effects of the GAGases, cynomolgus monkeys received intracameral injections of hyaluronidase (Streptomyces, 5 or 50 units, n=2) or chondroitinase ABC (0.05 or 0.25 units, n=2) biweekly for 8 months (4 months for each dose). IOP was measured at 3, 7, 10 and 14 days after each injection. Outflow facility (2-level constant pressure) was determined at 2, 4, 6, and 8 months. Monkeys were killed 6 days after the last injection. The changes in the distribution of hyaluronic acid and sulfated proteoglycans in the outflow pathway were examined using substrate-specific histochemical techniques. The acute effects of these enzymes on outflow facilities (30 min or 2 hr after enzyme) were determined in another group of animals (n=4 for each time enzyme-1). IOP and outflow facility were unchanged compared to controls (heat inactivated enzyme) at any time in the chronically or acutely treated monkeys. Hyaluronic acid staining was absent in the outflow pathways of eyes treated chronically with hyaluronidase compared with control eyes, while collagen-associated sulfated proteoglycans were decreased but not completely removed by the chronic chondroitinase ABC treatment. Chronic loss of these glycosaminoglycans from the trabecular meshwork does not appear to contribute to the IOP elevation and decrease in outflow facility that accompanies open-angle glaucoma. Most importantly, no increase in outflow facility was found with acute hyaluronidase or chondroitinase treatment.


Adult respiratory distress syndrome due to pulmonary and extrapulmonary causes: CT, clinical, and functional correlations.
To assess the differences in CT appearance between adult respiratory distress syndrome due to pulmonary disease (ARDSP) and that due to extrapulmonary disease (ARDSEXP) and determine whether the variable appearances of ARDS are due, in part, to the initial pulmonary and systemic causes. Thirty-three patients, 22 with ARDSP and 11 with ARDSEXP, underwent helical CT shortly after intubation. Two readers evaluated images for the type, extent, and distribution of pulmonary opacities; secondary findings; and correlation with survival and physiologic parameters. In both ARDSP and ARDSEXP, approximately 80% of the lung was abnormal. In ARDSP, ground-glass opacification and consolidation were equally prevalent, whereas in ARDSEXP ground-glass opacification was dominant. Ground-glass opacification was evenly distributed, whereas consolidation tended to be dorsal and caudal. ARDSP often caused asymmetric consolidation, whereas ARDSEXP caused symmetric ground-glass opacification. Air bronchograms were almost universal. Pleural effusions were present in one-half of the patients, and Kerley B lines and pneumatoceles were uncommon. Lung consolidation correlated with the ratio of mean partial pressure of arterial oxygen to fraction of inspired oxygen, shunt fraction, and pulmonary arterial pressure. The patients who died tended to have more consolidation and asymmetric disease. ARDSP tends to be asymmetric, with a mix of consolidation and ground-glass opacification, whereas ARDSEXP has predominantly symmetric ground-glass opacification. In both groups, pleural effusions and air bronchograms are common, and Kerley B lines and pneumatoceles are uncommon.


Impact of conversion to critical access hospital status for Oklahoma's rural hospitals.
The Medicare Rural Hospital Flexibility Grant Program established a new hospital category, the Critical Access Hospital, designed to provide financial stability to small rural hospitals that were losing money after changes in the Prospective Payment System implemented by Medicare. This article describes the impact of conversion to Critical Access Hospital (CAH) status for 15 small rural hospitals in Oklahoma. Objectives of the study were to identify how conversion to CAH affected hospital utilization and finances for the first year after conversion. A telephone survey was used to collect information from hospital administrators. Fifteen of 16 eligible hospitals participated in the study. In general, services and patient census declined slightly with conversion to CAH. All 15 hospitals had reported losses prior to conversion, totaling $6,985,033. Ten hospitals reported losses after conversion. After converting to CAH status, the hospitals reported total losses of $3,094,547. The hospitals had a net change of $4,293,040. Most of the 15 study hospitals greatly improved their financial situation in the first year after conversion to CAH status, but in aggregate still operated at a loss.


Semantic representation and naming in young children.
Children's semantic representations and semantic naming errors were the focus of this study. In Experiment 1, 25 normally developing children (mean age = 5 years 4 months) named, drew, and defined 20 age-appropriate objects. The results suggested that functional and physical properties are core aspects of object representations in the semantic lexicon and that these representations are often organized and accessed according to a taxonomic hierarchy. Results of a new procedure, comparative picture naming/picture drawing, suggested that the degree of knowledge in the semantic lexicon makes words more or less vulnerable to retrieval failure. Most semantic naming errors were associated with limited semantic knowledge, manifested as either lexical gaps or fragile representations. Comparison of definitions for correctly named and semantically misnamed objects provided converging evidence for this conclusion. In Experiment 2, involving 16 normally developing children (mean age = 5 years 5 months), the comparative picture naming/picture drawing results were replicated with a stimulus set that allowed a priori matching of the visual complexity of items drawn from correct and semantic error pools. Discussion focuses on the dynamic nature of semantic representations and the relation between semantic representation and naming during a period of slow mapping. The value of comparative picture naming/picture drawing as a new method for exploring children's semantic representations is emphasized.


Prospective study of apolipoprotein E genotype and functional outcome following ischemic stroke.
The apolipoprotein E (APOE) epsilon 4 allele is a marker of adverse outcome following head injury and intracerebral hemorrhage. Transgenic animal data in a focal cerebral ischemia model suggest that the epsilon 4 allele increases infarct size and functional impairment. To determine if APOE genotype is associated with functional recovery from ischemic stroke. Prospective study. Stroke service at a university teaching hospital. Patients with clinical and neuroimaging findings (computed tomography or magnetic resonance imaging) compatible with an acute ischemic stroke. Functional outcome by Barthel index (BI) and modified Rankin scale (mRS) was compared for epsilon 3/epsilon 3 patients vs epsilon 4 carriers and vs epsilon 2 carriers at 1 and 3 months. Univariate predictors of 3-month outcome were examined in a multivariate analysis. One hundred eighty nine patients were enrolled: 100 women, 89 men (mean +/- SD age, 69.4 +/- 11.0 years). There were 25 epsilon 2 alleles (frequency, 0.07), 292 epsilon 3 alleles (0.77), and 61 epsilon 4 alleles (0.16). Baseline National Institutes of Health Stroke Scale scores and Oxfordshire Community Stroke Project classifications were similar in all groups (epsilon 3/epsilon 3, epsilon 4, and epsilon 2 carriers). One-month (BI, P = .64; mRS, P = .59) and 3-month (BI, P = .87; mRS, P = .73) outcomes were not associated with possession of either epsilon 4 or the epsilon 2 allele. Baseline National Institutes of Health Stroke Scale scores (P < .001) and age (P = .002) were significant predictors of 3-month BI and mRS outcomes in multivariate analyses. Although there is a robust influence of APOE polymorphism on functional recovery after some types of brain injury in humans, it does not exert a major influence on injury severity or functional recovery following ischemic stroke. Arch Neurol. 2000;57:1480-1484


Overexpression of Grapevine VvIAA18 Gene Enhanced Salt Tolerance in Tobacco.
In plants, auxin/indoleacetic acid (Aux/IAA) proteins are transcriptional regulators that regulate developmental process and responses to phytohormones and stress treatments. However, the regulatory functions of the Vitis vinifera L. (grapevine) Aux/IAA transcription factor gene VvIAA18 have not been reported. In this study, the VvIAA18 gene was successfully cloned from grapevine. Subcellular localization analysis in onion epidermal cells indicated that VvIAA18 was localized to the nucleus. Expression analysis in yeast showed that the full length of VvIAA18 exhibited transcriptional activation. Salt tolerance in transgenic tobacco plants and Escherichia. coli was significantly enhanced by VvIAA18 overexpression. Real-time quantitative PCR analysis showed that overexpression of VvIAA18 up-regulated the salt stress-responsive genes, including pyrroline-5-carboxylate synthase (NtP5CS), late embryogenesis abundant protein (NtLEA5), superoxide dismutase (NtSOD), and peroxidase (NtPOD) genes, under salt stress. Enzymatic analyses found that the transgenic plants had higher SOD and POD activities under salt stress. Meanwhile, component analysis showed that the content of proline in transgenic plants increased significantly, while the content of hydrogen peroxide (H2O2) and malondialdehyde (MDA) decreased significantly. Based on the above results, the VvIAA18 gene is related to improving the salt tolerance of transgenic tobacco plants. The VvIAA18 gene has the potential to be applied to enhance plant tolerance to abiotic stress.


Graphene for the Building of Electroanalytical Enzyme-Based Biosensors. Application to the Inhibitory Detection of Emerging Pollutants.
Graphene and its derivates offer a wide range of possibilities in the electroanalysis field, mainly owing to their biocompatibility, low-cost, and easy tuning. This work reports the development of an enzymatic biosensor using reduced graphene oxide (RGO) as a key nanomaterial for the detection of contaminants of emerging concern (CECs). RGO was obtained from the electrochemical reduction of graphene oxide (GO), an intermediate previously synthesized in the laboratory by a wet chemistry top-down approach. The extensive characterization of this material was carried out to evaluate its proper inclusion in the biosensor arrangement. The results demonstrated the presence of GO or RGO and their correct integration on the sensor surface. The detection of CECs was carried out by modifying the graphene platform with a laccase enzyme, turning the sensor into a more selective and sensitive device. Laccase was linked covalently to RGO using the remaining carboxylic groups of the reduction step and the carbodiimide reaction. After the calibration and characterization of the biosensor versus catechol, a standard laccase substrate, EDTA and benzoic acid were detected satisfactorily as inhibiting agents of the enzyme catalysis obtaining inhibition constants for EDTA and benzoic acid of 25 and 17 mmol·L-1, respectively, and a maximum inhibition percentage of the 25% for the EDTA and 60% for the benzoic acid.


Integrins: structure and functions.
Integrins are cell surface transmembrane glycoproteins which perform receptor functions in cell interactions with the extracellular matrix and cell-cell contacts. Another function of integrins is transduction of signals which mediate the effects of the matrix on the physiological activity of cells (motility, proliferation, differentiation, etc.). Contemporary views on the molecular structure of integrins and their involvement in the organization of the cytoskeleton and control of gene activity are presented. Biochemical mechanisms, by which the signal functions of integrins are realized, are discussed. The data on the participation of integrins in oncogenic cell transformation, metastasis, and apoptosis are analyzed.


Human chorionic gonadotrophin: embryonic secretion is a time-dependent phenomenon.
Of 48 spare human pre-embryos achieving the expanded blastocyst stage, 22 (45.6%) secreted significant amounts of human chorionic gonadotrophin (HCG) (> 5 IU/l/day). Of these, nine remained intrazonal, seven partially hatched and six fully hatched. Embryonic production of HCG in vitro appeared to be time-dependent, starting after a certain minimum time (approximately 160 h post-insemination) and rising exponentially, with maximal HCG production around day 10. Hatching was not a prerequisite for HCG secretion, since similar amounts were produced by intrazonal blastocysts. Blastocysts derived from abnormally fertilized oocytes also began secreting HCG exponentially but secretion was delayed and the upper limit of maximum HCG secretion rate was comparatively low. The actual amount of HCG is thought to reflect the number of viable trophectoderm cells producing the hormone. HCG doubling times for blastocysts in vitro were rapid when compared to implanting blastocysts of a similar age in vivo, with 19/22 (86.4%) blastocysts having a doubling time of < 10 h. Provided a pre-embryo can secrete HCG and maintain an adequate doubling time, sufficient HCG should be produced for initial stages of embryonic recognition in vivo. Since intrazonal blastocysts are capable of fulfilling both of these criteria, the limiting factor in realizing their full potential may be escaping from the zona pellucida.


Proteome- and microarray-based expression analysis of lymphoma cell lines identifies a p53-centered cluster of differentially expressed proteins in mantle cell and follicular lymphoma.
We used a standardized electrophoresis protocol to identify differentially expressed proteins based on a sample pooling approach comparing three follicular lymphoma and three mantle cell lymphoma-derived cell lines. One hundred and seventy-five consistently differentially expressed proteins were identified out of more than 1600 protein spots per gel. Of these 175 protein spots, 38 of the 41 most highly expressed proteins were identified by MS analysis (MALDI-TOF), involving different cellular programs such as DNA repair (Rad50), cell cycle control (Mad1L1), transcription (SAFB), and apoptosis (Luca-15 protein). Expression data were confirmed by Western blot analysis of identified proteins and 2-D gel hybridization of proteins with known overexpression (G1/S-specific cyclin-D1, apoptosis regulator Bcl-2). Comparison of proteome analysis to RNA expression array data revealed only a modest correlation of RNA and protein level emphasizing the relevance of post-translational regulation in lymphomagenesis (p = 0.36). Most interestingly, additional data bank search identified 13 out of 17 referenced proteins (76%) as members of a TP53-dependent network of cell regulation.


The acquisition of arithmetic in normal children: assessment by a cognitive model of dyscalculia.
Developmental dyscalculia (DC) is a learning disability affecting the acquisition of arithmetic skills. The authors studied the normal developmental of arithmetic function of 200 normal children in grades 3 to 6. Number comprehension and production were well established by grade 3. With age and experience, children became more adept at addition, subtraction, multiplication and division for both number facts and complex exercises. The scores did not display a Gaussian distribution, but were negatively skewed. Low scores on the battery correlated well with the teacher's rating of the child's arithmetic knowledge, but not with reading ability. An estimate of the fifth centile was calculated. On the basis of these results, the authors conclude that this battery assesses number knowledge. Use of the fifth centile as a cut-off point will aid in identification of children with DC.


Efficacy of Bacteroides measurements for reducing the statistical uncertainty associated with hydrologic flow and fecal loads in a mixed use watershed.
This paper presents an analysis of the occurrence and uncertainty of source-specific Bacteroides and Escherichia coli in a stream in a mixed land-use watershed with human, cattle, and wildlife fecal inputs located in a karstic geologic region during baseflow conditions. The objectives of the study were to evaluate the occurrence, hydrologic significance, and source of fecal mass in the stream using assays for total Bacteroides (AllBac) and bovine-specific Bacteroides (BoBac), and then to compare these measurements with E. coli densities and loads. Samples were collected during baseflow conditions over several months at seven different main channel sites in the Stock Creek watershed, a 49.3 km2 basin located in Knoxville, TN (USA). We determined instantaneous loads for total fecal loads, bovine fecal loads, and E. coli from measured flow rates and the representative Bacteroides fecal masses and/or E. coli densities. The study indicated a strong correlation between total fecal load (kg d(-1)), bovine fecal load (kg d(-1)), E. coli load rate (CFU d(-1)), 7-d antecedent precipitation, and turbidity. The various datasets were used to establish parameter correlations and spatial dependencies throughout the watershed. The data analysis demonstrated two prevalent patterns throughout the watershed: (i) a runoff-dominated transport and occurrence; and (ii) potential groundwater-dominated transport and occurrence.


Reversing defaunation: restoring species in a changing world.
The rate of biodiversity loss is not slowing despite global commitments, and the depletion of animal species can reduce the stability of ecological communities. Despite this continued loss, some substantial progress in reversing defaunation is being achieved through the intentional movement of animals to restore populations. We review the full spectrum of conservation translocations, from reinforcement and reintroduction to controversial conservation introductions that seek to restore populations outside their indigenous range or to introduce ecological replacements for extinct forms. We place the popular, but misunderstood, concept of rewilding within this framework and consider the future role of new technical developments such as de-extinction.


Striatal and hippocampal involvement in motor sequence chunking depends on the learning strategy.
Motor sequences can be learned using an incremental approach by starting with a few elements and then adding more as training evolves (e.g., learning a piano piece); conversely, one can use a global approach and practice the whole sequence in every training session (e.g., shifting gears in an automobile). Yet, the neural correlates associated with such learning strategies in motor sequence learning remain largely unexplored to date. Here we used functional magnetic resonance imaging to measure the cerebral activity of individuals executing the same 8-element sequence after they completed a 4-days training regimen (2 sessions each day) following either a global or incremental strategy. A network comprised of striatal and fronto-parietal regions was engaged significantly regardless of the learning strategy, whereas the global training regimen led to additional cerebellar and temporal lobe recruitment. Analysis of chunking/grouping of sequence elements revealed a common prefrontal network in both conditions during the chunk initiation phase, whereas execution of chunk cores led to higher mediotemporal activity (involving the hippocampus) after global than incremental training. The novelty of our results relate to the recruitment of mediotemporal regions conditional of the learning strategy. Thus, the present findings may have clinical implications suggesting that the ability of patients with lesions to the medial temporal lobe to learn and consolidate new motor sequences may benefit from using an incremental strategy.


Changes in Australian prescription opioid use following codeine rescheduling: A retrospective study using pharmaceutical benefits data.
In February 2018, Australia up-scheduled the 'weak' opioid codeine to a prescription only medication. This study aimed to analyse the change in prescribing trends for codeine and other commonly prescribed opioids in Australia following this policy change to determine if removal of over-the-counter codeine resulted in an increase in opioid prescribing. Data was obtained through the Australian Government Department of Human Services statistics website, and contained monthly data about subsidised national prescription numbers for codeine, oxycodone, oxycodone-naloxone, tapentadol, tramadol, morphine, and fentanyl, from January 2016 to December 2018. Segmented linear regression accounting for autocorrelation was used to assess the effect of codeine rescheduling on the supply trends of these opioids. Rescheduling codeine to remove over-the-counter (non-prescription) supply does not appear to have had an immediate effect on the prescription rates of codeine, and there is no significant change in these rates in the months following. Analysis of data showed decreasing trends for codeine and most other schedule 8 prescription opioids, with no increase in any prescribed opioids associated with codeine up scheduling. Despite concerns, substitution of over-the-counter codeine with higher strength prescribed codeine has not been observed at a population level, nor has a shift to other prescribed opioids occurred. Overall, opioid prescribing in Australia has been decreasing since 2016, both for strong and weak opioids.


Impaired skeletal muscle function in patients with congestive heart failure. Relationship to systemic exercise performance.
In patients with congestive heart failure (CHF), the poor relationship between systemic exercise performance and cardiac function, together with morphologic and metabolic abnormalities in skeletal muscle, raises the possibility that skeletal muscle function may be impaired and limit systemic exercise performance. We assessed strength and endurance of the knee extensors during static and dynamic exercise in 16 patients with Class I-IV CHF and eight age-matched sedentary controls and related these measurements to systemic exercise performance. To assess skeletal muscle function independent of peripheral blood flow, endurance was repeated under ischemic conditions. Strength was not significantly different in the two groups. Dynamic endurance, quantified as the decline in peak torque during 15 successive isokinetic knee extensions, was significantly reduced in the patients compared to controls during aerobic (peak torque 65 vs. 86% of initial for exercise at 90 deg/s and 60 vs. 85% for exercise at 180 deg/s; P less than 0.002 for both), and during ischemic exercise (56 vs. 76% of initial torque; P less than 0.01). Static endurance, defined as the time required for force during a sustained maximal voluntary contraction to decline to 60% of maximal, was reduced in the patients compared to controls (40 +/- 14 vs. 77 +/- 29 s; P less than 0.02). There were highly significant relationships between systemic exercise performance and skeletal muscle endurance at 90 and 180 deg/s in the patients with CHF (r = 0.90 and 0.66, respectively). These findings indicate that skeletal muscle endurance is impaired in patients with CHF, that this abnormality is in part independent of limb blood flow, and that these changes may be important determinants of systemic exercise performance.


Fracture of a sirolimus-eluting stent in renal artery stenosis.
Atherosclerosis accounts for most adult cases of renal artery stenoses (RAS). Revascularization of atherosclerotic RAS is beneficial in terms of improved blood pressure control, preservation of renal function and overall decrease of cardiovascular risk, in particular in mononephric patients. Endovascular stenting has been proven superior to percutaneous transluminal angioplasty (PTA) alone in terms of initial success and restenosis rates in atherosclerotic RAS. In this case report revascularization of atherosclerotic RAS in a mononephric patient is presented and the long-term follow-up and complications of PTA and stenting are illustrated. Our case is the first report of fracture of a sirolimus-eluting stent overlapped to a previously implanted Palmaz-Schatz in the left renal artery. The mechanisms and possible remedies of this complication are discussed.


Toward a Next-Generation Digital Chest Tube.
Chest tubes in patients who have undergone pulmonary resection with pleural air leak are painful, impair ventilatory mechanics, and increase hospital length of stay and costs. Despite these well-documented concerns, current protocols for chest tube management in this setting are not well supported by evidence. Excessive suction applied to chest tubes has been associated with prolonged air leak due to alveolar over-distension, and most practitioners intuit that suction should be minimized to the lowest level needed to maintain desired pleural apposition. Unfortunately, there is no evidence-based protocol for the establishment of minimal adequate suction. Digital suction devices in current clinical use can identify air leak resolution preventing the delay of chest tube removal but cannot guide suction minimization while an air leak persists. We recently described a monitor of lung expansion in a porcine model of pleural air leak that could detect loss of pleural apposition continuously in real-time based on electrical impedance readings obtained directly from the surface of the lung via chest tube-embedded electrodes. The value of the impedance signal was "in-range" when pleural apposition was present but became abruptly "out-of-range" when pneumothorax due to inadequate suction developed. These findings suggested that a digitally controlled suction pump system could be programmed to recognize the development of pneumothorax and automatically identify and set the minimum level of suction required to maintain pleural apposition. We present here preliminary proof of concept for this system.


Helicobacter pylori infection and diabetes mellitus: the 2013 state of art.
It is well-known the role of Helicobacter pylori (H. pylori) infection in the development of gastroduodenal diseases. From two decades literature has suggested the potential relationship of the bacterium with extragastric manifestations. Aim of the present review was to analyze the consistency of a potential involvement of H. pylori infection in the pathogenesis of diabetes mellitus (DM) as well as in the gastric abnormalities associated with this disease. Several studies reported a higher prevalence of H. pylori infection in diabetic patients with or without dyspeptic symptoms than in controls and a positive association with insulin resistance (IR) has been shown. However, DM has a multifactorial pathogenesis and the detection of the role of each agent is difficult. The different factors implicated in the development of DM as well as of IR include inflammation, autoimmunuty, stimulation of innate immune system, trigger to platelet activation and platelet-leukocyte aggregation, action on leptin and ghrelin regulation, alterated lipid metabolism and insulin sensitivity. Effectiveness of H. pylori eradication results significantly lower in diabetic patients than in controls, most likely because of the large use of antibiotic in DM subjects, causing selection of resistant H. pylori strains. Finally, re-infection after bacterial eradication, although rarely observed in the general population, seems to be more frequent in diabetic patients than in controls.


The European List of Essential Medicines for Medical Education: a modified Delphi study.
Rational prescribing is essential for the quality of health care. However, many final-year medical students and junior doctors lack prescribing competence to perform this task. The availability of a list of medicines that a junior doctor working in Europe should be able to independently prescribe safely and effectively without supervision could support and harmonize teaching and training in clinical pharmacology and therapeutics (CPT) in Europe. Therefore, our aim was to achieve consensus on such a list of medicines that are widely accessible in Europe. For this, we used a modified Delphi study method consisting of three parts. In part one, we created an initial list based on a literature search. In part two, a group of 64 coordinators in CPT education, selected via the Network of Teachers in Pharmacotherapy of the European Association for Clinical Pharmacology and Therapeutics, evaluated the accessibility of each medicine in his or her country, and provided a diverse group of experts willing to participate in the Delphi part. In part three, 463 experts from 24 European countries were invited to participate in a two-round Delphi study. In total, 187 experts (40%) from 24 countries completed both rounds and evaluated 416 medicines, 98 of which were included in the final list. The top three Anatomical Therapeutic Chemical code groups were (1) cardiovascular system (n = 23), (2) anti-infective (n = 21), and (3) musculo-skeletal system (n = 11) This European List of Essential Medicines for Medical Education could be a starting point for country-specific lists and could be used for the training and assessment of CPT.


Daily interferon therapy for hepatitis C virus infection in liver transplant recipients.
Hepatitis C virus infection persists after liver transplantation and causes recurrent liver injury in the majority of patients. Standard dose interferon therapy has been largely unsuccessful for hepatitis C in transplant recipients. Twelve patients, at least 7 months posttransplant, with detectable hepatitis C virus RNA in serum and features of hepatitis C on liver biopsy were randomized to interferon-alpha2a, 3 mU daily for 12 months (n=8) or no treatment (n=4). The tolerability of daily interferon dosing in liver transplant recipients was evaluated and effects on hepatitis C virus RNA level, quasispecies evolution, and liver histology were studied. Treated patients had an improvement in histological activity index at the end of therapy relative to controls (median reduction of 2 versus median increase of 1.5) (P=0.04). Four treated patients had a virological response (all bDNA negative, one qualitative polymerase chain reaction negative) compared with none of the untreated patients. Only two of six treated patients tested had evidence of quasispecies diversification on therapy. Seven of eight patients in the treatment group required dose reduction for fatigue and/or depression. They tolerated 1.5 mU of interferon-alpha2a daily. Two treated patients developed graft dysfunction, one of who had histological evidence of rejection and subsequent graft loss. Low daily doses of interferon were tolerated by liver transplant recipients and provided histological benefit without associated quasispecies diversification in most cases. These findings provide a rationale to study low dose daily or pegylated interferon maintenance therapy for the management of hepatitis C posttransplant.


Comparison of glyburide and insulin in women with gestational diabetes mellitus and associated perinatal outcome: a randomized clinical trial.
Insulin is currently the drug of choice in treating patients with gestational diabetes mellitus but insulin is expensive, inconvenient to store and use and probably associated with more risks of asymptomatic hypoglycemia in comparison with some oral agents. This randomized clinical trial was conducted to evaluate the efficacy and safety of glyburide in patients with gestational diabetes mellitus in comparison with insulin therapy. Pregnant women aged between 18-45 years with singleton pregnancies and in their 24-36 weeks of gestation were assessed for eligibility. Women with gestational diabetes mellitus were randomly allocated to two insulin and glyburide groups and compared with maternal and neonatal outcome. Ninety-six women with gestational diabetes mellitus enrolled in the study. At screen and treated fasting and post-prandial blood glucose levels were similar in both groups. Time for beginning the treatment to control the glycemic index was 28.30 (±20.60) days in the insulin group and 22.56 (±18.86) in the glyburide group. There was no statistically significant difference in time-to-control the blood glucose level in two studied group. Time, between beginning the treatment of GDM and delivery, was 53.22 (±28.96) days in the insulin group and 56.67 (±30.47) in the glyburide group. There was no statistically significant difference between the times of treatment-to-delivery in two studied groups. There were no statistically significant differences between maternal and neonatal outcomes in two studied groups. Glyburide can effectively and safely control the glycemic index in women with gestational diabetes mellitus in comparison with insulin.


Search for the smallest random forest.
Random forests have emerged as one of the most commonly used nonparametric statistical methods in many scientific areas, particularly in analysis of high throughput genomic data. A general practice in using random forests is to generate a sufficiently large number of trees, although it is subjective as to how large is sufficient. Furthermore, random forests are viewed as "black-box" because of its sheer size. In this work, we address a fundamental issue in the use of random forests: how large does a random forest have to be? To this end, we propose a specific method to find a sub-forest (e.g., in a single digit number of trees) that can achieve the prediction accuracy of a large random forest (in the order of thousands of trees). We tested it on extensive simulation studies and a real study on prognosis of breast cancer. The results show that such sub-forests usually exist and most of them are very small, suggesting they are actually the "representatives" of the whole random forests. We conclude that the sub-forests are indeed the core of a random forest. Thus it is not necessary to use the whole forest for satisfying prediction performance. Also, by reducing the size of a random forest to a manageable size, the random forest is no longer a black-box.


A double blind randomized placebo controlled phase I/II study assessing the safety and efficacy of allogeneic bone marrow derived mesenchymal stem cell in critical limb ischemia.
Peripheral vascular disease of the lower extremities comprises a clinical spectrum that extends from no symptoms to presentation with critical limb ischemia (CLI). Bone marrow derived Mesenchymal Stem Cells (BM- MSCs) may ameliorate the consequences of CLI due to their combinatorial potential for inducing angiogenesis and immunomodulatory environment in situ. The primary objective was to determine the safety of BM- MSCs in patients with CLI. Prospective, double blind randomized placebo controlled multi-center study was conducted in patients with established CLI as per Rutherford classification in category II-4, III-5, or III-6 with infra-inguinal arterial occlusive disease and were not suitable for or had failed revascularization treatment. The primary end point was incidence of treatment - related adverse events (AE). Exploratory efficacy end points were improvement in rest pain, increase in Ankle Brachial Pressure Index (ABPI), ankle pressure, healing of ulcers, and amputation rates. Twenty patients (BM-MSC: Placebo = 1:1) were administered with allogeneic BM-MSCs at a dose of 2 million cells/kg or placebo (PlasmaLyte A) at the gastrocnemius muscle of the ischemic limb. Improvement was observed in the rest pain scores in both the arms. Significant increase in ABPI and ankle pressure was seen in BM-MSC arm compared to the placebo group. Incidence of AEs in the BM-MSC arm was 13 vs. 45 in the placebo arm where as serious adverse events (SAE) were similar in both the arms (5 in BM-MSC and 4 in the placebo group). SAEs resulted in death, infected gangrene, amputations in these patients. It was observed that the SAEs were related to disease progression and not related to stem cells. BM-MSCs are safe when injected IM at a dose of 2 million cells/kg body weight. Few efficacy parameters such as ABPI and ankle pressure showed positive trend warranting further studies. NIH website (http://www.clinicaltrials.gov/ct2/show/NCT00883870).


Return to Play After an Anterior Cruciate Ligament Injury: Prioritizing Neurological and Psychological Factors of the Decision-Making Algorithm.
Soccer players recovering from anterior cruciate ligament (ACL) injuries have better options for treatment today than they did 25 years ago. Surgical techniques have improved, and rehabilitation protocols have evolved considerably. Although the rehabilitation community is doing a better job of treating this patient population, the evidence does demonstrate that both re-injury and return- to-play (RTP) rates are still suboptimal. Most protocols focus on normalizing strength and range of motion (ROM) and achieving limb symmetry with soccer-specific movements. While these factors are certainly prerequisites for returning to the field, their inclusion does not provide a complete picture of the athlete's presentation. An additional factor that should be prioritized with this patient population is the central nervous system (CNS). Advanced imaging has shown that peripheral deafferentation does occur with musculoskeletal injuries; this ultimately results in cortical reorganization, which makes movement planning more difficult for the player, since simpler tasks must now be processed at higher levels in the CNS. The evidence also shows that the CNS demonstrates plasticity in these cases, so that through focused neuromotor rehabilitation techniques, it is possible to bring movement planning back down to a sub-cortical level. Cognitive issues may also be a factor in preventing the player from returning. Fear of re-injury and diminished confidence can influence the way the player moves on the field, and diminish ability to demonstrate protective kinematics with all soccer-specific tasks. We believe that an approach incorporating traditional musculoskeletal rehabilitation, CNS neuro-motor training, and consideration for cognitive factors, may define an improved paradigm for treating the soccer player and assessing readiness for RTP following ACL injury.


Trastuzumab Deruxtecan in Patients With HER2-Mutant Metastatic Non-Small-Cell Lung Cancer: Primary Results From the Randomized, Phase II DESTINY-Lung02 Trial.
Trastuzumab deruxtecan (T-DXd) 5.4 and 6.4 mg/kg showed robust antitumor activity in multiple cancer indications; however, T-DXd 5.4 mg/kg has not been evaluated in patients with previously treated human epidermal growth factor receptor 2-mutant (HER2m; defined as single-nucleotide variants and exon 20 insertions) metastatic non-small-cell lung cancer (mNSCLC). DESTINY-Lung02, a blinded, multicenter, phase II study, investigated T-DXd 5.4 mg/kg once every 3 weeks for the first time in previously treated (platinum-containing therapy) patients with HER2m mNSCLC and further assessed T-DXd 6.4 mg/kg once every 3 weeks in this population. The primary end point was confirmed objective response rate (ORR) per RECIST v1.1 by blinded independent central review. One hundred fifty-two patients were randomly assigned 2:1 to T-DXd 5.4 or 6.4 mg/kg once every 3 weeks. As of December 23, 2022, the median duration of follow-up was 11.5 months (range, 1.1-20.6) with 5.4 mg/kg and 11.8 months (range, 0.6-21.0) with 6.4 mg/kg. Confirmed ORR was 49.0% (95% CI, 39.0 to 59.1) and 56.0% (95% CI, 41.3 to 70.0) and median duration of response was 16.8 months (95% CI, 6.4 to not estimable [NE]) and NE (95% CI, 8.3 to NE) with 5.4 and 6.4 mg/kg, respectively. Median treatment duration was 7.7 months (range, 0.7-20.8) with 5.4 mg/kg and 8.3 months (range, 0.7-20.3) with 6.4 mg/kg. Grade ≥ 3 drug-related treatment-emergent adverse events occurred in 39 of 101 (38.6%) and 29 of 50 (58.0%) patients with 5.4 and 6.4 mg/kg, respectively. 13 of 101 (12.9%) and 14 of 50 (28.0%) patients had adjudicated drug-related interstitial lung disease (2.0% grade ≥ 3 in each arm) with 5.4 and 6.4 mg/kg, respectively. T-DXd demonstrated clinically meaningful responses at both doses. Safety profile was acceptable and generally manageable, favoring T-DXd 5.4 mg/kg.


Role of vesicle-associated syntaxin 5 in the assembly of pre-Golgi intermediates.
Syntaxins are thought to function during vesicular transport as receptors on the target membrane and to contribute to the specificity of membrane docking and fusion by interacting with vesicle-associated receptors. Here, syntaxin 5 (Syn5) was shown to be an integral component of endoplasmic reticulum-derived transport vesicles. This pool, but not the target, Golgi-associated Syn5 pool, was essential for the assembly of vesicular-tubular pre-Golgi intermediates and the delivery of cargo to the Golgi. The requirement for vesicle-associated Syn5 in transport suggests a reevaluation of the basis for operation of the early secretory pathway.


Structural, biochemical and immunochemical characterization of an acidic phospholipase A2 from Lachesis acrochorda (Viperidae: Crotalinae) venom.
Viperids of the genus Lachesis, also known as bushmasters, are capable of injecting great amounts of venom that cause severe envenomation incidents. Since phospholipases type A2 are mainly involved in edema and myonecrosis within the snakebite sites, in this work, the isolation, amino acid sequence and biochemical characterization of the first phospholipase type A2 from the venom of Lachesis acrochorda, named Lacro_PLA2, is described. Lacro_PLA2 is an acidic aspartic 49 calcium-dependent phospholipase A2 with 93% similarity to the L. stenophrys phospholipase. Lacro_PLA2 has a molecular mass of 13,969.7 Da and an experimental isoelectric point around 5.3. A combination of N-terminal Edman degradation and MS/MS spectrometry analyses revealed that Lacro_PLA2 contains 122 residues including 14 cysteines that form 7 disulfide bridges. A predicted 3D model shows a high resemblance to other viperid phospholipases. Nevertheless, immunochemical and phospholipase neutralization tests revealed a notorious level of immunorecognition of the isolated protein by two polyclonal antibodies from viperids from different genus, which suggest that Lacro_PLA2 resembles more to bothropic phospholipases. Lacro_PLA2 also showed significantly high edema activity when was injected into mice; so, it could be an alternative antigen in the development of antibodies against toxins of this group of viperids, seeking to improve commercial polyclonal antivenoms.


Plane warts under spontaneous regression. Immunopathologic study on cellular constituents leading to the inflammatory reaction.
Immunohistologically, cellular infiltrates in regressing plane warts were mainly composed of lymphocytes and mononuclear phagocytes. There were many infiltrating T lymphocytes. Immunoelectron microscopic observation demonstrated that both helper/inducer and suppressor/cytotoxic phenotypes of T lymphocytes infiltrated in the lesions. OKT6-positive cells were observed in the dermis as well as in the epidermis. Moreover, as noted in allergic contact dermatitis, the apposition of T lymphocytes to Langerhans' cell-like cells could be seen. Lymphocytes and a small number of mononuclear phagocytes were found adjacent to damaged keratinocytes in the epidermis, the picture of which has been described as satellite cell necrosis, a hallmark of cytotoxic reaction by aggressors. These findings suggest that specific cell-mediated immunity against virus-infected keratinocytes takes place in the process of regressing plane warts.


[Delayed symptoms of hypoxic brain damage after temporary improvement].
A shipyard worker was doing welding work inside a pipe with argon as cover gas. After taking a break he fainted while controlling the weld, also inside the pipe. He was rescued after 15-20 minutes and regained consciousness after a few hours. Two days later he was discharged from hospital in apparently good health. After a week he returned to the hospital suffering from confusion, failing memory, aphasia, apraxia and urinary incontinence. MR showed elevated signal intensity bilaterally in the caudate nucleus. He improved gradually and six months later was given less demanding work. The course of his illness is consistent with delayed neuropsychiatric sequelae after hypoxia, probably due to displacement of oxygen by argon. Delayed symptoms are caused by demyelination in the subcortical white matter.


SNRNP200 Mutations Cause Autosomal Dominant Retinitis Pigmentosa.
The small nuclear ribonucleoprotein 200 kDa (SNRNP200) gene plays a key role in the maturation of pre-message RNA (pre-mRNA) splicing with the indication for the etiology of retinitis pigmentosa (RP). Gene recognition can facilitate the diagnosis of these patients for better clinical management, treatment and counseling. This study aimed to outline the causative mutation in a Chinese family and the pathogenic mechanism of this SNRNP200 mutation in RP. Eighteen individuals from the affected family underwent a complete ophthalmic examination. Whole exome sequencing (WES) was conducted to identify the pathogenic variant in the proband, which was then confirmed by Sanger sequencing. Expression of the SNRNP200 transcript in zebrafish was identified via whole mount in situ hybridization. Morpholino oligonucleotide (MO) and SNRNP200 wild and mutant mRNA were injected into zebrafish embryos followed by analyses of the systemic changes and retinal phenotypes using immunofluorescence. Heterozygous SNRNP200c.C6088T (p.Arg2030Cys) mutation was ascertained in two members of this family: the proband and his father (II-2). Overexpression of SNRNP200Arg2030Cys, but not SNRNP200WT caused systemic deformities in the wild-type zebrafish embryos with the retina primarily injured, and significantly increased death rates in the morphant embryos, in which the orthologous zebrafish SNRNP200 gene was blocked. In conclusion, this study reports a novel heterozygous SNRNP200c.C6088T mutation, which is evidenced to cause RP via a dominant-negative effect.


[THE NEED FOR HELP OF FAMILY CAREGIVERS OF PERSONS WITH MENTAL ILLNESS IN A UNIQUE SERVICE FOR FAMILIES AT THE BEER SHEVA MENTAL HEALTH CENTER].
Limited research attention has been given to the needs of family caregivers of persons with mental illness in psychiatric hospitals despite the stressors and difficulties they experience. In light of the recognition of the significance of helping family caregivers, a new model of consultation and support centers for family caregivers, called Meital, has been developed. To examine the needs of family caregivers who receive help in Meital, at the Beer Sheva Mental Health Center. Eighty-five family caregivers participated in the research. They completed a structured questionnaire constructed for this research two weeks after they started receiving services from Meital. The questionnaire included four areas of needs for help. These areas examined the extent of the need for help with respect to each of the items in the instrument. The mean of the extent of need for help of the items in the 'information and knowledge' subscale was the highest. Average to high means of the items of the subscales were found in the subscales relating to 'difficulties stemming from the impact of the situation of the person with mental illness on the function of the family caregiver receiving help,' 'on the function of other family members' and 'difficulties coping with the person with mental illness.' The mean of the items of the subscale 'relationships with professionals and informal systems' was the lowest. An examination of the items within the subscales indicated that items relating to the 'impact of the situation of the person with mental illness on the family caregiver who receives help' were ranked higher than the items relating to the 'impact on the function of other family caregivers.' Items relating to 'relationships with professionals' were ranked higher than items relating to 'relationships with informal systems.' This research emphasizes the importance of implementing the family-centered approach, the basis of the Meital Model, in psychiatric institutions. The focus of this approach is on the need for help of family caregivers beyond the help needed for them to function as a resource of help for the ill person. The findings also illuminate the importance of making information and knowledge accessible for family caregivers.


Renin and renin blockade have no role in complement activity.
Renin, an aspartate protease, regulates the renin-angiotensin system by cleaving its only known substrate angiotensinogen to angiotensin. Recent studies have suggested that renin may also cleave complement component C3 to activate complement or contribute to its dysregulation. Typically, C3 is cleaved by C3 convertase, a serine protease that uses the hydroxyl group of a serine residue as a nucleophile. Here, we provide seven lines of evidence to show that renin does not cleave C3. First, there is no association between renin plasma levels and C3 levels in patients with C3 Glomerulopathies (C3G) and atypical Hemolytic Uremic Syndrome (aHUS), implying that serum C3 consumption is not increased in the presence of high renin. Second, in vitro tests of C3 conversion to C3b do not detect differences when sera from patients with high renin levels are compared to sera from patients with normal/low renin levels. Third, aliskiren, a renin inhibitor, does not block abnormal complement activity introduced by nephritic factors in the fluid phase. Fourth, aliskiren does not block dysregulated complement activity on cell surfaces. Fifth, recombinant renin from different sources does not cleave C3 even after 24 hours of incubation at 37 °C. Sixth, direct spiking of recombinant renin into sera samples of patients with C3G and aHUS does not enhance complement activity in either the fluid phase or on cell surfaces. And seventh, molecular modeling and docking place C3 in the active site of renin in a position that is not consistent with a productive ground state complex for catalytic hydrolysis. Thus, our study does not support a role for renin in the activation of complement.


Space Flight-Associated Neuro-ocular Syndrome.
New and unique physiologic and pathologic systemic and neuro-ocular responses have been documented in astronauts during and after long-duration space flight. Although the precise cause remains unknown, space flight-associated neuro-ocular syndrome (SANS) has been adopted as an appropriate descriptive term. The Space Medicine Operations Division of the US National Aeronautics and Space Administration (NASA) has documented the variable occurrence of SANS in astronauts returning from long-duration space flight on the International Space Station. These clinical findings have included unilateral and bilateral optic disc edema, globe flattening, choroidal and retinal folds, hyperopic refractive error shifts, and nerve fiber layer infarcts. The clinical findings of SANS have been correlated with structural changes on intraorbital and intracranial magnetic resonance imaging and in-flight and terrestrial ultrasonographic studies and ocular optical coherence tomography. Further study of SANS is ongoing for consideration of future manned missions to space, including a return trip to the moon or Mars.


[Lingual status and mandibular morphology].
Is the balance of the hyo-lingual musculature very important about mandibular morphology and orientation of mandibular growth? The purpose of this study is to detect the existence of possible reports between hyo-lingual position and mandibular morphogenesis. The selected population is made of fifty children from eight and a half to ten and a half years old, who are thumb sucking and mouth breathing. The means of study are norma lateralis teleradiographs made when facial musculature is at rest position. The statistical analysis implies a descriptive and correlative analysis of 82 variables, and a comparative study with an other sample. In spite of anatomic bonds, it seems to be a relative independence between lingual morphology and orientation of mandibular growth. Perhaps head posture is it a more important mechanism!


Arteriohepatic dysplasia. II. Hepatobiliary morphology.
Five children were noted to have arteriohepatic dysplasia (Alagille's syndrome) between 3 and 7 months of age. Prior to diagnosis, four underwent Kasai procedures after intraoperative cholangiograms failed to demonstrate patency of the extrahepatic bile ducts. In three patients, a focal proximal hypoplasia of the common hepatic duct was demonstrated with fibrosis and increased vascularity. Hypoplasia of the gallbladder occurred in two patients. Changes were observed in the porta hepatis. Eighty of 208 micrometers bile ducts were associated with peripherally located gland-like structures. These changes are indistinguishable from those in fibrous remnants of extrahepatic biliary atresia. Hepatic features of sequential liver biopsies obtained in the five patients were divided into early and late changes. From birth to 3 months of age, the pathology consisted of cholestasis and bile duct destruction. After 3 months of age, there was persistent cholestasis, paucity of interlobular bile ducts, and portal fibrosis. Ductular proliferation was not an intrinsic change. When present, it was related to a recent episode of cholangitis.


Pulmonary artery dilatation: an overlooked mechanism for angina pectoris.
Dilatation of the pulmonary artery may lead to the compression of adjacent structures. Of those, the extrinsic compression of the left main coronary artery is the most worrisome. We present the case of a 48-year-old woman who was diagnosed with pulmonary artery dilatation due to severe, thromboembolic pulmonary hypertension. She also had angina and coronary angiography revealed a 70% ostial stenosis of the left main coronary artery. The presence of this isolated lesion in a young woman without risk factors for atherosclerosis suggests extrinsic compression of the left main coronary artery by the dilated pulmonary artery as the likely mechanism. The patient underwent direct stenting of the left main coronary stenosis with a good result.


Elevated seawater pCO2 affects reproduction and embryonic development in the pygmy squid, Idiosepius pygmaeus.
The oceans are absorbing additional carbon dioxide (CO2) from the atmosphere and projected future CO2 levels and ocean acidification could have negative implications for many marine organisms, especially during early life stages. Cephalopods are ecologically important in marine ecosystems, yet the potential effects of increased partial pressure of CO2 (pCO2) in seawater on cephalopod reproduction and embryonic development are little studied. We allowed adult two-toned pygmy squid (Idiosepius pygmaeus) to breed in ambient control (~445 μatm; ~8.05 pHT) or elevated pCO2 conditions (~940 μatm; ~7.78 pHT) and compared reproductive traits in adults and developmental characteristics of their eggs, which remained in control or elevated pCO2 treatments until hatching. Breeding pairs at elevated pCO2 produced clutches with 40% fewer eggs, vitelli that were 14% smaller directly after spawning, embryos that were 5% smaller upon hatching, and eggs with an 8% increase in late-stage egg swelling compared with pairs at control conditions. Elevated pCO2 did not affect fertility, time to hatch, or hatching success. Eggs were laid 40% closer together in elevated pCO2 compared with control conditions, indicating a possible effect of elevated pCO2 on reproductive behaviour. These results show that elevated pCO2 can adversely affect reproduction and embryonic development of the two-toned pygmy squid. As the potential for adaptation is influenced by reproductive success, testing the capacity for squid to adapt to future ocean conditions should be a priority for future research.


Family and School Relationship during COVID-19 Pandemic: A Systematic Review.
Education systems worldwide have been affected by a sudden interruption in classroom learning because the coronavirus pandemic forced both the closure of all schools in March 2020 and the beginning of distance learning from home, thus compelling families, schools, and students to work together in a more coordinated fashion. The present systematic review was carried out following PRISMA guidelines. The main objective was to present critical information on the relationship between the family and the school in the face of the imposed distance learning scenario caused by COVID-19. A total of 25 articles dealing with the relationships established during the pandemic of any of the three agents involved (family, students, and school) were analysed. The results showed that the relationships between the three groups involved must be improved to some extent to meet the needs that have arisen as a result of distance learning. In conclusion, the educational scenario during the pandemic has been one of the most significant challenges experienced in the recent history of education.


Reduced connexin40 protein expression in the right atrial appendage of patients bearing the minor connexin40 allele (-44 G --> A).
The occurrence of connexin40 (Cx40) minor polymorphism (-44 G → A) was increased in patients with idiopathic atrial fibrillation (AF), although its effect on atrial Cx40 protein expression is unknown. We aimed to evaluate whether alterations in Cx40 are directly linked to the development of AF, we studied the effect of this polymorphism on Cx40 expression and distribution in patients without any history of AF and in patients who developed post-operative AF. Hundred and eight patients (mean age 67 ± 9 years), without a history of AF or conditions that predispose to AF, were included. During heart surgery, 10 cc blood was collected for DNA genotyping and the right atrial appendage was partly excised. Ten patients (9%) were homozygous for the minor allele (AA, Group 1), 30 (28%) were heterozygous (AG, Group 2), and 68 (63%) were non-carriers (GG, Group 3). Ten age- and sex-matched tissue samples per group were analysed for Cx40 expression by: (i) real-time quantitative polymerase chain reaction (Q-PCR), (ii) western blotting, and (iii) immunohistochemistry on cryosections. Real-time quantitative polymerase chain reaction showed no significant differences of Cx40 mRNA among the groups. Western blot analysis, however, revealed a reduction in Cx40 protein in Groups 1 (-36.4%) and 2 (-39.5%) as compared with Group 3. Immunohistochemistry confirmed this reduction but indicated an unaltered subcellular distribution of the remaining Cx40. Incidence of post-operative AF (28%) was age-dependent but unrelated to the presence of the polymorphism or fibrosis. Presence of the Cx40 minor allele (-44 G → A) results in a uniform down-regulation of right atrial appendage Cx40 protein which was not significantly related to development of post-operative AF.


Consistency in decision-making between survey teams and the decision-making body in a professional education program accrediting agency.
This article was migrated. The article was marked as recommended. Credibility of an education program accreditor is dependent on consistency in decision-making among program reviews. The use of peer-review accreditation survey teams is a potential source of inconsistency in the review of individual programs, especially so when the agency employs non-prescriptive accreditation standards. The accrediting agency that is the subject of this study utilized multiple steps to ensure consistency in the recommendations from peer survey teams. Analysis of the agreement between survey team recommendations and final decisions by the decision-making body revealed a coefficient of agreement of 0.927, indicated a high degree of agreement. The results suggest the accrediting agency's processes are effective in ensuring that peer review survey teams are applying the accreditation standards consistently between peer teams and the accrediting agency.


A square-planar osmium(ii) complex.
Reduction of the pincer complex [OsIIICl2(PNP)] (PNP = N(CHCHPtBu2)2) affords the isolation and full characterization of an osmium(ii) complex with square-planar coordination geometry, i.e. [OsIICl(PNP)]. Spectroscopic, structural and magnetic data in combination with multireference computations indicate strong temperature independent paramagnetism, which arises from an energetically well separated ground state that mixes with excited states through spin-orbit coupling.


Hormonal control of a developing neuromuscular system. I. Complete Demasculinization of the male rat spinal nucleus of the bulbocavernosus using the anti-androgen flutamide.
Prenatal treatment of male rats with the anti-androgen, flutamide (FL), demasculinizes the sexually dimorphic spinal nucleus of the bulbocavernosus (SNB) by reducing the number of SNB neurons, the size of the somas and nuclei of SNB neurons, and the size of their target muscles in adulthood. However, FL does not affect mounting or the traditional, postural measure of intromission, indicating that the SNB system does not play a major role in the mediation of these particular behaviors. Postnatal testosterone propionate (TP) treatment of male rats castrated on the day of birth results in more male copulatory behaviors in adulthood and masculinizes all measures of the SNB system. The postnatal masculinization by TP is more pronounced in males treated prenatally with FL, for morphological but not behavioral measures. The combined treatment of prenatal FL and day 1 castration without TP therapy results in a male with a completely demasculinized SNB system. Specifically, such males have SNB neurons that are as scarce and as small as those of females and, like females, they lack the target muscles of the SNB. These results support the hypothesis that perinatal androgens normally direct the sexually dimorphic development of the SNB and its target muscles.


pfcrt is more than the Plasmodium falciparum chloroquine resistance gene: a functional and evolutionary perspective.
Genetic, physiological and pharmacological studies are gradually revealing the molecular basis of chloroquine resistance (CQR) in the malaria parasite, Plasmodium falciparum. Recent highlights include the discovery of a key gene associated with resistance, pfcrt (Plasmodium falciparum chloroquine resistance transporter; PfCRT), encoding a novel transporter, and the characterization of global selective sweeps of haplotypes containing a K76T amino acid change within this protein. Little is known about the cellular mechanism by which resistant parasites escape the effects of chloroquine (CQ), one of the most promising drugs ever deployed, due in part to an unresolved mechanism of action. The worldwide spread of CQR argues that investigations into these mechanisms are of little value. We propose, to the contrary, that the reconstruction of the evolutionary and molecular events underlying CQR is important at many levels, including: (i) its potential to assist in the development of rational approaches to thwart future drug resistances; (ii) the stimulation of the use of CQ-like compounds in drug combinations for new therapeutic approaches; and (iii) the consideration of how the CQ-selected genome will function as the context in which current and future drugs will act, particularly in light of the many reports of multidrug resistance. The purpose of this review is to highlight, discuss and in some cases challenge the interpretations of recent findings on CQR. We consider the natural function of the PfCRT protein, the role of multiple genes and "genetic background" in the CQR mechanism, and the evolution of CQR in parasite populations. Genetic transformation techniques are improving in P. falciparum and continue to provide important insight into CQR. Here, we also discuss more subtle, yet important pharmacological approaches that may have been overlooked in a traditional "gene for drug resistance" way of thinking.


Cooperation of Antiporter LAT2/CD98hc with Uniporter TAT1 for Renal Reabsorption of Neutral Amino Acids.
Background Reabsorption of amino acids (AAs) across the renal proximal tubule is crucial for intracellular and whole organism AA homeostasis. Although the luminal transport step is well understood, with several diseases caused by dysregulation of this process, the basolateral transport step is not understood. In humans, only cationic aminoaciduria due to malfunction of the basolateral transporter y+LAT1/CD98hc (SLC7A7/SLC3A2), which mediates the export of cationic AAs, has been described. Thus, the physiologic roles of basolateral transporters of neutral AAs, such as the antiporter LAT2/CD98hc (SLC7A8/SLC3A2), a heterodimer that exports most neutral AAs, and the uniporter TAT1 (SLC16A10), which exports only aromatic AAs, remain unclear. Functional cooperation between TAT1 and LAT2/CD98hc has been suggested by in vitro studies but has not been evaluated in vivoMethods To study the functional relationship of TAT1 and LAT2/CD98hc in vivo, we generated a double-knockout mouse model lacking TAT1 and LAT2, the catalytic subunit of LAT2/CD98hc (dKO LAT2-TAT1 mice).Results Compared with mice lacking only TAT1 or LAT2, dKO LAT2-TAT1 mice lost larger amounts of aromatic and other neutral AAs in their urine due to a tubular reabsorption defect. Notably, dKO mice also displayed decreased tubular reabsorption of cationic AAs and increased expression of y+LAT1/CD98hc.Conclusions The LAT2/CD98hc and TAT1 transporters functionally cooperate in vivo, and y+LAT1/CD98hc may compensate for the loss of LAT2/CD98hc and TAT1, functioning as a neutral AA exporter at the expense of some urinary loss of cationic AAs. Cooperative and compensatory mechanisms of AA transporters may explain the lack of basolateral neutral aminoacidurias in humans.


Intracellular localization of GBF proteins and blue light-induced import of GBF2 fusion proteins into the nucleus of cultured Arabidopsis and soybean cells.
The G-box is an important regulatory element found in the promoters of many different genes. Four members of an Arabidopsis gene family encoding basic leucine zipper proteins (GBFs) which bind the G-box have previously been cloned. To study GBFs, a polyclonal antibody was raised against GBF1 expressed in bacteria. This antibody also recognized GBF2 and GBF3. Immunoblot analysis of nuclear and cytoplasmic fractions from Arabidopsis and soybean (SB-M) cell cultures indicated that over 90% of proteins detected with anti-GBF1 were cytoplasmic. Electrophoretic mobility shift assays indicated that over 90% of G-box binding activity was cytoplasmic. DNA affinity chromatography demonstrated that each protein detected with anti-GBF1 specifically bound the G-box. To study individual GBFs, DNA constructs fusing GBF1, GBF2 and GBF4 to GUS were made and assayed by transient expression in SB-M protoplasts. Of GUS:GBF1 proteins, 50-62% were localized in the cytoplasm under all conditions tested, while 97% of GUS:GBF4 was localized in the nucleus. By contrast, whereas about 50% of GUS:GBF2 was found in the cytoplasm of dark-grown cells, over 80% of this protein was found in the nucleus in cells cultured under blue light. Deletion analysis of GBF1 identified a region between amino acids 112 and 164 apparently required for cytoplasmic retention. These results suggest the intriguing possibility that limitation of nuclear access may be an important control on GBF activity. In particular, GBF2 is apparently specifically imported into the nucleus in response to light.


[Determination of acetone in urine by headspace gas chromatography-mass spectrometry].
Objective: To establish a method for determination of acetone in urine by headspace gas chromatography-mass spectrometry. Methods: From March to June 2021, the 3.0 ml urine sample was placed in a headspace bottle with 4.0 g of anhydrous sodium sulfate and sealed. Equilibration time was 30 min at 85 ℃. The separation was carried out on a DB-5MS column. The urine sample was detected by mass spectrometry and quantified by external standard method. Results: The method for the determination of acetone in urine by headspace gas chromatography-mass spectrometry had good linearity in the range of 51.2-1024.0 μg/L, and the correlation coefficient was 0.9995. The detection limit and the lower limit of quantification of acetone were 16.4 μg/L and 54.6 μg/L. The average recoveries of samples ranged from 94.9% to 96.8%. The intra-assay precision and inter-assay precision were both less than 10%. Samples can be stored at least 7 d at 4 ℃ or -20 ℃. Conclusion: This method has simple sample preparation and high sensitivity. It can be used for monitoring and evaluation of urinary acetone in the general population and occupationally exposed populations.


Community pharmacists' perspectives on cardiovascular disease pharmaceutical care in the United Arab Emirates: a questionnaire survey-based analysis.
Background: Community pharmacists play an intermediary role between prescribing physicians and patients in the United Arab Emirates (UAE) and thus are responsible for ensuring that patients receive optimal cardiovascular disease (CVD) pharmaceutical care. Methods: we used a cross-sectional design to assess the perceptions and practices of community pharmacists concerning pharmaceutical care for patients with CVD. A trained researcher visited randomly selected community pharmacies and used a structured questionnaire to conduct in-person interviews with pharmacists. The questionnaire collected demographic data and information on perceptions and practices regarding CVD pharmaceutical care. Results: Five hundred and fifty-one participants were recruited. The average participant age (mean ± SD) was 35 ± 2.7 years. The average perception score regarding CVD prevention and management was 75.6% (95% confidence interval [CI] 77.1%-74.2%), and the average practice score for CVD prevention and management was 87.1% (95% CI 76.5%-79.6%). Bivariate analysis revealed that gender (p = 0.001), education level (p < 0.001), pharmacy position (p = 0.004), work experience (p < 0.001), number of patients served per day (p < 0.001) and being trained on CVD prevention and management (p < 0.001) were significantly associated with perceptions about the prevention and management of CVD. Better practice scores were seen among older participants (OR 1.01; 95% CI 1-1.019), postgraduates (OR 1.77; 95% CI 1.66-1.89), workers at chain pharmacies (OR 1.24; 95% CI 1.11-1.39), pharmacists in charge (OR 1.22; 95% CI 1.01-1.47), pharmacists with >10 years of experience (OR 11.3; 95% CI 6.01-15.62), pharmacists with 6-10 years of experience (OR 4.42; 95% CI 3.90-5) and pharmacists trained on CVD prevention and management (OR 1.29; 95% CI 1.15-1.46). Conclusion: Pharmacy practitioners working in community pharmacies in the UAE actively engage in delivering pharmaceutical care to patients, playing a role in CVD management and prevention. However, they showed low levels of involvement in other healthcare services, specifically in screening and measuring patients' weight, glucose levels, and blood pressure, monitoring treatment responses, maintaining medical records, and reviewing medication refill histories. Activities such as educating patients, providing medication counseling, offering support for treatment adherence, and fostering collaborative relationships with other healthcare providers should be encouraged among UAE community pharmacists to ensure the provision of high-quality patient care.


Deep biosphere-related bacteria within the subsurface of tidal flat sediments.
Biogeochemical and microbiological processes in the upper sediment layers of tidal flats were analysed in many investigations, while deeper zones remained largely unexplored. Therefore, denaturant gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene fragments along the depth profile of up to 5.5 m-long sediment cores was performed in comparison with lithological and geochemical parameters. The investigation revealed that different compartments of the sediment columns were characterized by specific microbial communities. These compartments were analysed by sequencing of 113 DGGE bands. The upper layers down to 160-200 cm were dominated by gamma- and delta-Proteobacteria representing more than 60% of the total number of phylotypes. Underneath, a striking shift in community composition was observed, as the Proteobacteria were replaced by Chloroflexi with more than 60% of all sequences. As sulfate was still available as an electron acceptor in these layers, the abundance of Chloroflexi might be promoted by the electron donor or the quality of the carbon source. The dominance of this group, previously known as green non-sulfur bacteria, indicates the presence of a typical deep-biosphere microbial community in relatively young subsurface sediments. Thus, tidal flats might offer a convenient possibility to study and understand certain aspects of the deep biosphere in general.


Evaluation and characterization of micronuclei induced by the antitumour agent ASE [3beta-hydroxy-13alpha-amino-13, 17-seco-5alpha-androstan-17-oic-13, 17-lactam-p-bis(2-chloroethyl)amino phenylacetate] in human lymphocyte cultures.
3beta - Hydroxy - 13alpha - amino - 13, 17 - seco - 5alpha - androstan - 17 -oic-13,17-lactam-p-bis(2-chloroethyl)amino phenylacetate (ASE) is a homo-aza-steroidal ester of p-bis(2-chloroethyl) amino phenyl acetic acid and has been shown to display antineoplastic, mutagenic and genotoxic activity. In the present study an effort has been made to evaluate the ability of ASE to induce micronuclei (MN) in human lymphocytes treated in vitro using the cytokinesis-block assay. Lympocytes were treated with different concentrations of ASE (0.1, 0.25, 0.5, 1, 2.5, 5, 10 and 20 microg/ml) at two different cell culture times, 21 and 41 h after culture initiation. ASE treatment lasted until cell harvest, for 51 and 31 h, respectively. Two types of cultures were used, whole blood and isolated lymphocyte cultures. The content of induced MN was identified by FISH analysis, using an alpha-satellite DNA probe, in binucleate cells. Our results suggest that ASE is capable of increasing MN frequencies in human lymphocytes under both culture conditions. This increase is related to the concentration in a linear dose-dependent manner and is also dependent on the duration of treatment. FISH analysis has shown that the induced MN resulted mainly from breakage events. Additionally, a weak aneugenic effect was found at the higher concentrations in whole blood cultures as well as in isolated lymphocyte cultures. Cytotoxic effects of ASE were observed under both cell culture conditions with a linear dose-dependent relationship according to CBPI evaluation and were more pronounced in isolated lymphocyte cultures.


CYP450 Mediates Reactive Oxygen Species Production in a Mouse Model of β-Thalassemia through an Increase in 20-HETE Activity.
Oxidative damage by reactive oxygen species (ROS) is one of the main contributors to cell injury and tissue damage in thalassemia patients. Recent studies suggest that ROS generation in non-transfusion-dependent (NTDT) patients occurs as a result of iron overload. Among the different sources of ROS, the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase family of enzymes and cytochrome P450 (CYP450) have been proposed to be major contributors for oxidative stress in several diseases. However, the sources of ROS in patients with NTDT remain poorly understood. In this study, Hbbth3/+ mice, a mouse model for β-thalassemia, were used. These mice exhibit an unchanged or decreased expression of the major NOX isoforms, NOX1, NOX2 and NOX4, when compared to their C57BL/6 control littermates. However, a significant increase in the protein synthesis of CYP4A and CYP4F was observed in the Hbbth3/+ mice when compared to the C57BL/6 control mice. These changes were paralleled by an increased production of 20-hydroxyeicosatetraenoic acid (20-HETE), a CYP4A and CYP4F metabolite. Furthermore, these changes corroborate with onset of ROS production concomitant with liver injury. To our knowledge, this is the first report indicating that CYP450 4A and 4F-induced 20-HETE production mediates reactive oxygen species overgeneration in Hbbth3/+ mice through an NADPH-dependent pathway.


Effects on discrimination performance of selective attention to tactile features.
This study examined selective attention to tactile dimensions by combining a selective cueing paradigm with a test of integrality. In Experiment 1, subjects selectively attended to changes in the frequency or duration of pairs of vibrotactile stimuli and identified the higher frequency or longer duration stimulus. In Experiment 2, using surface gratings in an identical experimental procedure, subjects identified the rougher or longer duration stimulus. In both experiments, greater performance accuracy was found on trials where the cue correctly (valid) predicted the changing dimension, vs incorrectly (invalid) cued or no-cue (neutral) trials. More errors on the invalidly vs neutrally cued trials show the cost of focal attention. Increases in performance on validly vs neutrally cued trials show a benefit of filtering irrelevant stimuli in the cued conditions. Results effectively demonstrate focal attention to tactile features. Tests of integrality, in terms of the effects of correlated change in both dimensions, showed no redundancy gain for either vibrotactile or grating tasks, suggesting that frequency and roughness are separable from stimulus duration. Interference of negative correlated change for frequency but not roughness discriminations may be explained by differences in task difficulty.


Multiple hospital admissions in a calendar year.
Hospital in-patient workload is routinely measured as episodes of care. We report on the extent to which counts of episodes of care differ from counts of patients treated in different specialties and in different age groups. Linked records of hospital care in a population of 1.9 million people, collected over an 11-year period (1976-1986), were analysed. The all-ages multiple admission ratio (the number of admissions per 100 people admitted in the same specialty and year) varied between specialties from 102 to 171. Medical specialties tended to have higher ratios than surgical ones. The influence of age on multiple admission ratios varied between specialties, although in general the ratios increased with increasing age. There were progressive but small increases in multiple admission ratios over the period studied in a number of specialties but, by and large, stability over time was more striking than any change. The information presented could be used to estimate person-based admission rates from available episode-based data where the former are not available. This should be helpful both in managing hospital resources and in purchasing care on behalf of resident populations. Purchasers in particular should be aware of numbers of people being treated as well as the numbers of episodes of care provided.


Enhanced expression of fibronectin by cmd/cmd chondrocytes and its modulation by exogenously added proteoglycan.
Cartilage matrix deficiency (cmd/cmd) in mice is a genetic disorder associated with the failure of chondrocytes to synthesize the core protein of cartilage proteoglycan monomer (cartilage-PG). Immunohistochemical and biochemical analyses revealed enhanced accumulation of fibronectin in culture of cmd/cmd chondrocytes as well as in mutant cartilage tissue in vivo. Purified cartilage-PG, when added exogenously to a culture of cmd/cmd chondrocytes, caused a reduction in abnormal accumulation of fibronectin over several subsequent days of culturing. Using a fibronectin cDNA probe, we showed that, on the basis of total RNA, the fibronectin mRNA level was four or eight times higher in cmd/cmd chondrocytes than in the normal. The level of fibronectin mRNA in cmd/cmd cells was lowered by culturing the cells in the presence of added cartilage-PG. These findings suggest that the abnormal accumulation of fibronectin in the mutant cell culture is primarily due to elevation of fibronectin mRNA level, and that cartilage-PG in the extracellular matrix may affect the regulation of fibronectin biosynthesis at the steady-state level of mRNA.


The role of adrenergic receptors in the increased glucose uptake by the canine gut.
The study was carried out on fasted anaesthetized dogs. A vein draining a segment of the upper jejunum was cannulated for blood flow measurement. Arterial blood and venous blood from the upper jejunal segment, were obtained for measurement of glucose content. Glucose uptake was calculated as the product of jejunal blood flow and the difference between arterial and venous glucose levels. Prazosin, (0.2 mg/kg) had no effect on the glucose uptake induced by insulin and glucagon injections and glucose infusion. However, prazosin reduced the glucose uptake by about 50% during adrenaline-induced hyperglycaemia. Pretreatment of the animal with propranolol (0.5 mg/kg) significantly reduced the increased glucose uptake caused by adrenaline, glucagon and glucose hyperglycaemia. Propranolol also reduced by half the negative glucose uptake during insulin-hypoglycaemia. Since both the alpha and beta adrenergic receptor blocking agents did not abolish the induced glucose uptake, it was concluded that the effects of adrenaline, glucagon, glucose and insulin on intestinal glucose uptake are mediated in part by beta adrenergic receptors.


Bottom-up tissue engineering.
Recapitulating the elegant structures formed during development is an extreme synthetic and biological challenge. Great progress has been made in developing materials to support transplanted cells, yet the complexity of tissues is far beyond that found in even the most advanced scaffolds. Self-assembly is a motif used in development and a route for the production of complex materials. Self-assembly of peptides, proteins and other molecules at the nanoscale is promising, but in addition, intriguing ideas are emerging for self-assembly of micron-scale structures. In this brief review, very recent advances in the assembly of micron-scale cell aggregates and microgels will be described and discussed.


Psychophysiological determinants and concomitants of deficient decision making in pathological gamblers.
Psychophysiological responses are considered to be a mediating factor in the development of pathological gambling (PG) and PG has been associated with differential arousal levels during gambling. Yet little is known about the specific psychophysiological responses to wins and losses in PG. This study investigated heart rate (HR) and skin conductance responses (SCRs) during the Iowa Gambling Task (IGT) in an adult PG group (n=46) and a normal control (NC) group (n=47). Anticipatory psychophysiological reactions to disadvantageous and advantageous choices during the IGT and psychophysiological responses to wins and losses were measured. The PG group performed worse than the NC group on the IGT and exhibited lower anticipatory SCRs and HR decreases when pondering choices of disadvantageous card decks during the IGT. The PG group showed a decrease in HR after losses and wins, whereas the NC group showed a decrease in HR after losses, but an increase in HR after wins. Reward and punishment sensitivity as measured by the self-report BIS/BAS scale influenced IGT performance and psychophysiological responses, but in general these effects were similar for the PG group and the NC group. Lower anticipatory psychophysiological responses to disadvantageous choices in PG suggest impaired risk assessment in this group. Absence of a HR increase after wins possibly implies that reward sensitivity is decreased in PG. Because levels of reward and punishment sensitivity were associated with differential anticipatory HR responses to advantageous and disadvantageous decks, it would be advisable to include this taxonomy in studies on psychophysiological responses to rewards and losses.


Factors influencing the commercialization of inertial fusion energy.
Managing the IFE pathway to fusion electricity will involve management of commericalization scope, schedule, cost and risk. The technology pathway to economical fusion power comprises the commercialization scope. Industry assumes commercialization risk in fielding its own pre-pilot plant research programme for this compact-fusion pathway without the benefit of a federally coordinated IFE research and development programme. The cost of commercializing the mass-production of inexpensive targets and insisting on high reliability, availability, maintainability and inspectability has a major impact on the economics of commercializing fusion power plants. Schedule vulnerability for inertial fusion energy arises from the sensitivity of time-based roadmap stages to uncertainties in the pace of scientific understanding and technology development, as well as to unexpected and inexplicable changes of the budgeting process. Rather than rely on a time-based roadmap, a milestone-based roadmap is maximally appropriate, especially for industry and investors who are particularly well-suited to taking the risks associated with reaching the target milestones provided by the government. Milestones must be identified and optimally sequenced and the necessary resources must be delineated. Progress on the above factors, since the outcomes of recent U.S., U.K. and EUROfusion roadmapping exercises were released, are reported. This article is part of a discussion meeting issue 'Prospects for high gain inertial fusion energy (part 2)'.


The effects of dauricine and verapamil as calcium channel blockers on the plasma concentration of 6-keto-prostaglandin F1 alpha and thromboxane B2 during cardiopulmonary bypass.
Three groups of ten dogs underwent 80 minutes cardiopulmonary bypass. Two of the three groups were administered dauricine, as a new calcium channel blocker, and verapamil, as a generally recognized calcium channel blocker, respectively, from 15 minutes pre-bypass to the end of the bypass procedure (a period of 95 minutes). By means of radioimmunoassay plasma 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) and thromboxane B2 (TxB2) as the stable metabolites of prostacyclin and thromboxane A2 were surveyed. During the aortic clamping the hemodynamic changes were recorded. The dauricine and verapamil groups showed markedly inhibited generation of excessive 6-keto-PGF1 and TxB2, a narrowed ratio of TxB2 to 6-keto-PGF1 alpha, as well as a reduced total systemic peripheral resistance within the perfusion period. These results suggest that dauricine and verapamil have a salutary effect in extracorporeal circulation. The possible mechanisms are discussed.


Sustained Virologic Response at 24 Weeks after the End of Treatment Is a Better Predictor for Treatment Outcome in Real-World HCV-Infected Patients Treated by HCV NS3/4A Protease Inhibitors with Peginterferon plus Ribavirin.
Direct-acting antiviral agents against HCV with or without peginterferon plus ribavirin result in higher eradication rates of HCV and shorter treatment duration. We examined which is better for predicting persistent virologic response, the assessment of serum HCV RNA at 12 or 24 weeks after the end of treatment for predicting sustained virologic response (SVR12 or SVR24, respectively) in patients treated by HCV NS3/4A protease inhibitors with peginterferon plus ribavirin. In all, 149 Japanese patients infected with HCV genotype 1b treated by peginterferon plus ribavirin with telaprevir or simeprevir were retrospectively analyzed: 59 and 90 patients were treated with telaprevir- and simeprevir-including regimens, respectively. HCV RNA was measured by TaqMan HCV Test, version 2.0, real-time PCR assay. SVR12 or SVR24, respectively, was defined as HCV RNA negativity at 12 or 24 weeks after ending treatment. Total SVR rates were 78.0% and 66.7% in the telaprevir and simeprevir groups, respectively. In the telaprevir group, all 46 patients with SVR12 finally achieved SVR24. In the simeprevir group, 60 (93.8%) of the total 64 patients with SVR12 achieved SVR24, with the other 4 patients all being previous-treatment relapsers. SVR12 was suitable for predicting persistent virologic response in almost all cases. In simeprevir-including regimens, SVR12 could not always predict persistent virologic response. Clinicians should use SVR24 for predicting treatment outcome in the use of HCV NS3/4A protease inhibitors with peginterferon plus ribavirin for any group of real-world patients chronically infected with HCV.