Flow cytometric analysis of circulating platelet-monocyte aggregates in whole blood: methodological considerations.
Platelet-monocyte aggregates are increasingly being used to quantify platelet activation. The variables that influence platelet-monocyte aggregates have not been well defined. We sought to determine the effect of blood collection, handling and processing techniques on detected levels of platelet-monocyte aggregates using a flow cytometric assay. Whole blood was labelled with anti-CD14-PE and anti-CD42a-FITC. Thereafter, samples were fixed and red cells lysed. Analysis was performed with the flow cytometer initially triggering on light scatter and then on FL-2 to identify CD14-PE positive monocytes. Platelet-monocyte aggregates were defined as monocytes positive for CD42a. The effect of collection, handling and processing techniques on this assay were assessed. Anticoagulation with heparin (20.1 +/- 2.0%), PPACK (16.8 +/- 1.9%), sodium citrate (12.3 +/- 1.6%) and EDTA (9.5 +/- 1.0%) resulted in markedly different levels of platelet-monocyte aggregation (P < 0.0001). Platelet-monocyte aggregation was higher in samples obtained from intravenous cannulae compared to those obtained by venepuncture (20.9 +/- 3.9% vs.13.8 +/- 2.4%, P = 0.03). For every 10 minutes of delay prior to processing platelet-monocyte aggregates increased by 2.8% (P = 0.0001) in PPACK anticoagulated blood and 1.7% (P = 0.01) in citrate anticoagulated blood. Erythrocyte lysis together with fixation does not affect platelet-monocyte aggregation. Platelet-monocyte aggregates remained stable over 24 hours when fixed and stored at 4 degrees C. Multiple handling and processing factors may affect platelet-monocyte aggregation. We recommend the measurement of platelet-monocyte aggregates on samples collected by direct venepuncture, using a direct thrombin inhibitor as the anticoagulant and minimising the time delay before sample fixation.


Biomarkers of heatstroke-induced organ injury and repair.
What is the topic of this review? The status and potential role of novel biological markers (biomarkers) that can help identify the patients at risk of organ injury or long-term complications following heatstroke. What advances does it highlight? Numerous biomarkers were identified related to many aspects of generalized heatstroke-induced cellular injury and tissue damage, and heatstroke-provoked cardiovascular, renal, cerebral, intestinal and skeletal muscle injury. No novel biomarkers were identified for liver or lung injury. Classic and exertional heatstroke cause acute injury and damage across numerous organ systems. Moreover, heatstroke survivors may sustain long-term neurological, cardiovascular and renal complications with a persistent risk of death. In this context, biomarkers, defined as biological samples obtained from heatstroke patients, are needed to detect early organ injury, and predict outcomes to develop novel organ preservation therapeutic strategies. This narrative review provides preliminary insights that will guide the development and future utilization of these biomarkers. To this end, we have identified numerous biomarkers of widespread heatstroke-associated cellular injury, tissue damage and repair (extracellular heat shock proteins 72 and 60, high mobility group box protein 1, histone H3, and interleukin-1α), and other organ-specific biomarkers including those related to the cardiovascular system (cardiac troponin I, endothelium-derived factors, circulation endothelial cells, adhesion molecules, thrombomodulin and von Willebrand factor antigen), the kidneys (plasma and urinary neutrophil gelatinase-associated lipocalin), the intestines (intestinal fatty acid-binding protein 2), the brain (serum S100β and neuron-specific enolase) and skeletal muscle (creatine kinase, myoglobin). No specific biomarkers have been identified so far for liver or lung injury in heatstroke. Before translating the identified biomarkers into clinical practice, additional preclinical and clinical prospective studies are required to further understand their clinical utility, particularly for the biomarkers related to long-term post-heatstroke health outcomes.


Epidemlology of exercise-related transient abdominal pain at the Sydney City to Surf community run.
A questionnaire was administered to 848 participants (76% runners, 24% walkers) at the conclusion of the 14 km City to Surf community run in order to investigate their experience of exercise-related transient abdominal pain (ETAP). Twenty-seven percent of respondents reported experiencing ETAP during the event, with the condition reported more frequently (p< 0.01) by runners (30%) than walkers (16%). ETAP was mostly described as well-localised (88%) and of an aching (25%), sharp (22%) or cramping (22%) sensation. The most commonly-reported sites of the pain were the right (46%) and left lumbar (23%) regions of the abdomen. Forty-two percent of the respondents who experienced ETAP reported that the pain was detrimental to their performance. Reports of ETAP decreased with age (r= -0.23, p< 0.01) but were unrelated to gender, body mass index or the time taken to complete the event. Among respondents who ran, those who consumed a large mass of food relative to body weight in the time interval 1-2 hr before the event were more likely to develop symptoms of ETAP (p < 0.05). The nutritional content of the pre-event meal did not influence the experience of ETAP. Sufferers of ETAP were more likely to experience nausea (r = 0.12, p< 0.01) and report shoulder tip pain (r= 0.14, p< 0.01). The results indicate that ETAP is a commonly experienced problem and provide insights into the cause of the complaint.


The role of bacterial adhesion in cystic fibrosis including the staphylococcal aspect.
The bacteriology of cystic fibrosis shows a unique and predictable progression of colonizing micro-organisms. The reason for this sequence is still not known, but thought must be given to the idea that it may be related to the genetic disorder in some way. If this were to be true, an understanding of the colonization mechanisms at all stages in this progression could provide valuable insights for the development of novel therapies. As far as can be ascertained from published studies, mucus is the site of colonization in cystic fibrosis. While there is no doubt that the major pathogen, Pseudomonas aeruginosa, adheres to injured cells more avidly than to intact cells, the overwhelming evidence indicates that it also attaches more avidly to mucus than to intact airway cells by means of specific adhesin-receptor mechanisms. Studies with Staphylococcus aureus, the other major pathogen, are also in progress. These indicate that this organism also has an affinity for mucus. At this time the studies suggest a lesser affinity than P. aeruginosa, at least with adult mucins. These two organisms do not however appear to share the same receptor. In addition to these two major pathogens, Haemophilus influenzae and Streptococcus pneumoniae, pathogens of lesser importance also adhere to mucus. Therefore adhesion to mucus or mucins may be a recurring theme in all airway colonization. A knowledge of the factors which control these tropisms ought to provide insights into the bacterial specificity seen in cystic fibrosis and other diseases.


Incidence of mycoflora and mycotoxins in some edible fruits and seeds of forest origin.
Twelve fungi namelyAlternaria alternata, Aspergillus flavus, A niger, A ochraceus, Actinomucor repens, Capnodoium spp., Curvularia lunata, Fusarium pallidoroseum, F solani, F verticillioides, Penicillium citrinum and Rhizopus stolonifer were recorded from samples ofAegle marmelos, Aesculus indica, Buchanania lanzan andPinus gerardiana. In case ofPrunus amygdalus only Rstolonifer was recorded. A significant variation in pattern of mycoflora incidence was observed in terms of source and season. Fungal infestation in most of the substrates was found to be highest during monsoon. Aflatoxins were the most common mycotoxins elaborated by different isolates ofA flavus obtained fromA marmelos, B lanzan andP gerardiana. The amount of aflatoxins produced by the toxigenic isolates ofA flavus was in the range of traces to 0.9-26.0 μg/ml inA marmelos, 0.8-17.5 μg/ml inP gerardiana and 0.65-13.2 μg/ml inB lanzan. The percentage toxigenicity was comparatively lower in the isolates of other mycotoxigenic fungi. Aflatoxins were detected almost in all the samples analyzed for mycotoxin contamination. However, traces of zearalenone were detected inA marmelos. The concentration of aflatoxin B1 was in the range of 0.13-0.75 μg/g inA marmelos, 0.09-0.60 μg/g inP gerardiana and 0.01-0.20 ug/g inB lanzan. Mycotoxins were not detected inAesculus indica andPrunus amygdalus.


Retrograde traffic in the biosynthetic-secretory route: pathways and machinery.
In the secretory pathway, the forward (anterograde) membrane flow is compensated by retrograde transport of proteins and lipids. Membrane recycling is required for the maintenance of organelle homeostasis and the re-use of components of the transport machineries for the generation of new transport intermediates. However, the molecular mechanisms and other cellular functions of retrograde traffic are still poorly understood. In recent years, a multitude of protein factors that function in the secretory pathway have been discovered, most of them originally suggested to play a role in forward trafficking. However, in many cases subsequent studies have revealed that these proteins participate (also) in retrograde traffic. It is likely that this shift will continue, reflecting the fact that the two pathways are intimately connected.


The evolution of strategy variation: will an ESS evolve?
Evolutionarily stable strategy (ESS) models are widely viewed as predicting the strategy of an individual that when monomorphic or nearly so prevents a mutant with any other strategy from entering the population. In fact, the prediction of some of these models is ambiguous when the predicted strategy is "mixed", as in the case of a sex ratio, which may be regarded as a mixture of the subtraits "produce a daughter" and "produce a son." Some models predict only that such a mixture be manifested by the population as a whole, that is, as an "evolutionarily stable state"; consequently, strategy monomorphism or polymorphism is consistent with the prediction. The hawk-dove game and the sex-ratio game in a panmictic population are models that make such a "degenerate" prediction. We show here that the incorporation of population finiteness into degenerate models has effects for and against the evolution of a monomorphism (an ESS) that are of equal order in the population size, so that no one effect can be said to predominate. Therefore, we used Monte Carlo simulations to determine the probability that a finite population evolves to an ESS as opposed to a polymorphism. We show that the probability that an ESS will evolve is generally much less than has been reported and that this probability depends on the population size, the type of competition among individuals, and the number of and distribution of strategies in the initial population. We also demonstrate how the strength of natural selection on strategies can increase as population size decreases. This inverse dependency underscores the incorrectness of Fisher's and Wright's assumption that there is just one qualitative relationship between population size and the intensity of natural selection.


Awareness of Palestinians about lung cancer symptoms: a national cross-sectional study.
The majority of lung cancer (LC) cases are diagnosed at an advanced stage. Poor awareness of LC symptoms is a contributor to late diagnosis. This study aimed to assess the awareness of LC symptoms among Palestinians, and to examine the factors associated with displaying good awareness. Participants were recruited from hospitals, primary healthcare centers and public spaces using convenience sampling. A translated-into-Arabic version of the validated LC awareness measure was used to assess recognition of 14 LC symptoms. One point was given for each recognized symptom. The total score was calculated and categorized based on the number of symptoms recognized: poor (0-4), fair (5-9), and good (10-14). Multivariable logistic regression was used to examine the association between participant characteristics and having good awareness. The multivariable analysis adjusted for age-group, gender, education, monthly income, occupation, residence, marital status, any chronic disease, knowing someone with cancer, smoking history, and site of data collection. Of 5174 potential participants approached, 4817 completed the questionnaire (response rate = 93.1%) and 4762 were included in the final analysis. Of these, 2742 (56.9%) were from the West Bank and Jerusalem (WBJ) and 2020 (43.1%) were from the Gaza Strip. Participants from the WBJ were older, had higher monthly income but lower education, and suffered from more chronic diseases. The most recognized respiratory LC symptom was 'worsening in an existing cough'(n = 3884, 81.6%) while the least recognized was 'a cough that does not go away for two or three weeks'(n = 2951, 62.0%). The most recognized non-respiratory LC symptom was 'persistent tiredness or lack of energy'(n = 3205, 67.3%) while the least recognized was 'persistent shoulder pain'(n = 1170, 24.6%). A total of 2466 participants (51.8%) displayed good awareness of LC symptoms. Participants from both the Gaza Strip and the WBJ had similar likelihoods to have good awareness levels. Factors associated with a higher likelihood to display good awareness included female gender, having post-secondary education, being employed, knowing someone with cancer, and visiting hospitals and primary healthcare centers. About half of the study participants displayed a good level of awareness of LC symptoms. Further improvement in public awareness of LC symptoms by educational interventions might reduce LC mortality by promoting early diagnosis.


Extradural metastatic breast carcinoma detected on a bone scan.
A patient with an untreated breast carcinoma and a clinically minimally symptomatic, extradural spinal cord compression secondary to metastatic disease is described. The observation of an abnormally distended urinary bladder on a bone scan led to the prompt diagnosis and successful treatment of a total spinal cord block.


Habitual fat intake and basal fat oxidation in obese and non-obese Caucasians.
To examine the relationship between habitual fat intake and basal fat oxidation in obese and non-obese Caucasian men and women. Habitual fat intake was assessed by 7-day weighed dietary records and resting fat oxidation was determined after an overnight fast in 132 weight stable non-diabetic subjects (38 males, 94 females). All subjects were characterized for weight, height, waist-to-hip ratio, physical activity, plasma glucose and insulin response to an oral glucose load, plasma catecholamine and leptin levels. Under-reporters, defined according to plausibility of the relationship between energy expenditure and energy intake, were excluded from the analyses. The mean age was 53.1+/-10.6 y (19-72 y) and mean body mass index (BMI) was 30.7+/-5.8 kg/m(2) (19.4-45.8 kg/m(2)). Sixty-eight subjects were obese (BMI>30 kg/m(2)). Univariate regression analysis revealed a significant, albeit modest, relationship between absolute fat intake and BMI (r(2)=0.06; P<or=0.05) but not between fat intake and fat mass (r(2)=0.026; P=0.08). However, multiple regression analysis revealed significant effects of body fat mass (FM) and sex on basal fat oxidation (bFO) explaining 33% of the variation of bFO (P<or=0.0001; radical s.e.=18.0 g/24 h). In univariate regression analysis, habitual fat intake was significantly related to adjusted fat oxidation, explaining 12% of the variation (P<or=0.0001; radical s.e.=11.7 g/24 h). Habitual fat intake has a significant, albeit modest, effect on basal fat oxidation, even when adjusted for sex and body FM. The rather modest effect of habitual fat intake on fat oxidation may in part explain the increased propensity to gain FM on a high-fat diet.


Familial oligoasthenoteratozoospermia: evidence of autosomal dominant inheritance with sex-limited expression.
To report the familial occurrence of severe oligoasthenoteratozoospermia in a man and five male relatives related through their mothers. Case report. University medical center. Six affected family members. Blood and semen samples were collected from all affected males and some of their healthy male relatives. Pedigree analysis and exclusion of X-linked disorder were done. Analysis suggested that familial nonsyndromic male factor infertility was present. The family described in this report suggests the existence of an autosomal dominant trait of male infertility with sex-limited expression.


Uptake and metabolism of [3H]choline by the rat phrenic nerve-hemidiaphragm preparation.
A whole nerve-muscle preparation (about 160 mg) or an end-plate preparation (about 25 mg) of the rat phrenic nerve-hemidiaphragm were incubated with [3H]choline, to investigate choline uptake and choline metabolism. Choline uptake was measured from the disappearance of choline from the incubation medium during the loading period and from the retention of tritium in the tissue after the loading and washout period. Based on the results obtained with both methods the end-plate preparation takes up three times as much choline than the whole nerve-muscle preparation or a small muscle strip that was cut outside the end-plate region and had a similar size as the end-plate preparation. Choline uptake was not markedly affected by the degree of nerve activity or by a chronic denervation. However, hemicholinium-3 significantly reduced (50%) the choline uptake by the end-plate preparation. Most of the choline (70-88%) taken up was metabolized and incorporated into membrane structures. Phosphatidylcholine was the predominant metabolite in both preparations. The ratio of phosphatidylcholine/lysophosphatidylcholine in the end-plate preparation (16) was significantly lower than in the whole nerve-muscle preparation (31). This might indicate a higher metabolism of phosphatidylcholine in the end-plate preparation. It is suggested that choline uptake by the rat phrenic nerve-hemidiaphragm occurs mainly by the muscle fibres. The innervated part of the muscle fibres can accumulate more choline than the peripheral part outside the end-plate region, probably because of a very active choline phospholipid metabolism within the end-plate region.


Serum concentrations of small dense low-density lipoprotein cholesterol and lipoprotein(a) are related to coronary arteriostenosis in Takayasu arteritis.
Serum small dense low-density lipoprotein cholesterol (sdLDL-C) and lipoprotein(a) [Lp(a)] levels are related to coronary disease, but their specific associations with coronary arteriostenosis in Takayasu arteritis (TA) have not been ascertained. This study explored the correlations between serum sdLDL-C and Lp(a) levels and coronary arteriostenosis in TA patients as well as the degree of artery stenosis. This retrospective study included 190 TA patients and 154 healthy subjects. TA patients were divided into three categories based on the degree of coronary stenosis: Group I, stenosis >50%; Group II, stenosis 1%-50%; and Group III, stenosis 0%. Independent risk factors for coronary arteriostenosis in TA were identified by logistic regression, followed by receiver operating characteristic curve analysis to determine the specificity and sensitivity of risk factors and Youden's Index score calculation to determine the cutoff points. Takayasu arteritis patients had significantly higher serum levels of sdLDL-C and Lp(a) than healthy controls (p < 0.0001). The total cholesterol, triglyceride, LDL-C, sdLDL-C, and Lp(a) levels and the sdLDL-C/LDL-C ratio in Group I were significantly higher than those in Groups II and III (p < 0.05). An elevated serum sdLDL-C level elevated the risk of coronary arteriostenosis by 5-fold (cutoff value, 0.605 mmol/l). An increased serum Lp(a) level increased the risk of coronary arteriostenosis by 3.9-fold (cutoff value, 0.045 g/l). An elevated sdLDL-C/LDL-C ratio increased the risk of coronary arteriostenosis by 2.1-fold (cutoff value, 0.258). Serum sdLDL-C and Lp(a) levels and sdLDL-C/LDL-C ratio may be used as diagnostic factors for coronary arteriostenosis in TA patients.


The rate of dasotraline brain entry is slow following intravenous administration.
Drugs that rapidly increase dopamine levels have an increased risk of abuse. Dasotraline (DAS) is a dopamine and norepinephrine reuptake inhibitor characterized by slow oral absorption with low potential for abuse. However, it remains unclear whether intravenous (i.v.) administration would facilitate the rapid elevation of dopamine levels associated with stimulant drugs. To assess the kinetics of DAS across the blood-brain barrier and time to onset of dopamine transporters (DAT) inhibition. We compared the onset of DAT occupancy and the associated elevation of synaptic dopamine levels in rhesus monkey following i.v. administration of DAS or methylphenidate (MPH) using positron emission tomography (PET). Brain entry times were estimated by reductions in [18F]-FE-PE2I binding to DAT in rhesus monkeys. Elevations of synaptic dopamine were estimated by reductions in [11C]-Raclopride binding to D2 receptors. Intravenous administration of DAS (0.1 and 0.2 mg/kg) resulted in striatal DAT occupancies of 54% and 68%, respectively; i.v. administered MPH (0.1 and 0.5 mg/kg) achieved occupancies of 69% and 88% respectively. Brain entry times of DAS (22 and 15 min, respectively) were longer than for MPH (3 and 2 min). Elevations in synaptic dopamine were similar for both DAS and MPH however the time for half-maximal displacement by MPH (t = 23 min) was 4-fold more rapid than for DAS (t = 88 min). These results demonstrate that the pharmacodynamics effects of DAS on DAT occupancy and synaptic dopamine levels are more gradual in onset than those of MPH even with i.v. administration that is favored by recreational drug abusers.


Alpha 1-antitrypsin, alpha 1-antichymotrypsin, and alpha 2-macroglobulin in human gastric carcinomas: a retrospective immunohistochemical study.
One hundred twenty-six gastric carcinomas (68 advanced cancers and 58 early cancers) were examined immunohistochemically for alpha 1-antitrypsin (AAT), alpha 1-antichymotrypsin (ACT), and alpha 2-macroglobulin (AMG) within tumor cells. The incidence of these three protease inhibitors was markedly higher in advanced than in early cancers, regardless of the histologic type of gastric carcinoma. In advanced cancers the incidence of both AAT and AMG was significantly higher in well-differentiated adenocarcinomas than in poorly differentiated adenocarcinomas, but no difference was observed in the expression of ACT between these two types of advanced carcinomas. Eighty per cent of the AAT-positive advanced carcinomas had ACT, and 40 per cent of these tumors also contained AMG. The two-year survival rates clearly indicated that well-differentiated adenocarcinomas with AAT have worse prognoses than well-differentiated adenocarcinomas without AAT, but there was no relation between the expression of ACT or AMG and prognosis. These results strongly suggest that the presence of protease inhibitors in gastric carcinomas is related to the invasive growth of the tumors and that AAT is a tissue tumor marker of well-differentiated adenocarcinomas of the stomach. It may also serve as a biologic marker of high malignancy in patients with these gastric cancers.


[Correlations between studies with plain radiography and with cineventriculography in subjects with prior myocardial infarct].
In 86 subjects wiht healed myocardial infarction the reliability of cardiothoracic ratio (C/T), of the left half-diameter (EDTS/m2) and of the overall subjective evaluation of a standard chest X-Ray plate has been tested by comparison to quantitative data drawn by selective cineventriculography. C/T and EDTS/m2 are scarcely reliable, whereas subjective evaluation allowed a separation among different groups of patients, though with wide overlap. Right anterior oblique and lateral views, and the evaluation of the third aortic arch were more useful than left anterior oblique and lowered apex.


Differential arginine dependence and the selective cytotoxic effects of activated macrophages for malignant cells in vitro.
Normal and neoplastic cells from 4 species (man, rat, mouse and hamster) were examined for their dependence on exogenous L-arginine in tissue culture. The malignant cells required a higher concentration of L-arginine in the medium than their normal counterparts (with similar doubling times) to maintain optimal proliferation. Complete arginine deprivation resulted in equal growth inhibition of normal and malignant cells, but more rapid cytolysis of the malignant cell. Deprivation of L-arginine, followed 24 h later by rescue with L-arginine, allowed normal cells to proliferate, but the reproductive capacity of the malignant cells was irreversibly impaired. Since the cytotoxic activity of LPS-activated macrophages was associated with the release of arginase and was abrogated by excess L-arginine, it is suggested that the biological basis for the selective effects of such macrophages may reside in the L-arginine dependence of the target cells.


Redox sensitive miR-27a/b/Nrf2 signaling in Cr(VI)-induced carcinogenesis.
Hexavalent chromium [Cr(VI)] is a well-known carcinogen that can cause several types of cancer including lung cancer. NF-E2-related factor 2 (Nrf2), the redox sensitive transcription factor, can protect normal cells from a variety of toxicants and carcinogens by inducing the expression of cellular protective genes and maintaining redox balance. However, Nrf2 also protects cancer cells from radio- and chemo-therapies and facilitates cancer progression. Although Cr(VI) treatment has been demonstrated to upregulate Nrf2 expression, the mechanisms for Nrf2 regulation upon chronic Cr(VI) exposure remain to be elucidated. We found that Nrf2 was upregulated in BEAS-2B cells exposed to Cr(VI) from 1 to 5 months, and also in Cr(VI)-induced transformed (Cr-T) cells with Cr(VI) treatment for 6 months. We showed that KEAP1, the classic negative regulator of Nrf2, was downregulated after Cr(VI) exposure for 4 months, suggesting that Nrf2 induction by Cr(VI) treatment is through KEAP1 decrease at late stage. To further decipher the mechanisms of Nrf2 upregulation at early stage of Cr(VI) exposure, we demonstrated that miR-27a and miR-27b were redox sensitive miRNAs, since reactive oxygen species (ROS) scavengers induced miR-27a/b expression. After Cr(VI) exposure for 1 month, the expression levels of miR-27a/b was dramatically decreased. The changes of miR-27a/b and their target Nrf2 were confirmed in vivo by mouse model intranasally exposed to Cr(VI) for 12 weeks. Nrf2 was a direct target of miR-27a/b, which acted as tumor suppressors in vitro and in vivo to inhibit tumorigenesis and cancer development of Cr-T cells. The results suggested that the inhibition of miR-27a/b was responsible for Nrf2 upregulation at both early stage and late stage of Cr(VI) exposure. This novel regulation of Nrf2 upon chronic Cr(VI) exposure through redox-regulated miR-27a/b will provide potential targets for preventing and treating Cr(VI)-induced carcinogenesis in the future.


Variation in host susceptibility among and within populations ofPlantago lanceolata L. infected by the fungusPhomopsis subordinaria (Desm.) Trav.
Susceptibility toPhomopsis stalk disease ofPlantago lanceolata genotypes, sampled in three different populations with a variable degree of infection by the fungusPhomopsis subordinaria, was determined under greenhouse conditions. Susceptibility of the host varied within, but not among populations. No relationship between the intensity of the disease in the field and the mean susceptibility of the host genotypes sampled at those locations could be established. Host susceptibility appeared to be composed of the host genotypes sampled at those locations could be established. Host susceptibility appeared to be composed of different (uncorrelated) plant characteristics. Determining whether host genotypes are highly or slightly susceptible can only be achieved by field trials, where the plants are exposed to the whole set of disease inducing factors. The relevance of host susceptibility to the intensity of disease in the field is discussed in relation to the variation in pathogenicity of the fungus and the variation in environmental factors prevailing inP. lanceolata populations underP. subordinaria pathogen pressure.


Tooth-size ratio for patients requiring 4 first premolar extractions.
The relationship between total mesiodistal widths of the maxillary and mandibular teeth is an important factor in orthodontic treatment planning. The purposes of this article are to report a mathematical tooth-size ratio specifically designed for patients needing the extraction of 4 first premolars and to compare the anterior "6" and overall "12" ratio values reported by Bolton with the calculated anterior "6" and overall "10" ratio values obtained from data in this study. This study was conducted in 3 phases. In the first 2 phases, we used the peer assessment rating and ideal cephalometric norms to select 53 ideal posttreatment models of patients who had had 4 premolars extracted. In the third phase, the mean overall "10" ratio and the mean anterior "6" ratio were calculated for the selected models. Bolton's mean overall "12" (91.3%) and anterior "6" ratios (77.2%) were compared statistically with calculations derived from this study by using 1-sample t test. The mean overall "10" ratio and the mean anterior "6" ratio were found to be 89.28 +/- 1.07% and 77.68 +/- 1.12%, respectively. Although the difference in anterior ratio was not significantly different from Bolton's anterior "6" ratio, there was a statistically significant difference between Bolton's study and our study in overall ratio. The mathematical tooth size overall ratio of 89.28% was determined for patients requiring the extraction of 4 first premolars and is recommended for use in diagnosis and treatment planning.


Parametric study and global sensitivity analysis for co-pyrolysis of rape straw and waste tire via variance-based decomposition.
In present study, co-pyrolysis behavior of rape straw, waste tire and their various blends were investigated. TG-FTIR indicated that co-pyrolysis was characterized by a four-step reaction, and H2O, CH, OH, CO2 and CO groups were the main products evolved during the process. Additionally, using BBD-based experimental results, best-fit multiple regression models with high R2-pred values (94.10% for mass loss and 95.37% for reaction heat), which correlated explanatory variables with the responses, were presented. The derived models were analyzed by ANOVA at 95% confidence interval, F-test, lack-of-fit test and residues normal probability plots implied the models described well the experimental data. Finally, the model uncertainties as well as the interactive effect of these parameters were studied, the total-, first- and second-order sensitivity indices of operating factors were proposed using Sobol' variance decomposition. To the authors' knowledge, this is the first time global parameter sensitivity analysis has been performed in (co-)pyrolysis literature.


Parathyroid carcinoma arising from auto-transplanted parathyroid tissue after Total Parathyroidectomy in chronic kidney disease patient: a case report.
Secondary hyperparathyroidism is a common complication in patients with chronic kidney disease that requires vigilant treatment due to its high mortality rate. Pharmacologic therapy is recommended as an initial treatment; if there is no response, a total parathyroidectomy is performed. In some cases, surgery is accompanied by auto-transplantation of parathyroid tissue. The patient was diagnosed with chronic kidney disease and received a kidney transplant. However, due to rejection of the transplanted kidney, medical nephrectomy was carried out and routine hemodialysis was initiated and observed. At this time, secondary hyperparathyroidism with elevated parathyroid hormone and hyperphosphatemia developed and pharmacologic treatment was applied. However, there was no response to pharmacologic treatment; therefore, total parathyroidectomy with auto-transplantation was performed. Eight years after surgery, a growing mass was observed in the transplantation site, accompanied by an elevation of parathyroid hormone. A complete resection of the mass was performed, and the patient was diagnosed with parathyroid carcinoma. Additional adjuvant radiation therapy was ordered, and the patient is being monitored. This is a rare but remarkable case of parathyroid carcinoma arising from auto-transplanted parathyroid tissue after total parathyroidectomy in a patient with secondary hyperparathyroidism. We suggest caution should be taken when choosing to auto- transplant parathyroid tissue and that careful postoperative observation should be performed.


Intrathecal umbilical cord mesenchymal stem cells injection alleviates neuroinflammation and oxidative stress in the cyclophosphamide-induced interstitial cystitis rats through the Sirt1/Nrf2/HO-1 pathway.
Neuroinflammation in the spinal dorsal horn (SDH) region plays an important role in the pathogenesis of interstitial cystitis (IC)/bladder pain syndrome (BPS). Oxidative stress is an important etiological factor for inflammatory diseases. This study aimed to investigate the therapeutic effects of umbilical cord mesenchymal stem cells UMSCs on neuroinflammation and oxidative stress in IC and the underlying mechanisms. Rats were intraperitoneally injected with cyclophosphamide (50 mg/kg bodyweight) to establish the IC animal model. Additionally, rats were intrathecally injected with a Sirt1-specific agonist (SRT1720; 8 μg/rat) or inhibitor (EX527; 8 μg/rat). Furthermore, rats were intrathecally injected with human UMSCs (hUMSCS; 8 × 105 cells/rat). Rat behavior was examined using the mechanical allodynia test, novel object recognition test, sucrose preference test, and urodynamics analysis. Neuroinflammation and oxidative stress the SDH region were examined using western blotting, immunofluorescence, enzyme-linked immunosorbent assay, and commercial kits. The Sirt1/Nrf2/HO-1 pathway was downregulated in IC rats. Sirt1 activation and inhibition differentially affected the behavior of IC rats. hUMSCs effectively mitigated the upregulation of oxidative stress, proinflammatory cytokines, and glial activation in the SDH region. Additionally, hUMSCs suppressed mechanical allodynia, dysregulated urodynamics, memory deficits, and depressive-like behavior in IC rats. hUMSCs exerted therapeutic effects through the Sirt1/Nrf2/HO-1 pathway. intrathecal hUMSCs injection alleviated behavioral deficits of IC rats by mitigating neuroinflammation and oxidative stress through the Sirt1/Nrf2/HO-1 pathway and can be potentially an effective therapeutic strategy for IC.


Evaluation of administered dose using portal images in craniospinal irradiation of pediatric patients.
This study aimed to assess the administered dose based on portal imaging in craniospinal pediatric irradiation by evaluating cases in which portal images did or did not account for the total administered dose. We also intended to calculate the mean increase in total administered dose. Data were collected from General University Hospital Gregorio Marañón; we evaluated the total dose administered, total dose planned, number of portal images per treatment and corresponding monitor units of two different groups: one in which the dose from portal images is deducted from the total administered dose (D), and another in which it was not (N). We used descriptive statistics to analyze the collected data, including the mean and respective standard deviation. We used the Shapiro-Wilk and Spearman rank correlation coefficient tests and estimated the linear regression coefficients. Patients in group D received a mean dose of 29.00 ± 10.28 cGy based on the verification portal images, a quantity that was deducted from the planned dose to match the total administered dose. Patients in group N received a mean dose of 41.50 ± 30.53 cGy, which was not deducted from the planned dose, evidencing a mean increase of 41.50 ± 30.55 cGy over the total administered dose. The acquisition of the set-up verification portal images, without their inclusion in the total administered dose, reflects an average increase in total dose for craniospinal irradiation of pediatric patients. Subtraction of the monitor units used to acquire the verification images is recommended.


A critical review of the literature on ratings scales for perceived exertion.
The study of human performance and perceived exertion during physical activity has been an area of considerable interest and research for over 50 years. This review considers the evidence of many investigators who have been researching the physiological basis as well as non-physiological basis for the ratings of perceived exertion. During low levels of activity, physical perception in the working muscles appears to be the primary stimulus for effort perception. When work intensity exceeds the lactate threshold, incremental elevations in blood lactate complement peripheral input from the neuromuscular mechanisms. Once a critical absolute ventilatory threshold is reached, central input also contributes to effort perception. In most instances, peripheral input predominates over central cues, although it has been shown that pronounced central cues may dominate the perception of effort. Central (heart rate, VE, VO2) or local (muscle and blood lactate, adenosine triphosphate, creatine phosphokinase, glycogen) cues highlighted in these studies demonstrate both the complexity of effort perception, and the need for better understanding of the physiological components upon which it is based. Athletes have been shown to have a greater tendency to reduce perceptual ratings than their non-active counterparts. In view of these observations, it is apparent that a theoretical framework based upon physiological and psychological considerations may exist to support the concept of training-induced alterations in perceived exertion. This appears to be particularly true in higher ranges of exercise intensity. Part of the problem in reaching a conclusion on the issue of perceptual ratings trainability centres upon the agreement on what should be recognised as a significant decrement in perceived exertion. It is concluded that there is considerable variation in the findings of the literature and that any reported variations in performance may well be greatly influenced by intersubject variability, the type of exercise, and nutritional status of subject. Further research is required to understand this issue better.


Practice with persons living their dying: a human becoming perspective.
The purpose of this article is to discuss nursing practice with persons who are living their dying, from the perspective of Parse's theory of human becoming. Quality of life from the person's own perspective is the goal of practice guided by Parse's theory; hence, it is particularly relevant for nurses in palliative care settings, where quality of life is paramount. The practice methodology is explained and illustrated with an example drawn from experiences with a woman in a hospice in South Korea. As this woman lived her dying, suffering, joy, and sorrow emerged in cocreated relationships with important others.


An All-Inside Repair for Full Radial Posterior Lateral Meniscus Tears.
There is controversy about the treatment for unstable full radial posterior lateral meniscus tears, particularly that involving the posterior root. Some surgeons have advocated repairing these types of meniscus tears using various techniques, but their methods are somewhat technical. We developed the technique for an all-inside repair for full radial posterior lateral meniscus tears using the Meniscal Viper (Arthrex, Naples, FL). A doubled thread is passed through 1 edge of the radial tear by the Meniscal Viper and is kept in place without tying the knot. The Meniscal Viper is used again to set a new thread, repeating the same procedure to another edge of the tear. At this step, 2 doubled threads are passed through each stump of the tear, and both a loop end and 2 free ends of each thread are located outside of the joint. Then, 2 doubled threads pass the third thread into its own loop, pulling it out. Finally, the third thread becomes the mattress suture over the radial tear site and is fastened by sliding knot techniques. This procedure makes it easy to strictly, smoothly, and less invasively shorten the gap by drawing each stump of the meniscus in the direction of the circumference.


HER3 targeting augments the efficacy of panobinostat in claudin-low triple-negative breast cancer cells.
Patients with triple-negative breast cancer (TNBC) have a poor prognosis and high relapse rate due to limited therapeutic options. This study was conducted to determine the mechanisms of action of panobinostat, a pan-inhibitor of histone deacetylase (HDAC) and FDA-approved medication for multiple myeloma, in TNBC and to provide a rationale for effective drug combinations against this aggressive disease. RNA sequencing analyses of the claudin-low (CL) TNBC (MDA-MB-231) cells untreated or treated with panobinostat were performed to identify the differentially expressed genes. Adaptive alterations in gene expression were analyzed and validated in additional CL TNBC cells. Tumor xenograft models were used to test the in vivo antitumor activity of panobinostat alone or its combinations with gefitinib, an EGFR-tyrosine kinase inhibitor (TKI). Panobinostat potently inhibited proliferation and induced apoptosis in all TNBC cells tested. However, in CL TNBC cells, this HDAC inhibitor markedly enhanced expression of HER3, which interacted with EGFR to activate both receptors and Akt signaling pathways. Combinations of panobinostat and gefitinib synergistically suppressed CL TNBC cell proliferation and promoted apoptosis in vitro and in vivo. Upregulation of HER3 compromises the efficacy of panobinostat in CL TNBC. Inactivation of HER3 combined with panobinostat represents a practical approach to combat CL TNBC.


Modeling motor vehicle crashes for street racers using zero-inflated models.
Motor vehicle crashes are a leading cause of death for young people in the United States. Assessing which drivers are at a high risk of experiencing a crash is important for the implementation of traffic regulations. Illegal street racing has been associated with a high rate of motor vehicle crashes. In this study, we link Utah statewide driver license citations and motor vehicle crash data to evaluate the rate of crashes for drivers with street racing citations relative to other drivers. Using a zero-inflated negative binomial model we found that drivers with no citations are approximately three times more likely to be at zero risk of a crash compared to drivers with street racing citations. Moreover, among drivers at non-negligible risk of crash, cited street racers are more likely to be involved in a crash compared to drivers without citations or those cited for violations other than street racing. However, drivers with increased numbers of non-street-racing citations experience crash risks approaching those of the cited street racers.


Silencing of fat-1 transgene expression in sheep may result from hypermethylation of its driven cytomegalovirus (CMV) promoter.
The fat-1 gene was isolated from roundworm Caenorhabditis elegans, and built into pIRES2-EGFP expression vectors driven by cytomegalovirus (CMV) promoter or cytomegalovirus enhancer and chickenβ-actin (CAG) promoter. Both CMV- and CAG-driven expression vectors were transfected to sheep fetal fibroblast cells. Positive transfected cells were used as donors for somatic cell nuclear transfer (SCNT) and the cloned embryos were transferred into the oviducts of synchronized recipient sheep. Two lambs derived from CMV vector and three lambs derived from CAG vector developed to term. Although Southern analyses using tissues from the two lambs derived from CMV vectors indicated integration of fat-1 gene into the genome, fat-1 mRNAs were not detected by RT-PCR. However, there was fat-1 expression (detected by RT-PCR) in tissues from transgenic lambs driven by CAG vectors. To investigate potential mechanisms involved in the two transgene models, methylation state of the vector promoters were examined. In CMV-driven transgenics, CMV promoters had almost no methylation in transfected cells and the resultant cloned embryos, whereas high methylations were detected in tissues and organs in transgenic lambs. In the CAG-driven transgenics, there were almost no methylations in transgenic cells and transgenic cloned embryos, and cloned lambs expressed fat-1 mRNA (detected by RT-PCR). Moreover, although SV40 promoters which drove neo/kan marker gene in CMV vectors were highly methylated in tissues from transgenic lambs, they were without methylation in cells and embryos. Therefore, we concluded that highly methylated CMV promoters induced the silence of fat-1 transgene expression in sheep. Furthermore, CAG promoter, but not CMV promoter was suitable for generation of fat-1 transgenic sheep.


Activity of hypothalamic neurons in conscious rats decreased by hyperbaric environment.
Single-unit activity of neurons in rat hypothalamus, prefrontal cortex (PFC) and central medial thalamus (CM) was recorded through chronically implanted electrodes. Upon being subjected to 7 atmospheres absolute (ATA) hyperbaric air (709 kPa) environment, about two-thirds of the hypothalamic neurons tested reversibly decreased their firing rate to 71 +/- 12% (mean +/- SD, n = 25) compared to the precompression control activity, and the remaining one-third did not change their firing rate (100 +/- 6%, n = 13). It is suggested that the decrease in firing rate was attributable to increased partial pressure of both oxygen and nitrogen. In contrast to the hypothalamus, the firing rates of most neurons in the PFC and CM did not decrease (99 +/- 9% for PFC, 107 +/- 12% for CM). In separate acute experiments, 3 out of 5 hypothalamic neurons tested in the isolated forebrain also decreased their firing rates in hyperbaric environment. The results suggest that the hyperbaric air acted directly on the forebrain and depressed more than half of the hypothalamic neurons.


[Management of deep-vein thrombosis: A 2015 update].
Deep vein thrombosis (DVT) is a frequent and multifactor disease, with two major complications, post thrombotic syndrome and pulmonary embolism. Both transient (surgery, plaster immobilization, bed rest/hospitalization) and chronic/persistent (age, cancer, clinical or biological thrombophilia…) risk factors modulate treatment duration. Diagnostic management relies on clinical evaluations, probability followed by laboratory tests or imaging. So far, compression ultrasound is the diagnostic test of choice to make a positive diagnosis of DVT. Anticoagulants at therapeutic dose for at least 3 months constitute the cornerstones of proximal (i.e. involving popliteal or more proximal veins) DVT therapeutic management. The arrival of new oral anticoagulants should optimize ambulatory management of DVT.


Collaborative complementation study of temperature-sensitive mutants of herpes simplex virus types 1 and 2.
Twenty-three complementation groups of herpes simplex virus type 1 (HSV-1) and 20 of HSV-2 were identified by qualitative and quantitative complementation analysis from among 43 temperature-sensitive (ts) mutants of HSV-1 and 29 ts mutants of HSV-2 which had been isolated independently in 10 laboratories.


Discrete choice as a method for exploring education preferences in a Danish population of patients with type 2 diabetes.
To determine preferences among patients with type 2 diabetes for content and format of patient education. Using discrete choice methods, we surveyed patients about their preferences for patient education. We investigated preferred content and format regarding education on living well with diabetes, preventing complications, healthy eating, exercising, and psychosocial issues related to diabetes. We obtained usable responses from 2187 patients with type 2 diabetes. Acquiring competencies to live a fulfilling life with diabetes, adjust diet and exercise habits, and prevent complications was significantly more highly valued than was simply being informed about these topics. Patients preferred to be involved in the planning of their diabetes care and valued individually tailored content higher than prescheduled content. Women and younger patients found diet and exercise significantly more important than did men, and patients with poorly controlled diabetes valued all education and support more highly than did patients in better control. Patients with type 2 diabetes prefer to be actively involved in educational activities, to develop competencies to prevent and manage complications, and to involve their social network in supporting them. Future patient education should enhance participation and competence development and include relatives.


The development of sustained attention in children might be related to the maturation of frontal cortical functions.
The relationship between the development of sustained attention and the ability to inhibit responding to external stimuli was investigated in seventy five children, 8, 10 and 12 years old. An important period in the development of both functions is between 8 and 10 years of age. A positive correlation was found between the number of signals detected (sustained attention) and the number of successfully suppressed reflexive saccades to the peripherally located target (the ability to inhibit responding to external stimuli).


Comparative analysis of the volatile components in cut tobacco from different locations with gas chromatography-mass spectrometry (GC-MS) and combined chemometric methods.
A combined approach of subwindow factor analysis and orthogonal projection resolution was used to analyze the volatile components of cut tobacco samples from different sources. After extracted with simultaneous distillation and extraction method, the volatile components in cut tobacco from five different locations were detected by GC-MS. Then, the qualitative and quantitative analysis of the volatile components of cut tobacco from Changde area was completed with the help of subwindow factor analysis resolving two-dimensional original data into pure mass spectra and chromatograms. One hundred and two volatile components among 138 separated peaks were identified and quantified, accounting for about 88.90% of the total content. Finally, orthogonal projection method was used to extract the common peaks from different locations. Among the identified components, there were 74 components coexisting in five studied samples although the relative content of each component showed difference to some extent. The results showed a fair consistency in their GC-MS fingerprints. It was the first time to apply orthogonal projection method to compare different cut tobacco samples, and it reduced the burden of qualitative analysis as well as the subjectivity. The obtained results proved the combined approach powerful for the analysis of complex cut tobacco samples. The developed method can be used to compare the sameness and differences of cut tobacco from different sources and for quality control of cigarette production and materials.


Cross-sectional study of antimicrobial use and treatment decision for preweaning Canadian dairy calves.
Antimicrobials should be used prudently in farm animals to prevent the development of resistant bacteria in both humans and animals. The objective of this study was to investigate Canadian dairy producers' practices for antimicrobial use in the treatment of disease in preweaning dairy calves. In-person questionnaires were administered to 144 dairy producers across 5 provinces in Canada between July 2019 and August 2020. Almost all (96%) producers used antimicrobials to treat calves with respiratory disease, but only 27% indicated they had a written treatment protocol for respiratory disease. Most (95%) of these protocols for respiratory disease were developed with input from the herd veterinarian. Seventy-four percent of producers used antimicrobials to treat calf diarrhea, with 37% of producers having a written treatment protocol for calf diarrhea with input from the herd veterinarian. The combinations of signs adopted by the producers for antimicrobial treatment in calf respiratory disease and diarrhea were evaluated based on findings from other studies. More than half (56%) of producers who used antimicrobials for calf respiratory disease decided to use antimicrobials by evaluating multiple clinical signs. Eighty-two percent of producers who used antimicrobials for calf diarrhea made decisions based on systemic signs of disease, presence of bloody stool, no response to previous treatment, or on the recommendation from the herd veterinarian. Producers with a written treatment protocol had 3 to 7 times greater odds of using antimicrobials based on multiple signs or systemic signs of disease compared with those without a protocol. Further research may investigate other calf management practices related to decision-making by producers in using antimicrobials to improve antimicrobial stewardship on dairy farms.


The future role of the anaesthesiologist in pain management.
Relieving pain during and after surgery and trauma has always been a basic duty of anaesthesiologists. With their skills and expertise in regional analgesia and pharmacological analgesia, anaesthesiologists have improved management of severe cancer pain. Will there be a place for anaesthesiologists in multidisciplinary pain clinics managing chronic non-cancer pain patients in the future? This is a personal review of the development of pain management as a growing part of the responsibilities of anaesthesiologists during the past three decades and the importance of continued involvement of anaesthesiologists in this interesting and challenging aspect of clinical medicine. Optimal management of pain during and after surgery is a prerequisite for successful short- and long-term rehabilitation after surgery. After surgery, reducing dynamic pain with prolonged optimal epidural analgesia and regional blocks facilitates mobilization and reduces chronic pain. The expertise of well-trained anaesthesiologists in skillfully using regional analgesia and pharmacological pain relief continues to be in demand in palliative care. Some interventional techniques are useful in relieving chronic non-cancer pain in selected patients. Well-trained anaesthesiologist-pain clinicians can perform interventional treatments safely. No doubt, anaesthesiologists will continue to have important roles in pain management in the future.


3-O-Sulfation of Heparan Sulfate Enhances Tau Interaction and Cellular Uptake.
Prion-like transcellular spreading of tau in Alzheimer's Disease (AD) is mediated by tau binding to cell surface heparan sulfate (HS). However, the structural determinants for tau-HS interaction are not well understood. Microarray and SPR assays of structurally defined HS oligosaccharides show that a rare 3-O-sulfation (3-O-S) of HS significantly enhances tau binding. In Hs3st1-/- (HS 3-O-sulfotransferase-1 knockout) cells, reduced 3-O-S levels of HS diminished both cell surface binding and internalization of tau. In a cell culture, the addition of a 3-O-S HS 12-mer reduced both tau cell surface binding and cellular uptake. NMR titrations mapped 3-O-S binding sites to the microtubule binding repeat 2 (R2) and proline-rich region 2 (PRR2) of tau. Tau is only the seventh protein currently known to recognize HS 3-O-sulfation. Our work demonstrates that this rare 3-O-sulfation enhances tau-HS binding and likely the transcellular spread of tau, providing a novel target for disease-modifying treatment of AD and other tauopathies.


Effect of hydrothermal treatment on physicochemical properties of amorphous granular potato starch (AGPS).
Using restructuring technology, A- or B-type crystalline granular potato starch was produced from amorphous granular potato starch (AGPS). AGPS was prepared using ethanol-heat processing, and hydrothermal treatments were performed with different moisture contents (18, 29, 200% d.b.) and temperatures (4, 25, 40, 60, 80 °C) for 3 weeks. AGPS showed no endothermic peak in a DSC thermogram, while hydrothermally treated AGPS (HAGPS) revealed endothermic peaks. In X-ray diffraction, AGPS displayed an amorphous pattern, and HAGPS displayed A- or B-type crystalline patterns depending on treatment temperature and moisture content. Neither AGPS nor HAGPS had typical RVA pasting curves, and their viscosities gradually increased. Raman spectroscopy and FT-IR confirmed that ordered structure and crystalline regions increased in HAGPS. Resistant starch (RS) and slowly digestible starch (SDS) contents of HAGPS increased but rapidly digestible starch (RDS) content decreased compared to AGPS. These results elucidated that hydrothermal treatment could change the physicochemical properties of AGPS and produce an identical material, such as granular potato starch with A-type and B-type crystalline granular potato starch.


Novel oral anticoagulants in non-valvular atrial fibrillation.
Atrial fibrillation (AF) confers a significant risk of stroke or systemic thromboembolism. Oral anticoagulation is the most effective therapy for AF-related stroke prevention. A decision to advise oral anticoagulation should be based upon the individual absolute risks of stroke and bleeding, and almost all AF patients with ≥1 stroke risk factors have a positive net clinical benefit of oral anticoagulation. The novel oral anticoagulants (NOACs) dabigatran, rivaroxaban and apixaban are more convenient, and are at least equally effective and safer (regarding bleeding complications) for stroke prevention compared with vitamin K antagonists (VKAs). Availability of NOACs and improved stroke and bleeding risks assessment should increase the number of AF patients who receive adequate thromboprophylaxis. In this review article, we present an overview of the clinical phase III trials with NOACs for stroke prevention and discuss the contemporary principles of thromboprophylaxis in AF patients with various stroke and bleeding risk profiles, as well as practical aspects of NOACs therapy.


Defining recurrent pregnancy loss: associated factors and prognosis in couples with two versus three or more pregnancy losses.
The definition of recurrent pregnancy loss (RPL) differs internationally. The European Society of Human Reproduction and Embryology (ESHRE) defines RPL as two or more pregnancy losses. Different definitions lead, however, to different approaches to care for couples with RPL. This study aimed to determine whether the distribution of RPL-associated factors was different in couples with two versus three or more pregnancy losses. If a similar distribution were found, couples with two pregnancy losses should be eligible for the same care pathway as couples with three pregnancy losses. This single-centre, retrospective cohort study investigated 383 couples included from 2012 to 2016 at the Leiden University Medical Center RPL clinic. Details on age, body mass index, smoking status, number of pregnancy losses, mean time to pregnancy loss and performed investigations were collected. The prevalence of uterine anomalies, antiphospholipid syndrome, hereditary thrombophilia, hyperhomocysteinaemia, chromosomal abnormalities and positive thyroid peroxidase antibodies were compared in couples with two versus three or more pregnancy losses. No associated factor was found in 71.5% of couples with RPL. This did not differ statistically between couples with two versus three or more pregnancy losses (73.6% versus 70.6%; P = 0.569). The distribution of investigated causes did not differ between the two groups. As the distribution of associated factors in couples with two versus three or more pregnancy losses is equal, couples with two pregnancy losses should be eligible for the same care pathway as couples with three. This study supports ESHRE's suggestion of including two pregnancy losses in the definition of RPL.


[Differentiated thyroid carcinoma. Surgery and significance of lymph node involvement].
Nodal treatment in papillary (PTC) and in follicular (FTC) thyroid carcinoma is still a subject of debate. 1974-95 therapeutic lymphadenectomy (30/95), 1996-1999 frequent prophylactic lymphadenectomy (32/57; P = 0.005) was used for PTC, with therapeutic lymphadenectomy for FTC (15/115). 131I was used selectively for pN1-tumours. PTC: The incidence of pN0-, but not of pN1-status increased significantly (P = 0.03). Nodal recurrence was observed in 5/89 (6%) with therapeutic, vs. 1/54 (2%) with prophylactic lymphadenectomy (P = NS), i.e. in 1/107 (0.9%) patients without evidence of nodal disease, vs. 5/36 (14%) of those with pN1-status (P = 0.0004). Survival at 25 yrs. in stages TNM I and II was 100%, i.e., independent of N-status. FTC: No nodal recurrence was observed. Occult untreated nodal disease represented no major clinical problem. Selective nodal treatment may offer optimal results; meticulous nodal dissection is indicated for N1-tumours.


Referral patterns to a district general hospital gastroenterology outpatient clinic: implications for the 'two-week target'.
We defined the pattern and appropriateness of GPs' new-patient referrals to a large district general hospital gastroenterology department, and assessed the implications for workload in the context of the recently introduced 'two-week target'. Prospective data were collected on all new referrals over a two-month period and 426 new appointments were included, from which data were available on 390. Only six referrals were deemed inappropriate. Sixty-nine patients had a functional disorder, and GPs were less likely to diagnose this group at referral than the gastroenterologist was after the initial consultation. Nineteen per cent of all GP referrals were classified as urgent and 6% of these had a malignancy. Fifty per cent of patients with malignancy were not perceived as meriting an urgent referral by the GP Gastroenterology outpatient facilities are already overstretched and the implementation of the two-week target will add considerable strain to the current resources, with little gain in identifying malignancy.


Glenoid subchondral bone density in osteoarthritis: A comparative study of asymmetric and symmetric erosion patterns.
Recent studies have shown variations in glenoid bone density in asymmetric wear patterns but have yet to analyze non-arthritic or concentrically worn glenoids. The purpose of this study is to characterize and compare subchondral glenoid bone densities in both non-arthritic and A1, A2, B1, B2 and B3 osteoarthritic glenoids, as well as to assess uniformity in symmetric and asymmetric erosion wear patterns. In all, 150 computerized tomography (CT) scans containing equal numbers of non-arthritic (N), A1, A2, B1, B2 and B3 glenoids were segmented semi-automatically. Each reconstructed glenoid was divided first into anterior and posterior quadrants, and then further subdivided into four quadrants. Volumes of interest (VOI) were defined at depths of 0-2.5mm (Zone A), 2.5-5mm (Zone B) and 5-7.5mm (Zone C). Average bone densities were measured at each VOI depth and in each quadrant. Osteoarthritic glenoids had higher mean bone densities than N glenoids. Mean bone densities were uniform amongst all quadrants for N glenoids, but not for osteoarthritic glenoids. In A1 glenoids, the antero-superior quadrant was less dense in Zone C. A2 glenoids had increased bone density measured posteriorly in Zones B and C. In B1 and B2 glenoids, Zones B and C demonstrated increased bone densities of posterior quadrants compared to anterior quadrants. B3 glenoids presented similar results as A1 and A2 glenoids. Cystic changes were more pronounced in anterior quadrants of A2, B1, B2 and B3 glenoids. This study demonstrates that osteoarthritic glenoids have greater bone density than non-arthritic glenoids, independent of depth of interest. It also confirms that N glenoids have uniform erosion wear patterns and that B1 and B2 glenoids have irregular wear patterns. It is the first study to reveal that A1, A2 and B3 glenoids, though geometrically symmetrical, have irregular bony densities similar to B2 glenoids. These findings have clinical implications for reaming the glenoid and implant fixation. Basic Science, Anatomy, Imaging.


Multimodal evaluation of central and peripheral alterations in Stargardt disease: a pilot study.
The clinical phenotype of Stargardt disease (STGD) can be extremely heterogeneous, with variable macular and peripheral retinal involvement. The study aim was to correlate peripheral ultrawide field (UWF) involvement with macular alterations, as assessed by structural optical coherence tomography (OCT) and OCT angiography (OCTA), in order to identify potentially different phenotypes. The study involved patients with STGD and healthy controls. We performed a complete ophthalmologic assessment and multimodal imaging, including OCT, OCTA, fundus autofluorescence and UWF imaging. Patients with STGD were subdivided according to the peripheral involvement. OCT and OCTA quantitative parameters were analysed. The main outcome of the study was the classification of UWF subtypes and the correlation between UWF subtypes and macular involvement. Seventy STGD eyes (19 male; mean age 41.3±13.2 years) and 70 healthy eyes (35 male; 50%; mean age 41.2±9.8 years) were included in the analyses. Mean best-corrected visual acuity was 0.60±0.45 LogMAR for the STGD group and 0.0±0.0 LogMAR for controls (p<0.01). All clinical and imaging findings proved to be statistically worse in patients with STGD than in the control subjects (p<0.01). UWF types were distributed as follows: type I (49%), type II (34%), type III (17%). Type III patients proved to be significantly worse in terms of visual function and OCT and OCTA imaging parameters. The UWF autofluorescence performed in the present study suggests that there exist three different STGD phenotypes. Each phenotype is associated with variable OCT and OCTA impairment. Further studies providing a better assessment of the peripheral retinal involvement in STGD are warranted.


Molecular mobility and microscopic structure changes in κ-carrageenan solutions studied by gradient NMR.
Changes in the molecular mobility of κ-carrageenan were observed by the pulsed field gradient stimulated echo (PGSTE) and Carr-Purcell-Meiboom-Gill (CPMG) methods for elucidating the molecular aspect of the sol-to-gel transition. The echo signal intensity of κ-carrageenan without a gradient, Ikap(0), decreased steeply near the sol-to-gel temperature (Tsg), suggesting that κ-carrageenan chains formed aggregates and a network structure. Below Tsg, the spin-spin relaxation time T2 and the diffusion coefficient of κ-carrageenan (Dkap) increased with decreasing temperature, indicating that the solute κ-carrageenan chains have a lower molecular weight Mw than chains involved in the aggregation. The diffusion coefficient of pullulan (Dpul) added as a probe molecule in κ-carrageenan solutions was measured, and the characteristic hydrodynamic screening length, ξ, was then estimated from the degree of diffusion restriction. Below a certain temperature, Dkap reached a higher value than that of Dpul, suggesting that the Mw of solute κ-carrageenan became lower than that of pullulan. GPC measurements confirmed the presence of κ-carrageenan chains with a lower Mw than that of pullulan. A simple physical model of the structural change in κ-carrageenan solution was proposed with a bimodal distribution of κ-carrageenan with higher and lower Mw than the pullulan probe. The higher Mw chains form the gel network restricting the probe's diffusion, and the lower Mw chains increase the effective viscosity. The concentration of the high Mw solute κ-carrageenan chains in 1%, 2% and 4% κ-carrageenan solutions was estimated from Ikap(0) and the total κ-carrageenan concentration, and the relation with pullulan diffusion was studied.


Influence of albumin leakage on glycated albumin in patients with type 2 diabetes undergoing hemodialysis.
Glycated albumin (GA) is recommended as a better glycemic indicator than HbA1c in patients undergoing hemodialysis, because the red blood cell lifespan is generally faster than that in normal subjects. However, GA can be also affected by protein loss in urine and hemodialysis fluid. Therefore, in this study, we investigated the effect of albumin leakage induced by hemodialysis on GA. Nine patients undergoing hemodialysis with a large or small amount of albumin leakage were observed for 9 months in a crossover manner. As a result, it was shown that albumin leakage could affect GA, but the effect was practically small considering the prescription of diabetic drugs. The correlations between HbA1c and blood glucose levels and between GA and blood glucose levels were similar in our study. In conclusion, GA was a reliable indicator, even with the change of hemodialysis modality. The influence of albumin leakage induced by hemodialysis on GA was negligible practically. We should recognize that the preferable glycemic indicator in patients undergoing hemodialysis depends on the hemoglobin and albumin metabolism of each patient.


Contribution to the genus Pararrhynchium de Saussure (Hymenoptera: Vespidae: Eumeninae), with description of two new species from Vietnam.
Two new species of the genus Pararrhynchium de Saussure are described as new to science: P. simsanum Nguyen and Tran, sp. nov. from Bat Xat, Lao Cai province, and P. sparsum Nguyen and Ljubomirov, sp. nov. from Ham Yen, Tuyen Quang province. An updated key to all known species of the genus is also presented.


Correction to: Genome-wide characterization of the cellulose synthase gene superfamily in Pyrus bretschneideri and reveal its potential role in stone cell formation.
The above article was published online with the same Figures 7 and 8.


Purification and properties of phosphoserine aminotransferase from bovine liver.
L-Phosphoserine aminotransferase was purified from bovine liver to apparent homogeneity as judged by nondenaturing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, analytical ultracentrifugation, and immunochemical analysis. The purification procedure described involves the specific elution of the enzyme from Cibacron blue-agarose by micromolar concentrations of its substrate, phosphohydroxypyruvate. The purified enzyme had a specific activity of approximately 13 mumol of phosphohydroxypyruvate formed min-1 mg-1 of protein at 38 degrees C. Determinations of the native molecular weight and the subunit molecular weight indicated that the phosphoserine aminotransferase from bovine liver was a dimer composed of two subunits with identical molecular weights of 43,000.


Allosensitization in bridge to transplant Novacor left ventricular assist device patients: analysis of long-term outcomes with regard to acute rejection and chronic allograft vasculopathy.
The true relevance of allosensitization in patients benefiting from left ventricular assist device (LVAD) as bridge to transplant (BTT) is still debated. Available registry data referred to numerous devices precluding LVAD-specific analysis. Therefore, we studied all patients with Novacor LVAD prior to transplantation. From 1985 to 2006, 37 Novacor LVADs were implanted as BTT, with 30 patients surviving to transplantation (81%). Post-LVAD sensitization was determined for anti-HLA-class I and class II IgGs. Study endpoints were overall survival and/or graft loss, > or =3A cellular rejection and chronic allograft vasculopathy (CAV). The results from LVAD patients were compared to non-LVAD primary heart transplant recipients (n=318). After LVAD insertion, 5 out of 27 patients available for analysis developed anti-HLA antibodies (18.5%). The mean anti-HLA titer after Novacor LVAD implantation was 14% [SD 31]. Actuarial 5- and 10-year patient/graft survival for LVAD and non-LVAD transplant recipients were 73% and 55%, and 70% and 55%, respectively (p=NS). Overall prevalence of rejection > or =3A was 23.3 % (LVAD group) and 18.9% (non-LVAD group) (p=NS). At follow-up, the respective incidence of CAV was 8% (LVAD group) and 32.4% (non-LVAD group) (p<0.01). However, mean follow-up was significantly different for LVAD and non-LVAD patients, 46 vs 90 months (p<0.001). In this study, allosensitization occurred infrequently after Novacor LVAD implantation. Secondly, analysis of outcome variables shows that Novacor-LVAD BTT patients can anticipate similar survival to non-LVAD patients, thus minimizing the impact of allosensitization after LVAD implantation.


[Biomarkers in rheumatology. Biomarkers and imaging for the diagnosis and stratification of rheumatoid arthritis and spondylarthritis in the ArthroMark network funded by the Federal Ministry of Education and Research].
The introduction of biologics has continuously increased the demand for biomarkers for early diagnosis and therapeutic stratification. ArthroMark, a research network funded by the Federal Ministry of Education and Research, aims to establish such biomarkers for rheumatoid arthritis and spondyloarthritides. Biobanks and previous work on genotyping, gene expression and autoreactivity profiling build the basis. Bioinformatic networks will help to harmonize the investigations and a clinical study with modern imaging techniques to characterize the functional relevance of the new biomarkers as effectively as possible. To validate the markers for diagnostic application the network aims to expand gradually.


Predator-mediated interactions through changes in predator home range size can lead to local prey exclusion.
The strength of indirect biotic interactions is difficult to quantify in the wild and can alter community composition. To investigate whether the presence of a prey species affects the population growth rate of another prey species, we quantified predator-mediated interaction strength using a multi-prey mechanistic model of predation and a population matrix model. Models were parametrized using behavioural, demographic and experimental data from a vertebrate community that includes the arctic fox (Vulpes lagopus), a predator feeding on lemmings and eggs of various species such as sandpipers and geese. We show that the positive effects of the goose colony on sandpiper nesting success (due to reduction of search time for sandpiper nests) were outweighed by the negative effect of an increase in fox density. The fox numerical response was driven by changes in home range size. As a result, the net interaction from the presence of geese was negative and could lead to local exclusion of sandpipers. Our study provides a rare empirically based model that integrates mechanistic multi-species functional responses and behavioural processes underlying the predator numerical response. This is an important step forward in our ability to quantify the consequences of predation for community structure and dynamics.


Stereotactic Body Radiation Therapy (SBRT) for Spinal Metastases: Real-world Outcomes From an International Multi-institutional SBRT Registry.
The objective of this study was to compare clinical outcomes following single fraction versus fractionated stereotactic body radiotherapy (SBRT) for spinal metastases. A multi-institutional registry was queried for patients with spinal metastases treated with single-fraction or fractionated SBRT. Potential predictive factors of local control (LC) and overall survival were evaluated. Pretreatment and posttreatment Visual Analog Scale scores were analyzed to examine initial and durable pain responses and complete response (CR) rates. Logistic regression was utilized to assess potential correlations between pain response, biologically effective dose (BED), and fractionation. Four hundred sixty-six patients with 514 lesions treated with SBRT were identified; 209 and 104 lesions had information on LC and pain, respectively. The median pain score of patients with symptoms was 6 (range: 3 to 10). The median follow-up was 8.9 months (range: 0.4 to 125.5 mo). Utilizing Karnofsky Performance Score, age, and primary site (lung and/or nonbreast), 1-year overall survival rates were 76.1%, 59.1%, 54.9%, 37.2%, and 23.5% for patients with 0 to 4 of these factors, respectively (P<0.0001). One- and 2-year LC rates were 79.9% and 73.6%, respectively. Eighty-six patients (82.7%) had an initial pain response with a median decline of 3.5 and a CR rate of 47.1%. Sixty-five patients (62.5%) had a durable pain response with a median decline of 2 and a CR rate of 20.2%. Higher initial CR rates were observed with BED10 ≥51 Gy10 (58.7% vs. 37.9%; P=0.04). Following SBRT, encouraging palliative responses with >80% and 60% of patients having initial and durable pain responses, respectively. Dose escalation may result in improved initial CR rates. Performance status, age, and primary histology are factors to consider in the absence of pain.


Lectin histochemistry of mammalian Brunner's glands.
Lectin histochemical study was performed on twenty-eight specimens of formalin-fixed paraffin embedded tissues of proximal duodenum from human, cat, dog and Rhesus (macaque) monkey to demonstrate the pattern of carbohydrate residues in submucosal glands of Brunner as compared to that of the duodenal absorptive and goblet cells. Ten different biotinylated lectins were used as probes, and avidin-biotin-peroxidase (ABC) or avidin-gold-silver (AGS) complexes were used as "visualants". Brunner's gland cells of the four species studied exhibited a similar lectin-binding pattern which differ from other duodenal cells. The epithelium of Brunner's gland stained intensely with Ricinus communis agglutinin-I (RCA-I), succinylated-WGA (S-WGA) and wheat-germ agglutinin (WGA), moderately with Bandeirea simplicifolia agglutinin-I (BS-I), Concanavalia ensiformis agglutinin (Con A) peanut agglutinin (PNA) and Ulex europaeus agglutinin-I (UEA-I) and occasionally with Dolichos biflorus agglutinin (DBA), Lens culinaris agglutinin (LCA) and soybean agglutinin (SBA). Desialylation with neuraminidase resulted in only a slight elevation in binding intensities of PNA, DBA and SBA, indicating that glycoconjugates of the Brunner's gland cells are rich in asialo-oligosaccharides, which differs from duodenal epithelial cells. In addition, these histochemical reagents were useful in localizing Brunner's gland elements in the duodenal mucosa.


Genotoxic and inflammatory effects of organic extracts from traffic-related particulate matter in human lung epithelial A549 cells: the role of quinones.
Traffic-related particulate matter (PM) is associated with adverse health effects. Quinones present in the traffic-related PM are hypothesized to contribute to these harmful effects through reactive oxygen species (ROS) generation. However, the impacts of the traffic-related PM and quinones on inflammatory processes and genotoxic damages are less well known. In present study we aimed to examine the genotoxic and inflammatory impacts of organic extracts from traffic-related PM (oTRP) in human lung epithelial A549 cells, and reveal the contributions from quinones. Significant cytotoxicity and DNA damage were caused by oTRP. The pro-inflammatory genes, interleukin-6 (Il-6), interleukin-8 (Il-8) and tumor necrosis factor (Tnf), and two aromatic hydrocarbon receptor-regulated genes, Cyp1a1 and 1b1, were significantly up-regulated by oTRP. A concomitant increase in ROS was observed, suggesting that oTRP may mediate genotoxic and inflammatory effects through oxidative stress pathway. Second, the effects from two typical airborne quinones, 9,10-anthraquinone (AQ) and 1,4-naphthroquinone (NQ) were compared. NQ, but not AQ, induced significant DNA damage in A549 cells. NQ up-regulated Il-8, Tnf, and Mcp-1 genes, while AQ induced the expression of Rantes gene. These results suggest that the NQ and AQ may participate in the pro-inflammatory responses through releasing different types of cytokines/chemokines.


Labour productivity loss caused by premature deaths associated with breast cancer: results from Spain over a 10-year period.
Breast cancer is one of the most common cancers among women worldwide, and is the most common cause of cancer deaths in them. Given that such disease has a significant impact on middle-aged women, it can result in considerable labour productivity losses. The main objective of this study is therefore to analyse labour productivity losses and years of potential working life lost as a result of premature deaths from breast cancer in Spain over a 10-year period (2005-2014). To this end, by combining data on average salary (Salary Structure Survey), occupation (Active Population Survey) and deaths (death registration according to cause of death), a simulation model based on the human capital approach was constructed to estimate productivity losses associated with premature deaths caused by this disease. The results reveal that labour productivity losses in women are estimated to cost 2,137 million euros, as a result of the 22,716 deaths occurring during working age and the 292,848 years of potential productive life lost. This implies that breast cancer is responsible for 10.45% of all estimated female labour productivity losses caused by premature deaths in the reference year. The article concludes by highlighting the major economic impact associated with breast cancer deaths and by pointing out future research areas.


Clustering patients by depression symptoms to predict venlafaxine ER antidepressant efficacy: Individual patient data analysis.
To identify clusters of patients with major depressive disorder (MDD) based on the baseline 17-item Hamilton Rating Scale for Depression (HAM-D17) items and to evaluate the efficacy of venlafaxine extended release (VEN) vs placebo, and the potential effect of dose on efficacy, in each cluster. Cluster analysis was performed to identify clusters based on standardized HAM-D17 item scores of individual patient data at baseline from 9 double-blind, placebo-controlled studies of VEN for MDD. Change from baseline in HAM-D17 total score was analyzed using a mixed-effects model for repeated measures for each cluster; response and remission rates at week 8 were analyzed using logistic regression. Discontinuation rates were also evaluated in each cluster. In 2599 patients, 3 patient clusters were identified, characterized as High modified Core (mCore) Symptoms/High Anxiety (cluster 1), High mCore Symptoms/Medium Anxiety (cluster 2), and Medium mCore Symptoms/Medium Anxiety (cluster 3). Significant effects of VEN vs placebo were observed on change from baseline in HAM-D17 total score at week 8 for both clusters 1 and 2 (both P < 0.001), but not for cluster 3. In cluster 3, a significant treatment effect of VEN was observed at week 8 in the lower-dose subgroup but not in the higher-dose subgroup. All-cause discontinuation rates were significantly higher in placebo than VEN in each cluster. Three unique clusters of patients were identified differing in baseline mCore symptoms and anxiety. Cluster membership may predict efficacy outcomes and contribute to dose effects in patients treated with VEN. NCT01441440; other studies included in this analysis were conducted before the requirement to register clinical studies took effect.


Genetic studies on human thyroxine-binding globulin (TBG).
An enzyme immunoassay technique combined with Western blotting is described to demonstrate thyroxine-binding globulin (TBG) by isoelectric focusing in thin-layer polyacrylamide gels with 8 mol/l urea. Quantitative evaluation was by laser densitometry. No genetic charge variants of TBG were encountered in a sample of 840 unrelated individuals from southwestern Germany. There was no correlation between structural and quantitative variations in the TBG protein. Results from a family with quantitative TBG deficiency strongly support the postulated X-linked mode of inheritance. The method described can be considered as an additional diagnostic tool in thyroid evaluation.


Heritability and Reversibility of DNA Methylation Induced by in vitro Grafting between Brassica juncea and B. oleracea.
Grafting between tuber mustard and red cabbage produced a chimeric shoot apical meristem (SAM) of TTC, consisting of Layers I and II from Tuber mustard and Layer III from red Cabbage. Phenotypic variations, which mainly showed in leaf shape and SAM, were observed in selfed progenies GSn (GS = grafting-selfing, n = generations) of TTC. Here the heritability of phenotypic variation and its association with DNA methylation changes in GSn were investigated. Variation in leaf shape was found to be stably inherited to GS5, but SAM variation reverted over generations. Subsequent measurement of DNA methylation in GS1 revealed 5.29-6.59% methylation changes compared with tuber mustard (TTT), and 31.58% of these changes were stably transmitted to GS5, but the remainder reverted to the original status over generations, suggesting grafting-induced DNA methylation changes could be both heritable and reversible. Sequence analysis of differentially methylated fragments (DMFs) revealed methylation mainly changed within transposons and exon regions, which further affected the expression of genes, including flowering time- and gibberellin response-related genes. Interestingly, DMFs could match differentially expressed siRNA of GS1, GS3 and GS5, indicating that grafting-induced DNA methylation could be directed by siRNA changes. These results suggest grafting-induced DNA methylation may contribute to phenotypic variations induced by grafting.


Endocrine aspects and tumoral markers in intracranial germinoma: an attempt to delineate the diagnostic procedure in 14 patients.
This work outlines the endocrine abnormalities associated with intracranial germinomas (14 patients before treatment). Diabetes insipidus of various intensity was present in all cases. Adipsia is often present as well. In six patients, the first neuroradiologic examination after the onset of DI was normal. Growth hormone secretion was deficient in 11 of 13 patients. Abnormalities of TSH response to TRH were present in nine of ten (four insufficient and five exaggerated responses to TRH). Anterior pituitary dysfunction could not be predicted by the tumor site as determined by radiologic criteria. Tumoral markers have also been studied. Elevated plasma tumoral markers were found in four patients of 11 studied. Tumoral germinoma cells were present in CSF in five patients of ten, in one of them before radiologic confirmation. Pertinent endocrine evaluation and search for tumoral markers was of great value in systematic follow-up of patients with central diabetes insipidus, and could lead to early diagnosis and treatment of the tumor.


Does de novo synthesis of lysophosphatidylcholine occur in rat lung microsomes?
Incubation of rat lung microsomes with CDP[Me-14C]choline resulted in formation of radioactive lysophosphatidylcholine and phosphatidylcholine. Evidence is provided which suggests that lysophosphatidylcholine formation cannot be ascribed completely to phospholipase. A degradation of phosphatidylcholine. Lysophosphatidylcholine production can be stimulated by addition of monoacylglycerol or diacylglycerol. It is suggested that diacylglycerol is partly hydrolyzed to monoacylglycerol and subsequently converted to lysophosphatidylcholine. A direct transfer of phosphocholine from CDPcholine to monoacylglycerol is demonstrated by equimolar incorporation of 1(3)-[9,10-3H2]palmitoylglycerol and phospho[Me-14C]choline into lysophosphatidylcholine.


Integrated network pharmacology and intestinal flora analysis to determine the protective effect of Xuanbai-Chengqi decoction on lung and gut injuries in influenza virus-infected mice.
Xuanbai-Chengqi decoction (XBCQ) is a traditional Chinese medicine (TCM) compound used in the treatment of pulmonary infection in China. Despite the popular usage of XBCQ, its underlying protective roles and the associated molecular mechanisms with the gut-lung axis in influenza remain unclear. We aimed to explore the protective effects and the underlying mechanism of XBCQ efficacy on lung and intestine injuries induced by influenza A virus as well as to identify the main active components through integrated network pharmacology, intestinal flora analysis and pathway validation. The potential active components and therapeutic targets of XBCQ in the treatment of influenza were hypothesized through a series of network pharmacological strategies, including components screening, targets prediction and bioinformatics analysis. Inflammatory cytokines and pathway proteins were assayed to validate the results of network pharmacology. Then the mechanism of XBCQ alleviating lung and intestine injuries was further explored via intestinal flora analysis. The important role of Rhubarb in the formula was verified by removing Rhubarb. XBCQ could significantly improve the survival rate in IAV-infected mice. The network pharmacology results demonstrated that JUN, mitogen-activated protein kinase (MAPK), and tumor necrosis factor (TNF) are the key targets of XBCQ that can be useful in influenza treatment as it contains the core components luteolin, emodin, and aloe-emodin, which are related to the pathways of TNF, T-cell receptor (TCR), and NF-κB. Verification experiments demonstrated that XBCQ could significantly alleviate the immune injury of the lungs and the gut of the mice, which is attributable to the inhibition of the release of inflammatory cytokines (such as TNF-α, IL-6, and IL-1β), the downregulation of the protein expression levels of Toll-like receptors-7 (TLR7), MyD88, and p-NF-κB65, and the reduction in the relative abundance of Enterobacteriaceae and Proteus, while an increase in that of Firmicutes and Lachnospiraceae. The overall protective role of XBCQ contributing to the treatment of the lungs and the gut was impaired when Rhubarb was removed from XBCQ. Our results suggest that the efficacy of XBCQ is related to the inhibition of the immune injury and remodeling of the intestinal flora, wherein Rhubarb plays an important role, which cumulatively provide the evidence applicable for the treatment of viral pneumonia induced by a different respiratory virus with XBCQ.


Dissecting a complex neurosurgical illustration: step-by-step development.
Modern computer graphics software has enabled the medical illustrator to render very complex anatomy by composing many different layers of drawings simultaneously. This and the author's capacity to take an "editorial" approach to compress several chronological events into a single, comprehensive two-dimensional illustration are analyzed in a step-by-step process. Through a series of images, the article provides a visual synopsis of the development of an illustration for an extensive clinical case: total sacrectomy performed through an all-posterior approach. Originally given as a slide presentation at the American Association of Neurological Surgeons Theodore Kurze Lecture in April 2011, the article provides some detailed notes on the techniques the author used to develop a comprehensive neurosurgical illustration.


Uncovering targeting priority to yeast peroxisomes using an in-cell competition assay.
Approximately half of eukaryotic proteins reside in organelles. To reach their correct destination, such proteins harbor targeting signals recognized by dedicated targeting pathways. It has been shown that differences in targeting signals alter the efficiency in which proteins are recognized and targeted. Since multiple proteins compete for any single pathway, such differences can affect the priority for which a protein is catered. However, to date the entire repertoire of proteins with targeting priority, and the mechanisms underlying it, have not been explored for any pathway. Here we developed a systematic tool to study targeting priority and used the Pex5-mediated targeting to yeast peroxisomes as a model. We titrated Pex5 out by expressing high levels of a Pex5-cargo protein and examined how the localization of each peroxisomal protein is affected. We found that while most known Pex5 cargo proteins were outcompeted, several cargo proteins were not affected, implying that they have high targeting priority. This priority group was dependent on metabolic conditions. We dissected the mechanism of priority for these proteins and suggest that targeting priority is governed by different parameters, including binding affinity of the targeting signal to the cargo factor, the number of binding interfaces to the cargo factor, and more. This approach can be modified to study targeting priority in various organelles, cell types, and organisms.


Generation of a PDX1-EGFP reporter human induced pluripotent stem cell line, KSCBi005-A-3, using the CRISPR/Cas9 system.
PDX1 plays a crucial role in the development and maintenance of β-cells and directly regulates pancreatic β-cell-specific transcription factors by binding to the insulin gene. Here, we introduced an EGFP reporter into the C-terminus of PDX1 in KSCBi005-A human induced pluripotent stem cells through homologous recombination using CRISPR/Cas9 nuclease. The cells had a normal karyotype, expressed several pluripotency markers, and maintained their differentiation potential. KSCBi005-A-3 cells can be used to monitor PDX1 expression in live cells during β-cell differentiation; the cell line has been registered at the National Stem Cell Bank, Korea National Institute of Health.


Severe impairment in adenine metabolism with a partial deficiency of adenine phosphoribosyltransferase.
Among three unrelated patients with recurrent 2,8-dihydroxyadenine urolithiasis, two completely lacked adenine phosphoribosyltransferase (APRT) in both erythrocytes and proliferative T cells. The third patient possessed significant enzyme activities in both hemolysates and T-cell extracts at levels comparable to heterozygotes for complete APRT deficiency. Despite significant APRT activities in cell extracts, cultured T cells from the third patient were at least 100-fold more resistant than normal T cells to an adenine analog, 6-methylpurine, whose cytotoxicity is dependent on APRT. These data indicate that APRT activity in T cells from the third patient is positive in cell extracts, but apparently not operating in viable cells. Although the cells from the patients with complete APRT deficiency were as resistant to 6-methylpurine as the cells from the third patient, the cells from the heterozygotes for complete APRT deficiency were almost as sensitive as normal T cells. Therefore, adenine metabolism in the third patient but not in the heterozygotes seems to be as severely impaired as in the patients with complete APRT deficiency, which is quite consistent with the clinical manifestations in these individuals.


Qadirvirtide.
Qadirvirtide is a fusion inhibitor that may be used as prophylaxis or for the treatment of AIDS. It is a synthetic peptide that is composed of 36 amino acids. Qadirvirtide blocks the entry of HIV genome into human CD4 cells by binding to HR1 as the virus can not come close to the human cell membrane and ultimately fusion of the viral envelope with human cell membrane is prohibited.


A linear regime of hysteresis for calculating the dynamic contact angle under low capillary numbers with displacement experiments in microscale PDMS microchannels.
Determining capillary pressures and permeation resistances of multiphase seepage in porous-type reservoirs is crucial. The dynamic contact angle (CA) is one of the critical parameters used to calculate capillary pressures and permeation resistances. Under reservoir conditions, dynamic CAs are rate-independent of the triple-phase contact line velocity. Calculating rate-independent dynamic CAs remains challenging and is the main focus of this paper. An experimental system was designed to capture the dynamic CAs of liquid-fluid displacements in microscale polydimethylsiloxane (PDMS) microchannels. These microchannels were 20 × 80 μm or 20 × 40 μm in height and width. The capillary numbers (Ca) were controlled to satisfy the conditions of Ca < 4 × 10-5 for liquid-air displacements and Ca < 3.5 × 10-7 for liquid-liquid displacements. Since pinning forces were consistent at the receding interface and the advancing interface, energy barriers exhibited symmetry. However, CA hysteresis exhibited asymmetry. Based on our experiments, a linear regime of hysteresis was developed and verified to be consistent with others' experiments. The relation can be used to calculate hysteresis strengths and rate-independent CAs. This model was also compared with the fractal model. The hysteresis factor in this model can be derived using the Wenzel factor r and the Cassie fraction ϕs.


[Multiple heart valve diseases: differential therapeutic considerations].
After more than 40 years of heart valve surgery, the indication for double valve procedures still represents a problem in the presence of an obviously leading single valve disease. If the isolated valves do not already represent a clear indication for surgery then this situation is the best example for a good deal of thought in cardiac surgery and discussion with competent cardiologists.


Are negative symptoms really related to cognition in schizophrenia?
Previous studies have generally found a relationship between negative and cognitive symptoms in schizophrenia. The present study investigated the relationship between the 5 PANSS factors of a recent consensus model developed by NIMH researchers, and cognitive performance as assessed with the MATRICS Consensus Cognitive Battery (MCCB) in 80 patients with schizophrenia using correlation and regression analyses. The PANSS Cognitive factor showed a small to moderate significant association with MCCB Speed of processing, Working memory, Verbal learning, the Neurocognitive composite score, and the Overall composite score. Notably, however, no relationship was found between the PANSS Negative factor and any of the MCCB scores. The Positive, Excited and Depressed factors also did not show associations with the MCCB. These results highlight the need for refined assessment instruments and support the relative independence of cognition from other domains of psychopathology, including negative symptoms, in patients with schizophrenia.