Polarisation-sensitive OCT is useful for evaluating retinal pigment epithelial lesions in patients with neovascular AMD.
To examine the reproducibility of lesion dimensions of the retinal pigment epithelium (RPE) in neovascular age-related macular degeneration (AMD) with polarisation-sensitive optical coherence tomography (PS-OCT), specifically imaging the RPE. Twenty-six patients (28 eyes) with neovascular AMD were included in this study, and examined by a PS-OCT prototype. Each patient was scanned five times at a 1-day visit. The PS-OCT B-scan located closest to the macular centre presenting with RPE atrophy was identified, and the longitudinal diameter of the lesion was quantified manually using AutoCAD 2008. This procedure was followed for the identical B-scan position in all five scans per eye and patient. Reproducibility of qualitative changes in PS-OCT was evaluated. Interobserver variability was assessed. Results were compared with intensity-based spectral-domain OCT (SD-OCT) imaging. Mean variability of all atrophy lesion dimensions was 0.10 mm (SD±=0.06 mm). Coefficient of variation (SD±/mean) was 0.06 on average (SD±=0.03). Interobserver variability assessment showed a mean difference of 0.02 mm across all patients regarding RPE lesion size evaluation (paired t test: p=0.38). Spearman correlation coefficient was r=0.98, p<0.001. Results revealed a good overall reproducibility of ∼90%. PS-OCT specifically detected the RPE in all eyes compared with conventional intensity-based SD-OCT that was not capable to clearly identify RPE atrophy in 25 eyes (89.3%, p<0.01). PS-OCT offers good reproducibility of RPE atrophy assessment in neovascular AMD, and may be suitable for precise RPE evaluation in clinical practice. PS-OCT unambiguously identifies RPE changes in choroidal neovascularisation compared with intensity-based SD-OCT that does not identify the RPE status reliably.


Triggering Electron Transfer in Co(I) Dimers: Computational Evidences for a Reversible Disproportionation Mechanism.
An inner-sphere disproportionation mechanism of the Co(I) precursor CoCl(PPh3 )3 is described through a Density Functional Theory study. The essential role of oleylamine in this process is unravelled. A detailed analysis of the electronic structure of Cobalt dimers of the general formula Co2 Cl2 Ln (L=NH3 and PH3 ) demonstrates that electron transfer is triggered by asymetric coordination of amine and phosphine to stabilize a mixed-valence Co(II)-Co(0) dimer. This is consistent with the HSAB statement that both amine and phosphine ligands are required to stabilize the reaction products, respectively Co(II) and Co(0) centers. We propose a quasi-athermic multi-step disproportionation mechanism with low activation barriers where the electron transfer goes through simple ligand exchanges between Co.


Systematic review and meta-analysis of the effect of using percutaneous pedicle screw internal fixation for thoracolumbar fractures.
Thoracolumbar fractures are common in routine injuries. The best way to treat a thoracolumbar fracture is to repair the thoracic fracture with fixation, which is classified into percutaneous pedicle screw fixation and open pedicle screw fixation. The literature was searched in the English database PubMed, Ovid-Medline, Embase, and China Biology Medicine Disc (CBM) from the date of establishment of PubMed to June 2021, and keywords such as percutaneous pedicle screw and posterior percutaneous pedicle screw fixation were searched. A meta-analysis was performed using RevMan5.3, which was provided by Cochrane.com. A total of 9 articles were included. The results showed that the operative time of percutaneous pedicle screw fixation was shorter than that of open pedicle screw fixation. Percutaneous pedicle screw internal fixation had better Cobb angle restoration effect than open pedicle screw internal fixation. However, there were considerable differences in pain and screw misplacement rates. In addition, the pain after percutaneous pedicle screw fixation was smaller than that after open pedicle screw fixation, and patients had better compliance. Compared with open surgery, percutaneous pedicle screw fixation had a shorter operation time, better cobb angle recovery, and a lower pain sensation following surgery. The low rate of screw misplacement and postoperative infection suggested that percutaneous pedicle screw fixation was more effective than open surgery.


Microarray analysis of polychlorinated biphenyl mixture-induced changes in gene expression among Atlantic tomcod populations displaying differential sensitivity to halogenated aromatic hydrocarbons.
Several populations of fishes inhabiting contaminated Atlantic Coast estuaries exhibit resistance to early life-stage (ELS) toxicities induced by halogenated aromatic hydrocarbons such as coplanar polychlorinated biphenyls (PCBs). These toxicities include mortality, circulatory failure, edema, and craniofacial malformations. The mechanisms behind resistance to halogenated aromatic hydrocarbon toxicity in these populations are unknown. First and second generation Atlantic tomcod Microgadus tomcod embryos derived from the Hudson River ([HR]; New York, USA) population are highly resistant to PCB-induced cytochrome P4501A (CYP1A) expression and ELS toxicity when compared to embryos of Miramichi River ([MR]; New Brunswick, Canada) and Shinnecock Bay ([SB]; New York, USA) origin. The present study sought to identify novel genes involved in population differences in response to PCB exposure using custom microarrays. Microarray probes consisted of unsequenced inserts of randomly picked clones from a tomcod cardiac cDNA library. Tomcod embryos from three populations (HR, MR, and SB) were exposed to two doses of an environmentally relevant mixture of coplanar PCBs and screened for dose- and population-specific patterns of gene expression. Clones displaying significant differences between populations exposed to the high dose of PCBs were identified by DNA sequencing. Of the 28 identified nonribosomal protein clones, none displayed expression patterns highly similar to CYP1A (altered in MR and SB, but not in HR). However, several transcripts representing biomarkers of cardiomyopathy in mammals (cardiac troponin T2, cathepsin L, and atrial natriuretic peptide) were differentially altered among the three tomcod populations by PCBs. Although the present study did not identify any novel genes associated with PCB resistance in tomcod, several potential molecular biomarkers of PCB exposure were revealed.


Hepatocyte heterogeneity in uptake and metabolism of malate and related dicarboxylates in perfused rat liver.
1. In isolated perfused rat liver a near-maximal net malate uptake of about 120 nmol g-1 min-1 was observed at influent malate concentrations above 100 mumol l-1 and a half-maximal uptake at about 50 mumol l-1 in influent. 14CO2 production from added [U-14C]malate paralleled hepatic net malate uptake, however, 14CO2 production exceeded net malate uptake by 20-25%. This was observed in antegrade as well as in retrograde perfusions and regardless of whether NH4Cl was added to the influent perfusate. Stimulation of glutamine synthesis by NH4Cl only slightly affected net malate uptake and 14CO2 production, but resulted in a marked stimulation of [14C]glutamine release from the liver. 2. Because [U-14C]malate uptake by the liver (reflecting the influent/effluent concentration difference of labeled malate) could at least in part involve a malate/malate exchange mechanism, net malate uptake (as determined from the influent/effluent concentration difference of enzymatically assayable malate) may underestimate hepatic [U-14C]malate uptake. On the other hand, during metabolic steady states 14CO2 production from added [U-14C]malate can be considered as an upper limit estimate of [U-14C]malate uptake by the liver. Assuming that 14CO2 production equals [U-14C]malate uptake by the liver, extrapolation studies suggest that during maximal rates of NH4Cl-stimulated glutamine synthesis 80-110% of the [U-14]malate taken up by the liver was used for glutamine synthesis. This was true for retrograde and antegrade perfusions. Similar data, i.e. a 100-130% incorporation regardless of the direction of perfusion, were obtained when [U-14C]malate uptake was assumed to equal net malate uptake by the liver. 3. Substitution of Na+ in the perfusion fluid by choline abolished net malate uptake by the liver and inhibited 14CO2 production from [U-14C]malate by more than 90%. 4. 2-Oxoglutarate inhibited [14C]malate uptake and [1-14C]oxoglutarate uptake by the liver was inhibited by malate, fumarate, succinate and oxaloacetate, but not by aspartate and glutamate. Inhibition of [1-14C]oxoglutarate uptake and of 14CO2 production from added labeled 2-oxoglutarate by malate and fumarate seemed largely competitive. Malate, fumarate and succinate not only inhibited [1-14C]oxoglutarate uptake, but also stimulated the release of unlabeled 2-oxoglutarate from the liver. 5. The data are consistent with a predominant uptake of vascular malate by perivenous glutamine synthetase containing hepatocytes when glutamine synthesis is stimulated to Vmax values by NH4Cl. Malate and other citric acid cycle dicarboxylates, but not aspartate and glutamate, may compete with 2-oxoglutarate for uptake into perivenous glutamine synthesizing hepatocytes.(ABSTRACT TRUNCATED AT 400 WORDS)


Analysis of phosphate and phosphate containing headgroups enzymatically cleaved from phospholipids of Bacillus subtilis by capillary electrophoresis.
A new, fast, selective, and reliable capillary electrophoresis method has been developed for analysis of selected phosphoesters (phosphoserine, phosphoethanolamine, phosphoglycerol) and phosphate. The method is based on separation of specific phosphate containing headgroups (phosphoesters) which are cleaved from the glycerol skeleton of a phospholipid by a regioselective enzyme (phospholipase C). Analysis of intact phospholipids with the same polar headgroup but different fatty acids shows that fatty acid composition has a high impact on separation of phospholipids, so analysis of separated polar headgroups, which avoids this influence, represents a much more suitable approach for phospholipid class research. Optimization of method parameters results in running buffers of relatively narrow pH interval (pH about 10) where all phosphoesters are separated. Further method validation has shown that direct UV detection has a sufficient detection limit for all analytes to perform suitable analyses of cell membrane lipids. The optimized method was tested on the lysate of cell membrane of Bacillus subtilis, where all analytes were determined.


Screening for Cognitive Impairment by Model-Assisted Cerebral Blood Flow Estimation.
Alzheimer's disease (AD) is a progressive and debilitating neurodegenerative disease; a major health concern in the ageing population with an estimated prevalence of 46 million dementia cases worldwide. Early diagnosis is therefore crucial so mitigating treatments can be initiated at an early stage. Cerebral hypoperfusion has been linked with blood-brain barrier dysfunction in the early stages of AD, and screening for chronic cerebral hypoperfusion in individuals has been proposed for improving the early diagnosis of AD. However, ambulatory measurements of cerebral blood flow are not routinely carried out in the clinical setting. In this study, we combine physiological modeling with Holter blood pressure monitoring and carotid ultrasound imaging to predict 24-h cerebral blood flow (CBF) profiles in individuals. One hundred and three participants [53 with mild cognitive impairment (MCI) and 50 healthy controls] underwent model-assisted prediction of 24-h CBF. Model-predicted CBF and neuropsychological tests were features in lasso regression models for MCI diagnosis. A CBF-enhanced classifier for diagnosing MCI performed better, area-under-the-curve (AUC) = 0.889 (95%-CI: 0.800 to 0.978), than a classifier based only on the neuropsychological test scores, AUC = 0.818 (95%-CI: 0.643 to 0.992). An additional cohort of 25 participants (11 MCI and 14 healthy) was recruited to perform model validation by arterial spin-labeling magnetic resonance imaging, and to establish a link between measured CBF that predicted by the model. Ultrasound imaging and ambulatory blood pressure measurements enhanced with physiological modeling can improve MCI diagnosis accuracy.


Maternal caregiving and strategies used by inexperienced mothers of young infants with complex health conditions.
To describe maternal caregiving and related strategies used by first-time mothers of young infants with complex health conditions (CHC) in the first 6 months after discharge. Grounded theory. Data were collected in participants' homes in the Northeast United States. Eight first-time mothers of infants age 6 months or younger with CHC. Purposive and theoretical sampling were used. Semistructured interviews were completed at 2-month intervals, beginning 2 weeks after their infants' discharge. Analysis of 28 interviews was done with the constant comparative method. A grounded theory of maternal caregiving was conceptualized from the data. This time-and-experience-mediated process involved three phases of increasing confidence and expertise, developing in the context of decision-making responsibility. Related maternal strategies included appraising, normalizing, organizing, assessing, practicing, validating, experimenting, nurturing, and negotiating. Mothering became predictable and integrated in everyday life by about 6 months after the infant's discharge home. Findings can help clinicians and researchers better understand what happens over time as new mothers care for infants with CHC. Exploration of these patterns in a more diverse group of mothers of children with CHC can support the development of targeted interventions for this specialized population.


MRI findings in bucket-handle tears of the triangular fibrocartilage complex.
The triangular fibrocartilage complex (TFCC) is an intricate ligamentous and cartilaginous structure that helps transmit axial load across the wrist, and provide stability to the ulnocarpal and distal radioulnar joints (DRUJ). Because the blood supply to the TFCC varies depending on location, certain types of tears are more amenable to surgical repair than others. Since Palmer proposed his classification system of TFCC tears in 1989, only 1 case of a "bucket-handle" type tear has been reported. In this article, we describe two new cases of bucket-handle tears of the TFCC. In both cases, the torn fragment was displaced into a previously undescribed location (intra-articular DRUJ and prestyloid recess). Because this type of injury pattern has not been previously well characterized in the literature and such cases rarely reported, MRI findings have not been fully described and its implications on clinical management have largely yet to be determined.


The efficacy of montelukast monotherapy in moderate persistent asthmatic children.
Although current guidelines place single leukotriene receptor antagonists use as a therapeutic option in mild persistent asthma, there is still uncertainty about its place. In this study efficiency of montelukast monotherapy in children with moderate persistent asthma was evaluated. Children (age 6 to 16) with persistent moderate asthma were treated prospectively with budesonide combined with montelukast (n=20), or only montelukast (n=15) during six months. Asthma symptoms and exacerbations were obtained from diary data. This study suggests that both treatments were effective and well tolerated. It could be concluded that sole montelukast treatment in children with moderate persistent asthma is effective.


A method for the synthesis of substituted quinolines via electrophilic cyclization of 1-azido-2-(2-propynyl)benzene.
A new and efficient strategy for the synthesis of substituted quinolines via electrophilic cyclization is developed. The intramolecular cyclization of 1-azido-2-(2-propynyl)benzene 1 proceeds smoothly in the presence of electrophilic reagents (I(2), Br(2), ICl, NBS, NIS, and HNTf(2)) in CH(3)NO(2) at room temperature or in the presence of catalytic amounts of AuCl(3)/AgNTf(2) in THF at 100 degrees C to afford the corresponding quinolines 2 in good to high yields. In the case of the electrophilic reagents, E of 2 is either I, Br, or H, depending on the reagent type, while E of 2 is H in the case of the electrophilic catalyst.


A radioreceptor assay in which iodocyanopindolol is used to determine propranolol and its active metabolites in unextracted serum or plasma.
This rapid, sensitive, simple radioreceptor assay (RRA) for l-propranolol and its active metabolites in unextracted samples requires 5 microL of sample, a beta-adrenergic antagonist, 125-l-labeled (-)cyanopindolol (125ICYP), and turkey erythrocyte membrane receptors (Kd = 40 pmol/L). Equal volumes (100 microL) of diluted sample and 125ICYP are incubated with 500 microL of erythrocyte membranes for 30 min. Cold isotonic saline (2.5 mL) is added, and the mixture is centrifuged. The concentration of drug that inhibited receptor binding by 50% (IC50) was l-propranolol, 1.5 micrograms/L; d-propranolol, 243 micrograms/L; and 4-hydroxypropranolol, 8.8 micrograms/L. Analytical recovery of propranolol added to serum was 93 to 120%; results for 138 clinical samples tested by this method and by liquid chromatography correlated well (r = 0.96). Results correlated strongly (r = 0.98) for 23 clinical samples tested by RRA and analyzed for both propranolol and 4-hydroxypropranolol by liquid chromatography. The sensitivity of this RRA was 0.3 micrograms/L, and both intra- and interassay CVs were less than 12%. Modifications of this method could permit testing of other nonselective beta-blockers.


Inhibition of leukotriene C4 and B4 release by human eosinophils with the new 5-lipoxygenase inhibitor 6-hydroxy-2(4-sulfamoylbenzylamino)-4,5,7-trimethylbenzothiazo le hydrochloride.
Eosinophils generate and release leukotrienes C4 and B4 (LTC4, LTB4) and platelet activating factor (PAF), all of which have the capacity to cause inflammation and tissue injury in the airways. This study has examined the effects of a new 5-lipoxygenase inhibitor, 6-hydroxy-2-(4-sulfamoylbenzylamino)-4,5,7-trimethylbenzothiazo le hydrochloride (CAS 120164-49-0, E6080) on the release of LTC4, LTB4 and PAF by human eosinophils, Eosinophils stimulated by 1 mumol/l calcium ionophore A23187 for 15 min released 37.5 +/- 2.2 ng, 2.3 +/- 0.3 ng and 4.0 +/- 0.3 pmol per 10(6) cells of immunoreactive LTC4, LTB4 and PAF, respectively (mean +/- SEM, n = 4). LTC4 and LTB4 releases were inhibited dose-dependently by the addition of E6080 to the cell suspension. The IC50 values were 0.26 mumol/l for LTC4 and 0.23 mumol/l for LTB4. PAF release was not inhibited. These results suggest that E6080 is a potent inhibitor of LTC4 and LTB4 release from eosinophils and may provide a protective effect against bronchoconstriction during late-phase asthmatic responses.


Assistive technology and handwriting problems: what do occupational therapists recommend?
Handwriting difficulties for students are a common reason for referral to occupational therapy. Little research evidence is available concerning the factors guiding technology recommendations for these children. The objective of this survey research was to describe the technology-related recommendations and factors involved in the decisions made by Canadian occupational therapists for these students. More therapists recommended the use of keyboard-based strategies (93%) than dictation-based strategies (72%). Experienced therapists were more likely to prescribe technology tools. Dictation to a scribe (93%) and desktop computers (89%) were the strategies most frequently recommended. Equipment cost and availability of funding, and the availability of support in the school for the student were the most influential factors, respectively, on the keyboard and dictation strategy type prescribed. The results confirmed that occupational therapists prescribe a range of technology solutions. Factors influencing these recommendations differ depending on the nature of the technology, the person, environment or occupation. Knowing the factors guiding occupational therapist technology recommendations will help provide valuable information about the practical implications of the available technologies.


Dynamic tensile failure mechanics of the musculoskeletal neck using a cadaver model.
Although the catapult phase of pilot ejections has been well characterized in terms of human response to compressive forces, the effect of the forces on the human body during the ensuing ejection phases (including windblast and parachute opening shock) has not been thoroughly investigated. Both windblast and parachute opening shock have been shown to induce dynamic tensile forces in the human cervical spine. However, the human tolerance to such loading is not well known. Therefore, the main objective of this research project was to measure human tensile neck failure mechanics to provide data for computational modeling, anthropometric test device development, and improved tensile injury criteria. Twelve human cadaver specimens, including four females and eight males with a mean age of 50.1+/-9 years, were subjected to dynamic tensile loading through the musculoskeletal neck until failure occurred. Failure load, failure strain, and tensile stiffness were measured and correlated with injury type and location. The mean failure load for the 12 specimens was 3100+/-645 N, mean failure strain was 16.7+/-5.4%, and mean tensile stiffness was 172+/-54.5 N/mm. The majority of injuries (8) occurred in the upper cervical spine (Oc-C3), and none took place in the midcervical region (C3-C5). The results of this study assist in filling the existing void in dynamic tensile injury data and will aid in developing improved neck injury prevention strategies.


Studies on the synthesis of pectenotoxin II: synthesis of a C(11)-C(26) fragment precursor via [3 + 2]-annulation reactions of chiral allylsilanes.
[see reaction]. A synthesis of tetracycle 2 corresponding to the C(11)-C(26) fragment of pectenotoxin II is described. The synthesis features two highly stereoselective [3 + 2]-annulation reactions of chiral allylsilanes, generated via allylboration of aldehydes with the chiral gamma-silylallylborane 4 or the gamma-silylallylboronate 19, for construction of the highly substituted C and E rings.


Comparative study of intravenous ascorbic acid versus low-dose desferroxamine in patients on hemodialysis with hyperferritinemia.
In patients on hemodialysis (HD), parenteral iron improves the response to recombinant human erythropoietin (rhuEPO) therapy, but in some subjects it produces an iron overload, increasing their morbidity and mortality rates. In these cases, iron administration must be discontinued. This study aimed to investigate the efficiency of treatment with ascorbic acid (AA) or desferroxamine (DFO) to mobilize and reduce iron stores, and to determine the effect of these compounds on erythropoiesis. We performed a prospective and randomized trial over 6 months, which included 27 patients with serum ferritin levels >800 ng/mL, TSAT >30% and stabilized hemoglobin (Hb) and rhuEPO doses. All patients had previously received parenteral iron (Ferlecit). Nine patients received 200 mg of intravenous (i.v.) AA 3 times/week and nine patients received 1 mg/Kg/week of DFO; the remaining nine patients were the control group. There were no significant differences in iron loss or mobilization due to dialysis. When Ferlecit was discontinued, functional iron did not vary and the epoetin resistance index (rhuEPO dose/Hb) was reduced by 21% in the i.v. AA group. In the DFO and control groups, functional iron levels fell. In the DFO group the epoetin resistance index increased by 20%, with no modifications in the control group. There was a positive correlation between transaminases and serum ferritin. In HD patients with an iron overload, neither i.v. AA administration or low-dose DFO increased iron mobilization or iron loss due to dialysis. I.v. AA administration allows elimination of iron from stores without any drop in the functional iron produced by discontinuing parenteral maintenance iron; it also improves the response to rhuEPO. DFO did not elicit any positive effects on erythropoiesis.


Effects of epidermal growth factor on the growth of human gastric cancer cell and the implanted tumor of nude mice.
Epidermal growth factor (EGF) plays an important role in the regulation of gastrointestinal tissue growth and development, and it can stimulate epithelial proliferation, cell differentiation and growth. It has been established that the EGF can promote gastric cytoprotection and ulcer healing. But the potential ability of EGF to regulate the gastric cancer growth is unknown. This study is to investigate the influence of EGF on human gastric cancer cell and the implanted tumor growth of nude mice. The cell growth rates of human gastric adenocarcinoma cell lines MKN-28, MKN-45, SGC-7901 and normal human gastric epithelial cells 3T3 were assessed when incubated with recombinant human EGF (rhEGF, 0.05, 0.1, 0.5, 1.0, 10, 50, 100 mg.L(-1)) using MTT method. The cells of MKN-28, MKN-45, SGC-7901 (gastric cancer tissue 1.5mm(3)) were implanted in the BALB/cA nude mice for 10 days. The EGF was given intraperitoneally (15, 30, 60 microg.kg(-1)) for 3 weeks. The body weights of the tumor-bearing animals and their tumor mass were measured afterwards to assess the mitogenic effect of rhEGF in the nude mice. Within the concentration range of 0.05-100mg.L(-1), rhEGF could increase the cell growth of normal 3T3 cells (cell growth rate 100% vs 102.8%, P<0.05), but partially restrain the gastric cancer cell growth. The latter effect was related to cell differentiation. In 15-60 microg/kg rhEGF groups, the mean implanted tumor mass of MKN-28 cell were 1.75 g, 1.91 g, 2.08 g/NS group 1.97 g (P>0.05), the mean tumor mass of SGC-7901 cell were 1.53 g, 1.07 g, 1.20 g/NS group 1.07 g (P>0.05), and for MKN-45 cell, the tumor mass were respectively 1.92 g, 1.29 g, 1.77 /NS group 1.82 g (P>0.05). So rhEGF had no obvious effect on implanted MKN-28, SGC-7901 and MKN-45 tumor growth. EGF has no stimulating effect on the human gastric cancer cell growth neither in vitro nor in vivo.


The Influence of Butter and Oils on Oxidative Reactions during In Vitro Gastrointestinal Digestion of Meat and Fish.
Oxidative reactions during cooking and gastrointestinal digestion of meat and fish lead to the formation of various lipid- and protein oxidation products, some of which are toxic. In the present study, it was investigated how the addition of 3% butter or oils affect lipid- and protein oxidation during cooking and in vitro digestion of meat (chicken thigh, chicken breast, beef) and fish (mackerel, cod). These muscle foods were selected based on their differences in heme-Fe and PUFA contents, and n-6/n-3 PUFA ratio, and therefore varying potential to form oxidation products during digestion. Without additional fat, mackerel digests displayed the highest n-3 PUFA oxidation (4-hydroxy-2-hexenal, propanal, thiobarbituric reactive acid substances), and chicken digests the highest n-6 PUFA oxidation (4-hydroxy-2-nonenal, hexanal), whereas both lipid- and protein oxidation (protein carbonyl compounds) were low in cod and beef digests. Lipid oxidative reactions were generally not altered by the addition of butter to any muscle matrix, whereas the addition of fish oil and safflower oil in different ratios (3:0, 2:1, 1:2, 0:3) as n-3 PUFA and n-6 PUFA source respectively, stimulated oxidative reactions, especially during digestion of beef. Since beef was considered the muscle matrix with the highest potential to stimulate oxidation in the added fat substrate, in a second experiment, beef was cooked and digested with 3% butter or seven commercial vegetable oils (sunflower-, maize-, peanut-, rapeseed-, olive-, rice bran- or coconut oil), all labeled 'suitable for heating'. No relevant oxidative reactions were however observed during digestion of beef with any of these commercial vegetable oils.


Case Report: Accidental firearm injury during trophy hunting and the role of paramedics in managing such cases at rural health posts in Nepal.
Possession of a firearm without a certified valid license is against Nepalese law. Following a decade-long civil war, Nepal government issued stringent laws not allowing public to possess firearms without a valid reason, despite bearing a license. However, there are people who possess and use firearms for hunting purposes. The present case reports an accidental death of a teenage boy who used a musket for hunting. The present case highlights the fact that despite these stringent laws, illegal possession of arms for trophy hunting is still prevalent in rural Nepal. Furthermore, this study aims to highlight the importance of paramedics in early intervention, stabilization and transport of the sick and injured to the hospital in emergency situations. Also, recruitment of paramedics in the ambulance service might have prevented untimely death in this particular case.


Papovaviral sialoadenitis in athymic nude rats.
A wasting disease was found in 32 athymic nude rats. The rats had parotid sialoadenitis with intranuclear inclusion bodies in ductal and acinar epithelial cells. Other common lesions included bronchitis, bronchiolitis and secondary bacterial pneumonia. Less commonly, rhinitis and Harderian adenitis were seen. Intranuclear inclusions were also seen in bronchial epithelium of 1 rat, Harderian gland acini of 1 rat and laryngeal glands of 2 rats. Viral particles, averaging 45 nm in diameter, sometimes in crystalline arrays, were found in the nucleus of parotid epithelial cells. By the use of the avidin-biotin-peroxidase complex (ABC) immunoperoxidase technique, antibodies to disrupted SV40 virus (the group specific antigen of the polyomavirus (miopapovavirus) genus of the papovavirus family) reacted with intranuclear inclusions and cytoplasm of parotid epithelium and inclusions in lung and Harderian gland. The viral antigen did not cross react with antibodies to mouse polyoma, mouse K or disrupted bovine papilloma viruses.


A prospective study of 86 new patients with social anxiety disorder.
To determine the prognosis in 86 new patients with social anxiety disorder. Untreated subjects with social anxiety were recruited by advertising in Stockholm, randomized to 3 months treatment with paroxetine or placebo, and then offered continued specialist care. Metabolizing capacity was determined by genotyping CYP2D6 in the subjects on paroxetine. After a mean 32 months all were contacted for a personal interview. Of the 92 evaluable subjects, 86 (93%) were interviewed. A favourable prognosis was seen in the subjects randomized to paroxetine who chose to continue with serotonergic medication. The least favourable prognosis was in those given placebo who chose not to be treated after the trial. Twenty-four subjects were still symptomatic and dysfunctional and had not sought treatment. Drug-induced adverse effects caused treatment termination in six subjects, one of whom had a poor metabolizing genotype. Due to their condition, some subjects with social anxiety refrain from effective treatments. The efficacy of serotonergic medication was maintained and augmented after a mean period of 32 months.


Characterization of a sucrase gene from Staphylococcus xylosus.
The Staphylococcus xylosus gene scrB, encoding a sucrase, has been isolated from a genomic library of S. xylosus constructed in Escherichia coli. The gene was detected by its ability to confer utilization of the glucose and fructose residues of raffinose in an E. coli strain that is not able to metabolize galactose. It was found to reside within a 1.8-kb DNA fragment, the nucleotide sequence of which was determined. One large open reading frame, which is preceded by a ribosome binding site, is encoded on the fragment. Its deduced amino acid sequence yields a protein with a molecular mass of 57.377 kDa which shows significant homology with bacterial sucrose-6-phosphate hydrolases and sucrases. Overexpression of scrB in E. coli by the bacteriophage T7 polymerase promoter system resulted in the production of a protein with an apparent molecular mass of 58 kDa. Disruption of the scrB gene in the S. xylosus genome rendered S. xylosus unable to utilize sucrose. Thus, the ScrB sucrase is essential for sucrose metabolism in S. xylosus.


Monofunctional and polyfunctional CD8+ T cell responses to human herpesvirus 8 lytic and latency proteins.
Human herpesvirus 8 (HHV-8) is the etiological agent of Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman's disease. It is postulated that CD8(+) T cell responses play an important role in controlling HHV-8 infection and preventing development of disease. In this study, we investigated monofunctional and polyfunctional CD8(+) T cell responses to HHV-8 lytic proteins gB (glycoprotein B) and K8.1 and latency proteins LANA-1 (latency-associated nuclear antigen-1) and K12. On the basis of our previous findings that dendritic cells (DC) reveal major histocompatibility complex (MHC) class I epitopes in gB, we used a DC-based system to identify 2 novel epitopes in gB, 2 in K8.1, 5 in LANA-1, and 1 in K12. These new HHV-8 epitopes activated monofunctional and polyfunctional CD8(+) T cells that produced various combinations of gamma interferon, interleukin 2, tumor necrosis factor alpha, macrophage inhibitory protein 1β, and cytotoxic degranulation marker CD107a in healthy HHV-8-seropositive individuals. We were also able to detect HHV-8-specific CD8(+) T cells in peripheral blood samples using HLA A*0201 pentamer complexes for one gB epitope, one K8.1 epitope, two LANA-1 epitopes, and one K12 epitope. These immunogenic regions of viral lytic and latency proteins could be important in T cell control of HHV-8 infection.


Cloning of the gene encoding a putative serine/threonine protein kinase which enhances spermine uptake in Saccharomyces cerevisiae.
Polyamine uptake in Saccharomyces cerevisiae was modulated by extracellular magnesium; in a magnesium-limited medium, polyamine, especially spermine, was overaccumulated into the interior, whose level was then toxic for the growth of this organism (Maruyama, T., Masuda, N., Kakinuma, Y., and Igarashi K. (1994) Biochim. Biophys. Acta 1194, 289-295). Here we isolated a mutant (strain YTM22-8) whose growth was tolerant to spermine in magnesium-limited medium. This mutant was defective in polyamine uptake and did not overaccumulate spermine. From a yeast genome library we cloned a gene (POT1) which restored the spermine uptake of this mutant and the sensitivity of the growth to spermine. The nucleotide sequence of the POT1 gene indicated that it encodes a putative serine/threonine protein kinase and is located on chromosome XI. The results suggest that spermine uptake by this organism is probably regulated by phosphorylation and dephosphorylation.


Effect of protein supplementation on development to the hatching and hatched blastocyst stages of cat IVF embryos.
The effects of protein supplementation in culture medium on development to the hatching and hatched blastocyst stages of cat in vitro-fertilized embryos were investigated. In the first experiment, presumptive zygotes derived from in vitro maturation and in vitro fertilization (IVF) were cultured in modified Earle's balanced salt solution (MK-1) supplemented with 0.4% bovine serum albumin (BSA) or 5% fetal bovine serum (FBS) for 9 days. There were no significant differences between the BSA and FBS groups with respect to the proportion of cleavage and development to the morula and blastocyst stages of zygotes. However, the presence of FBS in the medium enhanced development to the hatching blastocyst stage of zygotes compared with the BSA group (31.4% v. 7.8%). Moreover, 2.9% of zygotes cultured with FBS developed to the hatched blastocyst stage. The mean cell number of blastocysts derived from zygotes cultured with FBS was significantly higher (P<0.01) than that from zygotes cultured with BSA (136.6 v.101.5). In the second experiment, embryos at the morula orblastocyst stage, which were produced by culturing in MK-1 supplemented with 0.4% BSA after IVF, were subsequently cultured in MK-1 with 0.4% BSA or 5% FBS. Significantly more morulae developed to the blastocyst (P<0.05) and hatching blastocyst stages (P<0.01) in the FBS group than in the BSA group (71.5% and 53.6% v. 44.9% and 6.0%, respectively). Although none of the morulae cultured with BSA developed to the hatched blastocyst stage, 11.5% of morulae cultured with FBS developed to the hatched blastocyst stage. Moreover, the proportion of development to the hatching blastocyst stage of blastocysts was significantly higher (P<0.01) in the FBS group than in the BSA group (68.7% v. 9.8%). None of the blastocysts cultured with BSA developed to the hatched blastocyst stage, whereas 7.3% of blastocysts cultured with FBS developed to the hatched blastocyst stage. The results of the present study indicate that supplementation with FBS at different stages of early embryo development promotes development to the hatching and hatched blastocyst stages of cat IVF embryos.


Catalytic, Enantioselective 1,2-Difluorination of Cinnamamides.
The enantio- and diastereoselective synthesis of 1,2-difluorides via chiral aryl iodide-catalyzed difluorination of cinnamamides is reported. The method uses HF-pyridine as a fluoride source and mCPBA as a stoichiometric oxidant to turn over catalyst, and affords compounds containing vicinal, fluoride-bearing stereocenters. Selectivity for 1,2-difluorination versus a rearrangement pathway resulting in 1,1-difluorination is enforced through anchimeric assistance from a N- tert-butyl amide substituent.


Zebrafish models of alx-linked frontonasal dysplasia reveal a role for Alx1 and Alx3 in the anterior segment and vasculature of the developing eye.
The cellular and genetic mechanisms that coordinate formation of facial sensory structures with surrounding skeletal and soft tissue elements remain poorly understood. Alx1, a homeobox transcription factor, is a key regulator of midfacial morphogenesis. ALX1 mutations in humans are linked to severe congenital anomalies of the facial skeleton (frontonasal dysplasia, FND) with malformation or absence of eyes and orbital contents (micro- and anophthalmia). Zebrafish with loss-of-function alx1 mutations develop with craniofacial and ocular defects of variable penetrance, likely due to compensatory upregulation in expression of a paralogous gene, alx3. Here we show that zebrafish alx1;alx3 mutants develop with highly penetrant cranial and ocular defects that resemble human ALX1-linked FND. alx1 and alx3 are expressed in anterior cranial neural crest (aCNC), which gives rise to the anterior neurocranium (ANC), anterior segment structures of the eye and vascular pericytes. Consistent with a functional requirement for alx genes in aCNC, alx1; alx3 mutants develop with nearly absent ANC and grossly aberrant hyaloid vasculature and ocular anterior segment, but normal retina. In vivo lineage labeling identified a requirement for alx1 and alx3 during aCNC migration, and transcriptomic analysis suggested oxidative stress response as a key target mechanism of this function. Oxidative stress is a hallmark of fetal alcohol toxicity, and we found increased penetrance of facial and ocular malformations in alx1 mutants exposed to ethanol, consistent with a protective role for alx1 against ethanol toxicity. Collectively, these data demonstrate a conserved role for zebrafish alx genes in controlling ocular and facial development, and a novel role in protecting these key midfacial structures from ethanol toxicity during embryogenesis. These data also reveal novel roles for alx genes in ocular anterior segment formation and vascular development and suggest that retinal deficits in alx mutants may be secondary to aberrant ocular vascularization and anterior segment defects. This study establishes robust zebrafish models for interrogating conserved genetic mechanisms that coordinate facial and ocular development, and for exploring gene--environment interactions relevant to fetal alcohol syndrome.


Association Between Type-specific HPV Infections and hTERT DNA Methylation in Patients with Invasive Cervical Cancer.
There exists limited information on the role of hTERT methylation, and its association with type-specific HPV infections in cervical cancer. Eighty-seven frozen samples were analyzed for type-specific HPV infection using a GP5+/GP6+ PCR-RLB assay (RLB). hTERT DNA methylation analysis was performed using a newly developed PCR-RLB-hTERT. Ninety-three percent of samples were HPV-positive and fifteen different types were detected. hTERT methylation analysis of region 1 revealed no methylation in 78.8% of the samples and partial methylation in 21.2%. In region two, 68.2% showed no methylation and 31.8% showed a pattern of partial methylation. An association between the alpha 9 and alpha 7 species with a pattern of no methylation of hTERT in the region 1 was established (p=0.02 and p=0.03, respectively). Differences in patterns of methylation of the hTERT core promoter [region 1 (nt -208 to -1) and region 2 (nt +1 to +104) relative to first ATG] are related to the HPV species present.


Fluorescent probes for targeting endoplasmic reticulum: design strategies and their applications.
Advances in developing organic fluorescent probes and fluorescence imaging techniques have enhanced our understanding of cell biology. The endoplasmic reticulum (ER) is a dynamic structure that plays a crucial role in protein synthesis, post-translational modifications, and lipid metabolism. The malfunction of ER contributes to several physiological and pathological conditions. Therefore, the investigations on the imaging and role of ER have attracted a lot of attention. Due to their simplicity, synthetic tunability, photostability, high quantum yields, easier cellular uptake, and lower cytotoxicity, organic fluorophores offer invaluable tools for the precision targeting of various cellular organelles and probe ER dynamics. The precision staining is made possible by incorporating specific functional groups having preferential and local organelle biomolecular interactions. For instance, functional moieties such as methyl sulfonamide, sulfonylurea, and pentafluorophenyl assist in ER targeting and thus have become essential tools to probe a deeper understanding of their dynamics. Furthermore, dual-function fluorescent probes that simultaneously image ER and detect specific physiological parameters or biological analytes were achieved by introducing special recognition or chemically reactive sites. This article attempts to comprehensively capture various design strategies currently employed by researchers utilizing small organic molecules to target the ER and detect specific analytes.


Copper-catalyzed aminobromination/elimination process: an efficient access to alpha,beta-unsaturated vicinal haloamino ketones and esters.
A novel copper-catalyzed aminobromination-elimination process has been developed, which provides an easy access to alpha,beta-unsaturated vicinal haloamindes derivatives from readily available alpha,beta-unsaturated ketones and esters in good to excellent yields. The isolated intermediate discloses that the current system proceeds through the aminobromination process.


Human immune response to cationized proteins. I. Characterization of the in vitro response to cationized diphtheria toxoid.
Cationization of proteins, i.e., increasing net positive charge by the substitution of carboxyl groups with positively charged residues, has been reported to enhance protein immunogenicity in animal model systems. In the present study, we have investigated the effect of cationization on the in vitro cell-mediated immune response of human mononuclear cells to diphtheria toxoid. A series of cationized DT preparations were generated by covalent modification with ethylenediamine, with pIs ranging from 4.6 to > 9.3, and tested for their ability to induce proliferation of normal human peripheral blood mononuclear cells. Cationized DT (cDT) was found to induce an antigen-specific, augmented proliferative response, relative to native antigen, which was directly proportional to the degree of cationization. Further characterization of the response to cDT demonstrated that (1) proliferative responses could be detected considerably earlier, and typically at much lower antigen concentrations, than the response to native DT; (2) the response was dependent on HLA-DR; (3) production of a number of cytokines, sp. IL-1 beta, IL-2, and IFN-gamma, was also elevated in cDT-stimulated cultures; and (4) the enhanced proliferative response to cDT could be attributed to CD4+ helper T cells. These results demonstrate that cationization of proteins enhances the ability to generate a cell-mediated immune response in humans and suggest that cationization may have utility in the design of more effective carrier proteins for human vaccines.


UVA/riboflavin-induced apoptosis in mouse cornea.
A mouse model of combined UVA/riboflavin irradiation to eliminate stromal cells and other antigen-presenting cells in the cornea provides the basis for a probably low risk of corneal transplantation. After abrasion of the epithelium, the central corneas of mouse eyes were treated with UVA/riboflavin in vitro. Histological studies of hematoxylin-eosin and immunohistochemical staining with caspase 3 were performed. Dissected mouse corneas were analyzed by Western blot. Apoptotic cells were shown on the central corneal stroma; a cell-free zone was displayed in the cornea. Numbers of dead cells increased according to cultivation time. However, the endothelium survived due to the adjustment of the irradiation dose. A cell-free zone in the stroma of the mouse cornea was produced by UVA/riboflavin irradiation in vitro. The technique makes possible to prevent or reduce immunological reactions and the risk of graft rejection by pretreatment of the donor cornea, ultimately prolonging graft survival.


Characterising those with incident polymyalgia rheumatica in primary care: results from the PMR Cohort Study.
The aim was to characterise the sociodemographic, general health and polymyalgia rheumatica (PMR)-specific features of participants in a large inception cohort of patients with PMR diagnosed in UK primary care. Patients (n = 739) with a new diagnosis of PMR were referred into the study and mailed a questionnaire detailing their general health and sociodemographic characteristics in addition to the symptoms of and treatment for PMR. Characteristics of responders and non-responders were compared and descriptive statistics were used to characterise the health of the cohort. A total of 654 individuals responded to the questionnaire (adjusted response 90.1 %). Responders and non-responders were similar in age, gender and deprivation (based on postcode). The mean (standard deviation) age of the recruited cohort was 72.4 (9.3) years; 62.2 % were female. The sample reported high levels of pain and stiffness (8 out of 10 on numerical rating scales) and reported stiffness that lasted throughout the day. High levels of functional impairment, fatigue, insomnia and polypharmacy were also reported. Overall, women reported worse general and PMR-specific health than did men. This first primary care cohort of patients with incident PMR is similar in demographic terms to cohorts recruited in secondary care. However, the extent of symptoms, particularly reported stiffness, is higher than has been described previously. Given the majority of patients with PMR are exclusively managed in primary care, this cohort provides important information on the course of PMR in the community that will help clinicians managing this painful and disabling condition.


A cross-language comparison of /d/-/th/ perception: evidence for a new developmental pattern.
Previous studies have shown that infants perceptually differentiate certain non-native contrasts at 6-8 months but not at 10-12 months of age, whereas differentiation is evident at both ages in infants for whom the test contrasts are native. These findings reveal a language-specific bias to be emerging during the first year of life. A developmental decline is not observed for all non-native contrasts, but it has been consistently reported for every contrast in which language effects are observed in adults, In the present study differentiation of English /d-th/ by English- and French-speaking adults and English- and French-learning infants at two ages (6-8 and 10-12 months) was compared using the conditioned headturn procedure. Two findings emerged. First, perceptual differentiation was unaffected by language experience in the first year of life, despite robust evidence of language effects in adulthood. Second, language experience had a facilitative effect on performance after 12 months, whereas performance remained unchanged in the absence of specific language experience. These data are clearly inconsistent with previous studies as well as predictions based on a conceptual framework proposed by Burnham [Appl. Psycholing. 7, 201-240 (1986)]. Factors contributing to these developmental patterns include the acoustic properties of /d-th/, the phonotactic uniqueness of English /th/, and the influence of lexical knowledge on phonetic processing.


An unorthodox sensor protein (TorS) mediates the induction of the tor structural genes in response to trimethylamine N-oxide in Escherichia coli.
We isolated and characterized three spontaneous mutations leading to trimethylamine N-oxide (TMAO)-independent expression of the tor operon encoding the TMAO-reductase anaerobic respiratory system in Escherichia coli. The mutations lie in a new for regulatory gene, the torS gene, which probably encodes a sensor protein of a two-component regulatory system. One mutation, which leads to full TMAO-constitutive expression, is a 3-amino-acid deletion within the potential N-terminal periplasmic region, suggesting that this region contains the TMAO-detector site. For the other two mutations, a further induction of the tor operon is observed when TMAO is added. Both are single substitutions and affect the linker region located between the detector and the conserved transmitter domains. Thus, as proposed for other sensors, the TorS linker region might play an essential role in propagating conformational changes between the detector and the cytoplasmic signalling regions. The TorS histidine kinase is an unorthodox sensor that contains a receiver and a C-terminal alternative transmitter domain in addition to the domains found in most sensors. Previously, we showed that TMAO induction of the for operon requires the TorR response regulator and the TorT periplasmic protein. Additional genetic data confirm that torS encodes the sensor partner of TorR and TorT. First, insertion within torS abolishes tor operon expression whatever the growth conditions. Second, overexpressed TorR bypasses the requirement for torS, whereas the torT gene product is dispensable for tor operon expression in a torS constitutive mutant. This supports a signal-transduction cascade from TorT to TorR via TorS.


Variations in stable isotope fractionation of Hg in food webs of Arctic lakes.
Biotic and abiotic fractionation of mercury (Hg) isotopes has recently been shown to occur in aquatic environments. We determined isotope ratios (IRs) of Hg in food webs (zooplankton, chironomids, Arctic char) and sediments of 10 Arctic lakes from four regions and investigated the extent of Hg isotope fractionation. Hg IRs were analyzed by multicollector inductively coupled plasma mass spectrometry (MC-ICP/MS). Hg mass independent fractionation (MIF; Delta(199)Hg) and mass dependent fractionation (MDF; delta(202)Hg) were calculated and compared among samples. IRs of Hg in sediment were characterized mainly by MDF and low MIF (Delta(199)Hg -0.37 to 0.74 per thousand). However, all biota showed evidence of MIF, most pronounced in zooplankton (Delta(199)Hg up to 3.40 per thousand) and char (Delta(199)Hg up to 4.87 per thousand). Zooplankton takes up highly fractionated MeHg directly from the water column, while benthic organisms are exposed to sedimentary Hg, which contains less fractionated Hg. As evidenced by delta(13)C measurements, benthic chironomids make up a large proportion of char diet, explaining in part why MIF(char) < MIF(zooplankton) in lakes, where both samples were measured. Hg IRs in char varied among regions, while char from lakes from each region showed similar degrees of MIF. A MIF-offset was derived representing the mean MIF difference between sediment and fish, and indicated that fish in two regions retain sediment signatures altered by a consistent offset. Due to its minimal lake-to-catchment area and very high water retention time ( approximately 330 years), the meteor impact crater lake (Pingualuk) reflects a "pure" atmospheric Hg signature, which is modified only by aqueous in-lake processes. All other lakes are also affected by terrestrial Hg inputs and sediment processes.


Quantifying spatial variation in the uptake of microplastic by mussels using biodeposit traps: A field-based study.
Spatial uptake patterns of microplastics (MP) by marine species are largely unexplored under field conditions. A novel "biodeposit trap" that measure uptake and egestion of MP by suspension-feeders through the analysis of their biodeposits, was designed and used to estimate the spatial variation of these processes by mussels in field conditions. Traps containing wild or farmed mussels or control empty shells were deployed at three sites characterised by different MP concentrations and water flow conditions. A different MP dimensional composition was observed between MP pools present in biodeposit and control traps, with the latter shifted towards higher dimensional range (0.05-5 mm). Conversely, mussels accumulated small MP (0.02-0.05 mm) into their biodeposits without any significant difference between wild and farmed specimens. MP uptake rates were on average 4-5 times higher at the site where MP contamination was expected to be highest and where water flow conditions were considered moderate.


DNA repair gene X-ray repair cross complementing group 1 Arg194Trp polymorphism on the risk of lung cancer: a meta-analysis on 22 studies.
DNA repair gene X-ray repair cross complementing group 1 (XRCC1) Arg194Trp polymorphism has been investigated widely on lung cancer risk. However, the results are inconclusive. To derive a more precise estimation of the relationship between XRCC1 Arg194Trp polymorphism and lung cancer risk, we performed this meta-analysis. An electronic search of the database PubMed, Embase, and Chinese National Knowledge Infrastructure was performed. The odds ratio (OR) was pooled by STATA 10.1. Subgroup analyses by ethnicity, gender, smoking, and histologic types of lung cancer were performed. Twenty-two studies including 7515 cases and 9560 controls were identified ultimately. The pooled OR for total population showed that homozygous Trp/Trp variant genotype could increase lung cancer risk compared with the homozygous wild Arg/Arg genotype (OR = 1.22, 95% confidence interval [CI] = 1.04-1.44, p = 0.01); however, heterozygote Arg/Trp variant genotype could decrease lung cancer risk (OR = 0.91, 95% CI = 0.85-0.99, p = 0.02). Subgroup analyses by ethnicity confirmed the result that homozygous Trp/Trp variant genotype increased lung cancer risk in Asians (OR = 1.19, 95% CI = 1.01-1.41, p = 0.04) but not in whites. It was interesting to find that both the heterozygote Arg/Trp and the combined Trp/Trp + Arg/Trp variant genotypes could decrease the risk of lung cancer in whites (OR = 0.83, 95% CI = 0.72-0.96, p = 0.01; OR = 0.85, 95% CI = 0.74-0.98, p = 0.03, respectively) but not in Asians. Subgroup analyses by gender, smoking, and histologic types of lung cancer did not indicate any significant difference between cases and controls, excepted for male population, which carried heterozygote Arg/Trp variant genotype that could decrease lung cancer risk (OR = 0.54, 95% CI = 0.31-0.95, p = 0.03). Homozygous Trp/Trp variant genotype of XRCC1 Arg194Trp polymorphism could increase lung cancer risk in total population, especially in Asians. However, the heterozygote Arg/Trp variant genotype might decrease the risk of lung cancer, especially in whites.


Antigen specificity of circulating anti-neutrophil cytoplasmic antibodies in inflammatory bowel disease.
To characterize the antigen specificity of circulating anti-neutrophil cytoplasmic antibodies (ANCAs) in inflammatory bowel disease (IBD). Analysis of the prevalence of circulating ANCAs in patients with ulcerative colitis and Crohn's disease, by both non-specific methods (immunofluorescence against fixed neutrophil leukocytes) and specific antigen techniques (against purified neutrophil leukocyte constituents). Indirect immunofluorescence against fixed polymorphonuclear leukocytes, and solid-phase enzyme-linked immunosorbent assay (ELISA) against neutrophil constituents (alpha-granules, elastase, myeloperoxidase, cathepsin g, lysozyme and lactoferrin). Although results using immunofluorescence were typical of other studies (ulcerative colitis positive in 41%, Crohn's disease in 10%), ELISA studies showed antibody activity against neutrophil components in 69% of patients with ulcerative colitis and 39% of those with Crohn's disease. Antibodies in ulcerative colitis were commonly directed (in descending order) against lysozyme, cathepsin G, elastase, and lactoferrin, and in Crohn's disease against lysozyme. Correlation of indirect immunofluorescence data and ELISA results indicated that even this large panel of specific antigens fails to identify all the ANCA targets in IBD. The lack of correlation between the findings of ANCAs, either in general or versus a specific target, and disease extent or activity in ulcerative colitis supports the suggestion that ANCAs are unlikely to be of primary importance in pathogenesis.


Territory-Wide Study on Hospital Admissions for Asthma Exacerbations in the COVID-19 Pandemic.
Rationale: Patients with asthma were advised to avoid coronavirus disease (COVID-19) and comply with medication during the COVID-19 pandemic. Respiratory tract infection is a common cause of asthma exacerbations. There has not been evidence suggesting the link between COVID-19 and asthma exacerbation, especially in places with dramatic responses in infection control with universal masking and aggressive social distancing. Objectives: To assess the number for admissions of asthma exacerbations in January to April 2020 in Hong Kong with reference to admission in the past 5 years. Methods: Admission records of asthma exacerbations were retrieved from the Clinical Data Analysis and Reporting System. Patients aged 18 years or older with a known history of asthma admitted for asthma exacerbation were included. Log-linear was used to model count, with year and masking used as covariate and further analysis on ambient temperature and length of hospital stays. Fisher's exact test was used to compare the mortality rate and mechanical ventilation between the periods. Admissions for myocardial infarction, ischemic stroke, and gastric ulcer were included as controls. Results: The number of admissions for asthma exacerbations significantly decreased by 53.2% (95% confidence interval [CI], 50.4-55.8%) in 2020 compared with monthly average admission in 2015-2019, with a higher magnitude of decrease compared with control diagnoses. Admissions for asthma exacerbations decreased by 2.0% (95% CI, 1.8-2.2%) with every 1°C (1.8°F) increase in temperature and by 0.8% with every 1% increase in masking (95% CI, 0.8-0.9%). Conclusions: Hospitalization number for asthma exacerbations significantly decreased in early 2020, with similar length of stay. This was observed with concomitant practice of universal masking and social distancing during the COVID-19 pandemic in Hong Kong. We proposed that universal masking and social distancing reduced respiratory viral infection, leading to fewer hospital admissions for asthma exacerbations.


Distinct Amino Acid Availability-Dependent Regulatory Mechanisms of MepS and MepM Levels in Escherichia coli.
Peptidoglycan (PG) hydrolases play important roles in various aspects of bacterial physiology, including cytokinesis, PG synthesis, quality control of PG, PG recycling, and antibiotic resistance. However, the regulatory mechanisms of their expression are poorly understood. In this study, we have uncovered novel regulatory mechanisms of the protein levels of the synthetically lethal PG endopeptidases MepS and MepM, which are involved in PG synthesis. A mutant defective for both MepS and MepM was lethal in an amino acid-rich medium, whereas it exhibited almost normal growth in a minimal medium, suggesting the expendability of MepS and MepM in a minimal medium. Protein levels of MepS and MepM dramatically decreased in the minimal medium. Although MepM was revealed as a substrate of Prc, a periplasmic protease involved in the proteolysis of MepS, only the decrease in the MepS level in the minimal medium was affected by the prc depletion. Phenotypic and biochemical analyses showed that the presence of aromatic amino acids in the medium induced the accumulation of MepS, but not MepM, while the presence of glutamate increased the level of MepM, but not MepS. Together, these results demonstrate that the protein levels of the two major PG endopeptidases are regulated in an amino acid availability-dependent manner, but their molecular mechanisms and signaling are significantly distinct.


[Congenital long QT syndrome].
Long QT syndrome (LQTS) is a clinically and genetically heterogenous syndrome characterized by a lengthening of the QT interval on the surface ECG and a propensity to severe ventricular arrhythmias such as torsades de pointes and ventricular fibrillation, leading eventually to syncope and sudden death. This rare syndrome with a mendelian inheritance occurs in subjects with otherwise normal cardiac morphological examination. The potentially severe prognosis justifies a presymptomatic diagnosis. The genetic nature of the disease has been confirmed with the identification of at least six loci and five genes. This syndrome is a perfect illustration of an adrenergic-induced ventricular arrhythmia. The first-line treatment is a beta-blocking agent for all symptomatic patients. In addition, a number of drugs known to lengthen ventricular repolarization must be prohibited. In case of suspicion of LQTS, all family members should be tested both clinically with a surface ECG and genetically in order to diagnose presymptomatic patients.


Multivariate assessment of lipid parameters as predictors of coronary heart disease among postmenopausal women: potential implications for clinical guidelines.
Over the past decade, lipid measurements have been significantly improved and standardized. We evaluated the usefulness of multiple plasma lipid parameters in predicting coronary heart diseases (CHD) among women. Among 32,826 women from the Nurses' Health Study who provided blood samples at baseline, 234 CHD events were documented during 8 years of follow-up. In a nested study, these cases were matched to controls (1:2) for age, smoking, fasting status, and month of blood draw. We estimated the relative risk (RR) for each lipid parameter, adjusted for C-reactive protein, homocysteine, body mass index, family history, hypertension, diabetes, postmenopausal hormone use, physical activity, alcohol intake, and blood draw parameters. The RRs associated with an increase of approximately 1 SD (mg/dL) were as follows: HDL cholesterol (HDL-C) (RR=0.6 [0.5 to 0.8], SD=17), apolipoprotein B100 (apoB100) (RR=1.7 [1.4 to 2.1], SD=32), LDL cholesterol (LDL-C) (RR=1.4 [1.1 to 1.7], SD=36), total cholesterol (TC) (RR=1.4 [1.1 to 1.6], SD=40), and triglycerides (RR=1.3 [1.0 to 1.5], SD=80). Among the lipid indexes, the RRs were: apoB100/HDL-C (RR=1.7 [1.4 to 2.1], SD=1.0), TC/HDL-C (RR=1.6 [1.3 to 1.9], SD=1.3), LDL-C/HDL-C (RR=1.5 [1.3 to 1.9], SD=1.0), and non-HDL-C (RR=1.6 [1.3 to 1.9], SD=42 mg/dL). After simultaneous control for several lipid biomarkers, HDL-C was the primary contributor of the variation in multivariate models (P=0.01), followed by LDL-C (P=0.01), whereas triglycerides and apoB100 did not contribute further information. HDL-C-related ratios were the strongest contributors to predicting CHD (P<0.0001). Lower levels of HDL-C may be a key discriminator of higher CHD events among postmenopausal women. HDL-C-related ratios (such as TC/HDL-C) provide a powerful predictive tool independently of other known CHD risk factors.


Stillbirths in women with pre-gravid obesity.
Objective To determine the association between pre-gravid obesity and stillbirth. Methods A retrospective study of a population-based dataset of births at ≥34 weeks' gestation. We excluded fetal deaths due to lethal anomalies and intrapartum fetal deaths. We calculated the incidence of stillbirths, neonatal respiratory distress syndrome (RDS) and neonatal intensive care unit (NICU) admissions per ongoing pregnancies for each gestational week in the two body mass index (BMI) categories (≥30 vs.<30). Results Pre-pregnancy obesity (BMI≥30), pre-pregnancy diabetes, oligo- and polyhydramnios, being small for gestational age (SGA) and preeclampsia were significantly associated with stillbirth. However, the only pre-gravid factor that is amenable to intervention was obesity [adjusted odds ratio (OR) 2.0; 95% confidence interval (CI) 1.20, 3.3]. The rates of stillbirth seem to increase with gestational age in both BMI categories. RDS and NICU admission would be presented. Conclusion Birth near term might reduce stillbirths and decrease NICU admissions occurring in term and in post-term obese women. This presumable advantage might be offset by the potential risk of labor induction and cesarean section among obese women. Women of childbearing age with a BMI≥30 should be counseled about these risks of obesity during pregnancy and childbirth.


Fast imputation using medium or low-coverage sequence data.
Accurate genotype imputation can greatly reduce costs and increase benefits by combining whole-genome sequence data of varying read depth and array genotypes of varying densities. For large populations, an efficient strategy chooses the two haplotypes most likely to form each genotype and updates posterior allele probabilities from prior probabilities within those two haplotypes as each individual's sequence is processed. Directly using allele read counts can improve imputation accuracy and reduce computation compared with calling or computing genotype probabilities first and then imputing. A new algorithm was implemented in findhap (version 4) software and tested using simulated bovine and actual human sequence data with different combinations of reference population size, sequence read depth and error rate. Read depths of ≥ 8× may be desired for direct investigation of sequenced individuals, but for a given total cost, sequencing more individuals at read depths of 2× to 4× gave more accurate imputation from array genotypes. Imputation accuracy improved further if reference individuals had both low-coverage sequence and high-density (HD) microarray data, and remained high even with a read error rate of 16%. With read depths of ≤4×, findhap (version 4) had higher accuracy than Beagle (version 4); computing time was up to 400 times faster with findhap than with Beagle. For 10,000 sequenced individuals plus 250 with HD array genotypes to test imputation, findhap used 7 hours, 10 processors and 50 GB of memory for 1 million loci on one chromosome. Computing times increased in proportion to population size but less than proportional to number of variants. Simultaneous genotype calling from low-coverage sequence data and imputation from array genotypes of various densities is done very efficiently within findhap by updating allele probabilities within the two haplotypes for each individual. Accuracy of genotype calling and imputation were high with both simulated bovine and actual human genomes reduced to low-coverage sequence and HD microarray data. More efficient imputation allows geneticists to locate and test effects of more DNA variants from more individuals and to include those in future prediction and selection.


Bacterial lipoproteins and other factors released by Francisella tularensis modulate human neutrophil lifespan: Effects of a TLR1 SNP on apoptosis inhibition.
Francisella tularensis infects several cell types including neutrophils, and aberrant neutrophil accumulation contributes to tissue destruction during tularaemia. We demonstrated previously that F. tularensis strains Schu S4 and live vaccine strain markedly delay human neutrophil apoptosis and thereby prolong cell lifespan, but the bacterial factors that mediate this aspect of virulence are undefined. Herein, we demonstrate that bacterial conditioned medium (CM) can delay apoptosis in the absence of direct infection. Biochemical analyses show that CM contained F. tularensis surface factors as well as outer membrane components. Our previous studies excluded roles for lipopolysaccharide and capsule in apoptosis inhibition, and current studies of [14 C] acetate-labelled bacteria argue against a role for other bacterial lipids in this process. At the same time, studies of isogenic mutants indicate that TolC and virulence factors whose expression requires FevR or MglA were also dispensable, demonstrating that apoptosis inhibition does not require Type I or Type VI secretion. Instead, we identified bacterial lipoproteins (BLPs) as active factors in CM. Additional studies of isolated BLPs demonstrated dose-dependent neutrophil apoptosis inhibition via a TLR2-dependent mechanism that is significantly influenced by a common polymorphism, rs5743618, in human TLR1. These data provide fundamental new insight into pathogen manipulation of neutrophil lifespan and BLP function.


NSAID-Gut Microbiota Interactions.
Nonsteroidal anti-inflammatory drugs (NSAID)s relieve pain, inflammation, and fever by inhibiting the activity of cyclooxygenase isozymes (COX-1 and COX-2). Despite their clinical efficacy, NSAIDs can cause gastrointestinal (GI) and cardiovascular (CV) complications. Moreover, NSAID use is characterized by a remarkable individual variability in the extent of COX isozyme inhibition, therapeutic efficacy, and incidence of adverse effects. The interaction between the gut microbiota and host has emerged as a key player in modulating host physiology, gut microbiota-related disorders, and metabolism of xenobiotics. Indeed, host-gut microbiota dynamic interactions influence NSAID disposition, therapeutic efficacy, and toxicity. The gut microbiota can directly cause chemical modifications of the NSAID or can indirectly influence its absorption or metabolism by regulating host metabolic enzymes or processes, which may have consequences for drug pharmacokinetic and pharmacodynamic properties. NSAID itself can directly impact the composition and function of the gut microbiota or indirectly alter the physiological properties or functions of the host which may, in turn, precipitate in dysbiosis. Thus, the complex interconnectedness between host-gut microbiota and drug may contribute to the variability in NSAID response and ultimately influence the outcome of NSAID therapy. Herein, we review the interplay between host-gut microbiota and NSAID and its consequences for both drug efficacy and toxicity, mainly in the GI tract. In addition, we highlight progress towards microbiota-based intervention to reduce NSAID-induced enteropathy.


Study of the Suitability of Different Types of Slag and Its Influence on the Quality of Green Grouts Obtained by Partial Replacement of Cement.
This paper is part of a research line focused on the reduction of the use of cement in the industry. In this work, the study of work methodologies for the manufacture of green cementitious grout mixtures is studied. Grout is widely used in construction and it requires an important use of raw materials. On the other hand, the steel industry faces the problem of the growing generation of slag wastes due to the increase in steel manufacturing. The green grout aims to achieve the dual objective of reducing the demand for cement and improve the slag waste valorization. Slag is not introduced as an aggregate but through the direct replacement of cement and no additives. The research seeks a product where we can use steel slag intensively, guaranteeing minimum resistance and workability. Results with substitutions between a 25% to 50% and water/cement ratio of 1 are presented. In particular, the suitability of different slags (two Ladle Furnace Slag (LFS) and one Blast Furnace Slag (GGBS)) in the quality of the final product are analyzed. The feasibility of replacing cement with slag and the importance of the origin and pretreatment are highlighted.


A vision for collaborative training infrastructure for bioinformatics.
In biology, a missing link connecting data generation and data-driven discovery is the training that prepares researchers to effectively manage and analyze data. National and international cyberinfrastructure along with evolving private sector resources place biologists and students within reach of the tools needed for data-intensive biology, but training is still required to make effective use of them. In this concept paper, we review a number of opportunities and challenges that can inform the creation of a national bioinformatics training infrastructure capable of servicing the large number of emerging and existing life scientists. While college curricula are slower to adapt, grassroots startup-spirited organizations, such as Software and Data Carpentry, have made impressive inroads in training on the best practices of software use, development, and data analysis. Given the transformative potential of biology and medicine as full-fledged data sciences, more support is needed to organize, amplify, and assess these efforts and their impacts.


Binding of Harmine Derivatives to DNA: A Spectroscopic Investigation.
Harmine belongs to a group of β-carboline alkaloids endowed with antitumor properties. Harmine and its derivatives are thought to bind to DNA and interfere with topoisomerase activities. We investigated the base-dependent binding of harmine, and three of its synthetic anticancer-active derivatives to the genomic DNA from calf thymus and two synthetic 20-mer double helices, the poly(dG-dC)·poly(dG-dC) and the poly(dA-dT)·poly(dA-dT), by means of UV-Vis and circular dichroism (CD) spectroscopies. The data show that the DNA binding and stabilising properties of the investigated derivatives are base pair-dependent. These results could be used as a guide to design and develop further bioactive analogues.


Vascular imaging with MRI: inadequacy in Takayasu's arteritis compared with angiography.
Magnetic resonance imaging (MRI) and angiography were compared in a prospective and blinded fashion in 10 consecutive patients with Takayasu's arteritis to evaluate the ability of MRI to detect vascular abnormalities in the large arteries of the thorax and abdomen. Only the aorta and its major branches were studied. MRI was performed with a 0.5-T superconducting instrument and a body coil. Overlapping cuts, oblique views, and cardiac gating were used. Using angiography as the gold standard, MRI correctly identified all lesions in three patients, gave false-positive findings in two patients, and gave false-negative findings in five patients. In the latter five patients, MRI missed areas of major pathology in proximal arch vessels. MRI sensitivity was only 38% with a patient-by-patient analysis and 54% with a lesion-by-lesion analysis. The visualization of specific arteries with MRI was compared to visualization with angiography, and was markedly inferior in all vessels except the aorta, the innominate artery, and the common iliac arteries. It appears that with current equipment and technology, MRI is inadequate as a screening examination or for detailed evaluation of arterial disease other than the aorta except in highly selected cases.


The management of ischaemic and periampuation pain.
This article describes the development and pilot application of a management tool designed to support healthcare practitioners in the management of ischaemic and periamputation pain. The ischaemic management periamputation care (IMPAC) guidelines have been piloted in the authors' clinical area and early results suggest that there are positive benefits accruing to patients and ward staff.


Comorbid substance use disorder in schizophrenia: a selective overview of neurobiological and cognitive underpinnings.
Although individuals with schizophrenia show a lifetime prevalence of 50% for suffering from a comorbid substance use disorder, substance abuse usually represents an exclusion criterion for studies on schizophrenia. This implies that surprisingly little is known about a large group of patients who are particularly difficult to treat. The aim of the present work is to provide a brief and non-exhaustive overview of the current knowledgebase about neurobiological and cognitive underpinnings for dual diagnosis schizophrenia patients. Studies published within the last 20 years were considered using computerized search engines. The focus was on nicotine, caffeine, alcohol, cannabis and cocaine being among the most common substances of abuse. All drugs of abuse target dopaminergic, glutamatergic and GABAergic transmission which are also involved in the pathophysiology of schizophrenia. Current literature suggests that neurocognitive function might beless disrupted in substance-abusing compared to non-abusing schizophrenia patients, but in particular the neuroimaging database on this topic is sparse. Detrimental effects on brain structure and function were shown for patients for whom alcohol is the main substance of abuse. It is as yet unclear whether this finding might be an artifact of age differences of patient subgroups with different substance abuse patterns. More research is warranted on the specific neurocognitive underpinnings of schizophrenia patients abusing distinct psychoactive substances. Treatment programs might either benefit from preserved cognitive function as a resource or specifically target cognitive impairment in different subgroups of addicted schizophrenia patients.


Androgen-dependent expression of fibroblast growth factor-1 in submaxillary gland of mouse.
We have purified a 16,000 dalton protein that stimulates growth of human umbilical cord vein-derived endothelial cells (HUV-EC) from mouse submaxillary glands by using heparin-Sepharose affinity and C4 reverse phase chromatography. The purified molecule was identified as an FGF-1 on the basis of its biological activities, its affinity for heparin and its N-terminal amino-acid sequence. The concentrations of FGF-1 in the submaxillary gland of male or testosterone-treated female mice were about 12 times those of untreated females or castrated males. The 2.3 and 4.1 kb FGF-1 mRNAs were expressed in the glands of male mice older than 4 weeks but not in the glands of female mice. These results suggest that FGF-1 may have important functions for growth, differentiation and development of mouse submaxillary glands, and it may act as an endocrine hormone.


Genistein causes germ cell reduction in the genital ridges of Japanese quail Coturnix japonica embryo.
Genistein (GEN), an isoflavonoid phytoestrogen, is one of the potent estrogenic compounds derived from plants that can cause disrupting effects on sex organ development in non-mammalian and mammalian species. The present study revealed effect of genistein on germ cell number in the genital ridges during gonadogenesis. Genistein (16 and 24 μg/g egg) was injected into the egg yolk prior to incubation. Effect of genistein on quail-primordial germ cells (PGCs) number was examined by counting the number of Wisteria floribunda (WFA)-positive cells localized in both left and right genital ridges compared with the control group. Both concentrations of genistein resulted in significant decrease of PGC number compared with the control group. Percentages of the sterility rate of the embryo treated with 16 and 24 μg of genistein/g egg were 19% and 23%, respectively. These results provide evidence that genistein may be a germ cell toxicant causing sterility later in life of adult birds. This is the first report on the effect of genistein on PGC number in the genital ridges of the avian embryo.


The deletion of petG in Chlamydomonas reinhardtii disrupts the cytochrome bf complex.
The 4-kDa protein encoded by chloroplast petG copurifies with the cytochrome bf complex of spinach and is found in a number of other photosynthetic organisms, including the eukaryotic alga Chlamydomonas reinhardtii. To determine whether petG is involved in the function or assembly of the cytochrome bf complex, the gene was cloned from C. reinhardtii, excised from the DNA fragment, and replaced with a spectinomycin resistance cassette. A petG deletion strain of C. reinhardtii was then obtained by biolistic transformation. The resulting homoplasmic petG deletion strains are unable to grow photosynthetically, and immunoblot analysis shows markedly decreased levels of cytochrome b6, cytochrome f, the Rieske iron-sulfur protein, and subunit IV. To verify that this phenotype was due to the removal of petG, we also constructed a strain with a deletion in the open reading frame (ORF56), which is found 25 base pairs downstream of petG. The ORF56 deletion strain grew photosynthetically and had wild-type levels of the four major cytochrome bf subunits. We conclude that the absence of the PetG protein affects either the assembly or stability of the cytochrome bf complex in C. reinhardtii.


Transverse viscoelastic extension in nitella: I. Relationship to growth rate.
Transverse viscoelastic extensibility was measured directly in isolated walls of Nitella internode cells. Cell walls extended transversely exhibit a yield point which is approximately twice the yield point in the longitudinal direction. Walls from young, growing cells are four to seven times more extensible longitudinally than transversely, while walls from mature, nongrowing cells are only two times more extensible longitudinally. Although longitudinal extensibility decreases drastically with the decrease in the growth rate, lateral extensibility is constant through development. There is a discrepancy between the lateral growth rate and transverse creep, since the lateral growth rate is not constant. However, the degree of wall anisotropy observed is consistent with the view that the transversely oriented cellulose microfibrils act as a "reinforcing filler" in Nitella cell walls.


Classic steroid 11 beta-hydroxylase deficiency caused by a C-->G transversion in exon 7 of CYP11B1.
Steroid 11 beta-hydroxylase deficiency (11 beta OHD) is derived from mutations in the P45011 beta gene (CYP11B1) and inherited in an autosomal recessive manner. In the present study, we have performed a molecular genetic analysis of CYP11B1 in a Japanese patient clinically diagnosed as classic 11 beta OHD. Nucleotide sequencing of the PCR-amplified exons from the patient's genomic DNA reveals a unique C-->G transversion that converts codon 384 CGA (arginine) to GGA (glycine) in exon 7. Restriction fragment length polymorphism (RFLP) data demonstrate that the patient is homozygous for this mutation. When the full-length cDNA corresponding to CYP11B1 of the patient is transfected into COS-7 cells, no steroid 11 beta-hydroxylase activity is detectable in mitochondria of the cells. These results indicate that this point mutation completely abolishes P45011 beta activity and causes the classic 11 beta OHD.


Direct synthesis of bipyrroles using phenyliodine bis(trifluoroacetate) with bromotrimethylsilane.
[reaction: see text] The hypervalent iodine(III) reagent, phenyliodine bis(trifluoroacetate) (PIFA), mediates the unprecedented, oxidative coupling reaction of pyrroles to give alpha-linked bipyrroles selectively in the presence of bromotrimethylsilane. This straightforward synthesis could provide 2,3'-bipyrrole by the choice of a N-substituent of pyrrole. Mechanistic consideration of the present reaction is also described.


[Iodine resorption from PVP-iodine preparations after their use in humans].
The absorption of iodine from iodine-containing PVP preparations and the possibility of iodide being split off from the organic compound were tested in subjects with normal thyroid function after they had used PVP-iodine as mouth-antiseptic (15 subjects), vaginal gel (20 subjects) or liquid soap (20 subjects). Serum I-, T3, T4, TSH and urinary iodide excretion were measured, as an index of thyroid function, before and after the PVP application. Increase in iodine supply was up to 2 mg daily in the test subjects. The overwhelming proportion of the absorbed iodine (greater than 75%) was as organically bound iodine. In none of the subjects was there evidence of developing hyper- or hypothyroidism as a result of the additional iodine supply. Approximate values for the biological half-life and the distribution volume of total iodine in the body were also obtained.


Prostate-specific antigen response to withdrawal of megestrol acetate in a patient with hormone-refractory prostate cancer.
Clinically significant responses after withdrawal of flutamide in patients with hormone-refractory prostate cancer (HRPC) are well documented. Failure to recognize this syndrome of response results in potential morbidity due to salvage therapy, confusion in interpretation of disease state, and introduction of a possible source of error in clinical trials. In this case report, we describe a patient with HRPC whose prostate-specific antigen levels decreased substantially in response to withdrawal of megestrol acetate. Such a response should be considered when megestrol acetate is used in the treatment of HRPC.


Plasma free amino acids in Taiwan Chinese: the effect of age.
Values of plasma amino acids (AA) in Taiwan Chinese in different age groups were determined. AA analysis was performed by ion-exchange chromatography. The results correlated well with the reference values published by Shapira in 1989, indicating that plasma AA concentrations are not evidently affected by race and dietary habits. We also confirmed the effect of age on plasma free AA levels. More than two thirds of 35 amino acids showed statistically significant differences between age groups (p<0.05, one-way ANOVA test). Levels of taurine, hydroxyproline, threonine, serine, glutamic acid, proline, citrulline and alpha-amino-eta-butyric acid varied with age (p<0.01). Only a few AA, including alpha-amino-adipic acid, glycine, alanine, cystine, homocystine, isoleucine, tyrosine, gamma-aminobutyric acid and tryptophan, showed no statistically significant differences between the different age groups. When comparing the mean differences in individual AA among these three age groups using the post-hoc test, alanine, tyrosine, phenylalanine, glycine, glutamine, cystine, homocystine, isoleucine, gamma-aminobutyric acid and tryptophan appeared to be constant across all age groups. Glutamic acid and serine correlated significantly with age (p<0.05), with a definite decline in concentration from infancy to adulthood (r=0.396 and 0.243; p<0.01, respectively). We conclude that AA values are not evidently affected by race, but they do vary with age.


Dilatational and shear rheology of soluble and insoluble monolayers with a Langmuir trough.
The surface dilatational and shear moduli of surfactant and protein interfacial layers can be derived from surface pressures measured with a Wilhelmy plate parallel, ΔΠpar and perpendicular ΔΠperp to the barriers in a Langmuir trough. Applying area oscillations, A0+ ΔAeiωt, in a rectangular Langmuir trough induces changes in surface pressure, ΔΠpar and ΔΠperp for monolayers of soluble palmitoyl-lysophosphatidylcholine (LysoPC), insoluble dipalmitoylphosphatidylcholine (DPPC), and the protein β-lactoglobulin to evaluate Es∗+Gs∗=A0ΔΠparΔA and Es∗-Gs∗=A0ΔΠperpΔA. Gs∗ was independently measured with a double-wall ring apparatus (DWR) and Es∗ by area oscillations of hemispherical bubbles in a capillary pressure microtensiometer (CPM) and the results were compared to the trough measurements. For LysoPC and DPPC, A0ΔΠparΔA≅A0ΔΠperpΔA meaning Es∗≫Gs∗ and Es∗≅A0ΔΠparΔA≅A0ΔΠperpΔA. Trough values for Es∗ were quantitatively similar to CPM when corrected for interfacial curvature. DWR showed G∗ was 4 orders of magnitude smaller than Es∗ for both LysoPC and DPPC. For β-lactoglobulin films, A0ΔΠparΔA>A0ΔΠperpΔA and Es∗ and Gs∗ were in qualitative agreement with independent CPM and DWR measurements. For β-lactoglobulin, both Es∗ and Gs∗ varied with film age and history on the trough, suggesting the evolution of the protein structure.


Revealing the Frank-Evans "Iceberg" Structures within the Solvation Layer around Hydrophobic Solutes.
Using computer simulations, the structural properties of solvation water of three model hydrophobic molecules, methane and two fullerenes (C60 and C80), were studied. Systems were simulated at temperatures in the range of 250-298 K. By analyzing both the local ordering of the molecules of water in the solvation layers and the structure of hydrogen bond network, it is shown that in the solvation layer of hydrophobic molecules, ordered aggregates consisting of water molecules are formed. Even though it is difficult to define the exact structure of these aggregates, their existence alone is clearly noticeable. Moreover, these aggregates become more pronounced with the decrease of temperature. The existence of the ordered aggregates around the hydrophobic solutes complies with the concept of "icebergs" proposed by Frank and Evans.


Determination of site-specific modifications of glucose-6-phosphate dehydrogenase by 4-hydroxy-2-nonenal using matrix assisted laser desorption time-of-flight mass spectrometry.
Products of the reaction of 4-hydroxy-2-nonenal (4HNE) with native and heat-denatured Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase (G6PDH) were analyzed to determine the structure and position of the protein modifications. Matrix assisted laser desorption time-of-flight mass spectrometry was used to measure molecular weights of the modified proteins and determine mass maps of peptides formed by digestion with cyanogen bromide. The molecular weight data show that one to two 4HNE molecules add to each subunit of native enzyme while approximately nineteen 4HNE molecules add to each subunit of heat-denatured enzyme. Peptides are observed in the cyanogen bromide mass map of modified native G6PDH that are consistent with selective modification of two segments of the amino acid sequence. One modified segment contains Lysine-182 that has been found to be part of the enzyme active site. Peptides are observed in the cyanogen bromide mass map of modified heat-denatured enzyme that are consistent with extensive modification of several segments of the amino acid sequence. The magnitude of the mass differences between modified and unmodified peptides were approximately 156 Da, consistent with a 1,4-addition of 4HNE. These results support the conclusion that 4HNE inactivates G6PDH by selectively modifying only two or three sites in the protein by a 1,4-addition reaction and that some aspect of the tertiary structure of the enzyme directs those modification reactions.


Plastid Phylogenomic Analysis of Tordylieae Tribe (Apiaceae, Apioideae).
Based on the nrDNA ITS sequence data, the Tordylieae tribe is recognized as monophyletic with three major lineages: the subtribe Tordyliinae, the Cymbocarpum clade, and the Lefebvrea clade. Recent phylogenomic investigations showed incongruence between the nuclear and plastid genome evolution in the tribe. To assess phylogenetic relations and structure evolution of plastomes in Tordylieae, we generated eleven complete plastome sequences using the genome skimming approach and compared them with the available data from this tribe and close relatives. Newly assembled plastomes had lengths ranging from 141,148 to 150,103 base pairs and contained 122-127 genes, including 79-82 protein-coding genes, 35-37 tRNAs, and 8 rRNAs. We observed substantial differences in the inverted repeat length and gene content, accompanied by a complex picture of multiple JLA and JLB shifts. In concatenated phylogenetic analyses, Tordylieae plastomes formed at least three not closely related lineages with plastomes of the Lefebvrea clade as a sister group to plastomes from the Selineae tribe. The newly obtained data have increased our knowledge on the range of plastome variability in Apiaceae.


Lameness in Beef Cattle: UK Farmers' Perceptions, Knowledge, Barriers, and Approaches to Treatment and Control.
Lameness in the beef industry has received little attention in the UK, despite the fact that it is a well-recognised problem in the dairy industry. The aims of this study were to (i) compare UK beef farmers' estimates of lameness prevalence to that of researchers, (ii) explore beef farmers' attitudes towards lameness, and (iii) help identify farmer reported barriers to lameness control and treatment. Beef farmers (11 finishing units and 10 suckler farms) were recruited from England and Wales. Farmers were asked to estimate their lameness prevalence, before a researcher conducted locomotion scoring using a five point scale, and a Bland Altman analysis performed. Face to face interviews were also conducted using a semi structured interview script aimed at capturing information such as current approaches and protocols as well as their views of lameness importance. Interviews were recorded and transcribed. An inductive thematic analysis was performed. All but two farmers underestimated lameness prevalence on their farms when compared to the researcher. Farmers initially underestimated lameness prevalence compared to the researchers estimates, with a mean underestimate of 7% (95% CI 5-9%). This is an important barrier to lameness detection and treatment. Thematic analysis identified four major themes: (1). Perception of lameness prevalence, (2). Technical knowledge and skills, (3). Perception of the impact of lameness, and (4). Barriers to the treatment and control of lameness. This study highlights that some approaches to lameness treatment are likely to be causing harm, despite being done with the intention to help the animal. There were four key areas of concern identified: recognition of lameness, treatment approaches, the training provided to farmers and confusion over transport and slaughter options available to farmers. This suggests an urgent need for future work to quantify and address the problem, and to provide evidence to justify the role of prevention and potentially start to break down barriers to control and treatment of lameness.


Targeting SHP2 sensitizes differentiated thyroid carcinoma to the MEK inhibitor.
Pharmacologic targeting of components of the MAPK/ERK pathway in differentiated thyroid carcinoma (DTC) is often limited due to the development of adaptive resistance. However, the detailed mechanism of MEK inhibitor (MEKi) resistance is not fully understood. Here, MEKi-resistant models were constructed successfully, in which multiple receptor tyrosine kinases (RTKs) signaling pathways and Src-homology 2 domain-containing phosphatase 2 (SHP2) were activated in MEKi-resistant cells. Given the physiological role of SHP2 as the downstream target of many RTKs, we first found blockade of SHP2 enhanced the sensitivity to MEKi in constructed MEKi-resistant models. Interestingly, we also found that compared with MEKi treatment alone, MEKi in combination with an SHP2 inhibitor markedly suppressed the reactivation of the MEK/ERK pathway; thus, the addition of the SHP2 inhibitor significantly improved the antitumor effects of MEKi. The synergistic suppression of DTC upon treatment with both inhibitors was further confirmed in xenograft models and transgenic models. Thus, our data suggest that RTKs activation leads to reactivation of the MAPK pathway and resistance to MEKi in DTC, which is reversed by SHP2 blockade. As a novel active inhibitor of SHP2, SHP099 in combination with MEKi is a promising therapeutic approach for advanced DTC and MEKi-resistant one.


Effect of Asp 96 isomerization on the properties of a lens αB-crystallin-derived short peptide.
One of the major reasons for age-related cataract formation is an accumulation of insoluble lens proteins. In particular, higher-order α-crystallin aggregates, comprising αA and αB subunits, are insolubilized by the build up of various post-translational modifications over time. Although we previously found an exceptional amount of Asp96 isomerization in αB-crystallin from aged human lens, the biological effect remains unknown. To approximate the effect of Asp 96 isomerization in αB-crystallin, here residues 93-103 of αB-crystallin were chemically synthesized as peptides in which l-α-Asp was replaced with l-β-Asp, D-α-Asp, or D-β-Asp. The resulting peptides were then compared in a biological assay. The results showed that isomerization of Asp 96 altered both the local structure of peptide and its stability against enzymatic digestion. In addition, the synthesized peptides decreased the insoluble fraction of heated α-crystallin. The D-β-Asp-containing peptide further decreased heat-induced precipitation of α-crystallin, and a chaperone assay based on heated alcohol dehydrogenase implied differential interaction of the peptides with substrate depending on the Asp isomer present in each. Our results suggest that the formation of Asp isomers is likely to affect the higher-order oligomer structure of α-crystallin and thereby its chaperone functions in aged lens.