A model for transmission of partial resistance to anti-malarial drugs.
Anti-malarial drug resistance has been identified in many regions for a long time. In this paper we formulate a mathematical model of the spread of anti-malarial drug resistance in the population. The model is suitable for malarial situations in developing countries. We consider the sensitive and resistant strains of malaria. There are two basic reproduction ratios corresponding to the strains. If the ratios corresponding to the infections of the sensitive and resistant strains are not equal and they are greater than one, then there exist two endemic non-coexistent equilibria. In the case where the two ratios are equal and they are greater than one, the coexistence of the sensitive and resistant strains exist in the population. It is shown here that the recovery rates of the infected host and the proportion of anti-malarial drug treatment play important roles in the spread of anti-malarial drug resistance. The interesting phenomena of ''long-time" coexistence, which may explain the real situation in the field, could occur for long period of time when those parameters satisfy certain conditions. In regards to control strategy in the field, these results could give a good understanding of means of slowing down the spread of anti-malarial drug resistance.


FPMOD: a modeling tool for sampling the conformational space of fusion proteins.
Fusion proteins are an important class of proteins with diverse applications in biotechnology. They consist of 2 or more rigid domains joined by a flexible linker. Understanding the conformational space of fusion proteins conferred by the flexible linkers is important to predicting its behavior. In this paper, we introduce a modeling tool called FPMOD (Fusion Protein MODeller) which samples the conformational space of fusion proteins by treating all domains as rigid bodies and rotating each of them around their flexible linkers. As a demonstration, FPMOD was used to predict the fluorescence resonance energy transfer (FRET) efficiency of three different fusion protein biosensors. The simulation results of the FRET efficiency prediction were consistent with the in vitro experimental data, which verified that FPMOD is a valid tool to predicting the behavior of fusion proteins.


Importance and relevance of melatonin to human biological rhythms.
The pineal hormone melatonin is a remarkable molecule, with a conserved time-keeping function across species. It is extensively used as a self-administered remedy for sleep disturbance in countries where it is freely available, and to some extent when it is available on prescription, as in the UK. In some circumstances, notably free-running sleep disorder of the blind, it is the treatment of choice. It is also the marker rhythm of choice for the determination of circadian phase and period. This review outlines the current state of knowledge within a physiological perspective with emphasis on human biological rhythms.


Possible antineoplastic agents I.
A few thalidomide and glutarimide derivatives were synthesized. Several compounds possessed significant antineoplastic activity against Ehrlich ascites carcinoma in Swiss albino mice.


Hemorrhagic toxin from the venom of Agkistrodon bilineatus (common cantil).
1. Hemorrhagic toxin was isolated from Agkistrodon bilineatus (Common cantil) venom using a three-step purification procedure to obtain 32.8 mg of purified hemorrhagic toxin from 700 mg of crude venom. 2. The purified toxin was homogeneous by disc polyacrylamide gel electrophoresis at pH 8.3, and by isoelectric focusing. 3. Hemorrhagic toxin possessed lethal, hemorrhagic and proteolytic activities. These activities of this toxin were inhibited by ethylenediaminetetraacetic acid (EDTA) and ethyleneglycol-bis-(beta-aminoethylether)N,N'-tetraacetic acid (EGTA), but not by cysteine or soybean trypsin inhibitor (SBTI). 4. Its molecular weight was approximately 48 kDa and the isoelectric point was 4.2. 5. Purified preparation hydrolyzed the Asn(3)--Gln(4), His(10)--Leu(11), Ala(14)--Leu(15), Tyr(16)--Leu(17), Arg(22)--Gly(23) and Phe(24)--Phe(25) bonds of oxidized insulin B. chain. 6. The A alpha chain of fibrinogen was first split and B beta chain was cleaved later by this toxin. 7. Hemorrhagic toxin contains 1 mol of zinc and 2 mol of calcium per mol of protein.


Arteriovenous grafts for vascular access in haemodialysis.
The outcome of 68 arteriovenous grafts placed in 46 patients requiring haemodialysis was studied over a period of 3.5 years. The biological grafts included autogenous saphenous vein, modified bovine carotid artery and human umbilical cord vein allograft, whereas the synthetic grafts comprised Sparks Dacron mandril, expanded reinforced polytetrafluoroethylene and knitted Dacron velour. These subcutaneous grafts were arranged as looped or straight configurations in the forearm or thigh. Of 59 grafts evaluated in patients with end-stage renal failure, only 48 per cent of the forearm grafts performed well, compared with 85 per cent of the thigh grafts. Although only 38 per cent of the looped grafts were successful, 78 per cent of the straight grafts functioned satisfactorily. Synthetic grafts suffered less serious complications than the commercial biological grafts.


Epidemiological assessment of hypospadias by degree of severity.
Hypospadias is one of the most common congenital malformations, with reportedly increasing rates of prevalence but poorly defined etiological factors for the disease and the varying degrees of manifestation. We characterize the prevalence of hypospadias in Arkansas with consideration of the degree of hypospadias. Data from the Arkansas State Birth Defects Surveillance Program of the Arkansas Reproductive Health Monitoring System were used to identify hypospadias cases and all male births by women residing in Arkansas between 1998 and 2007. Categorization of hypospadias severity into first, second or third degree was determined by a common 6-digit birth defect coding system used by surveillance programs across the country. Prevalence rates were computed for maternal and infant characteristics, birth year and birth residence. The prevalence of hypospadias in Arkansas for births between 1998 and 2007 was 74.2 cases per 10,000 live births. Degrees of hypospadias were graded as first (coronal/distal) in 60.7% of cases, second (mid penile/subcoronal) in 18.8%, third (perineal/proximal penile) in 4.6% and hypospadias not otherwise specified in 16%. The prevalence of hypospadias increased during the surveillance period, with 66.9 per 10,000 live births between 1998 and 2002, and 81.0 per 10,000 live births between 2003 and 2007 (p = 0.0003). While the prevalence rates for every degree of hypospadias also increased, the prevalence of hypospadias not otherwise specified decreased from 14.8 to 9.0 during these periods. Hypospadias prevalence in Arkansas is increasing. Improvement in recognition and categorization of degrees of hypospadias may have an impact on the reported prevalence rates.


Recurrences during a 10-year follow-up after first renal stone episode.
Each of 92 patients in a Swedish district served by only one hospital had been treated for their first renal stone in 1977 and was evaluated 10 years later. Recurrent stone formation during the observation period was observed in 26% of the patients, with no difference between men and women. Of all the patients who had sought medical advice in 1977 because of urinary stone colic, 51% were experiencing their first stone episode. Ten years later 37% of the original patients were still classified as single stone formers. The recorded recurrence rate was lower than that previously reported in the literature.


[Lipid peroxidation in the liver of dogs and guinea pigs receiving chloditan].
The contents and the rates of the formation of lipid peroxides in the liver homogenates were studied. Dogs and guinea pigs were fed with chloditan in doses of 50 and 150 mg/kg body weight, respectively, for 20 days. In the dogs chloditan caused atrophy of the adrenal glands. The drug accumulation in the adrenal glands of the guinea pigs did not produce damage of the glands. The feeding with chloditan failed to change the level of malondialdehyde in the livers of the dogs and decreased it in the livers of the guinea pigs. In vitro spontaneous and NADP N-dependent lipid peroxidation in the liver homogenates of the dogs treated with chloditan was slowed. The decrease of malondialdehyde content in the guinea pig liver and the slowing of lipid peroxidation rate in the dog liver homogenate can be explained by the change in the activity of the enzymes detoxicating xenobiotics and products of lipid peroxidation.


Resurfacing of atrophic facial acne scars with a high-energy, pulsed carbon dioxide laser.
Treatment of atrophic acne scars has been limited to the use of such traditional treatments as dermabrasion and chemical peels for many years. Recently, the addition of high-energy, pulsed carbon dioxide (CO2) lasers to the treatment armentarium has created renewed enthusiasm for cutaneous resurfacing due to their ability to create specific thermal injury with limited side effects. To determine the effectiveness of a high-energy, pulsed CO2 laser in eliminating atrophic facial scars and to observe for side effects. Fifty patients with skin phototypes I-V and moderate to severe atrophic facial acne scars were included in the study. Each patient received one high-energy, pulsed CO2 laser treatment using identical laser parameters by the same experienced laser surgeon. Baseline and 1-, 4-, 8-, 12-, and 24-week postoperative photographs and clinical assessments were obtained in all patients. Textural analysis of skin before and after laser irradiation were obtained in 10 patients to confirm clinical impressions. Clinical evaluations were conducted independently by two blinded assessors. There was an 81.4% average clinical improvement observed in acne scars following laser treatment. Skin texture measurements of laser-irradiated scars were comparable to those obtained in normal adjacent skin. Side effects were limited to transient hyperpigmentation lasting an average of 3 months in 36% of patients. Prolonged erythema (2 months average) was usual and considered to be a normal healing response. No hypertrophic scarring was observed following laser treatment. High-energy, pulsed CO2 laser treatment can safely and effectively improve or even eliminate atrophic facial scars and provides many benefits over traditional treatment methods.


Subclinical delta status in the newborn--an unfavorable prognostic sign.
A hitherto undescribed, unusual EEG abnormality invariably associated with a poor prognosis was reported and was called subclinical delta status. It consisted of continuous discharges of high voltage monomorphic or polymorphic delta activity occurring diffusely but often asynchronously in all leads, without sleep cycles. These discharges when monomorphic may be confused with respiratory artefacts, and when polymorphic they may be mistaken for part of the background activity. Four neonates with this abnormality all had intracranial bleeding and were mechanically ventilated because of frequent apneic attacks. Staring or deviation of the eyes was frequently observed during the EEG recording. A small dose of intravenous diazepam temporarily abolished this activity. Three infants died and the other survived with neurological sequelae.


1H MRSI and social communication deficits in pediatric complex partial seizures.
To investigate relationships between regional brain metabolites, social communication deficits, and seizure frequency in children and adolescents with cryptogenic epilepsy with complex partial seizures (CPS). In 12 children and adolescents with CPS and 23 age- and gender-matched healthy controls, we acquired proton magnetic resonance spectroscopic imaging (MRSI) at 1.5 T and 30 ms echo-time from bilateral inferior frontal and superior temporal gyri, regions associated with social communication deficits. Videotaped speech samples of all the subjects were coded for social communication deficits and parents provided information on seizure frequency. Four MRSI findings emerged in right inferior frontal gyrus. N-acetyl-aspartate (NAA) plus N-acetyl-aspartyl-glutamate (NAAG)--together called "tNAA"--was 11.4% lower in patients with CPS than in controls. Choline-compounds (Cho) were 15.4% lower in CPS than in controls. Within CPS, higher tNAA was associated with more frequent seizures and abnormal social communication. Localization of findings to right inferior frontal cortex supports the involvement of this area in social communication deficits and may be related to atypical lateralization of expressive language in pediatric epilepsy. Lower levels of tNAA and Cho may indicate local neuronal or glial damage or underpopulation due to excitotoxicity or other causes. The sensitivity of tNAA to seizure frequency suggests effects of ongoing CPS on neuronal and glial function in this brain region.


Pharmacoscintigraphic evaluation of potential of lipid nanocarriers for nose-to-brain delivery of antidepressant drug.
Efficacy of antidepressants relies upon their continued presence at the site of action (brain) over a prolonged period of time. The BBB restricts the access of antidepressants to the brain on oral as well as intravenous administration. Direct delivery (by-passing the BBB) of antidepressant drugs can increase the CSF concentration with concomitant reduction in dose and side effects. Intranasal administration of nanostructured lipid carriers (NLC) containing antidepressant drug circumvent the BBB and maintain the prolonged release at the site of action. The aim of the present study was to evaluate the enhancement in brain uptake of NLC containing duloxetine (DLX) after intranasal administration. Duloxetine loaded NLC (DLX-NLC) was evaluated pharmacoscintigraphically for drug targeting potential (DTP), drug targeting efficiency (DTE) and biodistribution studies in different organs including brain. The radiolabeling efficiency of DLX and DLX-NLC was found to be 98.41 ± 0.96 and 98.87 ± 0.82 after 30 min, respectively. The biodistribution studies exhibited higher percentage of radioactivity/g for DLX-NLC formulations in brain as compared with the DLX. The higher DTP (86.80%) and DTE (757.74%) suggested that DLX-NLC formulation has a better brain targeting efficiency than DLX solution (DTP=65.12%; DTE=287.34%) when administered intranasally. Moreover, the intranasal administration exhibited about 8-times higher concentration of DLX in brain when compared with the intravenous administration of DLX solution. The intranasal NLC containing DLX can be employed as an effective method for the treatment of depression.


G protein-coupled receptors in gastrointestinal physiology. I. CCK receptors: an exemplary family.
The CCK and gastrin families of peptides act as hormones and neuropeptides on central and peripheral receptors to mediate secretion and motility in the gastrointestinal tract in the physiological response to a normal meal. Thus far, two CCK receptors have been molecularly identified to mediate the actions of CCK and gastrin, CCK-A and CCK-B receptors (CCK-AR and CCK-BR, respectively). The regulation of CCK-AR and CCK-BR affinity by guanine nucleotides and the receptor activation of G protein-dependent stimulation of phospholipase C and adenylyl cyclase suggested that they were guanine nucleotide-binding protein-coupled receptors [G protein-coupled receptors (GPCRs)]; however, the eventual cloning of their cDNAs revealed their heptahelical structure and confirmed their membership in the GPCR superfamily. The gastrointestinal system is a rich source of neuroendocrine hormones that interact with a large number of GPCRs to regulate the complex tasks of digestion, absorption, and excretion of a meal. This article focuses on the CCK family of GPCRs, and its activities in the gastrointestinal system.


Secretagogue-induced protein phosphorylation and chloride transport in Caco-2 cells.
The effects of vasoactive intestinal polypeptide (VIP), 16,16-dimethyl prostaglandin E2 (DMPGE2) and dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP) on protein phosphorylation were studied in relation to stimulation of chloride transport in cell suspensions of the human colon epithelial cell line Caco-2. In 36Cl-loaded cells, VIP and DMPGE2 within 1 min decreased cellular chloride content 35-40%, with half-maximal effects being elicited at 1.0 and 85 nM concentration, respectively. A similar effect on chloride content occurred after 10 min of treatment with 0.5 mM DBcAMP. For all three secretagogues, decreases in cellular chloride content were associated with increases in membrane permeability to chloride. DMPGE2 and VIP within 1 min, and DBcAMP within 10 min, increased the phosphorylation of an unidentified soluble protein of Mr = 42,000 and pI = 6.1, and of a protein of Mr = 20,200 and pI = 4.9 identified as myosin regulatory light chain. Between 10 and 30 min of stimulation, however, phosphorylation of the Mr = 42,000 protein and chloride transport activity remained elevated in DMPGE2- and DBcAMP-treated cells, whereas light chain phosphorylation returned to control level. No effect of secretagogues on phosphorylation was detected in the total particulate fraction or an integral membrane protein fraction. It is concluded that increased membrane permeability to chloride induced by cAMP-mediated secretagogues in Caco-2 is temporally associated with the increased phosphorylation of a Mr = 42,000 soluble protein.


Costs and clinical outcomes in individuals without known coronary artery disease undergoing coronary computed tomographic angiography from an analysis of Medicare category III transaction codes.
Multidetector coronary computed tomographic angiography (CCTA) demonstrates high accuracy for the detection and exclusion of coronary artery disease (CAD) and predicts adverse prognosis. To date, opportunity costs relating the clinical and economic outcomes of CCTA compared with other methods of diagnosing CAD, such as myocardial perfusion single-photon emission computed tomography (SPECT), remain unknown. An observational, multicenter, patient-level analysis of patients without known CAD who underwent CCTA or SPECT was performed. Patients who underwent CCTA (n = 1,938) were matched to those who underwent SPECT (n = 7,752) on 8 demographic and clinical characteristics and 2 summary measures of cardiac medications and co-morbidities and were evaluated for 9-month expenditures and clinical outcomes. Adjusted total health care and CAD expenditures were 27% (p <0.001) and 33% (p <0.001) lower, respectively, for patients who underwent CCTA compared with those who underwent SPECT, by an average of $467 (95% confidence interval $99 to $984) for CAD expenditures per patient. Despite lower total health care expenditures for CCTA, no differences were observed for rates of adverse cardiovascular events, including CAD hospitalizations (4.2% vs 4.1%, p = NS), CAD outpatient visits (17.4% vs 13.3%, p = NS), myocardial infarction (0.4% vs 0.6%, p = NS), and new-onset angina (3.0% vs 3.5%, p = NS). Patients without known CAD who underwent CCTA, compared with matched patients who underwent SPECT, incurred lower overall health care and CAD expenditures while experiencing similarly low rates of CAD hospitalization, outpatient visits, myocardial infarction, and angina. In conclusion, these data suggest that CCTA may be a cost-efficient alternative to SPECT for the initial coronary evaluation of patients without known CAD.


Equilibrium and kinetic studies of the interaction of the benzimidazole dye Hoechst 8208 with nucleic acids.
The interaction of the benzimidazole dye Hoechst 8208 (H8208) with nucleic acids has been studied by temperature-jump relaxation method. The dependence of relaxation times on the nucleic acid concentration indicates that H8208 binds to RNA by a single biomolecular reaction and to DNA by a bimolecular reaction, followed by a slow isomerization.


Bone mineral density in female athletes representing sports with different loading characteristics of the skeleton.
To address the hypothesis that osteogenic effect of physical loading increases with increasing strain rates and peak forces, we examined 59 competitive Finnish female athletes (representing three sports with different skeletal loading characteristics), physically active referents (they reported an average of five various types of exercise sessions per week), and sedentary referents (two sessions per week) using dual energy X-ray absorptiometry. The measured anatomic sites were at the lumbar spine, femoral neck, distal femur, patella, proximal tibia, calcaneus, and distal radius. The athlete group consisted of aerobic dancers (N = 27), squash players (N = 18), and speed skaters (N = 14). The squash players had the highest values for weight-adjusted bone mineral density (BMD) at the lumbar spine (13.8% p < 0.001 as compared with the sedentary reference group), femoral neck (16.8%, p < 0.001), proximal tibia (12.6%, p < 0.001) and calcaneus (18.5%, p < 0.001). Aerobic dancers and speed skaters also had significantly higher BMD values at the loaded sites than the sedentary reference group, the difference ranging from 5.3% to 13.5%. The physically active referents' BMD values did not differ from those of the sedentary referents at any site. The results support the concept that training, including high strain rates in versatile movements and high peak forces, is more effective in bone formation than training with a large number of low-force repetitions.


Regulation of shortening velocity by calponin in intact contracting smooth muscles.
To elucidate the function of calponin in intact contracting smooth muscle cells in vivo, we generated mice with a mutated basic calponin (h1) locus (Yoshikawa et al., Genes Cells 3, 685-695, 1998). Crossbridge cycling rates were estimated in aortic smooth muscle by the force redevelopment following an isometric step shortening as a function of time after K(+) depolarization. Evidence is presented that calponin is involved in the inhibition of shortening velocity in the tonic phase of contraction. The phosphorylation levels of myosin regulatory light chain and cytosolic calcium concentrations were not significantly different in paired comparisons between calponin-deficient (-/-) and wild-type (+/+) muscles at any time point after stimulation. The force-velocity relationships in vas deferens smooth muscle showed that the maximum shortening velocity of -/- muscle was significantly faster than that of +/+ muscle. There was no change in the length-force relationships in both -/- and +/+ muscles of aorta and vas deferens. The results suggest that calponin plays a role in regulation of the crossbridge cycling and that it may be responsible for reduced shortening velocity during a maintained contraction of mammalian smooth muscle.


Correlates of intrusive memories and avoidance of memories of the Holocaust.
Although intrusive memories and avoidance of memories are psychological phenomena frequently reported by Holocaust survivors, their correlates have not been systematically researched. The present cross-sectional study examined socio-demographic and health characteristics of elderly Holocaust survivors, as well as the survivors' experiences during the Holocaust. In addition, present distress related to the Holocaust and fear of dying were examined. Results showed that survivors frequently suffered from intrusive memories, but not often from avoidance of memories of the Holocaust. Intrusive memories were strongly associated with present distress from the Holocaust, as well as with self-perceived bad health and fear of dying. These findings suggest that intrusive memories influence survivors' perceptions of life and change the way in which they deal with aging.


[Efficacy and metabolic safety of long-term treatment with ethinyl oestradiol/cyproterone and desogestrel/ethinyl oestradiol tablets in women with polycystic ovary syndrome].
To evaluate the efficacy and metabolic safety of long-term treatment with ethinyl oestradiol/cyproteroneand desogestrel/ethinyl oestradiol tablets in women with polycystic ovary syndrome (PCOS). Women with PCOSfrom West China Second Hospital of Sichuan University enrolled between September, 2011 and August, 2013 were randomlyallocated to receive either ethinyl oestradiol/cyproterone tablets (Group A, n=355) or desogestrel/ethinyl oestradiol tablets(Group B, n=357) for a prospective observation period of 6 months. Women with insulin resistance also received metformin. Atbaseline, 3 months, and 6 months, the patients were evaluated for menstruation, acne score, body mass index (BMI), waist-tohip ratio (WHR), plasma levels of sex hormones, fasting blood glucose (FPG), HOMA-insulin resistance index (HOMA-IR), serum lipid, ovarian volume, and the number of ovarian follicles. All the patients had a regular menstrual cycle aftertreatments. Testosterone level, acne score, LH/FSH, ovarian volume, and the number of follicles decreased significantly afterthe treatments without significant differences between the two groups. Significant increases were noted in TG, TCh, LDL, HDL, and AIP, and HDL level in group A as compared with group B (P < 0.001). FPG decreased in both groups, and wassignificantly lower in group B at 6 months (P < 0.05). BMI and WHR decreased in all the patients with insulin resistance aftercombination treatment with metformin (P < 0.05), but increased significantly in patients without insulin resistance (P < 0.05). Ingroup A, HOMA- IR significantly increased in patientswithout insulin resistance at 3 months (P < 0.05), whereas asignificant increase was not observed until 6 months ingroup B (P < 0.05). Both ethinyl oestradiol/cyproterone tablets and desogestrel/ethinyl oestradioltablets can relieve the symptoms of PCOS, but it isadvisable to assess the risk of cardiovascular diseasebefore the treatments.


Cellular and molecular alterations in the beta adrenergic system with cardiomyopathy induced by tachycardia.
The aim was to examine the relationship between changes in myocyte function to changes in protein and mRNA content of components of the beta adrenergic system with tachycardia induced cardiomyopathy. Contractile function and beta adrenergic responsiveness were measured in isolated myocytes from control pigs (n = 6) and in pigs subjected to three weeks of pacing induced supraventricular tachycardia (n = 6). beta Receptor density and affinity, the relative content of the stimulatory (Gs) and inhibitory (Gi) subunits of the G protein complex, and adenylate cyclase activity were determined from sarcolemmal preparations. In order to determine whether these changes were accompanied by alterations in steady state mRNA levels for specific components of the beta adrenergic system, mRNA content for the beta 1 adrenergic receptor and the G alpha s and G alpha i2 subunits of the G protein complex was measured. Chronic supraventricular tachycardia caused a 36% increase in left ventricular end diastolic dimension and a 61% decrease in left ventricular fractional shortening compared to controls. The velocity of isolated myocyte shortening was 50% lower in myocytes from hearts with tachycardia cardiomyopathy than in control myocytes. In the presence of 50 nM isoprenaline or 2 microM forskolin, the velocity of myocyte shortening was 65% lower in the myopathic myocytes than in the controls. With the development of tachycardic cardiomyopathy, beta adrenergic receptor density fell by 25% with no change in affinity, Gs decreased by 35%, and Gi increased by over 50% compared to controls. Basal adenylate cyclase activity and isoprenaline and forskolin stimulated adenylate cyclase activity fell by over 50% with supraventricular tachycardia compared to controls. The relative content of G alpha i2 mRNA increased threefold with the development of tachycardic cardiomyopathy with no change in the relative abundance of mRNA for the beta 1 receptor or G alpha s when compared with controls. The changes in myocyte beta adrenergic responsiveness with the development of tachycardic cardiomyopathy are due to alterations in cellular mechanisms (decreased beta receptor and Gs density, increased Gi) and in molecular mechanisms (increased Gi mRNA content).


Probiotics, prebiotics and the gastrointestinal tract in health and disease.
The microbiome located in the human gastrointestinal tract (GIT) comprises the largest community (diverse and dense) of bacteria, and in conjunction with a conducive internal milieu, promotes the development of regulated pro- and anti-inflammatory signals within the GIT that promotes immunological and metabolic tolerance. In addition, host-microbial interactions govern GIT inflammation and provide cues for upholding metabolic regulation in both the host and microbes. Failure to regulate inflammatory responses can increase the risk of developing inflammatory conditions in the GIT. Here, we review clinical studies regarding the efficacy of probiotics/prebiotics and the role they may have in restoring host metabolic homeostasis by rescuing the inflammatory response. The clinical studies reviewed included functional constipation, antibiotic-associated diarrhoea, Clostridium difficile diarrhoea, infectious diarrhoea/gastroenteritis, irritable bowel syndrome, inflammatory bowel diseases and necrotizing enterocolitis. We have demonstrated that there was an overall reduction in risk when probiotics were administered over placebo in the majority of GIT inflammatory conditions. The effect size of a cumulative reduction in relative risk for the GIT conditions/diseases investigated was 0.65 (0.61-0.70) (z = 13.3); p < 0.0001 that is an average reduction in risk of 35 % in favour of probiotics. We also progress a hypothesis that the GIT comprises numerous micro-axes (e.g. mucus secretion, Th1/Th2 balance) that are in operational homeostasis; hence probiotics and prebiotics may have a significant pharmacobiotic regulatory role in maintaining host GIT homeostasis in disease states partially through reactive oxygen species signalling.


Are protein complexes made of cores, modules and attachments?
It has recently been proposed by Gavin et al. (Nature 2006, 440, 631-636) that protein complexes in the cell exist in different forms. The proteins within each complex were proposed to exist as three different classes, being core, module or attachment proteins. This study investigates whether the core-module-attachment classification of proteins within each complex is supported by other high-throughput protein data. Core proteins were found to have lower abundance, and shorter half-life as compared to attachment proteins, whilst the abundance and half-life of core and module proteins were similar. When the cell was perturbed, core proteins had smaller changes in abundance as compared to module and attachment proteins. Comparisons between six different pairwise interaction types of core, module and attachment proteins within a complex showed interaction types involving core or module proteins were more likely to be mediated by domain-domain interactions (DDIs) than interaction types involving attachment proteins. Interaction types that involve attachment proteins had a relatively higher ratio of abundance and ratio of half-life. So we conclude that, the core, module and attachment model of protein complexes is supported by data from these proteomic scale datasets, and describe a model for a typical protein complex that considers the above results.


[Epidemiology of dental caries in the Netherlands].
Both in 6- and 12-year-old Dutch children the prevalence of dental caries has declined remarkably between 1970 and 1985. However, in 6-year-olds the decline terminated after that period, while it continued in the 12-year-olds. In 1986 a national dental survey of the caries prevalence of older age categories was executed. As no comparable caries data are available from an earlier period of time, no certain conclusion can be drawn about changes in adult caries prevalence. A comparison of the national data mentioned to data from a country where the caries prevalence in children did not decline (former DDR), suggests that Dutch adults younger than 25 year have a relatively low caries experience at this moment.


Mucosal melanoma: clinical, histological and c-kit gene mutational profile of 86 French cases.
Mucosal melanomas are rare and highly aggressive tumours. Few studies evaluated mucosal melanomas of locations other than the head and neck region, and other than those of the Asian population. The objective of this study was to analyse the clinical and histological features, as well as the mutational status of c-kit and b-raf gene of mucosal melanoma in any localization in a French series. We investigated clinical (sex, age, performance status, survival, treatment of the patients and lack of pigmentation of the tumours) and histopathological features (ulceration, Breslow's index, mitotic rate), as well as the mutational status of c-kit and b-raf of 86 mucosal melanomas diagnosed in 15 years in four French University Hospitals. Most melanomas affected women (72%) and the genital region (46.5%). A fifth of melanomas were amelanotic. 81% of melanomas had a Breslow's index ≥1, whereas all glans melanomas, and most vulvar melanomas had a Breslow index ≤1 mm. Overall survival was 54% at 3 years; 11.6% of the 43 tested mucosal melanomas were c-kit-mutated while the 15 tested genital melanomas were not. The c-kit gene mutation did not influence the overall survival. Age ≥ 50, amelanotic type and performance status ≥1 were not poor prognostic factors in our series. This study confirmed that mucosal melanomas are rare and could be difficult to diagnose being often amelanotic and in hidden sites. Most melanomas were thick at the diagnosis, but glans and vulvar melanomas were thinner probably because of their greater visibility. The frequency of the c-kit mutation varied depending on the initial tumour site. In our series, the prognosis was poor, independently from c-kit mutations and the patient's general health and age. The presence of metastasis at diagnosis was associated with a worse prognosis indicating the importance of an early diagnosis.


Breakthrough invasive fungal disease in patients receiving posaconazole primary prophylaxis: a 4-year study.
Posaconazole (PSC) is currently recommended as primary prophylaxis in neutropenic patients with acute myeloid leukaemia (AML) and in allogenic haematopoietic stem cell transplantation (AHSCT) recipients with graft-versus-host disease (GVHD). Studies focusing on breakthrough invasive fungal disease (IFD) upon PSC prophylaxis show disparate results. In order to evaluate the incidence of IFD in patients on PSC prophylaxis and identify IFD risk factors, we carried out a retrospective study of all consecutive patients on PP from January 2007 to December 2010 in our hospital. Breakthrough IFDs were identified from the database of the central pharmacy and the French administrative database (PMSI), registering final medical diagnoses of hospitalized patients. Medical data were reviewed to study proven or probable IFD, according to EORTC/MSG definition. PSC plasma concentrations (PPC) were also retrieved. Poisson models were used for statistical analysis. Two hundred and seventy-nine patients received PSC prophylaxis for a median duration of 1.4 months (range 0.2-17.9). Proven (n=6) or probable (n=3) IFDs were diagnosed in nine cases (3.2%). IFD incidence rate per 100 person-month was 1.65 (95% CI, 0.79-2.97). IFDs were candidaemia (Candida glabrata, n=2), pulmonary invasive aspergillosis (n=3), disseminated fusariosis (n=2) and pulmonary mucormycosis (n=2). Seven deaths were reported, directly related to IFD in three patients (33.3%). First dosage of PPC under 0.3 mg/L was the single significant risk factor for IFD (RR, 7.77; 95% CI, 1.30-46.5; p 0.025). Breakthrough IFD in patients receiving PSC prophylaxis is rare but associated with a poor outcome. Low PSC plasma concentrations are associated with an increased risk of IFD.


Sex and hemispheric differences for rapid auditory processing in normal adults.
Previous research suggests that left hemisphere specialisation for processing speech may specifically depend on rate-specific parameters, with rapidly successive or faster changing acoustic stimuli (e.g. stop consonant-vowel syllables) processed preferentially by the left hemisphere. The current study further investigates the involvement of the left hemisphere in processing rapidly changing auditory information, and examines the effects of sex on the organisation of this function. Twenty subjects participated in an auditory discrimination task involving the target identification of a two-tone sequence presented to one ear, paired with white noise to the contralateral ear. Analyses demonstrated a right ear advantage for males only at the shorter interstimulus interval durations (mean = 20 msec) whereas no ear advantage was observed for women. These results suggest that the male brain is more lateralised for the processing of rapidly presented auditory tones, specifically at shorter stimulus durations.


Preliminary study of DNA microarray of human placenta gene expression profile.
To prepare human placenta genechip for analyzing differential gene expressions. The target gene amplified from Hybrid Hunter cDNA library was dotted onto the slides by the arrayer (PixSys 5500). Total RNA from K562 cells treated with or without arsenic trioxide was extracted, the mRNA purified and cDNA synthesized. Fluorescent labeling of the samples was performed by restriction display PCR, followed by hybridization with the microarray that was subsequently washed and scanned, with the derived signals analyzed by computer. A reliable and specific method for preparing and assessing gene expression profile microarray was established, with which a total of 45 differentially expressed gene fragments were isolated. The genechips can be helpful in the analysis of differential gene expressions.


An ultrasensitive iron(III)-complex based hydrogen peroxide electrochemical sensor based on a nonelectrocatalytic mechanism.
In this communication, the first nonelectrocatalysis-type hydrogen peroxide electrochemical sensor is reported. The electroactive iron(III) diethylenetriaminepentaacetic acid (DTPA-Fe(III)) complex is immobilized on the cysteamine (cys) modified nanoporous gold (NPG) films by covalent method. The immobilized DTPA-Fe(III) complex quickly communicates an electron with the electrode. Upon addition of hydrogen peroxide, however, hydrogen peroxide inhibits the direct electron transfer of the DTPA-Fe(III) complex due to the generation of nonelectroactive DTPA-Fe(III)-H2O2 complex. Based on quenching mechanism, the first hydrogen peroxide electrochemical sensor based on a nonelectrocatalytic mechanism is developed. The novel hydrogen peroxide electrochemical sensor has the ultralow detection limit (1.0×10(-14) M) and wide linear range (1.0×10(-13) to 1.0×10(-8) M) with excellent reproducibility and stability.


Serum tumor markers as predictors for survival in advanced non-small cell lung cancer patients treated with gefitinib.
Though the imaging-based response (IBR) is the most frequently used index of the therapeutic effect in cancer patients, an index based on serum tumor markers might prove to be useful, especially in patients without measurable tumors. Paired pre- and post-treatment serum tumor markers (CEA, CA-125, and CA-199) were measured in 89 of 100 registered non-small cell lung cancer (NSCLC) patients who received gefitinib. Correlation and agreement analyses between the IBR at the 8th week and the tumor marker response (TMR) at the 4th or the 8th week were performed in patients with measurable lesions (n=68). Analysis of survival in relation to TMR was performed in all patients and in patients with no measurable lesions (n=21). IBR was closely correlated with individual tumor marker responses and the response of all 3 markers combined at 4 weeks (P values ranged from <0.001 to 0.002). The agreements were also significant (P values ranged from 0.001 to 0.004). In the whole group, 4-week TMR was predictive for progression-free survival (P values ranged from <0.0001 to 0.0086). In patients without measurable lesions, differences in progression-free survival and overall survival were closely correlated with the 4-week CA-125 response, CA-199 response, and TMR(overall) (P values ranged from 0.0002 to 0.0399). However, the correlation between the 8-week TMR and either IBR or survival was not significant. In gefitinib-treated NSCLC patients, correlation was good between 4-week TMR and IBR. Four-week TMR was predictive for survival. TMR may serve as a tool for assessing the gefitinib response in patients without measurable lesions.


Surgical treatment of intramuscular, infiltrating lipoma.
Lipoma may occasionally be found within the muscle, which is the intramuscular, infiltrating lipoma. Intramuscular, infiltrating lipoma is an uncommon lesion, and local recurrence can be expected if the surgical margin is not clear. We report 8 patients in a 2-year period diagnosed with intramuscular, infiltrating lipoma who were treated by marginal excision combined with wide excision in the Department of Orthopaedic Surgery, Changhua Christian Hospital. The surgical results were satisfactory, and no local recurrence was noted in an average followup period of 40 months. Surgical excision is the best way to treat the symptomatic intramuscular, infiltrating lipoma. Careful preoperative evaluation and complete tumor excision with clear margins are most important to ensure successful surgical results.


Comparison of Poly(l-lactide-co-ɛ-caprolactone) and Poly(trimethylene carbonate) Membranes for Urethral Regeneration: An In Vitro and In Vivo Study.
Urethral defects are normally reconstructed using a patient's own genital tissue; however, in severe cases, additional grafts are needed. We studied the suitability of poly(l-lactide-co-ɛ-caprolactone) (PLCL) and poly(trimethylene carbonate) (PTMC) membranes for urethral reconstruction in vivo. Further, the compatibility of the materials was evaluated in vitro with human urothelial cells (hUCs). The attachment and viability of hUCs and the expression of different urothelial cell markers (cytokeratin 7, 8, 19, and uroplakin Ia, Ib, and III) were studied after in vitro cell culture on PLCL and PTMC. For the in vivo study, 32 rabbits were divided into the PLCL (n = 15), PTMC (n = 15), and control or sham surgery (n = 2) groups. An oval urethral defect 1 × 2 cm in size was surgically excised and replaced with a PLCL or a PTMC membrane or urethral mucosa in sham surgery group. The rabbits were followed for 2, 4, and 16 weeks. After the follow-up, urethrography was performed to check the patency of the urethra. The defect area was excised for histological examination, where the epithelial integrity and structure, inflammation, and fibrosis were observed. There was no notable difference on hUCs attachment on PLCL and PTMC membranes after 1 day of cell seeding, further, the majority of hUCs were viable and maintained their urothelial phenotype on both biomaterials. Postoperatively, animals recovered well, and no severe strictures were discovered by urethrography. In histological examination, the urothelial integrity and structure developed toward a normal urothelium with only mild signs of fibrosis or inflammation. According to these results, PLCL and PTMC are both suitable for reconstructing urethral defects. There were no explicit differences between the PLCL and PTMC membranes. However, PTMC membranes were more flexible, easier to suture and shape, and developed significant epithelial integrity.


Imaging diagnosis: magnetic resonance imaging of a cervical wooden foreign body in a dog.
This article describes the discovery of a chronic cervical wooden foreign body ventral to the left transverse processes of the cranial cervical spine using magnetic resonance imaging (MRI) in a dog that presented with chronic neck pain and lameness. The dog did not exhibit dysphagia or chronic draining tracts, the most common signs of the presumed cause, that of a penetrating oropharyngeal foreign body. The foreign body itself was represented on MR images as an oval straight-edged core within an inflammatory tissue reaction. The wood was slightly hyperintense on T2- and isointense on T1-weighted images relative to muscle. Surrounding this was a more conspicuous contrast-enhancing reactive tissue rim that was hyperintense on all pulse sequences. Adjacent musculature also exhibited diffuse edema and contrast enhancement that extended around the left cervical vertebral transverse processes and local intervertebral nerve roots. The foreign body was found to be a wooden stick upon surgical removal. MRI is an excellent method for visualizing the inflammatory tissue reactions associated with soft-tissue foreign bodies because of its contrast resolution and depiction of anatomy in multiple imaging planes.


Hay bailed out by student support..
I read with some relief the article by Robert Hay on nursing research (A nurse's place..., Viewpoint, March 30).


[Testosterone deficiency and the problem of normal values].
If a testosterone deficiency is demonstrated, the exact cause of the endocrine hypogonadism should be determined by appropriate endocrinological, imaging and other andrological diagnostic techniques, as such a deficiency can be the result of various illnesses. At present there are no satisfactory evidence based, generally accepted norms for testosterone levels recognised for the aging male. Laboratory analysis of testosterone must be carried out taking into consideration the physiological variability using methods validated in house, with strict internal and external quality control. For the diagnosis of reduced testosterone levels, satisfactorily established own normal values for the technique used are recommended.


SCD1 inhibition during 3T3-L1 adipocyte differentiation remodels triacylglycerol, diacylglycerol and phospholipid fatty acid composition.
The conversion of saturated fatty acids (FAs) palmitate (16:0) and stearate (18:0) into monounsaturated FAs palmitoleate (16:1n-7) and oleate (18:1n-9) is catalyzed by stearoyl-CoA desaturase 1 (SCD1). These FAs represent the dominant constituents of adipocyte triacylglycerols (TAGs) and phospholipids (PLs). Given the critical role of SCD1 in lipid metabolism and the notable increase in its expression during adipogenesis, reductions in SCD1 activity have the potential to compromise the adipocyte's ability to accumulate lipid. The current study used thin-layer and gas chromatography to examine the content and FA composition of TAGs, PLs, cholesteryl esters, diacylglycerols and free fatty acids in SCD1-inhibited differentiating 3T3-L1 adipocyte cells. SCD1 inhibition reduced total cellular PL and TAG content concurrent with the down-regulation of genes involved in TAG and PL biosynthesis; however, the relative amount of PL was unaltered. While total DAG levels were increased ~2.7-fold in SCD1-inhibited adipocytes, this did not induce JNK activation; however, phosphorylated (Ser473) AKT was significantly reduced. As expected, total SFA and MUFA content were increased (~1.3-fold) and decreased (~4.0-fold). Further, SCD1 inhibition caused a ~2.2-fold increase and a ~8.3-fold decrease in total cellular 18:0 and 16:1n-7 levels, respectively. Similar changes were also seen in other lipid fractions. The levels of other FAs, including polyunsaturated FAs, were also changed in SCD1-inhibited adipocytes. Together, these results add to the existing body of knowledge regarding SCD1 function in adipocytes and highlight its important role in regulating global adipocyte lipid composition.


Comparative interinstrument reliability of uniaxial and triaxial accelerometers in free-living conditions.
The aim of this study was to measure the inter-instrument reliability of the ActiGraph and RT3 accelerometers in free-living conditions. 15 healthy adults wore eight ActiGraph accelerometers and five RT3 accelerometers fastened to their back with an elastic belt and adjustable buckle in free-living conditions. For both accelerometers, the inter-instrument coefficient of variation (CV) decreased with increasing physical activity intensity. The inter-instrument CV of the ActiGraph (3% to 10.5%) was significantly lower than that of the RT3 (12.6% to 35.5%; p < 0.05). This finding demonstrates that the ActiGraph has higher inter-instrument reliability than the RT3. For both accelerometers, the inter-instrument reliability was higher for moderate and vigorous physical activity, the two intensities recommended for public health purposes. Studies using these devices can be compared with a low risk of misclassification of participants who meet the recommendation to perform moderate to vigorous physical activity.


An advisory statement from the Pediatric Working Group of the International Liaison Committee on Resuscitation.
The International Liaison Committee on Resuscitation (ILCOR), with representation from North America, Europe, Australia, New Zealand, Africa, and South America, was formed in 1992 to provide a forum for liaison between resuscitation organizations in the developed world. This consensus document on resuscitation extends previously published ILCOR advisory statements on resuscitation to address the unique and changing physiology of the newly born infant within the first few hours after birth and the techniques for providing advanced life support. After careful review of the international resuscitation literature and after discussion of key and controversial issues, consensus was reached on almost all aspects of neonatal resuscitation, and areas of controversy and high priority for additional research were delineated. Consensus on resuscitation for the newly born infant included the following principles: Personnel trained in the basic skills of resuscitation should be in attendance at every delivery. A minority (fewer than 10%) of newly born infants require active resuscitative interventions to establish a vigorous cry and regular respirations, maintain a heart rate > 100 beats per minute (bpm), and maintain good color and tone. When meconium is present in the amniotic fluid, it should be suctioned from the hypopharynx on delivery of the head. If the meconium-stained newly born infant has absent or depressed respirations, heart rate, or muscle tone, residual meconium should be suctioned from the trachea. Attention to ventilation should be of primary concern. Assisted ventilation with attention to oxygen delivery, inspiratory time, and effectiveness judged by chest rise should be provided if stimulation does not achieve prompt onset of spontaneous respirations and/or the heart rate is < 100 bpm. Chest compressions should be provided if the heart rate is absent or remains < 60 bpm despite adequate assisted ventilation for 30 seconds. Chest compressions should be coordinated with ventilations at a ratio of 3:1 and a rate of 120 "events" per minute to achieve approximately 90 compressions and 30 rescue breaths per minute. Epinephrine should be administered intravenously or intratracheally if the heart rate remains < 60 bpm despite 30 seconds of effective assisted ventilation and chest compression circulation. Common or controversial medications (epineprine, volume expansion, naloxone, bicarbonate), special resuscitation circumstances affecting care of the newly born, continuing care of the newly born after resuscitation, and ethical considerations for initiation and discontinuation of resuscitation are discussed. There was agreement that insufficient data exist to recommend changes to current guidelines regarding the use of 21% versus 100% oxygen, neuroprotective interventions such as cerebral hypothermia, use of a laryngeal mask versus endotracheal tube, and use of high-dose epinephrine. Areas of controversy are identified, as is the need for additional research to improve the scientific justification of each component of current and future resuscitation guidelines.


Dielectric and elastic properties of liquid crystals.
The structural properties, the static and relaxation dielectric coefficients [epsilon(j) and epsilon(j)(omega) (j= ||, perpendicular)], the rotational diffusion constants D( perpendicular) and D( ||), the orientational correlation times tau(1)(i0) (i=0,1), and the bulk elastic constants K(i) (i=1,2,3) are investigated for polar liquid crystals, such as 4-n-pentyl-4(')-cyanobiphenyl (5CB). epsilon(j) are calculated by a combination of the existing molecular theory and statistical-mechanical approach (SMA) that takes into account translational and orientational correlations as well as their coupling, whereas epsilon(j)(omega) are calculated by combining SMA and nuclear magnetic resonance relaxation theory, both based on a rotational diffusion model in which the reorientation of an individual molecule is assumed as stochastic Brownian motion in a potential of mean torque. Reasonable agreement between the calculated and experimental values of epsilon(j) and epsilon(j)(omega) for 5CB is obtained. The bulk Frank elastic constants K(i) (i=1,2,3), for splay, twist, and bend distortion modes, as well as their ratios K(3)/K(1) and K(2)/K(1) are also obtained.


Anomalous coherence peak in the microwave conductivity of c-axis oriented MgB2 thin films.
The temperature dependence of the real part of the microwave complex conductivity at 17.9 GHz obtained from surface impedance measurements of two c-axis oriented MgB2 thin films reveals a pronounced maximum at a temperature around 0.6 times the critical temperature. Calculations in the frame of a two-band model based on Bardeen-Cooper-Schrieffer (BCS) theory suggest that this maximum corresponds to an anomalous coherence peak resembling the two-gap nature of MgB2. Our model assumes there is no interband impurity scattering and a weak interband pairing interaction, as suggested by band structure calculations. In addition, the observation of a coherence peak indicates that the pi band is in the dirty limit and dominates the total conductivity of our films.


Outcome of mechanically ventilated patients who require a tracheostomy.
To estimate the prevalence of, the risk factors associated with, and the outcome of tracheostomy in a heterogeneous population of mechanically ventilated patients. Prospective, observational cohort study. A total of 361 intensive care units from 12 countries. A cohort of 5,081 patients mechanically ventilated for >12 hrs. None. A total of 546 patients (10.7%) had a tracheostomy during their stay in the intensive care unit. Tracheostomy was performed at a median time of 12 days (interquartile range, 7-17) from the beginning of mechanical ventilation. Variables associated with the performance of tracheostomy were duration of mechanical ventilation, need for reintubation, and neurologic disease as the primary reason of mechanical ventilation. The intensive care unit stay of patients with or without tracheostomy was a median of 21 days (interquartile range, 12-32) vs. 7 days (interquartile range, 4-12; p < .001), respectively, and the hospital stay was a median 36 days (interquartile range, 23-53) vs. 15 days (interquartile range, 8-26; p < .001), respectively. Adjusting by other variables, tracheostomy was independently related with survival in the intensive care unit (odds ratio, 2.22; 95% confidence interval, 1.72-2.86). Mortality in the hospital was similar in both groups (39% vs. 40%, p = .65). Tracheostomy is a common surgical procedure in the intensive care unit that is associated with a lower mortality in the unit but with a longer stay and a similar mortality in the hospital than in patients without tracheostomy.


The Prevalence and Management of Metabolic Acidosis of Chronic Kidney Disease
Aim Emerging evidence supports initiating oral sodium bicarbonate (OSB) at a serum bicarbonate (HCO3) level of less than 22mmol/L. We look to identify the prevalence of metabolic acidosis of chronic kidney disease (MA-CKD) and its management with OSB at a regional university hospital. Methods Retrospective data was collected using the national electronic renal database (eMED) to identify chronic kidney disease (CKD) patients with MA-CKD over a one-year period. Results One-hundred and forty-four patients were identified with CKD, of which 131 (89%) were tested for HCO3. MA-CKD was present in 44 patients (34%), all had eGFR< 30ml/min/1.73m2, 7 (16%) were prescribed OSB, 7(16%) OSB was contraindicated, and 37 (84%) patients managed with dietary input only. Mean HCO3 level at initiation in OSB group was 18.3±1mmol/L compared to 19.4±1.4mmol/L in dietary input only group which was statistically significant (p<0.05). Conclusion A high burden of advanced CKD was found in the regional nephrology centre, with one third of patients demonstrating MA-CKD. Majority had dietary input only. Further awareness and consensus need to be established on the benefits of treating MA-CKD with OSB.


[Studies on the beneficial effect of levocarnitine chloride (LC-80) on organic acidemias, especially propionic acidemia and methylmalonic acidemia].
The beneficial effect of LC-80 in the therapy for organic acidemias, especially propionic acidemia and methylmalonic acidemia, was compared with those of its optical isomers, d-carnitine chloride (d-isomer) and dl-carnitine chloride (dl-isomer) in rat liver mitochondria. LC-80 at concentrations of 5 and 10 mM did not inhibit the mitochondrial function, while the d-isomer at a concentration of 5 mM significantly reduced the respiratory control ratio (RCR) of mitochondria. In addition, the dl-isomer at concentrations of 10 and 20 mM also significantly reduced RCR in a concentration-dependent manner. Thus, it seems likely that the d-isomer inhibits the mitochondrial function. On the other hand, the inhibition of mitochondrial function induced by a preincubation with propionate (4.76 mM) was significantly reversed by LC-80 (5 and 10 mM) in a concentration-dependent manner, while the d-isomer (5 mM) had no effect on the inhibitory effect of propionate. Moreover, although the dl-isomer (10 and 20 mM) significantly reversed the inhibitory effect of propionate as compared with the d-isomer, its effect was significantly weaker as compared with the effect of LC-80. The substrate specificity of rat liver mitochondrial carnitine acetyltransferase (CAT) was more potent with propionyl CoA than with acetyl CoA. Kinetic studies indicate that the d-isomer is a competitive inhibitor of CAT. These results suggest that LC-80 is useful in the clinical treatment of organic acidemias, whereas the d-isomer has a harmful effect in clinical application.


Removal of viable sheets of conjunctival epithelium with dispase II.
A technique (based on Gipson and Grill, Invest Ophthalmol Vis Sci 23:269, 1982) has been developed to obtain pure, viable, intact sheets of rabbit conjunctival epithelium free of the underlying basement membrane. After preparing a full thickness eye wall resection, Dispase, grade II (neutral protease-Bacillus polymyxa), 1.2 Units/ml MEM is injected intrasclerally. The conjunctiva and sclera are pinned in agar and incubated in the dispase with MEM for 1 hour. A 2 X 3 mm sheet of conjunctival epithelium can be dissected bluntly. Light microscopy shows a two- to three-layered epithelium with many goblet cells. Transmission electron microscopy reveals blebbing at the freed basal epithelial cell membrane, intact desmosomes, and intact goblet cells. The conjunctival sheets were cultured on epithelial-scraped corneal stromal carriers in vitro. Numerous goblet cells were present up to 12 hours on 4-mm carriers and 24 hours on 1-mm carriers. With this technique, pure populations of conjunctival epithelium can be isolated for further characterization and tissue culture.


Fast Versus Slow Recruitment Maneuver at Different Degrees of Acute Lung Inflammation Induced by Experimental Sepsis.
Large tidal volume (VT) breaths or "recruitment maneuvers" (RMs) are used commonly to open collapsed lungs, but their effectiveness may depend on how the RM is delivered. We hypothesized that a stepped approach to RM delivery ("slow" RM) compared with a nonstepped ("fast" RM), when followed by decremental positive end-expiratory pressure (PEEP) titration to lowest dynamic elastance, would (1) yield a more homogeneous inflation of the lungs, thus reducing the PEEP obtained during post-RM titration; (2) produce less lung morphofunctional injury, regardless of the severity of sepsis-induced acute lung inflammation; and (3) result in less biological damage in severe, but not in moderate, acute lung inflammation. Sepsis was induced by cecal ligation and puncture surgery in 51 Wistar rats. After 48 hours, animals were anesthetized, mechanically ventilated (VT = 6 mL/kg), and stratified by PO2/fraction of inspired oxygen ratio into moderate (≥300) and severe (<300) acute lung inflammation groups. Each group was then subdivided randomly into 3 subgroups: (1) nonrecruited; (2) RM with continuous positive airway pressure (30 cm H2O for 30 seconds; CPAPRM or fast RM); and (3) RM with stepwise airway pressure increase (5 cm H2O/step, 8.5 seconds/step, 6 steps, 51 seconds; STEPRM or slow RM), with a maximum pressure hold for 10 seconds. All animals underwent decremental PEEP titration to determine the level of PEEP required to optimize dynamic compliance after RM and were then ventilated for 60 minutes with VT = 6 mL/kg, respiratory rate = 80 bpm, fraction of inspired oxygen = 0.4, and the newly adjusted PEEP for each animal. Respiratory mechanics, hemodynamics, and arterial blood gases were measured before and at the end of 60-minute mechanical ventilation. Lung histology and biological markers of inflammation and damage inflicted to endothelial cells were evaluated at the end of the 60-minute mechanical ventilation. Respiratory system mean airway pressure was lower in STEPRM than that in CPAPRM. The total RM time was greater, and the RM rise angle was lower in STEPRM than that in CPAPRM. In both moderate and severe acute lung inflammation groups, STEPRM reduced total diffuse alveolar damage score compared with the score in nonrecruited rats. In moderate acute lung inflammation, STEPRM rats compared with CPAPRM rats had less endothelial cell damage and angiopoietin (Ang)-2 expression. In severe acute lung inflammation, STEPRM compared with CPAPRM reduced hyperinflation, endothelial cell damage, Ang-2, and intercellular adhesion molecule-1 expressions. RM rise angle correlated with Ang-2 expression. Compared with CPAPRM, STEPRM reduced biological markers associated with endothelial cell damage and ultrastructural endothelial cell injury in both moderate and severe sepsis-induced acute lung inflammation.


Outbreak of cholera in Berasia block of Bhopal District in Madhya Pradesh.
Bhopal district, Madhya Pradesh has been considered to be free of cholera. In an outbreak of gastro-enteritis in Berasia block of Bhopal district, Cholera vibrio Inaba and Ogawa were isolated from 17.64 per cent of stool samples collected. Extreme insanitary conditions, large aggregation of uncontrolled population and probably consumption of infected ice candies were responsible for the outbreak of cholera.


[Alternative bone replacement substances for preoperative design of individual CAD/CAM skull implants].
The preoperative manufacturing of individual skull implants using computer aided design (CAD) and computer aided manufacturing (CAM) is based on the use of titanium, although the use of other materials is also potentially possible. The use of poly-(D,L-lactide) (PDLLA) as an implant material was investigated using an adult, formalin fixed sheep's head with a complex frontolateral defect. A standard individual titanium implant as well as a resection template made of aluminium were milled in order to allow bone resection and reconstruction within one operation. A mould was made of Teflon for the fabrication of the PDLLA implant using carbon dioxide at high pressure. This procedure allowed a critical comparison to be made of both implant materials and showed that the production of a biodegradable PDLLA implant is possible. At present the titanium implant is superior to the PDLLA implant, as PDLLA settled with slightly larger dimensions than the mould, although the structure itself was exact. The goal of the present research is the fabrication of a functionally graded material made of polylactide, polyglycolide, calcium phosphate and osteoinductive proteins using existing technology, which will meet all of the requirements for stability, resorption kinetics, biocompatibility, radiotranslucence and osteogenic potency of an ideal implant material.


Markers of inflammation and weight change in middle-aged adults: results from the prospective MONICA/KORA S3/F3 study.
We investigated associations of markers of inflammation such as albumin, fibrinogen, C-reactive protein (CRP), and white blood cell count (WBCC) with future weight gain and weight loss in middle-aged adults in order to further elucidate the relationship between subclinical inflammation and weight change. Data were derived from the third population-based MONICA (Monitoring of Trends and Determinants in Cardiovascular Diseases) Augsburg survey (S3) conducted as part of the multinational World Health Organization MONICA project in 1994-1995 and a follow-up examination, to which all eligible subjects from S3 were invited in 2004-2005 (F3). In total, 2,792 persons (1,391 men, 1,401 women) aged 25-74 years at baseline were analyzed. Subjects with elevated concentrations of inflammatory markers were more prone to gain weight during follow-up. The odds ratios (OR) for a large mean annual weight gain (i.e., on average 1.02 kg/year) was 1.73 (95% confidence interval (CI) 1.27, 2.35) in fully adjusted analyses for subjects in the highest compared to the lowest quartile of fibrinogen. The respective ORs were 1.45 (95% CI, 1.08, 1.94) and 1.37 (95% CI, 1.03, 1.82) for CRP and WBCC. Stratified analyses revealed that associations were strongest among subjects who quitted smoking during the follow-up period (new quitters). Associations of inflammatory markers with large mean annual weight loss were weaker and became nonsignificant after multivariable adjustment. In conclusion, elevated levels of inflammatory markers are independently associated with weight gain in middle-aged adults, particularly among new quitters. This suggests that inflammation plays a key role in the process of weight gain, especially after smoking cessation.


A reduced oxygen tension (5%) is not beneficial for maintaining human embryonic stem cells in the undifferentiated state with short splitting intervals.
Human embryos grow naturally in vivo in lower oxygen (O(2)) tension environments than atmospheric O(2) tension. Therefore, human embryonic stem cells (hESC), a derivative of embryos, will likely grow more favorably in a reduced O(2) tension. This study aimed to compare the behavior of hESC under reduced O(2) tension (5%) versus normoxia (21%). hESC lines were cultured in different O(2) tensions and then examined for morphology, apoptosis and gene expression profiles. hESC grown in 5% O(2) tension were not morphologically different from hESC grown in normoxia on day 7 of the first and fourth passages. However, after prolonged culture without splitting (10-14 days), hESC colonies were thinner and looked better morphologically in 5% O(2), but the cells proliferated more slowly and their sizes were larger. At most time points, the gene expression profiles in both O(2) tensions showed no major difference in representative stemness genes (Oct-3/4, Nanog and Cripto), differentiation genes (Desmin, Nestin, alpha-fetoprotein and GDF-9) and hypoxia-related genes (HIF-1alpha and VEGF). A lower level of cyclin-D1 mRNA (suggestive of less Wnt pathway signaling on day 7 of the fourth passage) and a higher level of Desmin (suggestive of more differentiation to mesoderm, at day 7 of the first passage) were detected in 5% O(2). This study suggests that for routine culture of hESC with a short splitting interval (7 days), a low O(2) tension (5% O(2)) probably does not provide significant advantages over the standard 21% O(2) tension for the maintenance of an undifferentiated state by the criteria used in this study.


Fitness attenuates the prevalence of increased coronary artery calcium in individuals with metabolic syndrome.
Background The association between cardiorespiratory fitness, physical activity and coronary artery calcium (CAC) is unclear, and whether higher levels of fitness attenuate CAC prevalence in subjects with metabolic syndrome is not fully elucidated. The present study aims to: a) investigate the independent association of fitness on the prevalence of CAC, after adjustment for moderate-to-vigorous physical activity and sedentary time, and b) study the possible attenuation of increased CAC by higher fitness, in participants with metabolic syndrome. Design Cross-sectional. Methods In total 678 participants (52% women), 50-65 years old, from the SCAPIS pilot study were included. Fitness (VO2max) was estimated by submaximal cycle ergometer test and moderate-to-vigorous physical activity and sedentary time were assessed using hip-worn accelerometers. CAC score (CACS) was quantified using the Agatston score. Results The odds of having a significant CACS (≥100) was half in participants with moderate/high fitness compared with their low fitness counterparts. Further consideration of moderate-to-vigorous physical activity, sedentary time and number of components of the metabolic syndrome did only slightly alter the effect size. Those with metabolic syndrome had 47% higher odds for significant CAC compared with those without metabolic syndrome. However, moderate/high fitness seems to partially attenuate this risk, as further joint analysis indicated an increased odds for having significant CAC only in the unfit metabolic syndrome participants. Conclusions Being fit is associated with a reduced risk of having significant CAC in individuals with metabolic syndrome. While still very much underutilized, fitness should be taken into consideration in everyday clinical risk prediction in addition to the traditional risk factors of the metabolic syndrome.


Deciphering the crystal ball in spine care: can preoperative patient variables predict postoperative functional outcomes and risks?
Meredith DS, Huang RC, Nguyen J, Lyman S. Obesity increases the risk of recurrent herniated nucleus pulposus after lumbar microdiscectomy. Spine J 2010;10:575-580 (in this issue). Yadla S, Malone J, Campbell PG, et al. Obesity and spine surgery: reassessment based on a prospective evaluation of perioperative complications in elective degenerative thoracolumbar procedures. Spine J 2010;10:581-587 (in this issue).


Gastric cancer. Clinico-biological updating and analysis of 400 operated cases.
Gastric cancer is a rather common disease worldwide. In Italy it still accounts for 15,000 deaths annually. A sharp drop in the incidence rate of Lauren's intestinal histotype has been reported, whereas the frequency of the diffuse histotype is relatively steady. If the histogenesis of the latter is still somewhat obscure, the intestinal type confirms the sequence: atrophic gastritis--intestinal metaplasia--dysplasia--neoplasia. These different stages of development can nowadays be singled out through a series of indicators, the most reliable of which are the pepsinogen I/pepsinogen II ratio, the presence of sulphomucins and Lewis antigens in the gastric juices and NOR (Nucleolar Organizer Regions), cell ploidy and oncogenes determination. The genes involved in the neoplastic transformation are mostly oncosuppressors, the most frequent alterations being those relative to the APC gene, p53 and c-myc. In addition to the by now indispensable pathological staging of the disease, the modern prognostic factors are arising great interest: the most significant are the immunohistochemical examination of the peritoneal washing, and cell ploidy. Surgery is still the only potentially curative treatment: the earlier surgery is performed in the course of disease, the greatest the curative potential. The Authors' experience, which includes 400 operated cases with complete follow-up records, is here reported. The resectability rate turned out to be 84%, overall operative mortality was 6.5% with that due to surgical causes along being 3.7%. Overall survival at 5 years was 36%, while that of the curative operations 47%. Good results were obtained with the association surgery + intraoperative radiotherapy which resulted in a significant decrease in local recurrences of the disease.


Gelation in mixtures of polymers and bidisperse colloids.
We investigated the effects of varying the volume fraction of large particles (r) on the linear rheology and microstructure of mixtures of polymers and bidisperse colloids, in which the ratio of the small and large particle diameters was α=0.31 or α=0.45. Suspensions formulated at a total volume fraction of ϕ_{T}=0.15 and a constant concentration of polymer in the free volume c/c^{*}≈0.7 contained solid-like gels for small r and fluids or fluids of clusters at large r. The solid-like rheology and microstructure of these suspensions changed little with r when r was small, and fluidized only when r>0.8. By contrast, dense suspensions with ϕ_{T}=0.40 and α=0.31 contained solid-like gels at all concentrations of large particles and exhibited only modest rheological and microstructural changes upon varying the volume fraction of large particles. These results suggest that the effect of particle-size dispersity on the properties of colloid-polymer mixtures are asymmetric in particle size and are most pronounced near a gelation boundary.


Stimulus-transcription coupling in focal cerebral ischemia.
Glutamate-mediated spreading depression is currently thought to be a key event in the pathogenesis of potential neuronal degeneration in the ischemic 'penumbra'. Glutamate receptor stimulation causes induction of transcription factors that belong to the class of immediate early genes (IEGs), thought to be involved in coupling neuronal excitation to target gene expression. Focal cerebral ischemia elicits a homogeneous expression of several IEGs, prominently in cortex. In the ischemic core, discrepancies are observed between mRNA and protein levels, due to a severe, persistent protein synthesis deficit, preventing the translation of IEG encoded mRNAs. Outside the ischemic core, widespread IEG expression occurs in the entire ipsilateral cortex at mRNA as well as at protein level. This homogeneous expression of transcription factors can be pinpointed to at least two different pathogenetic mechanisms by means of appropriate pharmacological antagonists. Prolonged IEG induction in the 'penumbra', an area in which neurons are metabolically compromised but not yet energy-depleted, cannot be suppressed by the administration of N-methyl-D-aspartate (NMDA) receptor antagonists. In contrast, short-lasting IEG induction in undamaged neurons remote from the ischemic territory, though also caused by ischemia-elicited spreading depression, can be blocked by NMDA receptor antagonists. In both areas, IEG expression identifies neurons destined to survive but is likely to be mediated by different signal transduction pathways, at the receptor, second messenger and/or the DNA level.


Sonographic diagnosis of hypertrophic pyloric stenosis.
In many pediatric centers, sonography has become the primary imaging method for the diagnosis of hypertrophic pyloric stenosis, but investigators have disagreed about which dimension of the gastric outlet is most useful and accurate. We analyzed 326 consecutive sonograms in 319 infants who presented with vomiting in order to further evaluate the sonographic criteria for the diagnosis of hypertrophic pyloric stenosis. Four observers retrospectively measured three parameters: (1) the thickness of the muscle in the wall of the gastric outlet, (2) the length of that muscle, and (3) the length of the pyloric channel. In the patients with hypertrophic pyloric stenosis, muscle thickness was 4.8 +/- 0.6 mm, muscle length was 2.1 +/- 0.3 cm, and channel length was 1.8 +/- 0.3 cm. In the patients without hypertrophic pyloric stenosis, muscle thickness was 1.8 +/- 0.4 mm, muscle length was 1.3 +/- 0.3 cm, and channel length was 1.1 +/- 0.3 cm. The mean measurement for each parameter was significantly larger in patients who had hypertrophic pyloric stenosis than in those who did not (p less than .01). However, histograms of the three parameters show significant overlap in the measurements of muscle length and channel length between the patients with and those without hypertrophic pyloric stenosis. The thickness of the muscle is the most discriminating and accurate criterion for hypertrophic pyloric stenosis.


Sleep patterns of co-sleeping and solitary sleeping infants and mothers: a longitudinal study.
Controversies exist regarding the impact of co-sleeping on infant sleep quality. In this context, the current study examined: (a) the differences in objective and subjective sleep patterns between co-sleeping (mostly room-sharing) and solitary sleeping mother-infant dyads; (b) the predictive links between maternal sleep during pregnancy and postnatal sleeping arrangement; (c) the bi-directional prospective associations between sleeping arrangement and infant/maternal sleep quality at three and six months postpartum. The sample included 153 families recruited during pregnancy. Data were obtained in home settings during the third trimester of pregnancy and at three and six months postpartum. Mothers were asked to monitor their own sleep and their infants' sleep for five nights using actigraphy and sleep diaries. Questionnaires were used to assess sleeping arrangements, feeding methods, socio-demographic characteristics, and maternal depressive and anxiety symptoms. Mothers of co-sleeping infants reported more infant night-wakings than mothers of solitary sleeping infants. However, none of the objective sleep measures was significantly different between co-sleeping and solitary sleeping infants, after controlling for feeding techniques. Co-sleeping mothers had significantly more objective and subjective sleep disturbances than mothers in the solitary sleeping group. Moreover, poorer maternal sleep during pregnancy and at three months postpartum predicted higher levels of co-sleeping at six months. Mothers of co-sleeping infants report more infant night-wakings, and experience poorer sleep than mothers of solitary sleeping infants. The quality of maternal sleep should be taken into clinical consideration when parents consult about co-sleeping.


[Electrocochleography using Transtympanic, Ear Drum and Ear Canal Electrode in Diagnosis of Morbus Menière].
Aim of this study was to explore the clinical value of non-invasive recordings of electrocochleography in comparison to the use of invasive electrodes for the diagnosis of endolymphatic hydrops in patients with suspicious Morbus Menière. 15 patients with clinical suspicion of Morbus Menière underwent electrocochleography with transtympanic needle-electrode as well as tympanic membrane electrode. 10 patients received electrocochleography with transtympanic needle-electrode and ear canal electrode. A control group of 12 healthy adults was evaluated using tympanic membrane electrode. The negative electrode was placed on the ipsilateral mastoid, the ground electrode in the middle of the forehead. For stimulation broadband click stimulus as well as long tone bursts of 1, 2 and 4 kHz were applied. In addition the impedance of the electrode was registered. While transtympanic electrocochleography gave reliable results in 100% of these cases, extratympanic recordings often manifested poor wave morphology. The electrocochleographic measures following stimulation with 1 and 2 kHz long tone bursts were the best measures to identify endolymphatic hydrops. This was the case with transtympanic as well as with tympanic membrane electrodes. There was no correlation between impedance and wave morphology. For the evaluation of Morbus Menière non-invasive electrocochleography showed good results using a tympanic canal electrode with 1 and 2 kHz tone bursts. Yet in unclear cases transtympanic electrocochleography should be preferred.


Sequential and selective localized optical heating in water via on-chip dielectric nanopatterning.
We study the use of nanopatterned silicon membranes to obtain optically-induced heating in water. We show that by varying the detuning between an absorptive optical resonance of the patterned membrane and an illumination laser, both the magnitude and response time of the temperature rise can be controlled. This allows for either sequential or selective heating of different patterned areas on chip. We obtain a steady-state temperature of approximately 100 °C for a 805.5nm CW laser power density of 66 µW/μm2 and observe microbubble formation. The ability to spatially and temporally control temperature on the microscale should enable the study of heat-induced effects in a variety of chemical and biological lab-on-chip applications.


A prospective comparison of continuous wave versus high pulse repetition frequency Doppler echocardiography for quantifying transvalvular pressure gradients in adults with aortic stenosis.
To evaluate the relative accuracy of continuous wave (CW) and high pulse repetition frequency (HPRF) Doppler for estimating aortic transvalvular pressure gradients, Doppler examinations with both devices were obtained in 87 consecutive patients with aortic valve disease. Thirty-seven (43%) also underwent catheterization and direct determination of the pressure gradient. The correlation between CW Doppler and catheterization measurements was high (r = 0.91) and significantly superior to HPRF Doppler (r = 0.72; p less than 0.005). Disparities between the Doppler measurements predominantly occurred in patients with gradients exceeding 50 mm Hg in whom underestimation by the HPRF instrument was frequent. These data suggest that continuous wave Doppler is the preferred modality for the estimation of transvalvular pressure gradients in adult patients with moderate or severe aortic stenosis.


An Element of Life.
While it has been known for decades that the essential function of selenium was in the form of its incorporation as selenocysteine into selenoproteins-including the enzyme glutathione peroxidase-4-now, Ingold et al. (2018) reveal the precise role of selenolate-based catalysis by this enzyme.


ZEA exerts toxic effects on reproduction and development by mediating Dio3os in mouse endometrial stromal cells.
Zearalenone (ZEA) and imprinted long noncoding RNAs (lncRNAs) are both closely related to reproduction and development. However, whether they have connections in regulating reproduction and development is not clear yet. The aim of this research is to investigate their relationship. lncRNA microarray was performed to analyze differentially expressed genes, and real-time quantitative polymerase chain reaction (PCR) was used to verify the accuracy of microarray analysis. Meanwhile, the technologies of rapid amplification of cDNA ends, RNA fluorescence in situ hybridization and bioinformatics were adopted to characterize the selected lncRNA. Analysis of lncRNA microarray showed lncRNAs and messenger RNAs related to reproduction and development were significantly differently expressed, and Dio3os was probably the target lncRNA. Then, the experiment of real-time quantitative PCR verified the accuracy of microarray data. Characterization of Dio3os showed Dio3os, an antisense lncRNA with 2312 bp and 15 open reading frames, was enriched in the cytoplasm. Our findings suggest ZEA probably exerts toxic effects on reproduction and development by mediating Dio3os.


14-3-3beta binds to and negatively regulates the tuberous sclerosis complex 2 (TSC2) tumor suppressor gene product, tuberin.
TSC2, or tuberin, is the product of the tuberous sclerosis tumor suppressor gene TSC2 and acts downstream of the phosphatidylinositol 3-kinase-Akt signaling pathway to negatively regulate cellular growth. One mechanism underlying its function is to assemble into a heterodimer with the TSC1 gene product TSC1, or hamartin, resulting in a reduction in phosphorylation, and hence activation, of the ribosomal subunit S6 kinase (S6K). We identified a novel interaction between TSC2 and 14-3-3beta. We found that 14-3-3beta does not interfere with TSC1-TSC2 binding and can form a ternary complex with these two proteins. Association between 14-3-3beta and TSC2 requires phosphorylation of TSC2 at a unique residue that is not a known Akt phosphorylation site. The overexpression of 14-3-3beta compromises the ability of the TSC1-TSC2 complex to reduce S6K phosphorylation. The antagonistic activity of 14-3-3beta toward TSC is dependent on the 14-3-3beta-TSC2 interaction, since a mutant of TSC2 that is not recognized by 14-3-3beta is refractory to 14-3-3beta. We suggest that 14-3-3 proteins interact with the TSC1-TSC2 complex and negatively regulate the function of the TSC proteins.


Relationship between two serotypes of avian mycoplasmas by serological and DNA-DNA hybridization tests.
Genetic relatedness among eight strains of avian mycoplasmas representing two serotypes, C-O and D-P, was determined using DNA-DNA hybridization tests. One strain from each serotype was found to be misclassified. The average percent homology between the two serotypes was 34.1% at 56 C and 18.6% at 66 C. The average homology among the four C-O organisms was 99.6% at 56 C and 93.5% at 66 C, while among the four D-P strains the average homology was 91.6% at 56 C and 92.2% at 66 C. These relationships correlated with antigenic cross-reactions observed in complement fixation (CF) and fluorescent antibody (FA) tests. Thermal denaturation of the pure DNA from each strain resulted in base compositions of 25.0 to 28.2 moles percent guanosine plus cytosine (GC). Thermal denaturation of selected hybrid DNA revealed a linear relationship between percent homology and delta melting temperature (1 C delta Tm for 15% difference in percent homology). Thirty-four percent homology between the organisms of C-O and D-P serotypes indicates a minor antigenic relationship; thus, it is recommended that these two serotypes be retained as separate groups.


Molecular analysis of non structural rotavirus group A enterotoxin gene of bovine origin from India.
The rotavirus enterotoxin NSP4 (nonstructural protein 4), plays a pivotal role in viral morphogenesis as well as pathogenesis. In this study, the NSP4 gene of rotavirus group A (RVA) isolates of bovine origin isolated in several states of India from 2008 to 2011 were characterized. The complete open reading frame of 23 RVA strains were sequenced and analyzed phylogenetically. Genotype E1 was detected for the first time in bovines from India, in addition to the more common bovine genotype E2. Sequence similarity analysis of the E1 sequences showed a close genetic relatedness to human strains. Six of the bovine E2 genotypes strains clustered near bovine and unusual human strains (possible human animal reassortant) from Thailand, while the remaining E2 sequences clustered with Indian bovine strains. Analysis pointed out one positively selected site (154aa), believe to be part of an antigenic region and 123 negatively selected sites. Unexpectedly, a pentameric NSP4 structure of the coiled coil domain in the E1 carrying strains and a monomeric NSP4 in RVA strain P14 (E2) was predicted based on homology modeling, potentially affecting the biological properties of NSP4. The close relationship between bovine and human rotavirus strains further highlights the complex interaction among rotaviruses of different species.


Hyper-selective plasmonic color filters.
The subwavelength mode volumes of plasmonic filters are well matched to the small size of state-of-the-art active pixels in CMOS image sensor arrays used in portable electronic devices. Typical plasmonic filters exhibit broad (> 100 nm) transmission bandwidths suitable for RBG or CMYK color filtering. Dramatically reducing the peak width of filter transmission spectra would allow for the realization of CMOS image sensors with multi- and hyperspectral imaging capabilities. We find that the design of 5 layer metal-insulator-metal-insulator-metal structures gives rise to multi-mode interference phenomena that suppress spurious transmission features and give rise to single transmission bands as narrow as 17 nm. The transmission peaks of these multilayer slot-mode plasmonic filters (MSPFs) can be systematically varied throughout the visible and near infrared spectrum, leading to a filter that is CMOS integrable, since the same basic MSPF structure can operate over a large range of wavelengths. We find that MSPF filter designs that can achieve a bandwidth less than 30 nm across the visible and demonstrate experimental prototypes with a FWHM of 70 nm, and we describe how experimental structure can be made to approach the limits suggested by the model.


Coevolution of female ovulatory signals and male-male competition in primates.
Visual signals of ovulation vary among primate species. Although slight ovulatory signals are considered primate ancestral traits of which some species still exhibit, some show prominent swelling of their perineal skin (exaggerated sexual swellings) and others do not exhibit any signals of ovulation (concealed ovulation). These signals strongly affect male mating behaviors. I develop an evolutionary model of female ovulatory signals and male-male competition. I assume that each male allocates his effort between attraction of females and male-male competition for dominance. Each female gains a benefit if she is fertile and free from the alpha male who always guards one of the most fertile females in the group, but suffers a cost if she expresses a different ovulatory signal from an ancestral trait. I show that various types of ovulatory signals may evolve and can be multistable. Male-male competition becomes intense when the signal honestly indicates ovulation. Ovulatory signals may evolve to be less exaggerated in unimale groups than in multimale groups and monogamy is more likely to evolve when ovulation is concealed. These results may partly explain why various types of primate ovulatory signals evolved and how they have affected primate societies.


Several adaptor proteins promote intracellular localisation of the transporter MRP4/ABCC4 in platelets and haematopoietic cells.
The multidrug resistance protein 4 (MRP4/ABCC4) has been identified as an important transporter for signalling molecules including cyclic nucleotides and several lipid mediators in platelets and may thus represent a novel target to interfere with platelet function. Besides its localisation in the plasma membrane, MRP4 has been also detected in the membrane of dense granules in resting platelets. In polarised cells it is localised at the basolateral or apical plasma membrane. To date, the mechanism of MRP4 trafficking has not been elucidated; protein interactions may regulate both the localisation and function of this transporter. We approached this issue by searching for interacting proteins by in vitro binding assays, followed by immunoblotting and mass spectrometry, and by visualising their co-localisation in platelets and haematopoietic cells. We identified the PDZ domain containing scaffold proteins ezrin-binding protein 50 (EBP50/NHERF1), postsynaptic density protein 95 (PSD95), and sorting nexin 27 (SNX27), but also the adaptor protein complex 3 subunit β3A (AP3B1) and the heat shock protein HSP90 as putative interaction partners of MRP4. The knock-down of SNX27, PSD95, and AP3B1 by siRNA in megakaryoblastic leukaemia cells led to a redistribution of MRP4 from intracellular structures to the plasma membrane. Inhibition of HSP90 led to a diminished expression and retention of MRP4 in the endoplasmic reticulum. These results indicate that MRP4 localisation and function are regulated by multiple protein interactions. Changes in the adaptor proteins can hence lead to altered localisation and function of the transporter.


Absolute iron deficiency without anaemia in patients with chronic systolic heart failure is associated with poorer functional capacity.
Functional status is one of the main concerns in the management of heart failure (HF). Recently, the FAIR-HF and CONFIRM-HF trials showed that correcting anaemia using intravenous iron supplementation improved functional variables in patients with absolute or relative iron deficiency. Relative iron deficiency is supposed to be a marker of HF severity, as ferritin concentration increases with advanced stages of HF, but little is known about the impact of absolute iron deficiency (AID). To study the impact of AID on functional variables and survival in patients with chronic systolic HF. One hundred and thirty-eight non-anaemic patients with chronic systolic HF were included retrospectively. Patients were divided into two groups according to iron status: the AID group, defined by a ferritin concentration<100μg/L and the non-AID group, defined by a ferritin concentration≥100μg/L. Functional, morphological and biological variables were collected, and survival was assessed. Patients in the AID group had a poorer 6-minute walking test (342 vs. 387m; P=0.03) and poorer peak exercise oxygen consumption (13.8 vs. 16.0mL/min/kg; P=0.01). By multivariable analysis, ferritin<100μg/L was associated with impaired capacity of effort, assessed by peak exercise oxygen consumption. By multivariable analysis, there was no difference in total mortality between groups, with a mean follow-up of 5.1±1.1 years. The poorer functional evaluations in iron-deficient patients previously reported are not caused by the merging of two different populations (i.e. patients with absolute or relative iron deficiency). Our study has confirmed that non-anaemic HF patients with AID have poorer peak oxygen consumption. However, AID has no impact on the survival of these patients.


Patents in therapeutic recombinant protein production using mammalian cells.
The industrial production of recombinant proteins preferentially requires the generation of stable cell lines expressing proteins in a quick, relatively facile, and a reproducible manner. Different methods are used to insert exogenous DNA into the host cell, and choosing the appropriate producing cell is of paramount importance for the efficient production and quality of the recombinant protein. This review addresses the advances in recombinant protein production in mammalian cell lines, according to key patents from the last 30 years.


Nocebo as headache trigger: evidence from a sham-controlled provocation study with RF fields.
A large proportion of the population in Norway has experienced headache in connection with mobile phone use, but several double-blind provocation studies with radiofrequency (RF) and sham exposures have shown no relation between headache and mobile phone RF fields. To investigate the type and location of headache experienced by participants in one provocation study in order to gain insight into possible causes and mechanisms of the headaches. Questionnaire about headache, indication on figure of location of headache after exposure, interview with neurologist about headache features to make headache diagnoses. The 17 participants went through 130 trials (sham or RF exposure). No significant difference existed in headache type, laterality or location between the headaches experienced with the two exposures types. In most participants, the headache was compatible with tension-type headache. As participants experienced their typical 'mobile phone headache' both with and without RF exposure, and since the experiment did not involve the stress or the arm/head position of mobile phone use, the most likely explanation is that the headache in this situation is caused by negative expectations (nocebo). This and other similar studies indicate that headache occurring in connection with mobile phone use is not related to RF fields, and that a nocebo effect is important for this and possibly other headache triggers.


Effect of cyclosporine on the antigen-presenting function of human and murine accessory cells.
Recent reports have challenged the belief that accessory cells are resistant to cyclosporine. Such a tenet was based on the observation that several functions of accessory cells, such as IL-1 production and phagocytosis, are resistant to the drug. On the other hand, when a less primitive, more refined function of accessory cells was examined--i.e., the capacity to take up, process, and present antigen in an MHC-restricted fashion to antigen-specific T lymphocytes, CsA proved to be an effective inhibitor. In contrast to this finding, when antigen was provided in the form of an immune complex prepared with a monoclonal antibody, uptake of antigen--likely mediated by the Fc receptors--and subsequent processing and presentation were not affected by CsA. These results suggest that, depending on whether the antigen is taken up by constitutive or by receptor-mediated endocytosis, accessory cells can be functionally defined as resistant or sensitive to CsA.


Primary antiphospholipid antibody syndrome presenting as venous infarct and deep vein thrombosis.
We report the case of a young lady with primary antiphospholipid antibody syndrome, who had two spontaneous abortions and cerebral venous thrombosis and subsequently deep vein thrombosis of the leg veins. Three classes of antiphospholipid antibodies (IgG, IgM, IgA) were elevated. There was no clinical or laboratory evidence for other autoimmune or systemic illnesses. We are presenting the case due to the rarity of the same.


Failure of trimethoprim/sulfamethoxazole prophylaxis for Pneumocystis carinii pneumonia with concurrent leucovorin use.
Pneumocystis carinii is a common cause of pneumonia in patients with AIDS, however, the incidence has dropped with the availability of effective prophylactic regimens. First-line treatment for both acute Pneumocystis pneumonia and chronic prophylaxis is trimethoprim/sulfamethoxazole (TMP/SMX). This combination can cause hypersensitivity reactions as well as myelosuppression. The simultaneous administration of leucovorin during acute treatment has been shown to reduce the incidence of neutropenia, but may interfere with the efficacy of TMP/SMX. We report a case of P. carinii pneumonia in a patient with AIDS who failed TMP/SMX prophylaxis while taking leucovorin.