Activation of 5-HT1C/2 receptors depresses polysynaptic reflexes and excitatory amino acid-induced motoneuron responses in frog spinal cord.
Sucrose gap recordings from the ventral roots of isolated, hemisected frog spinal cords were used to evaluate the effects of high concentrations of serotonin (5-HT) and alpha-methyl-5-HT (alpha-Me-5-HT) on the changes in motoneuron potential produced by dorsal root stimulation and by excitatory amino acids and agonists. Bath application of 5-HT in concentrations of 10 microM or greater produced a concentration-dependent motoneuron depolarization. Polysynaptic ventral root potentials evoked by dorsal root stimuli were reduced in both amplitude and area by 5-HT or alpha-Me-5-HT (both 100 microM). This may result from a reduction of the postsynaptic sensitivity of motoneurons to excitatory amino acid transmitters because 5-HT significantly depressed motoneuron depolarizations produced by addition of L-glutamate and L-aspartate to the superfusate. Similarly, 5-HT reduced depolarizations produced by the excitatory amino acid agonists N-methyl-D-aspartate (NMDA), quisqualate, alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid (AMPA), and kainate. alpha-Me-5-HT reduced NMDA depolarizations. Tetrodotoxin (TTX) did not affect the ability of 5-HT to attenuate NMDA or kainate depolarizations, but did eliminate the 5-HT-induced attenuation of quisqualate and AMPA depolarizations. The glycine receptor site associated with the NMDA receptor did not appear to be affected by 5-HT because saturation of the site by excess glycine did not alter the 5-HT-induced depression of NMDA responses. The 5-HT1C/2 antagonist ketanserin and the 5-HT1A/2 antagonist spiperone significantly attenuated the 5-HT-induced depression of NMDA-depolarizations.(ABSTRACT TRUNCATED AT 250 WORDS)


Normal reference values of three-dimensional speckle-tracking echocardiography-derived mitral annular dimensions and functional properties in healthy adults: Insights from the MAGYAR-Healthy Study.
There is a limited number of echocardiographic studies determining mitral annular (MA) dimensions in healthy subjects. The present study aimed to establish normal reference values of three-dimensional speckle-tracking echocardiography-derived MA dimensions and functional properties in healthy adults in relation with age and gender. The present study comprised 298 healthy adult subjects. From this population, 94 subjects were excluded due to inadequate image quality. Therefore the remaining group consisted of 204 subjects with the mean age of 33.88 ± 12.97 years (107 males). The population sample was further divided into age categories: 18-29 years (n = 105; mean age: 24.11 ± 2.98 years, 51 males), 30-39 years (n = 44; mean age: 33.80 ± 2.39 years, 31 males), 40-49 years (n = 19; mean age: 43.47 ± 3.18 years, 11 males) and ≥50 years of age (n = 36, mean age: 57.42 ± 6.11 years, 14 males). End-diastolic MA dimensions did not change significantly during the decades. End-systolic MA diameter, area, and perimeter were larger over the age of 50 years than in the 18-29 year-old group. MA fractional area change was found smaller over the age of 50 years than in 18-29-year-old group. While end-diastolic MA variables did not show gender-differences, end-systolic MA area and perimeter were lower in females in the 18-29-year-old group. End-systolic MA dimensions change over decades, resulting in a special pattern of MA functional properties with significant reduction over the age of 50 years.


Genetic variability in ABCB1, occupational pesticide exposure, and Parkinson's disease.
Studies suggested that variants in the ABCB1 gene encoding P-glycoprotein, a xenobiotic transporter, may increase susceptibility to pesticide exposures linked to Parkinson's Disease (PD) risk. To investigate the joint impact of two ABCB1 polymorphisms and pesticide exposures on PD risk. In a population-based case control study, we genotyped ABCB1 gene variants at rs1045642 (c.3435C/T) and rs2032582 (c.2677G/T/A) and assessed occupational exposures to organochlorine (OC) and organophosphorus (OP) pesticides based on self-reported occupational use and record-based ambient workplace exposures for 282 PD cases and 514 controls of European ancestry. We identified active ingredients in self-reported occupational use pesticides from a California database and estimated ambient workplace exposures between 1974 and 1999 employing a geographic information system together with records for state pesticide and land use. With unconditional logistic regression, we estimated marginal and joint contributions for occupational pesticide exposures and ABCB1 variants in PD. For occupationally exposed carriers of homozygous ABCB1 variant genotypes, we estimated odds ratios of 1.89 [95% confidence interval (CI): (0.87, 4.07)] to 3.71 [95% CI: (1.96, 7.02)], with the highest odds ratios estimated for occupationally exposed carriers of homozygous ABCB1 variant genotypes at both SNPs; but we found no multiplicative scale interactions. This study lends support to a previous report that commonly used pesticides, specifically OCs and OPs, and variant ABCB1 genotypes at two polymorphic sites jointly increase risk of PD.


Effects of the water extract of Gynura bicolor (Roxb. & Willd.) DC on physiological and immune responses to Vibrio alginolyticus infection in white shrimp (Litopenaeus vannamei).
Gynura bicolor (Roxb. & Willd.) DC is widely distributed in certain areas of Asia and is very popular in vegetarian cuisine in Taiwan. To investigate the regulatory roles of G. bicolor in various functions in crustaceans, we examined innate non-specific immune responses (including total hemocyte count (THC), phenoloxidase activity (PO), respiratory bursts (RBs), and superoxide dismutase (SOD) activity), physiological responses (including haemolymph glucose, lactate, and lipids), and gene expressions (including prophenoloxidase (proPO), lipopolysaccharide- and b-1,3-glucan-binding protein (LGBP), and peroxinectin (PE) mRNA transcripts) to the pathogen Vibrio alginolyticus in white shrimp (Litopenaeus vannamei) that were individually injected with the water extract from G. bicolor at 2, 4, and 8 μg g(-1). Results indicated that PO, RBs, SOD activity, proPO, LGBP, and PE mRNA transcripts of shrimps receiving the water extract of G. bicolor at 2, 4, and 8 μg g(-1) significantly increased after challenge with V. alginolyticus for 96 h. However, no significant difference in the THC was seen at any dose. L. vannamei injected with the water extract of G. bicolor at all doses respectively maintained lower glucose, lactate, and lipid levels in response to V. alginolyticus challenge at 12-36, 24-36, and 24-48 h. Survival rates at 24-72 h of L. vannamei that received G. bicolor at any dose was significantly higher than those of shrimp that received saline. It was concluded that the water extract of G. bicolor can maintain physiological homeostasis and enhance immunity against V. alginolyticus infection in L. vannamei.


Study of the RNA-degrading activities during the development of the insect Ceratitis capitata.
1. Three different RNA-degrading activities have been characterized in the insect C. capitata. Two of them are non dependent on divalent cations and show acid and alkaline optimum pH values respectively. For the third one, the maximum activity is observed at pH 8.5, being this enzyme inhibited by EDTA. 2. Distribution of the enzyme levels during the development of the insect is reported. Results are interpreted in terms of the functional role of these enzymes.


Implementation of multiuser MUMPS language database systems on microcomputers.
This paper outlines the problems involved in implementing multiuser MUMPS language database systems on microcomputers and discusses practical solutions. Examples of the techniques presented are derived from experience with a large database system, PTIN/PTQ/MEDAR, running on an IBM AT with up to 10 separate users.


Estimation of the use of dietary supplements in the National Diet and Nutrition Survey: people aged 65 years and Over. An observed paradox and a recommendation.
To compare the evidence derived from blood biochemical status indices with the evidence from a questionnaire and from a 4-day weighed dietary record of micronutrient supplement use in the British National Diet and Nutrition Survey (NDNS) of People Aged 65 Years and Over; to resolve some apparent incompatibility between nutrient intake and status estimates, and to recommend an approach towards supplement recording that should improve accuracy. The survey procedures described in the National Diet and Nutrition Survey Report (1998) included a health-and-lifestyle questionnaire, a 4-day weighed diet record, and fasting blood and urine sample for biochemical indices, including a wide range of micronutrients. Eighty randomly selected postcode sectors from mainland Britain during 1994-1995. Of 2060 people interviewed, 1467 provided a blood sample and 1217 provided both a blood sample, and a complete 4-day diet record. About 20% were living in institutions such as nursing homes, and the remainder were living in private households. After assigning the subjects to four categories by the use of dietary supplements (A, those not taking supplements (by questionnaire or by the 4-day record); B, those taking supplements (excluding prescribed ones) by questionnaire only; C, those taking supplements by 4-day record only; and D, those taking supplements by both questionnaire and 4-day record), these categories were then compared with respect to estimated total nutrient intakes and blood biochemical indices. Those in category B had estimated (4-day) nutrient intakes (from foods and supplements) that were indistinguishable from those in category A, but had biochemical indices that indicated significantly higher dietary intakes of several vitamins. CONCLUSIONS AND RECOMMENDATION: The 4-day weighed intake record may not have identified all of the subjects who were regularly taking micronutrient supplements in amounts sufficient to improve their biochemical status. Because survey respondents may use supplements irregularly or change their usual patterns of supplement use during a period of intensive diet-recording, it is important to design a dietary instrument that will minimise this potential source of inaccuracy. We therefore recommend that population surveys in which an accurate estimate of micronutrient intakes is required, from supplements as well as from food, should record supplement use for a period longer than 4-days. It is likely that a better estimate of long-term intakes can be achieved by combining a 4-day weighed diet record with a structured recall or several weeks of diary records, which focus specifically on the use of supplements.


Whipworms and intestinal threadworms.
Whipworms (genus Trichuris) parasitize the lower bowel of humans, domestic animals, and wild animals. The species of Trichuris that are important in small animal practice are T. vulpis in the dog and T. campanula and T. serrata in the cat. Intestinal threadworms (Strongyloides stercoralis in the dog and S. tumefaciens in the cat) parasitize the small intestine. The identification of whipworms and threadworms, their life cycles, and the diagnosis, treatment, and control of infection are discussed in this article.


Late recruitment of synapsin to nascent synapses is regulated by Cdk5.
Synapse formation is a complex process that involves the recruitment and assembly of a myriad of pre- and postsynaptic proteins. Despite being present at every synapse in the vertebrate CNS, little is known about the transport, recruitment, and stabilization of synapsin at nascent synapses during development. We examined the transport and recruitment of synapsin to nascent presynaptic terminals in vivo in the developing zebrafish spinal cord. Synapsin was transported in a transport packet independently of two other presynaptic organelles: synaptic vesicle (SV) protein transport vesicles (STVs) and Piccolo-containing active zone precursor transport vesicles (PTVs). During presynaptic assembly, recruitment of all three transport packets occurred in an ordered sequence: STVs preceded PTVs, which in turn preceded synapsin. Importantly, cyclin-dependent kinase 5 (Cdk5) specifically regulated the late recruitment of synapsin transport packets at synapses. These results point to additional layers of complexity in the established mechanisms of synaptogenesis.


Contributions of marginal offspring to reproductive success of Nazca booby (Sula granti) parents: tests of multiple hypotheses.
While obligate siblicide is a phylogenetically widespread behavior, known from plants, insects, birds, and other taxa, with important implications for life history evolution, comprehensive evaluations of its costs and benefits to parents are rare. We used 12 years of breeding and band resight data to evaluate the importance of several potential benefits that marginal offspring (the usual victims of obligate siblicide) could provide to parent Nazca boobies (Sula granti), a seabird. We found no evidence for the resource-tracking hypothesis: 99.95% of two-chick broods were reduced to one chick before fledging, and the single exceptional brood probably lost one chick between fledging and independence. Behavioral observations indicated that siblicidal aggression caused most mortality of marginal chicks, and at least contributed to the remainder. We also found no evidence that marginal offspring provide a food resource for other family members. Marginal chicks benefit parents via adoption into other families, and possibly also in the context of progeny choice, but these benefits are minor compared to the insurance that marginal chicks provide against early failure of core (first-hatched) offspring. Further evaluation of the Insurance Egg Hypothesis showed that marginal and core offspring are functionally equivalent in the absence of sibling interactions, and that core offspring incur no detectable costs from behaving siblicidally. Nazca boobies are truly obligate brood reducers, with parents receiving principally insurance benefits from marginal offspring, but many birds and other taxa exhibiting persistent, unconditional sibling aggression do not exhibit universal brood reduction. Insurance is only one of several potential benefits that marginal offspring can confer on parents, and a multi-hypothesis approach to decompose the different types of benefits is required to understand the evolution of clutch size in other obligately siblicidal species.


Pulmonary atresia with intact ventricular septum and right ventricular dependent coronary circulation through the "vessels of Wearn".
We present an autopsy case of a male baby born at 35 weeks of gestation with pulmonary atresia with intact ventricular septum (PAIVS), who had coronary blood flow that was dependent on outflow from the right ventricle through the vessels described by Wearn. At 7 weeks of age, he underwent single ventricle palliation consisting of ligation of a patent ductus arteriosus and placement of a modified Blalock-Taussig shunt. The patient experienced a perioperative myocardial infarction, requiring extracorporeal membrane oxygenation. Progressive hemodynamic decline resulted in death 8 days after surgery. Autopsy revealed acute and remote infarctions in both ventricles and fibromuscular dysplasia of the subepicardial and intramural coronary arteries. In 1926, Grant first reported the association between PAIVS and secondary dysplasia of the heart vasculature and hypothesized that the high pressure resulted in dilation of the myocardial sinusoids. Confusion secondary to the unmeritorious dismissal of the myocardial sinusoids has obscured the pathogenesis of PAIVS and led to several publications suggesting second heart field abnormalities as a disease model for PAIVS. We discuss the pathogenesis of PAIVS, the ventriculocoronary arterial connections and the sinusoidal relationship to the vessels described by Wearn, and we attempt to correct the solecism plaguing the nomenclature of myocardial vasculature.


Bitoric gas-permeable contact lens application in pellucid marginal corneal degeneration.
To evaluate the authors' experience fitting patients with pellucid marginal corneal degeneration (PMCD) with bitoric gas-permeable contact lenses. The records of 11 patients with a confirmed diagnosis of PMCD were retrospectively reviewed. The numbers of diagnostic contact lenses at the first visit, office visits, and lenses ordered to complete the fitting; uncorrected, spectacle-corrected, and contact lens-corrected Snellen visual acuities; contact lens success or failure; and complications encountered were all assessed. These data were compared to historic data for patients with other corneal irregularities (keratoconus, penetrating keratoplasty, and refractive surgery). The mean number of diagnostic contact lenses used at the first visit was 1.09 +/- 0.302, and the number of lenses ordered to complete the fitting was 1.82 +/- 1.33. The number of visits during 4 months was 6.18 +/- 2.14. Uncorrected visual acuity (logMAR) was 0.957 +/- 0.398 (20/181 in Snellen); the best-corrected spectacle visual acuity (logMAR) was 0.231 +/- 0.309 (20/34 in Snellen); and the best contact lens-corrected visual acuity (logMAR) was 0.0424 +/- 0.06275 (20/22 in Snellen). Corrected vision in eyes with PMCD improved approximately two lines with bitoric gas-permeable contact lenses compared to that achieved with spectacles. Ten (91) of the patients met the criteria of contact lens success during the 4-month follow-up period. Bitoric gas-permeable contact lenses achieved a comfortable and visually successful fit in most eyes with PMCD. Patients with PMCD used the fewest contact lenses and had the fewest lens exchanges, but required more office visits than other groups of patients with various corneal irregularities.


Voltammetric determination of catechol and hydroquinone at poly(murexide) modified glassy carbon electrode.
The electrochemical study of Catechol (CC) was studied in presence of Hydroquinone (HQ) using poly(murexide) modified glassy carbon electrode (MGCE) by cyclic voltammetric study. The modified electrode exhibited good affirmative response towards the electro chemical oxidation of CC in presence of HQ in 0.2 M phosphate buffer solution at the physiological of pH 7.4. The influence of scan rate, concentration and pH parameters were examined with the cyclic voltammetric method. The modified electrode showed good selectivity and sensitivity with an advantage of interference study and free determination of selected analytes in this experiment. Finally a simple, sensitive and selective method was developed for the determination of dihydroxybenzene isomeric compounds.


Ultrasound-contrast agent mediated naked gene delivery in the peritoneal cavity of adult rat.
Gene transfer into the peritoneal cavity by nonviral methods may provide an effective therapeutic approach for peritoneal diseases. Herein, we investigated the feasibility and the effectiveness of ultrasound-microbubble-mediated delivery of naked plasmid DNA into the peritoneal cavity in rats. Following the intraperitoneal or the intravenous administration of a mixture of plasmid DNA (100 microg) and ultrasound contrast agent microbubbles, an ultrasound transducer was applied on the abdominal wall. The reporter pTRE plasmid encoding Smad7 was used to evaluate transfection efficiency. Smad7 expression was induced by doxycycline in drinking water. We detected less than 10% apoptotic cells and no inflammatory reaction in peritoneal tissues following the ultrasound-microbubble-mediated transfection. More importantly, the insonation significantly improved the transfection efficiency in peritoneal tissues. The transfection efficiency by intraperitoneal delivery route was higher than the intravenous route. The reporter gene, pTRE-Smad7, was readily detected in the parietal peritoneum, mesentery, greater omentum and adipose tissue. The peak of transgene expression occurred 2 days after transfection and the transgene expression diminished in a time-dependent manner thereafter. Overall, the effectiveness and simplicity of the ultrasound-microbubble-mediated system may provide a promising nonviral means for improving gene delivery for treating peritoneal diseases in vivo.


High-grade glioma imaging volumes and survival: a single-institution analysis of 101 patients after resection using intraoperative MRI.
Intraoperative magnetic resonance imaging (iMRI) has been efficacious in maximizing resection of high-grade gliomas (HGGs). In this single-institution study of patients with HGGs who underwent resection using iMRI, the authors present a volumetric-based survival analysis to evaluate progression-free survival (PFS) and overall survival (OS), as well as the impact of additional resection on survival. This retrospective analysis included patients with HGGs who underwent resection using iMRI from 2011 to 2021. Volumetric analyses of T1-weighted contrast-enhancing (T1W-CE), T2-weighted (T2W), and T2W fluid-attenuated inversion recovery (FLAIR) MRI sequences were assessed at preoperative, intraoperative, immediate postoperative, and three-month postoperative timepoints. Statistical analyses were carried out using log-rank and multivariable Cox proportional hazard regression analyses. A total of 101 patients (median age 57.0 years) were treated. In keeping with prior studies, statistically significant associations between greater EOR and longer PFS and OS were seen (p = 0.012 and p = 0.006, respectively). The results demonstrated significant associations of lower preoperative T2W, 3-month postoperative T2W, and 3-month postoperative FLAIR volumes with longer PFS and OS (p = 0.045 and p = 0.026, p = 0.031 and p = 0.006, p = 0.018 and p = 0.004, respectively), as well as associations between lower immediate postoperative T2W and immediate postoperative FLAIR volumes with longer OS (p = 0.002 and p = 0.02). There was no observed association in either PFS or OS for patients undergoing additional resection after initial iMRI scan (p = 0.387 and p = 0.592). This study of 101 patients with new or recurrent HGGs shows three-month postoperative T2W and FLAIR imaging volumes were significant prognosticators with respect to PFS and OS.


Trigeminal disynaptic circuit mediating corneal afferent input to M. depressor palpebrae inferioris motoneurons in the pigeon (Columba livia).
Corneal afferent projections to the trigeminal brainstem nuclear complex (TBNC) and associated structures, as determined by transganglionic transport of various tracers, were found to be predominantly concentrated in two distinct patches in the dorsolateral medulla at periobex levels. One was in the external cuneate nucleus, and the other was in the ventralmost part of the ophthalmic division of the TBNC. The projections of putative second-order neurons in these regions, as determined by injections of wheat germ agglutinin conjugated to horseradish peroxidase into the dorsolateral medulla, were found to include the dorsal trigeminal motor nucleus (Vd), which innervates the M. depressor palpebrae inferioris. Electrical stimulation of Vd, which elicited lower eyelid movements, was then used to guide injections of tracer into Vd, which retrogradely labeled clusters of neurons in the corneal afferent recipient regions of the dorsolateral medulla. The lower eyelid of pigeons, unlike the nictitating membrane and upper lid, does not appear to be appreciably involved in either reflex blinking in response to relatively mild stimulation of the cornea (e.g., air puff), or in eye closure during the saccade-like head movements associated with walking, or in eye closure during pecking; but in response to a stimulus that makes corneal contact, an upward movement of the lower lid follows descent of the nictitating membrane and upper lid as part of a defensive eye-closing mechanism. The anatomical results thus appear to define a dedicated disynaptic trigeminal sensorimotor circuit for the control of lower eyelid motility in response to mechanical or noxious stimuli of the cornea. Injections of tracers into the lower and upper eyelids labeled palpebral sensory afferents that terminated predominantly in maxillary and ophthalmic portions, respectively, of the dorsal horn of upper cervical spinal segments. These terminal fields were therefore largely separate from those of corneal afferents. There were no specific corneal afferent projections upon accessory abducens motoneurons that innervate the two muscles controlling the nictitating membrane.


Influence of cytochrome P450 oxidoreductase genetic polymorphisms on CYP1A2 activity and inducibility by smoking.
Cytochrome P4501A2 (CYP1A2) presents a high interindividual variability in its activity and also in its inducibility by smoking. Cytochrome P450 oxidoreductase (POR) is an electron transfer protein that catalyzes the activity of several cytochromes P450. We aimed to study the influence of POR genetic polymorphisms on CYP1A2 activity while smoking and after smoking cessation, as well as on CYP1A2 inducibility. CYP1A2 activity was determined by the paraxanthine/caffeine ratio in 184 smokers and in 113 of these smokers who were abstinent during a 4-week period. Participants were genotyped for POR rs17148944G>A, rs10239977C>T, rs3815455C>T, rs2286823G>A, rs2302429G>A, and rs1057868C>T (POR*28) polymorphisms. While smoking, none of the tested POR polymorphisms showed a significant influence on CYP1A2 activity. After smoking cessation, significantly higher CYP1A2 activity was found in POR rs2302429A carriers (P=0.038) and in carriers of rs17148944G-rs10239977C-rs3815455T-rs2286823G-rs2302429A-rs1057868T haplotype (P=0.038), whereas carriers of POR rs2286823A (P=0.031) and of the rs17148944G-rs10239977C-rs3815455C-rs2286823A-rs2302429G-rs1057868C haplotype (P=0.031) had decreased CYP1A2 activity. In the complete regression model, only POR rs2302429G>A showed a significant effect (P=0.017). No influence of POR genotypes or haplotypes was observed on the inducibility of CYP1A2. POR genetic polymorphisms influence CYP1A2 basal but not induced activity and do not seem to influence CYP1A2 inducibility. Future work is warranted to identify other clinical and genetic factors that may explain the variability in CYP1A2 activity and inducibility by smoking.


Efficacy and Safety of a Transdermal Donepezil Patch in Patients with Mild to Moderate Alzheimer's Disease: Open-Label, Extension Study.
In Japan, only oral formulation of donepezil hydrochloride is approved for the treatment of Alzheimer's disease. To evaluate safety and efficacy of a donepezil patch 27.5 mg application for 52 weeks in patients with mild-to-moderate Alzheimer's disease; and to evaluate safety on switching from donepezil hydrochloride tablets. This 28-week, open-label study (jRCT2080224517) is an extension of a 24-week double-blind (donepezil patch 27.5 mg versus donepezil hydrochloride tablet 5 mg) noninferiority study. The patch group (continuation group) continued administration of the patch and the tablet group (switch group) switched to the patch in this study. A total of 301 patients participated (156 patients continued using patches; 145 patients switched). Both groups showed similar course on the Alzheimer's Disease Assessment Scale-cognitive component-Japanese version (ADAS-Jcog) and ABC dementia scales. At weeks 36 and 52, changes in ADAS-Jcog from week 24 [mean (standard deviation)] were 1.4 (4.8) and 2.1 (4.9) in the continuation group, and 1.0 (4.2), and 1.6 (5.4) in the switch group. The incidence of adverse events at application site in the continuation group over 52 weeks was 56.6% (98/173). Erythema, pruritus, and contact dermatitis at application site were observed in more than 10 patients each. There was no additional adverse event of clinical concern, and no increase in their incidence from the double-blind study. During the four weeks following switching, no patient discontinued or suspended administration due to adverse events. Application of the patch for 52 weeks was well tolerated and feasible, including switching from tablets.


Improving the immunogenicity of a trivalent Neisseria meningitidis native outer membrane vesicle vaccine by genetic modification.
Trivalent native outer membrane vesicles (nOMVs) derived from three genetically modified Neisseria meningitidis serogroup B strains have been previously evaluated immunologically in mice and rabbits. This nOMV vaccine elicited serum bactericidal activity (SBA) against multiple N. meningitidis serogroup B strains as well as strains from serogroups C, Y, W, and X. In this study, we used trivalent nOMVs isolated from the same vaccine strains and evaluated their immunogenicity in an infant Rhesus macaque (IRM) model whose immune responses to the vaccine are likely to be more predictive of the responses in human infants. IRMs were immunized with trivalent nOMV vaccines and sera were evaluated for exogenous human serum complement-dependent SBA (hSBA). Antibody responses to selected hSBA generating antigens contained within the trivalent nOMVs were also measured and we found that antibody titers against factor H binding protein variant 2 (fHbpv2) were very low in the sera from animals immunized with these original nOMV vaccines. To increase the fHbp content in the nOMVs, the vaccine strains were further genetically altered by addition of another fHbp gene copy into the porB locus. Trivalent nOMVs from the three new vaccine strains had higher fHbp antigen levels and generated higher anti-fHbp antibody responses in immunized mice and IRMs. As expected, fHbp insertion into the porB locus resulted in no PorB expression. Interestingly, higher expression of PorA, an hSBA generating antigen, was observed for all three modified vaccine strains. Compared to the trivalent nOMVs from the original strains, higher PorA levels in the improved nOMVs resulted in higher anti-PorA antibody responses in mice and IRMs. In addition, hSBA titers against other strains with PorA as the only hSBA antigen in common with the vaccine strains also increased.


Mapping rare and common causal alleles for complex human diseases.
Advances in genotyping and sequencing technologies have revolutionized the genetics of complex disease by locating rare and common variants that influence an individual's risk for diseases, such as diabetes, cancers, and psychiatric disorders. However, to capitalize on these data for prevention and therapies requires the identification of causal alleles and a mechanistic understanding for how these variants contribute to the disease. After discussing the strategies currently used to map variants for complex diseases, this Primer explores how variants may be prioritized for follow-up functional studies and the challenges and approaches for assessing the contributions of rare and common variants to disease phenotypes.


A case report of solitary fibrous tumor of the pleura: ultrasound diagnostic features.
Solitary fibrous tumor of the pleura is a rarely encountered clinical entity. Although the majority of these neoplasms have a benign course, the malignant form has also been reported. We describe the case of a 57-year-old male smoker with diabetes who was incidentally diagnosed on chest X-ray with a large solitary mass of the left hemithorax. The diagnostic tests included computed tomography, ultrasound, and contrast-enhanced ultrasound. Radical surgical resection was performed and histological examination confirmed a malignant solitary fibrous tumor of the pleura. The novelty of the case is the use of contrast-enhanced ultrasound in the diagnostic workup.


Influence of cognition, premorbid adjustment and psychotic symptoms on psycho-social functioning in first-episode psychosis.
The aim of this study is to evaluate the relationship between psycho-social functioning and symptoms, cognitive function, and premorbid adjustment, in patients with a first-episode psychosis. Clinical data were obtained from 90 patients, who were assessed with the Disability Assessment Scale (DAS-sv), the Positive and Negative Syndrome Scale (PANSS), the Premorbid Adjustment Scale (PAS-S) and with a battery of cognitive tests including Trail Making Tests A and B (TMTA- B), Continous Performance Test (CPT), some subscales of the Wechler Adult Intelligence Scale (WAIS), and the Verbal Learning Test España-Complutense (TAVEC). The results of the study suggest that psycho-social functioning in first-episode psychosis is significantly related to: positive, negative, excitative, affective and disorganized symptoms, social premorbid adjustment, cognitive flexibility, working memory, short term and long term memory. Of these, those which best explained psycho-social functioning are the positive and excitative symptoms, premorbid adjustment, flexibility and memory. These findings highlight the importance early intervention on cognitive and clinical variables to help provide a better psycho-social functioning in people with a first-episode of psychosis.


Assessment of femoral artery atherosclerosis at the adductor canal using 3D black-blood MRI.
To assess whether the three-dimensional (3D) black-blood motion-sensitized driven equilibrium (MSDE) prepared rapid gradient-echo sequence (3D MERGE) magnetic resonance imaging (MRI) sequence is sensitive enough to detect differences in atherosclerotic plaque size and morphology occurring in the adductor canal and the proximal bifurcation segment. Fifty pairs of adductor canal and bifurcation segments from 25 patients with intermittent claudication were examined using 3D MERGE. The two-dimensional (2D) transverse section showing the largest plaque burden in each segment was chosen for comparison. Wall and lumen boundaries were segmented from each 2D section and quantified using six metrics: wall area (WA), lumen area (LA), normalized wall index (NWI), maximum wall thickness (MaxWT), minimum wall thickness (MinWT), and eccentricity. The mean LA in the adductor region was significantly lower than that in the bifurcation segment (p < 0.0001). Mean NWI, MaxWT, and eccentricity in the adductor region were significantly higher than those at bifurcation (p < 0.0001, p < 0.0021, and p < 0.0045, respectively). Mean WA and MinWT of the two segments did not show a statistically significant difference. WA in both regions was positively correlated with eccentricity (p < 0.0049 and p < 0.0049, respectively). LA was negatively correlated with eccentricity (p < 0.0017), and NWI was positively correlated with eccentricity only in the adductor region (p < 0.0004). The results suggest that compensatory enlargement was limited in the adductor canal when compared to the proximal bifurcation segment. 3D MERGE, as a fast and non-invasive sequence, may assist the evaluation of femoral atherosclerosis by assessing the size and morphology of plaques, knowledge of which can guide clinical treatment.


Association of red cell distribution width/albumin ratio with intraoperative blood transfusion in cervical cancer patients.
Although minimally invasive surgical techniques have reduced intraoperative bleeding, the risk of transfusion exists. However, few studies have evaluated risk factors for transfusion in radical hysterectomy. We aimed to evaluate the association between preoperative red cell distribution width/albumin ratio (RDW/albumin) and transfusion in cervical cancer patients. We analyzed 907 patients who underwent radical hysterectomy between June 2006 and February 2015. Logistic regression and Cox regression analyses were performed to determine the risk factors for transfusion and mortality at 5-year and overall. Net reclassification improvement (NRI) and integrated identification improvement (IDI) analyses were performed to verify the improvement of the intraoperative transfusion model upon the addition of RDW/albumin. RDW/albumin was an independent risk factor for transfusion (odds ratio [OR]: 1.34, 95% confidence interval [CI]: 1.02-1.77, p = 0.035). Additionally, body mass index, operation time, laparoscopic surgery, total fluids, and synthetic colloid were risk factors for transfusion. RDW/albumin was an independent risk factor for 5-year mortality (hazard ratio [HR]: 1.51, 95% CI: 1.07-2.14, p = 0.020), and overall mortality (HR: 1.48, 95% CI: 1.06-2.07, p = 0.021). NRI and IDI analyses showed the discriminatory power of RDW/albumin for transfusion (p<0.001 and p = 0.046, respectively). RDW/albumin might be a significant factor in transfusion and mortality in cervical cancer patients.


Attenuated stress responsiveness in an animal model for neurodevelopmental psychopathological disorders.
Day 7 amygdala-lesioned (D7 AMX) rats have been proposed as a model for neurodevelopmental psychopathological disorders such as schizophrenia. Patients with schizophrenia are sensitive to stress and show an impaired hypothalamic-pituitary-adrenal response to certain stressful stimuli. Therefore, we investigated neuroendocrine and behavioral stress responses in the D7 AMX lesion model. Plasma concentrations of ACTH, corticosterone, and catecholamines were measured in response to foot shock and novelty in D7 and D21 lesioned (AMX) and non-lesioned (SHAM) animals. Behavior was recorded and analyzed afterwards. D7 AMX rats, unlike other rats, had a reduced ACTH response to foot shock and showed less active behavior in response to novelty. Neurodevelopmental dysfunction of target structures of the amygdala is associated with disturbed endocrine and behavioral responses to stress. These data accord with the notion that the D7 amygdala-lesioned rat can function as a neurodevelopmental model with relevance to schizophrenia.


Meiosis in the foetal mouse ovary. I. An analysis at the light microscope level using surface-spreading.
The identification and progression of the prophase stages of meiosis in the mouse foetal ovary are reported, from d 13 of gestation to d 1 postpartum. Air-dried Giemsa-stained oocyte preparations are compared with surface-spread silver-stained cells. The latter method allows a more detailed quantitative analysis of the pachytene stage. Numbers of synaptonemal complexes can be counted, and the degree of synapsis determined. The progression of cells appears to be relatively synchronous, in agreement with previous reports. The activity of nucleolar organisers, in particular one associated with the shortest synaptonemal complex (chromosome No. 19) is described. At late pachytene the lateral elements of the No. 19 bivalent desynapse precociously with apparent nucleolar involvement.


Management of advanced-phase chronic myeloid leukemia.
The management of chronic myeloid leukemia (CML) in accelerated or blast phase (advanced phase) remains a significant challenge despite the introduction of very effective tyrosine kinase inhibitors (TKIs). The biology of advanced-phase CML is complex and engages several pathways that are not optimally targeted by TKIs. Allogeneic stem cell transplantation remains the only potentially curative therapy, but the effectiveness of this conventional approach is limited. New strategies are required to improve the outlook for these patients.


Unsuitability of polioviruses as indicators of virological quality of water.
Polioviruses could not be detected in about 50% of sewage samples despite routine administration of live attenuated poliovirus vaccine to the infant population.


METTL3 promotes the proliferation and invasion of esophageal cancer cells partly through AKT signaling pathway.
Methyltransferase-like 3 (METTL3) is identified as a methyltransferase responsible for N6-methyladenosine (m6A) modification of mRNA, miRNA and lncRNA. Emerging evidences suggest that METTL3 is involved in tumorigenesis and progression of multiple tumor types. However, the functional role of METTL3 in esophageal cancer (EC) remains unclear. We used specific shRNA to down-regulate the METTL3 expression, and used pcDNA3.1-METTL3 cDNA plasmid to up-regulate the METTL3 expression in Eca-109 and KY-SE150 cells. Biological functions of METTL3 were performed by CCK-8, colony formation, apoptosis analysis, transwell and wound healing assays. Finally, an in-depth mechanism study was performed by an AKT inhibitor. METTL3 knockdown reduced the proliferation, clonality, migration and invasion of Eca-109 and KY-SE150 cells, and induced cell apoptosis, which may be mediated by activation of the mitochondrial apoptotic pathway. Further, METTL3 overexpression promoted the proliferation, clonality, migration and invasion of Eca-109 and KY-SE150 cells, and inhibited cell apoptosis. In addition, METTL3 regulated the expression of Wnt/β-catenin and AKT signaling pathway components. A double-effect inhibitor (BEZ235) inhibited AKT and mTOR phosphorylation and hindered the effect of METTL3 overexpression on the proliferation and migration of Eca-109 and KY-SE150 cells. Our data suggest that METTL3 plays a carcinogenic role in human EC progression partially through AKT signaling pathways, suggesting that METTL3 may serve as a potential therapeutic target for EC therapy.


Lentiviral vector-mediated transduction of murine CD34(-) hematopoietic stem cells.
Efficient gene transfer into murine hematopoietic stem cells (HSCs) provides a powerful tool for exploring hematopoietic stem cell biology. In this study, we evaluated the efficiency of lentiviral vector-mediated gene transfer into murine CD34(-/low)c-Kit(+)Sca-1(+)Lin(-) (CD34(-) KSL) cells that are highly enriched for HSCs. FACS-sorted CD34(-) KSL cells were transduced with the vesicular stomatitis virus G glycoprotein-pseudotyped HIV-1-based lentiviral vector containing the green fluorescent protein (GFP) gene under the control of the cytomegalovirus promoter, and then 50 transduced cells were transplanted into lethally irradiated mice. Transduction efficiency was assessed by FACS analysis for GFP expression in peripheral blood (PB) cells. FACS-sorted GFP(+) KSL bone marrow (BM) cells from primary recipients were used for secondary transplantation, and GFP expression in PB cells of reconstituted mice was analyzed by FACS. GFP expression was detected in PB cells of all primary recipients (n = 10) at an average of 40% (range 26-58%) when the lentiviral vector containing the woodchuck hepatitis virus posttranscriptional regulatory element was used. GFP(+) cells were found in multilineage cells in PB, BM, spleen, and thymus for at least 8 months posttransplantation. In secondary recipients, donor-derived GFP(+) KSL BM cells could reconstitute and GFP expression was detected in both myeloid and lymphoid cells in PB. Our results indicate that lentiviral vectors can efficiently transduce highly enriched murine HSCs and sustain long-term expression of the transgene in the multilineage differentiated progeny in reconstituted mice.


Impact of surgical wait times on oncologic outcomes in resectable pancreas adenocarcinoma.
Timely surgical resection in patients with suspected or diagnosed pancreas adenocarcinoma is an essential part of care. We hypothesized that longer surgical wait time was associated with worse oncologic outcomes. A retrospective cohort of patients (N = 144) with resectable pancreas adenocarcinoma was divided into four wait time groups (<4, 4-8, 8-12, and >12 weeks), defined from the time of diagnosis on cross-sectional imaging. Overall and recurrence-free survival were analyzed using the Kaplan-Meier method and Cox proportional hazards regression. A higher rate of conversion to palliative bypass in patients waiting over 4 weeks was observed and further analyzed using post-hoc multivariate regression. On multivariable analysis, longer wait time was associated with improved overall (HR 0.49, 95% CI: 0.28-0.85) and recurrence-free survival (HR 0.29, 95% CI: 0.15-0.56) in >12 weeks compared to <4 weeks group. On post-hoc analysis, longer wait time over 8 weeks was positively associated with palliative bypass (OR 5.33, 95% CI: 1.32-27.88). Wait time over 8 weeks was associated with a higher rate of palliative bypass. There was an improvement in overall and recurrence-free survival in patients who waited over 12 weeks, likely due to selection bias.


The movement speed-accuracy relation in space-time.
Two experiments investigated a new approach to decomposing the contributions of spatial and temporal constraints to an integrated single space-time performance score in the movement speed-accuracy relation of a line drawing task. The mean and variability of the space-time performance error score were lowest when the task space and time constraint contributions to the performance score were comparable (i.e., middle range of velocities). As the contribution of either space or time to the performance score became increasingly asymmetrical at lower and higher average velocities, the mean performance error score and its variability increased with a greater trade-off between spatial and temporal movement properties. The findings revealed a new U-shaped space-time speed-accuracy function for performance outcome in tasks that have both spatial and temporal demands. The traditional speed-accuracy functions for spatial error and temporal error considered independently map to this integrated space-time movement speed-accuracy function.


Lipoprotein receptor loss in forebrain radial glia results in neurological deficits and severe seizures.
The Alzheimer disease-associated multifunctional low-density lipoprotein receptor-related protein-1 is expressed in the brain. Recent studies uncovered a role of this receptor for the appropriate functioning of neural stem cells, oligodendrocytes, and neurons. The constitutive knock-out (KO) of the receptor is embryonically lethal. To unravel the receptors' role in the developing brain we generated a mouse mutant by specifically targeting radial glia stem cells of the dorsal telencephalon. The low-density lipoprotein receptor-related protein-1 lineage-restricted KO female and male mice, in contrast to available models, developed a severe neurological phenotype with generalized seizures during early postnatal development. The mechanism leading to a buildup of hyperexcitability and emergence of seizures was traced to a failure in adequate astrocyte development and deteriorated postsynaptic density integrity. The detected impairments in the astrocytic lineage: precocious maturation, reactive gliosis, abolished tissue plasminogen activator uptake, and loss of functionality emphasize the importance of this glial cell type for synaptic signaling in the developing brain. Together, the obtained results highlight the relevance of astrocytic low-density lipoprotein receptor-related protein-1 for glutamatergic signaling in the context of neuron-glia interactions and stage this receptor as a contributing factor for epilepsy.


Formulation and biochemical evaluation of designer diet enriched with botanicals for bone health.
The study designed to evaluate the role of sesame, fenugreek flax, and nuts seeds as best alternatives for therapeutic agents to improve bone mineral status. Effect of these plant seeds on proximate composition, antioxidant potential, mineral, fatty acid, and amino acid profile of supplements was studied. The sesame, fenugreek, and flaxseeds were incorporated with nuts at level of 35% to develop supplement. Improved contents of protein, fat, and NFE were recorded in range of 25.72-32.23, 18.92-23.15, and 34.25%-39.59%, correspondingly. The antioxidant potential of supplements is high (450.56 mg GAE/ 100g) prepared with fenugreek followed by flaxseeds and sesame seeds. Calcium as fundamental bone mineral found greater (525.15%-790.21%) in seed-based supplement. Enhanced contents of Oleic, linoleic, and linolenic acids ranged 17.32-41.78, 21.72-44.23, and 1.00%-47.12%, respectively. Nutritional enriched supplements may be best alternatives for improving bone health by lowering osteoclast and increasing osteoblast mechanism in bone development.


Two randomized controlled trials for colorectal cancer screening invitations developed using a behavioral science approach.
The effectiveness of behaviorally informed, targeted invitations to standard invitations and to no invitation (control arm, primary analysis) were compared in the context of an organized colorectal cancer (CRC) screening program. Two multi-arm, pragmatic randomized controlled trials in men (arms: male-specific, unisex, standard invitation, or no invitation) and in women (arms: unisex, standard invitation, or no invitation), were conducted in Ontario, Canada. Eligible persons aged 50-74, due for CRC screening, were randomized. Primary and secondary outcomes were completion of the guaiac fecal occult blood test (gFOBT) and uptake of any colorectal test, respectively, within 5 months of mailing. Impact of invitation type was assessed using logistic regression. Letters were mailed to 75,810 men and women; 38,673 males and 34,453 females were included in the analyses. Men who received the male-specific letter were most likely to screen with gFOBT compared to controls (odds ratio (OR) 7·24, 95% CI: 5·77, 9·09), followed by those receiving the unisex letter (OR 6·75, 95% CI: 5·37, 8·47) and the standard letter (OR 5·99, 95% CI: 4·76, 7·53). Women who received the unisex letter were most likely to be screened with gFOBT compared to controls (OR 7·07, 95% CI: 5·83, 8·59), followed by those receiving the standard letter (OR 6·76, 95% CI: 5·56, 8·21). In both trials, the findings were similar for the secondary outcome. Mailed invitations were effective for both men and women. With greater targeting using the behaviorally informed invitations, the magnitude of benefit relative to no invitation appeared to increase. (ClinicalTrials.gov, NCT02364895).


Glycosphingolipids in the cerebrospinal fluid of patients with multiple sclerosis.
Glycosphingolipids in cerebrospinal fluid (CSF) of individual patients with multiple sclerosis (MS) were analyzed using a glycolipid-overlay technique. The ganglioside composition of CSF of non-MS patients was characterized by an abundance of polysialo species, including GT1b and GQ1b. This pattern is completely different from that of human white or gray matter, in which mono- and disialogangliosides predominate. Increased levels of GM1, either associated with or without increases of other gangliosides, such as GD1a, were observed in 16% of the patients with MS (6 of 37 cases: 1 of 15 progressive progressive stage, 4 of 16 progressive stationary stage, and 1 of 6 relapsing stage). The concentration of GD3 was increased in 23% (3 of 13 cases), whereas 1 of 13 cases (8%) showed a dramatic increase of sulfoglucuronyl paragloboside (SGPG) associated with a high level of GD3. These changes may reflect the cellular changes associated with the known pathological lesions in MS, which are characterized by demyelination, gliosis, and/or remyelination with oligodendrocytic proliferation.


Effect of viable isolated hepatocytes or hepatocyte fractions on survival rate following galactosamine-induced acute liver failure.
Administration of galactosamine to rats results in an acute liver failure. The dose-effect relation appeared to be highly strain-dependent. Intraperitoneal injection of 1,000 mg/kg galactosamine resulted in a 90% lethal acute liver failure in Wag/Rij rats. Transplantation of viable isolated hepatocytes, administration of homogenate or cytosol of normal or regenerating liver tissue, or administration of cell-free liver cell culture supernatant did not improve the survival rate in this model of acute liver failure. These findings, which are in striking contrast with the data reported in the literature, are discussed in relation to the specific properties of this type of experimental model for acute hepatic failure.


The relationship between alertness and spatial attention under simulated shiftwork.
Higher and lower levels of alertness typically lead to a leftward and rightward bias in attention, respectively. This relationship between alertness and spatial attention potentially has major implications for health and safety. The current study examined alertness and spatial attention under simulated shiftworking conditions. Nineteen healthy right-handed participants (M = 24.6 ± 5.3 years, 11 males) completed a seven-day laboratory based simulated shiftwork study. Measures of alertness (Stanford Sleepiness Scale and Psychomotor Vigilance Task) and spatial attention (Landmark Task and Detection Task) were assessed across the protocol. Detection Task performance revealed slower reaction times and higher omissions of peripheral (compared to central) stimuli, with lowered alertness; suggesting narrowed visuospatial attention and a slight left-sided neglect. There were no associations between alertness and spatial bias on the Landmark Task. Our findings provide tentative evidence for a slight neglect of the left side and a narrowing of attention with lowered alertness. The possibility that one's ability to sufficiently react to information in the periphery and the left-side may be compromised under conditions of lowered alertness highlights the need for future research to better understand the relationship between spatial attention and alertness under shiftworking conditions.


Fos expression in the medullary dorsal horn of the rat after chronic constriction injury to the infraorbital nerve.
Chronic constriction injury to the rat's infraorbital nerve (IoN-CCI) induces asymmetric face grooming directed to the injured nerve territory and, beginning at 7-12 days postoperative, hyperresponsiveness to mechanical stimulation in this territory (B.P. Vos, A.M. Strassman, and R.J. Maciewicz, 1994, J. Neurosci. 14:2708-2723). To examine central mechanisms involved in these behavioral alterations, changes in nonevoked and mechanical stimulation-evoked fos-like immunoreactivity (fos-LI) following IoN-CCI were quantified in the medullary dorsal horn. Following the appearance of hyperresponsiveness in IoN-CCI rats, experimental and matched sham-operated rats were anesthetized with urethane and received either no stimulation or repeated stimulation with either a 2- or 15-g von Frey hair applied to the hairy skin between vibrissae B3-4/C3-4 on the operated side. Unstimulated IoN-CCI rats had increased fos-LI in laminae I-IV of the ipsilateral medullary dorsal horn. In both groups, mechanical stimulation produced a distinct pattern of fos-LI in the ipsilateral medullary dorsal horn, the quantity of which was related to stimulus intensity. For both stimulus intensities, the total amount and the rostrocaudal spread of evoked fos-LI were significantly larger in IoN-CCI rats. In IoN-CCI rats, stimulation-evoked increases in fos-LI were proportionally larger in laminae I-II than in III-IV. This laminar effect was also present in sham-operated rats but only for 15-g stimulation. Neither condition nor stimulus intensity affected fos-LI in the contralateral medullary dorsal horn. Positive correlations were found between the behavioral parameters of increased trigeminal nociceptive activity and the total amount of fos-LI in the ipsilateral medullary dorsal horn. The results demonstrate that IoN-CCI induces significant alterations in the central processing of afferent signals, which may underlie behavioral manifestations of increased nociceptive activity.


Characterization of a S1PR2 specific 11C-labeled radiotracer in streptozotocin-induced diabetic murine model.
Diabetes mellitus is a chronic progressive metabolic disorder that affects millions of people worldwide. Emerging evidence suggests the important roles of sphingolipid metabolism in diabetes. In particular, sphingosine-1-phosphate (S1P) and S1P receptor 2 (S1PR2) have important metabolic functions and are involved in several metabolic diseases. In diabetes, S1PR2 can effectively preserve β cells and improve glucose/insulin tolerance in high-fat diet induced and streptozotocin (STZ)-induced diabetic mouse models. We previously developed a group of potent and selective S1PR2 ligands and radioligands. In this study, we continued our efforts and characterized our leading S1PR2 radioligand, [11C]TZ34125, in a STZ-induced diabetic mouse model. [11C]TZ34125 was radiosynthesized in an automated synthesis module and in vitro saturation binding assay was performed using recombinant human S1PR2 membrane. In vitro saturation autoradiography analysis was also performed to determine the binding affinity of [11C]TZ34125 against mouse tissues. Type-1 diabetic mouse model was developed following a single high dose of STZ in C57BL/6 mice. Ex vivo biodistribution was performed to evaluate the distribution and amount of [11C]TZ34125 in tissues. In vitro autoradiography analysis was performed to compare the uptake of [11C]TZ34125 between diabetic and control animals in mouse spleen and pancreas. Our in vitro saturation binding assay using [11C]TZ34125 confirmed [11C]TZ34125 is a potent radioligand to recombinant human S1PR2 membrane with a Kd value of 0.9 nM. Saturation autoradiographic analysis showed [11C]TZ34125 has a Kd of 67.5, 45.9, and 25.0 nM to mouse kidney, spleen, and liver tissues respectively. Biodistribution study in STZ-induced diabetic mice showed the uptake of [11C]TZ34125 was significantly elevated in the spleen (~2 fold higher) and pancreas (~1.4 fold higher) compared to normal controls. The increased uptake of [11C]TZ34125 was further confirmed using autoradiographic analysis in the spleen and pancreases of STZ-induced diabetic mice, indicating S1PR2 can potentially act as a biomarker of diabetes in pancreases and inflammation in spleen. Future mechanistic analysis and in vivo quantitative assessment using non-invasive PET imaging in large animal model of diabetes is worthwhile. Overall, our data showed an increased uptake of our lead S1PR2-specific radioligand, [11C]TZ34125, in the spleen and pancreases of STZ-induced diabetic mice, and demonstrated [11C]TZ34125 has a great potential for preclinical and clinical usage for assessment of S1PR2 in diabetes and inflammation.


Effects of tonsillectomy on speech and voice.
The aim of this study was to evaluate changes in acoustic features of speech after tonsillectomy and to establish concepts of patient management and rational therapeutic approach. Before and 1 month after surgery, phonetically balanced sentences and sustained vowels a, e, i were carried out and digitalized with Multi-Dimensional Voice Program (Kay Elemetrics, Lincoln Park, NJ) in all the patients, as an evaluation of nasal resonance, speech articulation, and voice handicap index (VHI). These parameters were estimated: average of fundamental frequency, Jitter percent, Shimmer, noise-to-harmonics ratio, voice turbulence index, soft phonation index, degree of voiceless, degree of voice breaks, and peak amplitude variation. Our data showed that 1 month after tonsillectomy, improvements in all the acoustic parameters, a subjective decrease of hypernasality, and an improvement of speech articulation and VHI were achieved. These data suggest the reduction of the nasal resonance and highlight the role of tonsillectomy in the improvement of voice and speech quality. Furthermore, tonsillectomy should be performed before consideration of pharyngeal flap surgery. Our results highlight that objective evaluation of speech and voice helps the specialist to improve patient management and avoid unnecessary and dangerous surgical procedures.


Generation of Fcabs targeting human and murine LAG-3 as building blocks for novel bispecific antibody therapeutics.
The immunoglobulin superfamily protein lymphocyte-activation gene 3 (LAG-3) participates in immune suppression and has been identified as a suitable target for cancer therapies. In order to generate bispecific antibodies targeting LAG-3, Fcabs (Fc-region with antigen binding) targeting human and murine LAG-3 were generated from phage libraries. These Fcabs bind to LAG-3, inhibiting its interaction with MHC class II, and induce IL-2 production in a T cell assay. Bispecific antibodies, known as mAb2, were produced by replacing the Fc region of a monoclonal antibody with Fcab sequences in the CH3 domain. mAb2 containing anti-LAG-3 Fcabs have mAb-like biophysical characteristics and retain LAG-3 binding and functional activity. mAb2 can thus be generated using multiple Fabs to investigate bispecific parings and develop novel therapeutics.


Paraplegia after tethered cord surgery: an uncommon combined anomaly of spinal arteriovenous fistula and sacral lipoma--case report.
The coexistence of spinal arteriovenous malformation (AVM) with congenital abnormalities is relatively common. However, the association of a spinal AVM and lipoma is rare. We present an adult patient with this combined anomaly and discuss the clinical relevance of this case. A 42-year-old Caucasian man with progressive paraparesis initially underwent surgery for a tethered spinal cord. Postoperatively, he became paraplegic. He improved gradually over an interval of 8 months and, at that point, worsened again. Subsequent angiographic study revealed a spinal dural arteriovenous fistula located at S1-S2. Additionally, an occipital dural AVM was discovered near the transverse sinus. The spinal arteriovenous fistula was excised along with the sacral lipoma. The occipital arteriovenous fistula was embolized successfully at a later time. The patient had immediate improvement in sensory symptoms after surgery. At a 9 month follow-up examination, he had regained the ability to walk with crutches, but his bladder dysfunction persisted. Recognition of co-existing vascular anomalies, such as spinal AVMs, is important in patients with tethered cords. The mechanisms involved in this patient's worsening neurological condition after release of the tethered cord are discussed.


New insights into the pathophysiology of the tuberous sclerosis complex: Crosstalk of mTOR- and hippo-YAP pathways in cell growth.
Tuberous Sclerosis Complex (TSC) is a genetic disease causing uncontrolled growth of hamartomas involving different organ systems. In the last decade, dysregulation of the mTORC1 pathway was shown to be a main driver of tumor growth in TSC. Recently, a new crosstalk was detected between the mTORC1 and the Hippo-YAP pathway, another major cell signaling cascade controlling cell growth and organ size. Elucidating this connection is an important step in understanding the complexity of TSC, enabling new pharmacological targets and therapeutical options.


The novel HLA-DQB1*03:493 allele, the first with glutamic acid at position-10 in the leader peptide.
A nonsynonymous nucleotide substitution in exon 1 results in the novel HLA-DQB1*03:493 allele.


Experimental and techno-economic investigation of industrial para-xylene plant revamping to produce meta-xylene.
In this research, the feasibility study of revamping of simulated moving bed for paraxylene separation in ELUXYL process to produce meta-xylene using industrial Ba-faujasite exchanged adsorbent by changing operating condition (temperature and pressure) were examined experimentally and theoretically. Two different mixed-xylene feed cases (with and without presence of para-xylene) were considered. Different sets of temperature and pressure were evaluated with the help of equilibrium and dynamic experiments to obtain optimum operating condition in a favor of separation of meta-xylene. Results confirm that in the presence of para-xylene in a feed, selectivity of the adsorbent could not change towards meta-xylene. But, in the absence of para in some pressure and temperature meta-xylene was more selective than ortho and ethylbenzene. Finally, by the means of statistical experimental design method the results of all experiments were compered and an optimum temperature and pressure were found. Breakthrough experiment in optimum condition showed MX/OX and MX/EB selectivity as 1.83 and 1.15, respectively. Furthermore, the design and simulation of the real industrial SMB plant was performed in Aspen Chromatography and HYSYS software to evaluate the real performance of MX separation. Simulation results showed the final purification in SMB plant can be reached to 83.91%. At the end, for the aim of promoting purity by considering extra distillation towers the purity of meta-xylene was achieved by 96%. The economical investigation showed that by considering 700$/ton for feed supplied, the process can be satisfactory from economical point of view.


Biochemical and structural characterization of TLXI, the Triticum aestivum L. thaumatin-like xylanase inhibitor.
Thaumatin-like xylanase inhibitors (TLXI) are recently discovered wheat proteins. They belong to the family of the thaumatin-like proteins and inhibit glycoside hydrolase family 11 endoxylanases commonly used in different cereal based (bio)technological processes. We here report on the biochemical characterisation of TLXI. Its inhibition activity is temperature- and pH-dependent and shows a maximum at approximately 40 degrees C and pH 5.0. The TLXI structure model, generated with the crystal structure of thaumatin as template, shows the occurrence of five disulfide bridges and three beta-sheets. Much as in the structures of other short-chain thaumatin-like proteins, no alpha-helix is present. The circular dichroism spectrum of TLXI confirms the absence of alpha-helices and the presence of antiparallel beta-sheets. All ten cysteine residues in TLXI are involved in disulfide bridges. TLXI is stable for at least 120 min between pH 1-12 and for at least 2 hours at 100 degrees C, making it much more stable than the other two xylanase inhibitors from wheat, i.e. Triticum aestivum xylanase inhibitor (TAXI) and xylanase inhibitor protein (XIP). This high stability can probably be ascribed to the high number of disulfide bridges, much as seen for other thaumatin-like proteins.


Sex-Specific Associations of Smoking with Spontaneous Subarachnoid Hemorrhage: Findings from Observational Studies.
Some studies have reported that women are at higher risk for spontaneous subarachnoid hemorrhage (SAH) compared with men, and smoking is the most important lifestyle risk factor for spontaneous SAH. However, it is still unknown whether the risk of SAH from smoking and smoking status is differential for women and men. We performed a meta-analysis to estimate the effect of smoking on SAH in women compared with men. PubMed (January 1, 1966 to February 19, 2020) and EMBASE (January 1, 1980 to February 19, 2020) were systematically searched. Studies that estimated sex-specific relative risks (RRs) of SAH were selected. We pooled sex-specific RRs, comparing women with men using random-effects meta-analysis. Data from 20 observational studies that included 1,387,204 participants (563,898 women) and 7,838 SAHs (3,977 women) were analyzed. The combined women-to-men RRs of former smokers versus never smokers for SAH were 1.08 (95% confidence interval [CI] 0.62-1.89, p = 0.78). The pooled women-to-men RRs of current smokers versus never smokers were 1.39 (95% CI 1.05-1.83, p = 0.02). The combined women-to-men RRs of total smokers versus never smokers RRs were 1.15 (95% CI 0.88-1.52, p = 0.30). Our study shows there is not enough evidence to suggest that women who smoke have a greater risk for SAH than men; however, women who persistently smoke have a greater risk. Smoking seems to be more susceptible in the increased SAH risk in women.


Acoustical detection of coronary occlusions using neural networks.
A nonlinear neural network classifier was applied to noninvasive acoustic detection of coronary artery disease; the classifier included a feature vector, derived from diastolic heart sounds, and a multi-layered network trained by the backpropagation. The feature vector is based on the linear prediction coefficients of the autoregressive method after an adaptive line enhancement method was used as the input pattern to the neural network. One hundred and twelve recordings (70 abnormal, 42 normal) were studied and the network was trained on a randomly chosen set of six abnormal and six normal patients. It was tested on a database consisting of 100 recordings to which it had not been exposed. The network correctly identified 50 of the 64 patients with coronary artery disease and 32 of the 36 patients without any coronary artery occlusions. These results showed that this neural network is capable of distinguishing normal patients from abnormal patients. In addition, the diagnostic capability of this approach is much better than any other available noninvasive approach.


Acetoacetate protects hippocampal neurons against glutamate-mediated neuronal damage during glycolysis inhibition.
Glucose is the main substrate that fulfills energy brain demands. However, in some circumstances, such as diabetes, starvation, during the suckling period and the ketogenic diet, brain uses the ketone bodies, acetoacetate and beta-hydroxybutyrate, as energy sources. Ketone body utilization in brain depends directly on its blood concentration, which is normally very low, but increases substantially during the conditions mentioned above. Glutamate neurotoxicity has been implicated in neurodegeneration associated with brain ischemia, hypoglycemia and cerebral trauma, conditions related to energy failure, and to elevation of glutamate extracellular levels in brain. In recent years substantial evidence favoring a close relation between glutamate neurotoxic potentiality and cellular energy levels, has been compiled. We have previously demonstrated that accumulation of extracellular glutamate after inhibition of its transporters, induces neuronal death in vivo during energy impairment induced by glycolysis inhibition. In the present study we have assessed the protective potentiality of the ketone body, acetoacetate, against glutamate-mediated neuronal damage in the hippocampus of rats chronically treated with the glycolysis inhibitor, iodoacetate, and in hippocampal cultured neurons exposed to a toxic concentration of iodoacetate. Results show that acetoacetate efficiently protects against glutamate neurotoxicity both in vivo and in vitro probably by a mechanism involving its role as an energy substrate.


Urea as a promotive coupler of plant-herbivore interactions.
Growth responses of Kyllinga nervosa Steud., a sedge from the Serengeti short-grass plains, were examined in a factorial experiment which included clipped and unclipped plants, and nitrogen supplied as either urea or ammonium nitrate. Results were expressed in relation to three transfer processes: flow to grazers, flow to producers and flow to reproduction. Clipping increased biomass and nitrogen flow to grazers by significantly increasing nitrogen uptake, aboveground nitrogen flow, and the weights of and proportional allocation to green leaf production. This was at the expense of flow to vegetative and sexual reproduction, since the weights and proportional investments in roots, crowns and reproductive structures were reduced. Urea nutrition increased flow to grazers and plant reproduction through increases in green leaf weight, flower weight, allocation to green leaves, flowers and stems, and aboveground: belowground biomass ratios. Stimulation of aboveground productivity by urea was a consequence of increased tillering rates.Interactive responses of clipping and nitrogen source regulated plant growth, thus controlling flow to each transfer process. Combined effects of clipping and urea resulted in compensatory production of both green leaves and flowers, and maximized biomass and nitrogen flow to grazers. Both urea and clipping tightened herbivore-producer recycling by significantly reducing litter nitrogen and carbon masses. In contrast, when plants were unclipped and grown on NH4NO3, biomass allocation and weights of roots and crowns were increased at the expense of aboveground tissues, thus increasing flow to primary producers. Plant growth responses to experimental treatment combinations simulating nutritional status of grazed and ungrazed field plants indicate that urea represents a potential importance beyond it nitrogen contribution by introducing a positive feedback to herbivores.


Efficacy of transcranial magnetic stimulation in treatment-resistant depression.
The use of Transcranial Magnetic Stimulation (TMS) in the add-on treatment of patients with treatment-resistant depression (TRD) is becoming more common. This study aims to investigate the efficacy of TMS on depression and accompanying anxiety symptoms among patients with TRD. The current study was conducted with 38 patients diagnosed with TRD. The patients were randomly divided into two groups and received 20 sessions of high-frequency (10Hz) TMS and 20 sessions of sham TMS to the left dorsolateral prefrontal cortex in a double-blind and cross-over fashion without a change in their pharmacotherapy. In the clinical evaluation, Hamilton Depression Rating Scale (HAM-D) and Hamilton Anxiety Rating Scale (HAM-A) were carried out three times in total: before, crossover phase, and at the end of the treatment. A statistically significant decrease was found in the HAM-D and HAM-A in the group who were actively stimulated in the cross-over phase of the study. While there was a significant decrease in the HAM-A in the group who received sham stimulation, the decrease in the HAM-D was not statistically significant. Group comparisons revealed a statistically significant decrease in HAM-D in the group who were actively stimulated compared to the group receiving sham stimulation. At the end of the study, 63% of 38 patients responded to treatment, 15% partially responded, and 42% reached remission. This randomized, double-blind, sham-controlled, cross-over study revealed that TMS is superior to sham-TMS, provides clinically significant improvement when implemented besides pharmacotherapy among patients with treatment-resistant depression, and is beneficial for accompanying anxiety symptoms.


Effect of combined montelukast and desloratadine on the early asthmatic response to inhaled allergen.
The early asthmatic response (EAR) to inhaled allergen results from IgE-mediated release of multiple mast-cell mediators, including leukotrienes and histamine, both of which cause bronchoconstriction. Combination therapy directed at blocking the effects of both mediators might protect against the EAR better than either therapy alone. We sought to evaluate the effect of desloratadine and montelukast, administered alone and in combination, on the EAR to inhaled allergen. Ten adults with mild-to-moderate atopic asthma participated in a randomized, 4-way crossover study design comparing placebo, 5 mg of desloratadine, 10 mg of montelukast, and the combination administered at 26 hours and 2 hours before each allergen challenge conducted at least 7 days apart. The primary end point was the concentration of allergen that resulted in a 20% decrease in FEV1 (PC20). The geometric mean allergen PC20 (mean log +/- SEM) for combination therapy, montelukast, desloratadine, and placebo was 697 U/mL (2.8433 +/- 0.3253), 338 U/mL (2.5295 +/- 0.2979), 123 U/mL (2.0883 +/- 0.2102), and 104 U/mL (2.0166 +/- 0.2553), respectively (n = 9; P < .00001, ANOVA). Montelukast increased the allergen PC20 4.8-fold, and combination therapy increased the allergen PC20 8.9-fold. The effect of the combination was greater than that with montelukast alone (P < .02). Desloratadine treatment was no different than placebo. The early response to inhaled allergen was unchanged after desloratadine therapy and partially inhibited with montelukast therapy. The combination of desloratadine and montelukast provided superior efficacy to either blocker administered alone. Investigations into the possible mechanisms of the enhanced inhibition are necessary.


Cell Type Specific CAG Repeat Expansions and Toxicity of Mutant Huntingtin in Human Striatum and Cerebellum.
Brain region-specific degeneration and somatic expansions of the mutant Huntingtin (mHTT) CAG tract are key features of Huntington's disease (HD). However, the relationships between CAG expansions, death of specific cell types, and molecular events associated with these processes are not established. Here we employed fluorescence-activated nuclear sorting (FANS) and deep molecular profiling to gain insight into the properties of cell types of the human striatum and cerebellum in HD and control donors. CAG expansions arise in striatal medium spiny neurons (MSNs) and cholinergic interneurons, in cerebellar Purkinje neurons, and at mATXN3 in MSNs from SCA3 donors. CAG expansions in MSNs are associated with higher levels of MSH2 and MSH3 (forming MutSβ), which can inhibit nucleolytic excision of CAG slip-outs by FAN1 in a concentration-dependent manner. Our data indicate that ongoing CAG expansions are not sufficient for cell death, and identify transcriptional changes associated with somatic CAG expansions and striatal toxicity.


Myomectomy during pregnancy: A systematic review.
Uterine fibroids affect 2-10 % of pregnant women. Although usually asymptomatic, they may be associated with pregnancy complications. Myomectomy is preferably avoided antenatally, however, it has been reported in symptomatic cases that did not respond to conservative management. The aim of this study was to summarize the published literature and present the reported outcomes and associated risks of this procedure. A systematic research of the literature was conducted in PubMed/MEDLINE, Scopus and the Cochrane Library, including case reports and case series. An effort was made to numerically analyse all parameters included in the case reports. Overall, 54 relevant articles were identified, including 97 patients. The median gestational age at diagnosis was 13 (range 6-26) weeks, while the median age at myomectomy was 16 (range 6-26) weeks. Abdominal pain, not responding to medical treatment was the most common indication for surgery. The median number of fibroids removed per patient was one (range 1-5). Most of them were subserous pedunculated or subserous and fundal. Laparotomy (78.4 %) was the principal surgical approach, however, laparoscopic and vaginal operations were also reported. The median duration of surgery was 53 (range 20-150) min. The histopathology revealed necrosis and degeneration as the main findings of removed fibroids. The pregnancy outcome was favourable in most of the cases, with few complications reported. Based on the limited published data, myomectomy during pregnancy appears as a safe procedure in cases of symptomatic uterine fibroids not responding to conservative management and therefore it may be considered, following appropriate counselling regarding the associated risks.


The relation between body mass index, mental health, and functional disability: a European population perspective.
To examine the association between body mass, mental disorders, and functional disability in the general population of 6 European countries. Data (n = 21 425) were derived from the European Study on the Epidemiology of Mental Disorders (ESEMeD). The third version of the Composite International Diagnostic Interview was administered to assess mental disorders (mood, anxiety, and alcohol disorders) according to the Diagnostic Statistical Manual of Mental Disorders-fourth edition, body mass index (BMI) (kg/m(2), based on self-reported height and weight), and functional disability in the previous 30 days, assessed with the World Health Organization Disablement Assessment Scale-second version. About 3% of the respondents were underweight (BMI < 18.5 kg/m(2)), 53% had normal weight (BMI 18.5 to 24.9 kg/m(2)), 33% were overweight (BMI 25 to 29.9 kg/m(2)), and the remaining 12% met criteria for obesity (BMI >or= 30.0 kg/m(2)). Compared with individuals of normal weight, obese individuals were more likely to have mood (OR 1.3; 95%CI, 1.0 to 1.8) or more than one mental disorder (OR 1.4; 95%CI, 1.0 to 2.2). BMI had no impact on work loss days, whereas mental disorders had a considerable effect on work loss days. This is the first cross-national study investigating the role between BMI, mental disorders, and functional disability in the general population. Being overweight or obese is a common condition in the 6 ESEMeD countries. Although there is a moderate association between obesity and mental disorders, BMI did not independently influence functional disability.


Effect of titanium dioxide nanoparticles and co-composted biochar on growth and Cd uptake by wheat plants: A field study.
Cadmium (Cd) is a common toxic trace element found in agricultural soils which is mainly due to anthropogenic activities. Cadmium posed a significant risk to humans all around the world due to its cancer-causing ability. The current study demonstrated the effects of soil-applied biochar (BC) and foliar-applied titanium dioxide nanoparticles (TiO2 NPs) (at a rate of 0.5% and 75 mg/L respectively) alone or in combination on growth and Cd accumulation in wheat plants under field experiment. Soil applied BC and foliar TiO2 NPs, as well as BC coupled with TiO2 NPs, reduced Cd contents in grains by 32%, 47%, and 79%, than control respectively. The usage of NPs and BC boosted the plant height as well as chlorophyll contents by lowering oxidative injury and changing selected antioxidant enzyme activities in leaves than control plants. The combined use of NPs and BC prevented excess Cd accumulation in grains over the critical level (0.2 mg/kg) for cereals. The health risk index (HRI) due to Cd was reduced by 79% by co-composted BC + TiO2 NPs treatment than control. Although, HRI was lower than one for all treatments but this may exceed the limit if grains obtained from such field consumed over long periods. In conclusion, TiO2 NPs and BC amendments can be implemented in fields across the globe where excess Cd is present in soils. Additional studies on the use of such approaches in more precise experimental settings are needed in order to address this environmental problem at larger scale.


Limited value of balloon dilatation in calcified aortic stenosis in adults: direct observations during open heart surgery.
Percutaneous balloon dilatation was recently recommended as a treatment for management of calcified aortic valvular stenosis. This procedure was initially reserved for patients who were not considered surgical candidates; it is now regarded by some as an acceptable alternative for valve replacement. To investigate the validity of this postulate, balloon valvuloplasty was performed under direct vision in the operating room in 16 patients just before excision and replacement of their ossified aortic valve. Changes after valvuloplasty were evaluated by inspection as well as by geometric measurements. The authors found that balloon valvuloplasty did not make a detectable impact on valvular anatomy in about two-thirds of the patients and induced enlargement of the functional aortic orifice judged as "minimal" or "moderate" in one-third of the cases. In no patient was there significant increase in the functional orifice size. Other investigators have shown that hemodynamic and clinical improvement may be induced in some patients by small increases in the aortic orifice; based on the observations herein, such an improvement, if it occurs at all, would be short-lasting; the procedure should be offered only to those who present truly prohibitive risk by standards of modern surgery.


[Alteration in the phospholipid spectrum of blood serum and the state of hemocoagulation in experimental loading with phospholipids].
Phosphatidyl serine, phosphatidyl choline and sphingomyelin were administered into rats separately and simultaneously (at natural ratios, up to the final concentration exceeding 2-fold the normal level in blood). The phospholipid spectrum was estimated in blood serum by means of thin-layer chromatography and the state of haemocoagulation was monitored by means of autocoagulogramms. Sphingomyelin, reducing the content of some physiologically active phospholipids, did not alter haemocoagulation. Administration of phosphatidyl choline, phosphatidyl serine and mixtures of phospholipids resulted in an increase in blood serum of phosphatidyl serine that was responsible for hypocoagulation effect. The accelerating effect of phosphatidyl choline on phosphatidyl serine and importance of the phosphatidyl choline-phosphatidyl serine complex in haemocoagulation is discussed.


An adaptive group sequential phase II design to compare treatments for survival endpoints in rare patient entities.
For rare diseases, standard treatments are often not available and essential study parameters are difficult or impossible to obtain. Therefore, designs of clinical trials for these diseases are often based on little information. Adaptive designs allow such trials to be started and to gain information during the study. Motivated by a trial for a rare subtype of renal-cell carcinoma, we present a two-stage adaptive design for right-censored time-to-event data and a two-sided test. After the first stage, one can stop for futility or continue with reestimated sample size. The properties of such designs are analyzed by simulation studies.


Germline MUTYH gene mutations are not frequently found in unselected patients with papillary breast carcinoma.
MUTYH-associated polyposis (MAP) is an autosomal recessive disease, which predisposes to polyposis and colorectal cancer. There is a trend towards an increased risk of breast cancer in MAP patients, with a remarkable proportion of papillary breast cancers. To determine whether MUTYH mutations are associated with this specific and rare type of breast cancer, 53 unselected patients with papillary breast cancer were analyzed for founder mutations in the MUTYH gene. No germline mutations were identified, indicating that biallelic MUTYH mutations are not a frequent underlying cause for the development of papillary carcinomas of the breast.


GC balance in the internal transcribed spacers ITS 1 and ITS 2 of nuclear ribosomal RNA genes.
The internal transcribed spacer (ITS) 1 and 2, the 5.8S rRNA gene, and adjacent 18S rRNA and 25S rRNA coding regions of two Cucurbitaceae (Cucurbita pepo, zucchini, ITS 1: 187 bp, and ITS 2: 252 bp in length, and Cucumis sativus, cucumber, ITS 1: 229 bp, and ITS 2: 245 bp in length) have been sequenced. The evolutionary pattern shown by the ITSs of these plants is different from that found in vertebrates. Deletions, insertions, and base substitutions have occurred in both spacers; however, it is obvious that some selection pressure is responsible for the preservation of stem-loop structures. The dissimilarity of the 5' region of ITS 2 found in higher plants has consequences for proposed models on U3 snRNA-ITS 2 interaction in higher eukaryotes. The two investigated Cucurbitaceae species show a G + C content of ITS 1 that nearly equals that of ITS 2. An analysis of the ITS sequences reveals that in 19 out of 20 organisms published, the G + C content of ITS 1 nearly equals that of ITS 2, although it ranges from 20% to 90% in different organisms (GC balance). Moreover, the balanced G + C content of the ITSs in a given species seems to be similar to that of so-called expansion segments (ESs) in the 25/28S rRNA coding region. Thus, ITSs show a phenomenon called molecular coevolution with respect to each other and to the ESs. In the ITSs of Cucurbitaceae the balanced G + C composition is at least partly achieved by C to T transitions, via deamination of 5-methylcytosine. Other mutational events must be taken into account. The appearance of this phenomenon is discussed in terms of functional constraints linked to the structures of these spacers.


Multimodal image-guided folic acid targeted Ag-based quantum dots for the combination of selective methotrexate delivery and photothermal therapy.
Multifunctional quantum dots (QDs) with photothermal therapy (PTT) potential loaded with an anticancer drug and labelled with a targeting agent can be highly effective nano-agents for tumour specific, image-guided PTT/chemo combination therapy of cancer. Ag-chalcogenides are promising QDs with good biocompatibility. Ag2S QDs are popular theranostic agents for imaging in near-infrared with PTT potential. However, theranostic applications of AgInS2 QDs emitting in the visible region and its PTT potential need to be explored. Here, we first present a simple synthesis of small, glutathione (GSH) coated AgInS2 QDs with peak emission at 634 nm, 21% quantum yield, and excellent long-term stability without an inorganic shell. Ag2S-GSH QDs emitting in the near-infrared region (peak emission = 822 nm) were also produced. Both QDs were tagged with folic acid (FA) and conjugated with methotrexate (MTX). About 3-fold higher internalization of FA-tagged QDs by folate-receptor (FR) overexpressing HeLa cells than HT29 and A549 cells was observed. Delivery of MTX by QD-FA-MTX reduced the IC50 of the drug from 10 μg/mL to 2.5-5 μg/mL. MTX release was triggered at acidic pH, which was further enhanced with local temperature increase created by laser irradiation. Irradiation of AgInS2-GSH QDs at 640 nm (300 mW) for 10 min, caused about 10 °C temperature increase but did not cause any thermal ablation of cells. On the other hand, Ag2S-GSH-FA based PTT effectively and selectively killed HeLa cells with 10 min 808 nm laser irradiation via mostly necrosis with an IC50 of 5 μg Ag/mL. Under the same conditions, IC50 of MTX was reduced to 0.21 μg/mL if Ag2S-GSH-FA.


Phenotypic characterization of a DFNA6 family showing progressive low-frequency sensorineural hearing impairment.
Only three autosomal dominant hearing loss loci (DFNA1, DFNA6/14/38 and DFNA54) have been reported to be associated with predominantly low-frequency (<2kHz) sensorineural hearing impairment (LFSNHI). The DFNA6 locus was previously mapped to chromosome 4p16.3. It was showed that WFS1 is located in this region. This study presents a six-generation family from Hungary with nonsyndromic, post-lingual, bilateral, symmetric, progressive LFSNHI, that discloses positive linkage to the DFNA6 region. Eleven genetically affected family members have LFSNHI. The HI is started before the age of 25 years. The severity of HI varies from mild to moderate, related to age. Progression was mild but significant at all frequencies causing a flat type audiogram. High-resolution temporal bone CT scan showed normal external, middle and inner ear without any osseus malformations in the temporal bone. Studying genotype-phenotype correlations will enhance our understanding of normal and disturbed hearing process.


Correction: Genus-wide Leptospira core genome multilocus sequence typing for strain taxonomy and global surveillance.
[This corrects the article DOI: 10.1371/journal.pntd.0007374.].


Disordered eating behaviour in adolescents with type 1 diabetes on continuous subcutaneous insulin infusion; relation to body image, depression and glycemic control.
Disordered eating behaviour (DEB) represents a significant morbidity among people with type-1 diabetes (T1D). Continuous-subcutaneous insulin infusion (CSII) improves glycemic control and psychological wellbeing in those with T1D. However, its relation to DEB remains obscure. To compare DEB among adolescents with T1D on CSII versus basal-bolus regimen and correlate it with body image, HbA1C and depression. Sixty adolescents with T1D (30 on CSII and 30 on basal-bolus regimen), aged 12-17 years were studied focusing on diabetes-duration, insulin therapy, exercise, socioeconomic standard, hypoglycemic attacks/week and family history of psychiatric illness. Anthropometric measures, HbA1C, binge eating scale (BES), body image tool, patient health questionnaire-9 (PHQ9) and the Mini-KID depression scale were assessed. Among the studied adolescents with T1D, six had DEB (10%), 14 had poor body-image perception (23.3%), 42 had moderate body-image perception (70%) and 22 had depression (36.7%). Adolescents with T1D on CSII had significantly lower BES (p = 0.022), Mini-KID depression (p = 0.001) and PHQ9 (p = 0.02) than those on basal-bolus regimen. BES was positively correlated to depression (p < 0.001), HbA1C (p = 0.013) and diabetes-duration (p = 0.009) and negatively correlated to body-image (p = 0.003). DEB is a prevalent comorbidity among adolescents with T1D, with higher frequency in those on basal-bolus regimen than CSII.


Correlates of forearm bone mass among women during maximal bone mineralization.
The distribution and correlates of bone mass among women ages 20-35 were studied. During young adulthood, bone mass reaches its maximum level and appears to stabilize. Factors that establish the individual's relative position in this plateau period may influence the ultimate expression of bone loss with aging. Mid-distal radial bone mass was measured in a geographically defined population of 86 women in two rural, demographically similar communities in Iowa. The water supplies provided 55 and 375 mg/liter of elemental calcium, generating significantly different mean community dietary calcium intakes of 871 and 1,233 mg/day, respectively. Bone mass was measured by single-beam photon densitometry and correlated with data from physical measurements, medical history, and reported nutritional intake. In evaluating the joint effect of variables in a multiple regression procedure, forearm bone mass was negatively and significantly associated with alcohol consumption (P = 0.0327) and with a first pregnancy prior to age 20 (P = 0.0250). There was a trend for current calcium intake estimated from 24-hr recall to be positively associated with bone mass (P = 0.0816). Because the 24-hr recall is characterized by significant error due to daily variability of an individual's intakes, a more general calcium intake was estimated from food frequency. Women whose estimated intake of calcium from food frequency was greater than 800 mg/day, the Recommended Daily Allowance, had significantly greater bone mass than women whose intake was estimated to be less than 800 mg/day (P = 0.0053). No relationship was observed with oral contraceptive use, parity, breastfeeding practices, smoking behavior, or measures of physical activity.


Right-to-left interatrial shunt causing platypnea after pneumonectomy. A recent experience and diagnostic value of dynamic magnetic resonance imaging.
Shortness of breath after pneumonectomy is a common finding that has multiple causes. We report the cases of two patients with shortness of breath on assuming an upright posture (platypnea) that followed pneumonectomy; these individuals developed right-to-left shunt across a patent foramen ovale (PFO) with normal right-sided intracardiac pressures. Both contrast echocardiography and magnetic resonance imaging (MRI), including a recently introduced dynamic ultrafast imaging technique, proved helpful in diagnosing this condition noninvasively.


Accessing the Wrist: From Data to Tips and Tricks.
Over the past 2 decades, radial artery access has increasingly become the standard approach for coronary angiography and intervention. Compared with femoral arteries, transradial access is associated with better hemostasis. Transradial access has increased patient preference, facilitates early ambulation, and is cost-effective. An important limitation of transradial access is access site failure, and it carries a crossover rate of 3% to 7% in randomized prospective trials comparing radial with femoral artery access among experienced operators. Crossover rates for failed primary radial artery access can be reduced with ultrasonography guidance and increased familiarity with alternative access sites in the wrist.


Toxicity and genotoxic evaluations in African catfish Clarias gariepinus (Burchell 1822) exposed to Act Force Gold®, Butaforce®, and Atraforce®.
Act Force Gold®, Butaforce®, and Atraforce® are among the most commonly used pesticides in Nigeria. The lethal concentrations and the respective toxic units for the three pesticides were determined. The genotoxic effects of the three pesticides were investigated in the red blood cells of Clarias gariepinus using micronucleus (MN) assay. The 96 h LC50 was 4.75, 4.84, and 54.74 mg L-1 for Act Force Gold®, Butaforce®, and Atraforce®, respectively. The toxic units in ascending order of toxicity were 1.83, 20.66, and 21.05 for Act Force Gold®, Butaforce®, and Atraforce® respectively. The estimated safe levels based on NAS/NAE varied from 4.75 × 10-1-4.75 × 10-5 in Act Force Gold® through 4.64 × 10-1-4.85 × 10-5 in Butaforce® to 5.74-5.74 × 10-5 in Atraforce®. Fish specimens were exposed to the pesticides and sampling was done at regular intervals at days 1, 7, 14, and 21 and after another 7-day recovery period. The results obtained indicated concentration- and duration-dependent increase in % MN formation with maximum values of 3.40 ± 0.25 for Act Force Gold® on day 14 and 3.05 ± 0.36 and 2.35 ± 0.14 for Butaforce® and Atraforce® respectively on day 7 of exposure. The 7-day recovery period could not reverse the trend as the % MN values obtained were significantly different from the control. The results further support the use of MN assay in assessing the toxicity of aquatic pollutants and can be used in the monitoring of aquatic ecosystems.


Malignancy Associated with Low-Risk HPV6 and HPV11: A Systematic Review and Implications for Cancer Prevention.
High-risk human papillomavirus (HPV) is etiologically related to cervical cancer, other anogenital cancers and oropharyngeal carcinomas. Low-risk HPV, especially HPV6 and HPV11, cause genital warts and laryngeal papillomas. However, the accumulating data suggests that HPV6 and HPV11 may cause malignant lesions at non-cervical anatomic sites. This review aims to estimate the proportions of single and dual HPV6/11 infections in multiple cancers reported in the last 10 years in the Cochrane, Embasa and PubMed databases. Secondly, the genomes of HPV6/11 were compared with the most common high-risk genotype, HPV16, to determine the similarities and differences. A total of 11 articles were selected, including between one and 334 HPV+ cancer patients. The frequencies of single or dual HPV6/11 infections ranged between 0-5.5% for penile and 0-87.5% for laryngeal cancers and were null for vulvar, vaginal and oral cancers. The genomic similarities between HPV6/11 and HPV16 mainly involved the E7 gene, indicating a limited ability to block cell differentiation. The presence of single or dual HPV6/11 infections in variable proportions of penile and laryngeal cancers support the vaccination strategies that cover these genotypes, not only for preventing genital warts but also for cancer prevention. Other risk factors and co-carcinogens are likely to participate in epithelial carcinogenesis associated with low-risk HPV.


An Anionic Amido-Substituted Seven-Vertex Siliconoid Cluster.
The silanide [Si4 {N(SiMe3 )Dipp}3 ]- (1) transforms into the anionic siliconoid cluster [Si7 {N(SiMe3 )Dipp}3 ]- (2) with four unsubstituted silicon atoms as a contact ion pair with [K([18]crown-6)] in C6 D6 at room temperature within five weeks. Anion 2 was investigated by natural population analysis and visualization of intrinsic atomic orbitals. Magnetically induced current-density calculations of 2 revealed two distinct strong diatropic vortices that sum up in one direction and create a strongly shielded apical silicon atom in 2.