Pharyngeal and laryngeal symptoms and signs related to extraesophageal reflux in patients with heartburn in gastroenterology practice: a prospective study.
To describe pharyngeal and laryngeal symptoms and findings in correlation to extraesophageal reflux (EER) in patients with heartburn. A prospective study. Patients referred to Department of Gastroenterology, Karolinska University Hospital Huddinge. Forty-three patients with chronic heartburn as the primary symptom, subdivided on the basis of oesophageal manometry to have gastro-oesophageal reflux disease (GORD) (n = 25) or not (n = 18). Laryngeal video recordings of 43 patients were examined off-line. The occurrence of symptoms and laryngeal pathology were correlated to the results of 24-h double-probe pH monitoring. Pharyngeal or laryngeal symptoms occurred often or quite often in 72% (18 of 25) of the GORD patients and 61% (11 of 18) of the patients without GORD. EER occurred in 73% (16 of 22) of the GORD patients with laryngeal symptoms and in 38% (6 of 16) of those without GORD but with laryngeal symptoms (P = 0.047). In the multivariate analysis the odds for hoarseness was 17 times higher if the patient had heartburn more than five times a day, compared with patients with heartburn less than once a day (OR 17.8, CI 1.7-405.9; P = 0.01). Cohen's kappa for intrarater reliability was 0.851 (P = 0.001). Laryngeal pathology was found in 56% of the GORD patients and 44% patients without GORD. There was no significant difference in the occurrence of any specific symptom or finding between the patients with and without GORD, nor with and without EER. Although GORD together with laryngeal symptoms seem to be predictive of EER, there are no specific pharyngeal and laryngeal symptoms or findings that would help distinguishing patients with EER from those without. However, the more frequent the heartburn to the more frequent is hoarseness.


In vitro metabolic stability and metabolite profiling of TJ0711 hydrochloride, a newly developed vasodilatory β-blocker, using a liquid chromatography-tandem mass spectrometry method.
In this paper, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous analysis of metabolic stability and metabolite profiling of 1-[4-(2-methoxyethyl) phenoxy]-3-[[2-(2-methoxyphenoxy) ethyl]amino]-2-propanol hydrochloride (TJ0711 HCl), a new vasodilatory β-blocker. Multiple reaction monitoring (MRM) was used as a survey scan to quantify the parent compound and to trigger the acquisition of enhanced product ions (EPI) for the identification of formed metabolites. In addition, comparison between MRM-only and MRM-information dependent acquisition-EPI (MRM-IDA-EPI) methods was conducted to determine analytical variables, including linearity, limit of detection (LOD), lower limit of quantification (LLOQ), as well as intra-day and inter-day accuracy and precision. Results demonstrated that MRM-IDA-EPI quantitative analysis was not affected by the addition of EPI scans to obtain qualitative information during the same chromatographic run, compared to MRM-only method. Thereafter, metabolic stability and metabolite identification of TJ0711 HCl were investigated using human liver microsomes (HLM) by the MRM-IDA-EPI method. The in vitro metabolic stability parameters were calculated and t(1/2), microsomal intrinsic clearance (CL(int)), as well as hepatic CL, were 13.0 min, 106.5 μL/min/mg microsomal protein, and 1082.2 mL/min, respectively. The major formed metabolites were also simultaneously monitored and the metabolite profiling data demonstrated that this MRM-IDA-EPI method was capable of targeting a large number of metabolites, in which demethylation and hydroxylation were the principle metabolism pathways during the in vitro incubation with HLM.


Folate and Inflammatory Markers Moderate the Association Between Helicobacter pylori Exposure and Cognitive Function in US Adults.
Helicobacter pylori (H. pylori) infection is associated with cognitive deficits in humans, an association potentially mediated or moderated by folate concentration or inflammation. We used the National Health and Nutrition Examination Survey (NHANES) datasets to examine whether folate concentration or inflammation mediates or moderates the relationship between H. pylori and cognitive function. Models were performed using linear, Poisson, and zero-inflated Poisson regression, and we performed separate analyses for groups aged 20-59 and 60-90 years with sample sizes ranging from 700 to 1700. We did not find evidence of mediation in either age group. In the 20- to 59-year group, interactions between H. pylori and ferritin (p values ranging from .004 to .039) were associated with worse processing speed, better working memory, and worse reaction time. Interactions between H. pylori and fibrinogen (p values ranging from .023 to .045), C-reactive protein (CRP) (p = .023), and the inflammatory index (p = .045) were associated with worse processing speed. In 60- to 90-year-olds, H. pylori interacted with ferritin and the inflammatory index to predict fewer mathematical errors (p values of .036 and .023). Interactions with folate (p values of .016 and .006) and C-reactive protein (p values ranging from <.001 to .048) were inconsistent in directionality. In this dataset, representative of the US population, inflammation and folate concentrations moderated but did not mediate the association between H. pylori seropositivity and cognition.


Injecting practices during and after hepatitis C treatment and associations with not achieving cure among persons who inject drugs.
Persons who inject drugs (PWID) are a key population for hepatitis C virus (HCV) treatment. Study aims were to describe injection practices of PWID during HCV treatment with direct-acting antivirals (DAAs) and assess whether injection practices were associated with not achieving a sustained virologic response (SVR). Secondary analysis of the HERO Study (ClinicalTrials.gov, NCT02824640), a pragmatic randomized trial in 8 U.S. states to evaluate the effectiveness of HCV care models among active PWID seen in opioid treatment programs and community clinics. Frequency, sharing and reuse of injecting equipment were assessed at baseline, end-of-treatment (EOT) and quarterly visits up to 60 weeks post-treatment. Generalized Estimating Equations logistic regression models with linear spline were used to compare trends in injecting behaviors during vs. post-treatment. Multivariable logistic regression models explored associations between injecting behaviors during treatment and lack of SVR. Among 501 participants, 27% were female, 35% were non-white, mean age was 44 (SD 11.5) years and nearly half (49%) were unhoused. At baseline, 41% reported receptive sharing of injecting equipment, declining to 16% at EOT visit. Receptive sharing of cookers, rinses, or needles/syringes during treatment was associated with a nearly 5-fold increase in not achieving SVR (adjusted odds ratio (aOR)=4.83; 95% CI: 2.26, 10.28) as was reuse of one's own needles/syringes (aOR=2.37; 95% CI: 1.11, 4.92). PWID in the HERO study adopted safer injecting behaviors during DAA treatment; receptive sharing of injecting equipment and reuse of one's own equipment during treatment were associated with not achieving cure.


Pulmonary migration of a fragment of plastic coating sheared from a stylet.
In trauma patients, particularly with head immobilization, tracheal intubation without the use of a stylet may be impossible. To report a very rare but potentially fatal complication that may happen in any Emergency Department: fracture of the plastic sheath of an intubation stylet, reported only twice before in the literature. Two large plastic fragments detached from a stylet while intubating a trauma patient. One piece was removed from the endotracheal tube a few hours later in the operating room. The second fragment migrated asymptomatically into the pulmonary airway. It was successfully retrieved from the right bronchus 24 h later. This potentially life-threatening event may go unnoticed after intubation if the endotracheal tube is not obstructed by the fragment. Gentle withdrawal of the stylet from the tube is essential to avoid stylet fracture. Careful examination of the stylet after intubation may suggest a stylet fracture.


Down-regulation of the sucrose transporters HvSUT1 and HvSUT2 affects sucrose homeostasis along its delivery path in barley grains.
Sucrose transport and partitioning are crucial for seed filling. While many plasma-membrane-localised sucrose transporters (SUT1 family members) have been analysed in seeds, the functions of vacuolar SUT2 members are still obscure. In barley grains, expression of HvSUT1 and HvSUT2 overlap temporally and spatially, suggesting concerted functions to regulate sucrose homeostasis. Using HvSUT2-RNAi plants, we found that grains were also deficient in HvSUT1 expression and seemingly sucrose-limited during mid-to-late grain filling. Transgenic endosperms accumulated less starch and dry weight, although overall sucrose and hexose contents were higher. Comprehensive transcript and metabolite profiling revealed that genes related to glycolysis, the tricarboxylic acid cycle, starch and amino acid synthesis, grain maturation, and abscisic acid signalling were down-regulated together with most glycolytic intermediates and amino acids. Sucrose was increased along the sucrose delivery route in the nucellar projection, the endosperm transfer cells, and the starchy endosperm, indicating that suppressed transporter activity diminished sucrose efflux from vacuoles, which generated sugar deficiency in the cytoplasm. Thus, endosperm vacuoles may buffer sucrose concentrations to regulate homeostasis at grain filling. Transcriptional changes revealed that limited endosperm sucrose initiated sugar starvation responses, such as sugar recycling from starch, hemicelluloses and celluloses together with vacuolar protein degradation, thereby supporting formation of nucleotide sugars. Barley endosperm cells can thus suppress certain pathways to retrieve resources to maintain essential cell functions.


Investigation of substituted-benzene dopants for charge exchange ionization of nonpolar compounds by atmospheric pressure photoionization.
Atmospheric pressure photoionization (APPI) using a dopant enables both polar and nonpolar compounds to be analyzed by LC/MS. To date, the charge exchange ionization pathway utilized for nonpolar compounds has only been efficient under restrictive conditions, mainly because the usual charge exchange reagent ions--the dopant photoions themselves--tend to be consumed in proton transfer reactions with solvent and/or dopant neutrals. This research aims to elucidate the factors affecting the reactivities of substituted-benzene dopant ions; another, overriding, objective is to discover new dopants for better implementing charge exchange ionization in reversed-phase LC/MS applications. The desirable properties for a charge exchange dopant include low reactivity of its photoions with solvent and dopant neutrals and high ionization energy (IE). Reactivity tests were performed for diverse substituted-benzene compounds, with substituents ranging from strongly electron withdrawing (EW) to strongly electron donating (ED). The results indicate that both the tendency of a dopant's photoions to be lost through proton transfer reactions and its IE depend on the electron donating/withdrawing properties of its substituent(s): ED groups decrease reactivity and IE, while EW groups increase reactivity and IE. Exceptions to the reactivity trend for dopants with ED groups occur when the substituent is itself acidic. All told, the desirable properties for a charge exchange dopant tend towards mutual exclusivity. Of the singly-substituted benzenes tested, chloro- and bromobenzene provide the best compromise between low reactivity and high IE. Several fluoroanisoles, with counteracting EW and ED groups, may also provide improved performance relative to the established dopants.


Substance P in proliferative vitreoretinopathy: the significance of aqueous humor levels for evolution of the disease.
We detected aqueous humor levels of substance P in patients with various grades of proliferative vitreoretinopathy and with uncomplicated rhegmatogenous retinal detachment. To evaluate the significance of the concentration of substance P at the time of surgery for retinal detachment for subsequent development of proliferative vitreoretinopathy, the latter patients also underwent fundoscopic control examination. Using a highly specific and sensitive radioimmunoassay, the content of substance P in fresh samples of aqueous humor obtained by paracentesis was determined both in cataract controls and in patients with uncomplicated rhegmatogenous retinal detachment and with various grades of proliferative vitreoretinopathy. Retinal detachment patients underwent fundoscopic control examination 6 months after surgical reattachment. The mean concentration of substance P in cataract controls was 40.3 (+22.4) fmol/mg protein, in the retinal detachment group 61.9 (+/-13.9) fmol/mg protein and in proliferative vitreoretinopathy 335.2 (+/-24.8) fmol/mg protein, but no correlation between levels of the peptide and various grades of the disease was observed. Already at surgery for retinal detachment three in four patients who developed proliferative vitreoretinopathy presented with levels of substance P in the range of the disease. The concentration of substance P in aqueous humor is significantly high in patients with proliferative vitreoretinopathy in whom surgery is indicated. Furthermore, elevation of the peptide in retinal detachment that originates most obviously from a neurogenic mechanism may indicate initiation of processes associated with proliferative vitreoretinopathy, thus representing an indicator of significant risk for evolution of the disease at a very early time.


Human MRP3 transporter: identification of the 5'-flanking region, genomic organization and alternative splice variants.
In humans, at least six members of the multidrug resistance-associated protein (MRP) family are thought to exist. Here we report the molecular cloning of two splice variants of MRP3 from human liver. In addition, MRP3 genomic organization including the 5'-flanking region and a major portion of the MRP3 intron-exon organization are identified and characterized.


TGF-beta1 expression in EL4 lymphoma cells overexpressing growth hormone.
Our previous studies show that growth hormone overexpression (GHo) upregulates the expression of the IGF-1R and IGF-2R resulting in the protection of the EL4 lymphoma cell line from apoptosis. In this study, we report that GHo also increases TGF-beta1 protein expression measured by luciferase promoter assay, Western analysis, and ELISA. Further, the data show that antibody to TGF-betaR2 decreases TGF-beta1 promoter activity to the level of vector alone control cells. GHo cells treated with (125)I-rh-latent TGF-beta1 showed increased activation of latent TGF-beta1 as measured by an increase in the active 24kDa, TGF-beta1 compared to vector alone control cells. The ability of endogenous GH to increase TGF-beta1 expression is blocked in EL4 cells by antisense but not sense oligodeoxynucleotides or in cells cultured with antibody to growth hormone (GH). The data suggest that endogenous GH may protect from apoptosis through the IGF-1R receptor while limiting cellular growth through increased expression and activation of TGF-beta1.


Aphasic reading and writing: possible evidence for right hemisphere participation.
Aphasic patients performed two tasks, reading aloud and writing to dictation. In both tasks, success was positively correlated to the emotionality, the imageability and the frequency of the target words. In addition, the patients made more semantic Also, the relative difficulty of words was similar for aphasic patients to read and for normal male subjects to recognize in the left visual field. These results suggest that the performance of the aphasic patients on these language tasks were mediated in part by the right hemisphere.


Tear volume estimation using a modified Schirmer test: a randomized, multicenter, double-blind trial comparing 3% diquafosol ophthalmic solution and artificial tears in dry eye patients.
We aimed to evaluate the feasibility of using a modified Schirmer test to determine the increase in tear volume after administration of 3% diquafosol ophthalmic solution (diquafosol 3%) in dry eye patients. A randomized, multicenter, prospective, double-blind clinical study recruited 50 qualified subjects. They received diquafosol 3% in one eye and artificial tears in the other eye. The study protocol comprised a screening and treatment procedure completed within 1 day. The Schirmer test was performed on closed eyes three times a day. The primary efficacy end points were the second Schirmer test scores 10 minutes after the single dose. Secondary end points were the third Schirmer test scores 3 hours and 40 minutes after the single dose and the symptom scores prior to the second and third Schirmer tests. According to the Schirmer test, 10 minutes after administration, diquafosol 3% significantly increased tear volume compared to artificial tears. Diquafosol 3% and artificial tears both showed significant improvements in the symptom scores compared to baseline. However, there was no significant difference in the symptoms score between diquafosol 3% and artificial tears. The modified Schirmer test can detect a minute change in tear volume in dry eye patients. These findings will be useful in the diagnosis of dry eye, assessment of treatment benefits in daily clinical practice, and the development of possible tear-secreting compounds for dry eye.


The molecular structure and function of sorting nexin 10 in skeletal disorders, cancers, and other pathological conditions.
SNX10 is a member of the phox homology domain-containing family of phosphoinositide-binding proteins. Intracellularly, SNX10 localizes to endosomes where it mediates intracellular trafficking, endosome organization, and protein localization to the centrosome and cilium. It is highly expressed in bone and the gut where it participates in bone mineral and calcium homeostasis through the regulation of osteoclastic bone resorption and gastric acid secretion, respectively. Not surprisingly, patients harboring mutations in SNX10 mutation manifest a phenotype of autosomal recessive osteopetrosis or malignant infantile osteopetrosis, which is clinically characterized by dense bones with increased cortical bone into the medullary space with bone marrow occlusion or depletion, bone marrow failure, and anemia. Accordingly, SNX10 mutant osteoclasts exhibit impaired bone resorptive capacity. Beyond the skeleton, there is emerging evidence implicating SNX10 in cancer development, metabolic disorders, inflammation, and chaperone-mediated autophagy. Understanding the structural basis through which SNX10 exerts its diverse biological functions in both cell and tissue-specific manners may therefore inform new therapeutic opportunities toward the treatment and management of SNX10-related diseases.


Application of an amphibian (Silurana tropicalis) metamorphosis assay to the testing of the chronic toxicity of three rice paddy herbicides: simetryn, mefenacet, and thiobencarb.
We examined the chronic toxicity of three rice paddy herbicides (simetryn, mefenacet, and thiobencarb) using an amphibian (Silurana tropicalis) metamorphosis assay (a 28-day semistatic test under an individual-separated exposure system). Each herbicide was tested at two concentrations (1/100 and 1/10 of the 96-h LC50 value reported previously) with morphometric, gravimetric, and thyroid-histological endpoints. Simetryn caused significant retardation in growth and development at both test concentrations (0.04 and 0.40mg/L), as indicated by significantly shorter total body lengths and hind limb lengths, smaller wet body masses, and delayed developmental stages compared to those observed in the control tadpoles. However, no clear histopathology was observed in the thyroid glands of the tadpoles exposed to simetryn. These results suggest that simetryn can act as a chemical stressor retarding tadpole growth and development without disrupting thyroid functions, even at 1/100 of the 96-h LC50 value. In addition, scoliosis near the tail base was observed in the tadpoles exposed to 0.40mg/L of simetryn at a significantly high incidence (7/30=23.3%). Therefore, simetryn can also act as a teratogen inducing axial malformations at 1/10 of the 96-h LC50 value. During the 28 days of exposure, neither mefenacet (0.03 and 0.30mg/L) nor thiobencarb (0.008 and 0.080mg/L) induced any abnormalities, although the test concentrations measured immediately before the solution renewals decreased to nearly 50 percent of the nominal concentrations since day 14. Because the concentrations tested for simetryn are likely to occur in paddy water, wild anuran tadpoles in paddy water may therefore be adversely impacted by simetryn.


Contact-dependent delivery of IL-2 by dendritic cells to CD4 T cells in the contraction phase promotes their long-term survival.
Common γ chain cytokines are important for immune memory formation. Among them, the role of IL-2 remains to be fully explored. It has been suggested that this cytokine is critically needed in the late phase of primary CD4 T cell activation. Lack of IL-2 at this stage sets for a diminished recall response in subsequent challenges. However, as IL-2 peak production is over at this point, the source and the exact mechanism that promotes its production remain elusive. We report here that resting, previously antigen-stimulated CD4 T cells maintain a minimalist response to dendritic cells after their peak activation in vitro. This subtle activation event may be induced by DCs without overt presence of antigen and appears to be stronger if IL-2 comes from the same dendritic cells. This encounter reactivates a miniature IL-2 production and leads a gene expression profile change in these previously activated CD4 T cells. The CD4 T cells so experienced show enhanced reactivation intensity upon secondary challenges later on. Although mostly relying on in vitro evidence, our work may implicate a subtle programing for CD4 T cell survival after primary activation in vivo.


Impact of Parkinson's disease and levodopa on resting state functional connectivity related to speech prosody control.
Impaired speech prosody is common in Parkinson's disease (PD). We assessed the impact of PD and levodopa on MRI resting-state functional connectivity (rs-FC) underlying speech prosody control. We studied 19 PD patients in the OFF and ON dopaminergic conditions and 15 age-matched healthy controls using functional MRI and seed partial least squares correlation (PLSC) analysis. In the PD group, we also correlated levodopa-induced rs-FC changes with the results of acoustic analysis. The PLCS analysis revealed a significant impact of PD but not of medication on the rs-FC strength of spatial correlation maps seeded by the anterior cingulate (p = 0.006), the right orofacial primary sensorimotor cortex (OF_SM1; p = 0.025) and the right caudate head (CN; p = 0.047). In the PD group, levodopa-induced changes in the CN and OF_SM1 connectivity strengths were related to changes in speech prosody. We demonstrated an impact of PD but not of levodopa on rs-FC within the brain networks related to speech prosody control. When only the PD patients were taken into account, the association between treatment-induced changes in speech prosody and changes in rs-FC within the associative striato-prefrontal and motor speech networks was found.


Survival of patients with epilepsy: an estimate of the mortality risk.
To investigate the extent and causes of the differences in mortality found in studies on mortality in epilepsy based on a quantitative review of the literature. We used MEDLINE database and Cumulative Index Medicus for 1960-2001, Excerpta Medica for 1948-1965, and relevant journals and bibliographies. We selected comparative studies investigating mortality in epilepsy patients conducted in the last 100 years. The Standardized Mortality Ratio (SMR) was selected as primary outcome. Nineteen studies were included. Pooled estimates were precision weighted and tested for homogeneity. Sources of variability between risk estimates were explored by using multivariate fixed-effects models. SMRs ranged from 1.3 to 9.3. Risk estimates proved heterogeneous (chi-square test statistic: 1,177; df = 18; p < 0.001). The most important determinant was "source population," explaining half of the variance of the estimates (R(2), 0.47; p = 0.006). SMRs in community studies ranged from 1.3 to 3.1, and for institutionalized populations, from 1.9 to 5.1. Our results show that the mortality risk in patients with epilepsy is dependent on source population of patients. Within the different source populations, considerable unexplained variance remains. Hence no uniform summary estimate for the elevated mortality could be determined.


Abdominal and thoracoabdominal aortic aneurysm.
Most abdominal aortic aneurysms (AAA) and thoracoabdominal aortic aneurysms (TAAA) are asymptomatic and are found on physical exam or incidentally during radiological studies for other indications. These aneurysms are repaired primarily because their risk of rupture increases geometrically as the size exceeds 5 cm. The potential morbidity of intraoperative visceral and spinal ischemia involved with TAAA repair may be reduced with various adjunctive maneuvers.


Interspecies regulation of the SOS response by the E. coli lexA+ gene.
A plasmid-encoded E. coli lexA+ gene was introduced into 6 species of Enterobacteria. Ultraviolet light-sensitization occurred in all species except P. rettgeri, and 4 organisms showed reduced inducibility of RecA-like proteins. The mechanism of lexA+ control of the SOS response therefore appears common to several species.


Emergency Medical Care of People with Intellectual Disabilities: A Scoping Review.
The paper intends to do a scoping review of people with intellectual disabilities in emergency care where this group seems to face access barriers and discrimination. It analyses the conceptual and methodological framework for studies examining the former. A scoping review is conducted. The studies' quality is assessed via a checklist developed by the authors drawing on a compilation of common assessment tools for study quality. Fourteen quantitative studies fulfil the inclusion criteria for further analysis. Summary measures are extracted. Results are synthesized with Andersen's Behavioral Model of Health Service Use. Studies employ a combination of variables attributable to different aspects of population characteristics and health behavior. Most studies seek to quantify or predict emergency care overuse by people with intellectual disabilities. Future studies should also take patients' poor health or treatment outcomes and their perspectives into account.


Do sleep disturbances in depressed adolescents improve following psychological treatment for depression?
Persistent sleep disturbances are one of the most common symptoms of Major Depressive Disorder (MDD) in adolescence. These are not typically targeted in psychological treatments and it is not known if psychological treatment for depression improves sleep. Secondary analyses were conducted using data from a large, multi-centre, randomised controlled trial (Goodyer et al., 2017b). Young people aged 12-18 years (N = 465; 75% female) met diagnostic criteria for Major Depressive Disorder, based on the Kiddie-Schedule for Affective Disorders and Schizophrenia (K-SADS). They were randomised to one of three psychological treatments. Sleep difficulties were assessed at baseline, post-treatment (36 weeks) and one year follow up (86 weeks) with the K-SADS, and the Mood and Feelings Questionnaire (MFQ). At baseline, 92% of young people exhibited clinically significant sleep difficulties. Exploratory analyses suggest that sleep difficulties significantly decreased from baseline to end of treatment on self-report and interview-based measures, and this decrease was maintained at follow up. Reduction in sleep difficulties did not differ between the psychological treatments. Approximately, half of young people reported residual sleep difficulties at the end of treatment and at follow-up. This paper reports secondary data analyses and findings are exploratory. Tentative results suggest that psychological treatments for depression reduced sleep problems for some participants. However, young people with treatment-resistant sleep problems may benefit from adjunctive sleep interventions. Future work with a range of sleep measures is needed to determine those who have residual sleep problems at the end of treatment and post-treatment follow-up.


Time course for development of anticonvulsant tolerance and GABAergic subsensitivity after chronic diazepam.
The time courses for development of neuronal and behavioral tolerance to diazepam (DZ) were estimated in rats continuously exposed to low levels of DZ for 3, 7, 14 or 21 days. Microiontophoretic sensitivity of dorsal raphe neurons to gamma-aminobutyric acid (GABA) was initially facilitated after short-term exposure to DZ released from implanted capsules for up to 3 days but returned to control levels by 7 days postimplantation and continued to decrease thereafter. GABAergic sensitivity remained depressed for a minimum of 5 days following removal of DZ capsules. To obtain a behavioral measure of tolerance, the anticonvulsant activity of DZ against bicuculline-induced seizures was also assessed. Rats studied 3 days after capsule implantation showed a significant elevation in seizure threshold. Seizure liability returned to control levels ca. 7 days after chronic treatment was initiated. These results indicate that tolerance to anticonvulsant efficacy against bicuculline seizures are temporally related to the onset of reduced GABA sensitivity on dorsal raphe neurons during prolonged exposure to DZ.


Ipsi- and contralateral fibre transformations by cross-reinnervation. A principle of symmetry.
Cross-reinnervation of rabbit soleus muscle by the peroneal nerve induces a 90% transformation of slow into fast fibres. These changes are reflected in corresponding transformations of the enzyme activity pattern of energy metabolism, the isozyme pattern of lactate dehydrogenase and, in confirmation of previous results (Srihari et al. 1981), transitions from a slow to a fast type myosin light chain pattern. The transformation process appears to be complete after 6 months. Similar changes, although less extensive are also found in the soleus muscle of the contralateral leg. Fibre type transitions in the contralateral muscle are not accompanied by fibre type grouping, as seen in the cross-reinnervated muscle and therefore these changes appear to result from a transformation of the motor units themselves. This phenomenon is interpreted as a compensatory process in maintaining symmetry within the neuromotor system.


Evaluation of a liquid formulation of monensin to control bloat in pasture-fed milking cows.
To evaluate the effectiveness of a liquid formulation of monensin in reducing bloat score in milking cows grazing pasture. A Friesian x Jersey crossbred herd which had been genetically selected for high bloat susceptibility since 1973 was used in this study. Two trials were conducted, each involving two groups of 15 cows, randomly allocated to Treated or Control groups. Trial 1 involved twice-daily administration of a novel liquid formulation of monensin given to Treated cows at morning and afternoon milkings. Trial 2 involved once-daily administration of the same product to Treated cows at the morning milking only. Control cows received no preventative treatment. The total daily dose of monensin in each trial was 300 mg per cow, given in a total volume of 100 ml of liquid. Animals were scored for bloat twice-daily whilst grazing on white-clover/ryegrass or red-clover pastures. The scoring system used a scale of 0, 1, 1.5, 2, 2.5, or 3, representing scores of increasing bloat severity as assessed by palpation of abdominal pressure on the left and right sides of the cow. In Trial 1, severe bloat was recorded at 9 out of 23 scoring sessions. Twelve Control cows required therapeutic treatment for bloat on at least one day, compared to 3 cows in the Treated group (p=0.003). In Trial 2, severe bloat in 3 of 9 scoring sessions led to 8/15 Control animals requiring therapeutic treatment compared to 1/14 cows in the Treated group (p=0.02). Oral drenching with the liquid formulation of monensin tested was effective in reducing bloat score in milking cows grazed on pasture.


On the possible presence of a beta 2-microglobulin-like protein in extracts of livers from normal chickens and chickens with erythroblastosis--I. Recognition of a small-molecular weight (mol. wt 11,000) protein.
1. An extract from the livers of both normal chickens (N) and chickens infected with avian erythroblastosis virus (Eb) contains a small molecular weight protein (SMWP, mol. wt 11,000). 2. Double immunodiffusion studies with rabbit antiserum against fowl serum proteins shows a precipitin arc for SMWP in N and Eb extracts, which is continuous with one from one of the normal chicken serum proteins. 3. When treated with 60% saturated ammonium sulphate the SMWP in the liver extracts divides between the precipitate and the supernatant although the specific serological activity of Eb extracts (gag--or COFAL--determined antigenic activity) is restricted to the precipitated SMWP fraction. 4. The COFAL activity of Eb liver extracts could be associated with SMWP by its attachment to this protein, or this phenomenon of "association" could represent the result of changes in synthesis of SMWP or post-synthetic changes.


The Hawthorne effect, sponsored trials, and the overestimation of treatment effectiveness.
To determine if the results of rheumatoid arthritis (RA) clinical trials are upwardly biased by the Hawthorne effect. We studied 264 patients with RA who completed a commercially sponsored 3-month, open-label, phase 4 trial of a US Food and Drug Administration approved RA treatment. We evaluated changes in the Health Assessment Questionnaire disability index (HAQ) and visual analog scales for pain, patient global, and fatigue during 3 periods: pretreatment in the trial, on treatment at the close of the trial, and by a trial-unrelated survey 8 months after the close of the trial, but while the patients were receiving the same treatment. The HAQ score (0-3) improved by 41.3% during the trial, but only by 16.5% when the endpoint was the post-trial result. Similar results for the other variables were patient global (0-10) 51.9% and 34.6%, pain (0-10) 51.7% and 39.7%, fatigue (0-10) 45.6% and 24.6%. Worsening between the trial end and the first survey assessment was HAQ 0.29 units, pain 0.8 units, patient global 0.8 units, and fatigue 1.1 units. Almost half the improvement noted in the clinical trial HAQ score disappeared on entry to a non-sponsored followup study, and from 23% to 44% of improvements in pain, patient global, and fatigue also disappeared. These changes can be attributed to the Hawthorne effect. Based on these data, we hypothesize that the absolute values of RA outcome variables in clinical trials are upwardly biased, and that the treatment effect is less than observed.


Development and application of an indirect enzyme-linked immunosorbent assay for serological survey of Japanese encephalitis virus infection in dogs.
Japanese encephalitis virus (JEV) causes serious acute encephalitis in humans and horses. Although dogs are good sentinels for assessing the risk of JEV infection to humans, a virus neutralization test has been the only method available for measuring the levels of JEV antibody in dogs. In this study, an indirect enzyme-linked immunosorbent assay (ELISA) using purified viral particles as an antigen, was developed for serological survey of JEV infection in dogs. In dogs inoculated experimentally with JEV, the ELISA detected anti-JEV IgM 3 days after infection, with IgM levels peaking 7 days after infection. Anti-JEV IgG was detected 14 days after infection and peaked on 21-28 days after infection. Virus neutralization titers correlated with anti-JEV immunoglobulins measured by the ELISA. To test the utility of the new assay, the seroprevalence of JEV infection among 102 dogs in Kyushu, Japan, was examined by IgG ELISA and by virus neutralization. The correlation coefficient between the IgG ELISA and virus neutralization was 0.813 (p<0.001); comparison of the IgG ELISA and virus neutralization showed a sensitivity and specificity of 82% and 98%, respectively. The IgG ELISA was used to survey dogs in Bangkok, Thailand and 51% of these dogs were found seropositive for JEV. These data suggest that in the capital city of Thailand, the risk of infection with JEV remains high.


Atmospheric observations suggest methane emissions in north-eastern China growing with natural gas use.
The dramatic increase of natural gas use in China, as a substitute for coal, helps to reduce CO2 emissions and air pollution, but the climate mitigation benefit can be offset by methane leakage into the atmosphere. We estimate methane emissions from 2010 to 2018 in four regions of China using the GOSAT satellite data and in-situ observations with a high-resolution (0.1° × 0.1°) inverse model and analyze interannual changes of emissions by source sectors. We find that estimated methane emission over the north-eastern China region contributes the largest part (0.77 Tg CH4 yr-1) of the methane emission growth rate of China (0.87 Tg CH4 yr-1) and is largely attributable to the growth in natural gas use. The results provide evidence of a detectable impact on atmospheric methane observations by the increasing natural gas use in China and call for methane emission reductions throughout the gas supply chain and promotion of low emission end-use facilities.


The amphipathic helix concept: length effects on ideally amphipathic LiKj(i=2j) peptides to acquire optimal hemolytic activity.
In a minimalist approach to modeling lytic toxins, amphipathic peptides of LiKj with i=2j composition and whose length varies from 5 to 22 residues were studied for their ability to induce hemolysis and lipid vesicle leakage. Their sequences were designed to generate ideally amphipathic alpha helices with a single K residue per putative turn. All the peptides were lytic, their activities varying by more than a factor of 103 from the shortest 5-residue-long peptide (5-mer) to the longest 22-mer. However, there was no monotonous increase versus length. The 15-mer was as active as the 22-mer and even more than melittin which is used as standard. Partition coefficients from the buffer to the membrane increased in relation to length up to 12 residues, then weakly decreased to reach a plateau, while they were expected to increase monotonously with peptide length and hydrophobicity as revealed from HPLC retention times. Fluorescence labeling by a dansyl group at the N-terminus, or by a W near the CO-terminus, show that up to 12 residues, the peptides were essentially monomeric while longer peptides strongly aggregated in the solution. Lipid affinity was then controlled by peptide length and was found to be limited by folding and self-association in buffer. The lytic activity resulted both from lipid affinity, which varied by a factor of 20-fold, and from efficiency in disturbing the membrane when bound, the latter steeply and monotonously increasing with length. The 15-residue-long peptide, KLLKLLLKLLLKLLK, had the optimal size for highest lytic activity. The shallow location of the fluorescent labels in the lipids is further evidence for a model of peptides remaining flat at the interface.


Insects could exploit UV-green contrast for Landmark navigation.
Illumination-invariant detection of landmark features is a prerequisite for landmark navigation in insects. It is suggested that a contrast mechanism involving the UV and green receptors of insect eyes could guarantee a robust separation between natural objects as foreground and sky as background. Using a sensor with a UV and a green channel that in their spectral characteristics are close to the corresponding insect photoreceptors, data of natural objects and sky were collected. The data show that the two classes can be separated by a fixed threshold in the UV-green color space, offering an advantage over a purely UV-based separation that would require a dynamic threshold. Based on a numerical method, UV-green antagonism is shown to guarantee a more reliable discrimination than UV-blue antagonism.


In-situ infiltration performance of different permeable pavements in a employee used parking lot--A four-year study.
Permeable pavements are being adopted as a green solution in many parts of the world to manage urban stormwater quantity and quality. This paper reports on the measured in-situ infiltration performance over a four-year period since construction and use of three permeable parking sections (permeable pavers, permeable concrete and permeable asphalt) of an employee car parking lot. There was only a marginal decline in infiltration rates of all three pavements after one year of use. However, between years two to four, the infiltration rates declined significantly due to clogging of pores either by dry deposition of particles and/or shear stress of vehicles driving and degrading the permeable surfaces; during the last two years, a greater decline was also observed in driving areas of the parking lots compared to parking slots, where minimal wear and tear are expected. Maintenance strategies were employed to reclaim some of the lost infiltration rate of the permeable pavements to limited success. Despite this decline, the infiltration rates were still four to five times higher than average rainstorm intensity in the region. Thus, these permeable pavement parking lots may have significant ecological importance due to their ability to infiltrate rainwater quickly, reduce the runoff in the catchment area, and also dampen runoff peak flows that could otherwise enter the collection system for treatment in a combined sewer area.


Design of liposomal formulations for cell targeting.
Liposomes have gained extensive attention as carriers for a wide range of drugs due to being both nontoxic and biodegradable as they are composed of substances naturally occurring in biological membranes. Active targeting for cells has explored specific modification of the liposome surface by functionalizing it with specific targeting ligands in order to increase accumulation and intracellular uptake into target cells. None of the Food and Drug Administration-licensed liposomes or lipid nanoparticles are coated with ligands or target moieties to delivery for homing drugs to target tissues, cells or subcellular organelles. Targeted therapies (with or without controlled drug release) are an emerging and relevant research area. Despite of the numerous liposomes reviews published in the last decades, this area is in constant development. Updates urgently needed to integrate new advances in targeted liposomes research. This review highlights the evolution of liposomes from passive to active targeting and challenges in the development of targeted liposomes for specific therapies.


The potential of Burkholderia thailandensis E264 for co-valorization of C5 and C6 sugars into multiple value-added bio-products.
Despite known metabolic versatility of Burkholderia spp., sugar metabolism and end-product synthesis patterns in Burkholderia thailandensis have been poorly characterized. This work has demonstrated that B. thailandensis is capable of simultaneously uptaking glucose and xylose and accumulating up to 64% of its dry mass as poly(3-hydroxyalkanoate) (PHA) biopolymers, resulting in a PHA titer of up to 3.8 g L-1 in shake flasks. Rhamnolipids - mainly (68-75%) in the form of Rha-Rha-C14-C14 - were produced concomitantly with a titer typically in the range of 0.2-0.4 g L-1. Gluconic and xylonic acids were also detected in titers of up to 5.3 g L-1, and while gluconic acid appeared to be back consumed, xylonic acid formed as a major end product. This first example of co-production of three products from mixed sugars using B. thailandensis paves the way for improving biorefinery economics.


Progress in the diagnosis of deep venous thrombosis: the efficacy of real-time B-mode ultrasonic imaging.
Seventy-six limbs with clinically suspected acute deep venous thrombosis (DVT) were evaluated by means of ultrasonic imaging (UI) to define the ability of this technique to detect acute and chronic venous obstruction and to determine the origin and distribution of venous thrombi. UI was compared with ascending contrast phlebography in 46 limbs and was found to be 100% accurate in detecting both acute and chronic venous thrombosis. Overall, acute DVT was present in 63 of 76 limbs (83%) studied. Acute DVT was found in 24% and recurrent acute DVT in 76%. Our results indicate that although the calf veins are the most common site of involvement (89%), thrombi may frequently arise simultaneously in multiple anatomic sites. All limbs with recurrent acute DVT had evidence of previous calf thrombi but only 13% had previous proximal disease. This suggests that asymptomatic calf DVT is common and the prevalence of recurrent acute DVT is significantly greater than previously believed. We found UI is a practical, accurate, non-invasive method for investigating the pathogenesis of venous disease.


IgG pair formation on one antigenic molecule is the main mechanism of synergy between antibodies in complement-mediated lysis.
Two rat IgG2b monoclonal antibodies bound to different epitopes on a offgle RT1Aa antigen molecule synergize in their induction of complement-mediated lysis of the red cells. The mechanism of this synergistic action has been investigated by determining the C1q:IgG relationship on red cells using the fluorescence-activated cell sorter and making use of the considerable variation in antigen density between individual red cells. With a synergistic pair of antibodies, the C1q:IgG ratio was approximately 1 and independent of antibody density per cell, indicating that the antibody molecules are present as closely associated pairs, each pair binding two C1q molecules and thus forming a cyclic tetramer. When one of the antibodies was used alone, the number of C1q molecules bound relative to the amount of antibody was higher than expected from theoretical considerations based on a presumed random distribution of antigen-antibody complex. This result can be explained if there is some mobility of the complex within the membrane. The interpretation of the results gives strong support to the hypothesis that C1q must bind bivalently to rat IgG2b complexes.


Breast cancers arising in subjects with germline BRCA1 or BRCA2 mutations: Different biological and clinical entities with potentially diverse therapeutic opportunities.
Breast cancers (BCs) arising in carriers of germline BRCA1 and BRCA2 pathogenic variants (PVs) have long been considered as indistinguishable biological and clinical entities. However, the loss of function of BRCA1 or BRCA2 proteins has different consequences in terms of tumor cell reliance on estrogen receptor signaling and tumor microenvironment composition. Here, we review accumulating preclinical and clinical data indicating that BRCA1 or BRCA2 inactivation may differentially affect BC sensitivity to standard systemic therapies. Based on a different crosstalk between BRCA1 or BRCA2 and the ER pathway, BRCA2-mutated Hormone Receptor-positive, HER2-negative advanced BC may be less sensitive to endocrine therapy (ET) plus CDK 4/6 inhibitors (CDK 4/6i), whereas BRCA2-mutated triple-negative breast cancer (TNBC) may be especially sensitive to immune checkpoint inhibitors. If validated in future prospective studies, these data may have relevant clinical implications, thus establishing different treatment paths in patients with BRCA1 or BRCA2 PVs.


WITHDRAWN: The majority of T cells, including Treg cells, are developed from CD4-CD8-T progenitor cells without the involvement of the CD4+CD8+ stage in the thymus.
This article has been withdrawn at the request of the editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.


The role of cytolysin in pneumococcal ocular infection.
A highly purified preparation of pneumococcal cytolysin was responsible for the corneal damage that ensued upon intra-corneal injection of crude cell extract into the rabbit eye. The lysin may produce this pathologic finding by effecting activation or release of corneal degradative enzymes.


Relationship between cellular calcium and vitamin E metabolism during protection against cell injury.
The extent of chemically induced injury to isolated hepatocytes has been previously shown to depend on the content of alpha-tocopherol in the cells, the levels of which are influenced by the concentration of extracellular calcium. Investigations into the effect of calcium on the alpha-tocopherol content of nonchemically exposed cells demonstrated that incubation of isolated hepatocytes in a calcium-deficient medium decreased cell calcium content to 10% of initial levels, and resulted in the depletion of endogenous alpha-tocopherol. This loss in alpha-tocopherol was not accounted for by alpha-tocopherylquinone formation. After supplementation of the cell incubation medium with alpha-tocopheryl succinate, the decreased cell calcium content was associated with higher levels of cellular alpha-tocopherol than in calcium-adequate cells. This was the result of greater intracellular hydrolysis of the tocopheryl ester in the calcium-depleted cells, and not an effect of extracellular calcium concentration on the uptake of alpha-tocopheryl succinate into the cells or on the extracellular hydrolysis of the ester. Uptake studies indicated a much greater achievable level of alpha-tocopherol in hepatocytes after incubation with alpha-tocopherol than with the alpha-tocopheryl ester. These data provide substantial support for the hypotheses that the content of extracellular calcium per se is not the determinant in toxic injury to hepatocytes, but that cell calcium content affects the intracellular metabolism of alpha-tocopherol and its esters, which may subsequently govern the outcome of a toxic challenge.


Improving the cotton simulation model, GOSSYM, for soil, photosynthesis, and transpiration processes.
GOSSYM, a mechanistic, process-level cotton crop simulation model, has a two-dimensional (2D) gridded soil model called Rhizos that simulates the below-ground processes daily. Water movement is based on gradients of water content and not hydraulic heads. In GOSSYM, photosynthesis is calculated using a daily empirical light response function that requires calibration for response to elevated carbon dioxide (CO2). This report discusses improvements made to the GOSSYM model for soil, photosynthesis, and transpiration processes. GOSSYM's predictions of below-ground processes using Rhizos are improved by replacing it with 2DSOIL, a mechanistic 2D finite element soil process model. The photosynthesis and transpiration model in GOSSYM is replaced with a Farquhar biochemical model and Ball-Berry leaf energy balance model. The newly developed model (modified GOSSYM) is evaluated using field-scale and experimental data from SPAR (soil-plant-atmosphere-research) chambers. Modified GOSSYM better predicted net photosynthesis (root mean square error (RMSE) 25.5 versus 45.2 g CO2 m-2 day-1; index of agreement (IA) 0.89 versus 0.76) and transpiration (RMSE 3.3 versus 13.7 L m-2 day-1; IA 0.92 versus 0.14) and improved the yield prediction by 6.0%. Modified GOSSYM improved the simulation of soil, photosynthesis, and transpiration processes, thereby improving the predictive ability of cotton crop growth and development.


Inhibition of protein kinase C by H-7 potentiates the release of oleic, linoleic and arachidonic acids in A23187-stimulated human neutrophils.
This present report describes the effect of H-7, a protein kinase C inhibitor, on the release of oleic, linoleic and arachidonic acids in A23187-stimulated neutrophils. Surprisingly, the inhibitor potentiated the release of all three unsaturated fatty acids in neutrophils stimulated with A23187 alone. In contrast, released oleic acid, linoleic acid and arachidonic acid in phorbol 12-myristate 13-acetate-primed neutrophils were attenuated by 35, 47 and 33%, respectively, in the presence of H-7 (300 microM). Phorbol 12-myristate 13-acetate (PMA) had no effect on A23187-stimulated release of saturated fatty acids. Both PMA and H-7 when used alone had no effect on the release of saturated or unsaturated fatty acids. We, therefore, conclude that H-7 may have effects other than inhibiting PMA-primed responses including superoxide generation, degranulation and arachidonic acid release in human neutrophils.


Pulmonary Manifestations of Skin Disorders in Children.
Systemic diseases often manifest with cutaneous findings. Many pediatric conditions with prominent skin findings also have significant pulmonary manifestations. These conditions include both inherited multisystem genetic disorders such as yellow-nail syndrome, neurofibromatosis type 1, tuberous sclerosis complex, hereditary hemorrhagic telangiectasia, Klippel-Trénaunay-Weber syndrome, cutis laxa, Ehlers-Danlos syndrome, dyskeratosis congenita, reactive processes such as mastocytosis, and aquagenic wrinkling of the palms. This overview discusses the pulmonary manifestations of skin disorders.


Rational design of novel pyrrolidine derivatives as orally active neurokinin-3 receptor antagonists.
The rational design of a novel series of pyrrolidine derivatives as neurokinin-3 receptor antagonists is reported starting from a selective neurokinin-1 receptor antagonist. Typical representatives in this series showed in vivo efficacy after oral administration in a NK3 mediated functional assay. This series of NK3 antagonists shows promise to deliver a novel antipsychotic.


Identification of a nef allele that causes lymphocyte activation and acute disease in macaque monkeys.
Residues 17 and 18 in nef of SIVmac239 were changed from RQ to YE to create a translated sequence of SRPSGDLYERLLRARGETYGRLLGEVEDGYSQSP from residues 10-43. The YXXL motifs in this context match very well with consensus sequences for SH2 binding domains and are similar to ones present in nef of the acutely lethal pathogen SIVpbj14. The YE variant of SIVmac239, unlike SIVmac239 but like SIVpbj14, replicated well in resting peripheral blood mononuclear cell cultures, caused extensive T lymphocyte activation, and produced an acute disease in rhesus and pigtailed monkeys characterized by severe diarrhea, rash, and extensive lymphoid proliferation in the gastrointestinal tract. The YEnef gene transformed NIH 3T3 cells in culture. Both 239nef and YEnef were found to associate with src in cotransfected COS cells, and both 60 kDa src and 34 kDa nef were phosphorylated at tyrosine in these cells. The extent of tyrosine phosphorylation of 239nef was considerably less than that of YEnef in these assays. These findings identify an important determinant of the SIVpbj14 phenotype, and they provide evidence of a role for nef in signal transduction and cellular activation.


Meta-analysis on the performance of sonography for the diagnosis of carpal tunnel syndrome.
We aimed to review and pool recent large methodological studies evaluating the diagnosis performance of ultrasonography vs electrodiagnostic testing (EDX). Using the keywords: "carpal tunnel syndrome", "ultrasound", and "validity", recent articles evaluating ultrasonography compared with a reference including EDX were selected from 4 databases (PubMed, Embase, Web of science, and BDSP) and from previous review for older articles, after 2 rounds. Relevant data for different thresholds of cross-sectional area of the median nerve were extracted from the articles to calculate the pooled sensitivity, specificity, and likelihood ratios. Different analyses were also performed to study potential sources of heterogeneity, such as calculation of area under the curve, using summary receiver operating characteristic curve. Among the 189 articles found, 13 articles were included. A cross-sectional area of the median nerve between 9.5 and 10.5 mm(2) (study included once only), found for 11 studies, gave the pooled sensitivity as 0.84 [0.81 to 0.87] and the likelihood ratio for a negative test as 0.21 [0.17 to 0.27]. Specificity (0.78 [0.69-0.88]) and the likelihood ratio for a positive test (3.74 [2.30-6.10]) were heterogeneous. For a threshold at 7.0 to 8.5 mm(2), pooled sensitivity was 0.94 [0.87 to 1.00], and for 11.5 to 13.0 mm(2) specificity was 0.97 [0.91 to 1.00]. The only significant variable on potential sources of heterogeneity was the cross-sectional area of the median nerve threshold and area under the curve was 0.87 (asymmetric). Pooling recent articles seems to confirm that sonography using cross-sectional area of the median nerve could not be an alternative to EDX for diagnosis of carpal tunnel syndrome but could give complementary results.


Relationship between anxiety and concussion symptoms among adolescents enrolled in a randomized controlled trial of aerobic exercise.
Affective symptoms, specifically, anxiety, are often overlooked after sport-related concussion (SRC), and may contribute to prolonged recovery. To examine the impact of anxiety during clinical recovery among adolescents (13-18y) enrolled in a randomized trial of aerobic exercise for SRC. Patients at three sites were randomized into aerobic exercise or stretching arms, and enrolled in the 4-week intervention. The relationship between PROMIS Anxiety score at initial visit and time to symptom resolution was evaluated with survival analysis. The relationship between weekly PROMIS Anxiety score and Post-concussion Symptom Inventory (PCSI) score was evaluated with Linear Mixed Models. Analyses adjusted for study arm and baseline covariates. Among 54 adolescents (median age = 15.8y, initial visit PCSI score = 32, pre-injury PROMIS Anxiety score = 2), median time to symptom resolution was 10 days (25th-75th percentiles: 6-24) in the Low-PROMIS Anxiety group and 12 days (25th-75th percentiles: 5-21) in the High-PROMIS Anxiety group (p = 0.62). Each additional unit of PROMIS Anxiety score corresponded to a 1.52-unit higher PCSI total score (p < 0.01). Neither effect varied by aerobic exercise/stretching group. Higher initial PROMIS Anxiety score was not significantly associated with delayed symptom resolution. However, over time, PROMIS Anxiety score was significantly associated with elevated PCSI score, regardless of exercise/stretching group.


Arsenic exposure impairs intestinal stromal cells.
Arsenic is a toxicant commonly found in drinking water. Even though its main route of exposure is oral, little is known of the impact of in vivo arsenic exposure on small intestine. In vitro studies have shown that arsenic decreases differentiation of stem and progenitor cells in several different tissues. Thus, small intestinal organoids were used to assess if arsenic exposure would also impair intestinal stem cell differentiation. Unexpectedly, no changes in markers of differentiated epithelial cells were seen. However, exposing mice to 100 ppb arsenic in drinking water for 5 weeks impaired distinct populations of intestinal stromal cells. Arsenic reduced the width of the pericryptal lamina propria by 1.6-fold, and reduced Pdgfra mRNA expression, which is expressed in intestinal telocytes and trophocytes, by 4.2-fold. The height or extension of Pdgfra+ telopodes into the villus tip was also significantly reduced. Transcript expression of several other stromal cell markers, such as Grem1, Gli, CD81, were reduced by 1.9-, 2.3-, and 1.4-fold, respectively. Further, significant correlations exist between levels of Pdgfra and Gli1, Grem1, and Bmp4. Our results suggest arsenic impairs intestinal trophocytes and telocytes, leading to alterations in the Bmp signaling pathway.


Survey for g-proteins in the prokaryotic genomes: prediction of functional roles based on classification.
The members of the family of G-proteins are characterized by their ability to bind and hydrolyze guanosine triphosphate (GTP) to guanosine diphosphate (GDP). Despite a common biochemical function of GTP hydrolysis shared among the members of the family of G-proteins, they are associated with diverse biological roles. The current work describes the identification and detailed analysis of the putative G-proteins encoded in the completely sequenced prokaryotic genomes. Inferences on the biological roles of these G-proteins have been obtained by their classification into known functional subfamilies. We have identified 497 G-proteins in 42 genomes. Seven small GTP-binding protein homologues have been identified in prokaryotes with at least two of the diagnostic sequence motifs of G-proteins conserved. The translation factors have the largest representation (234 sequences) and are found to be ubiquitous, which is consistent with their critical role in protein synthesis. The GTP_OBG subfamily comprises of 79 sequences in our dataset. A total of 177 sequences belong to the subfamily of GTPase of unknown function and 154 of these could be associated with domains of known functions such as cell cycle regulation and t-RNA modification. The large GTP-binding proteins and the alpha-subunit of heterotrimeric G-proteins are not detected in the genomes of the prokaryotes surveyed.


The country-of-origin label impact on intention to consume insect-based food.
The global food consumption scenario and the influence of a country's image on consumer decision-making motivated this research. The global edible insect market is growing, and cultural issues, disgust, and low perception of quality are barriers to consumption. Through an online experiment (n = 194) Brazilian consumers demonstrate that the use of the country-of-origin label (COOL) can result in greater intention and quality expectation regarding a cricket flour. Consumers showed a preference for the flour produced by a positive image country label. Both intention and quality expectation were higher for the cricket flour produced in the United States. Furthermore, the mediation model indicates that quality expectation mediated the effect of COOL on the intention to consume. The results show that even though consumers have created a positive expectation concerning the product, a higher level of neophobia diminished their intention to consume the product and this relates to the aversion and disgust that can exist towards an edible-insect product. The neophobia level can weaken the effect of COOL on intention mediated by quality expectation. The study aimed to contribute to a better understanding of the consumer's perception of an insect-based food and provide insights into variables related to the intention to consume by verifying their contribution to the consumer's behavior.


The sulphydryl containing ACE inhibitor Zofenoprilat protects coronary endothelium from Doxorubicin-induced apoptosis.
Pediatric and adult cancer patients, following the use of the antitumor drug Doxorubicin develop cardiotoxicity. Pharmacological protection of microvascular endothelium might produce a double benefit: (i) reduction of myocardial toxicity (the primary target of Doxorubicin action) and (ii) maintenance of the vascular functionality for the adequate delivery of chemotherapeutics to tumor cells. This study was aimed to evaluate the mechanisms responsible of the protective effects of the angiotensin converting enzyme inhibitor (ACEI) Zofenoprilat against the toxic effects exerted by Doxorubicin on coronary microvascular endothelium. We found that exposure of endothelial cells to Doxorubicin (0.1-1μM range) impaired cell survival by promoting their apoptosis. ERK1/2 related p53 activation, but not reactive oxygen species, was responsible for Doxorubicin induced caspase-3 cleavage. P53 mediated-apoptosis and impairment of survival were reverted by treatment with Zofenoprilat. The previously described PI-3K/eNOS/endogenous fibroblast growth factor signaling was not involved in the protective effect, which, instead, could be ascribed to cystathionine gamma lyase dependent availability of H2S from Zofenoprilat. Furthermore, considering the tumor environment, the treatment of endothelial/tumor co-cultures with Zofenoprilat did not affect the antitumor efficacy of Doxorubicin. In conclusion the ACEI Zofenoprilat exerts a protective effect on Doxorubicin induced endothelial damage, without affecting its antitumor efficacy. Thus, sulfhydryl containing ACEI may be a useful therapy for Doxorubicin-induced cardiotoxicity.


Monoclonal antibodies reacting with isotypic and/or allotypic determinants of human IgG.
Mice were immunized with purified human IgG myeloma proteins and hybridomas were prepared using their spleen cells. 1817 of the hybridomas secreted anti-Ig antibodies. Several of them detected subclass-associated determinants. Eight different specificities could be distinguished. The number of hybridomas in each category were the following: 3 anti-IgG1 (1.8% of all anti-Ig clones) 5 anti-IgG2 (0.3% of all anti-Ig clones) 2 anti-IgG3 (2.5% of all anti-Ig clones) 3 anti-IgG4 (12% of all anti-Ig clones) 12 anti-IgG1, IgG2, IgG3 70 anti-IgG1, IgG2, IgG4 2 anti-IgG2, IgG4 7 anti-IgG2, IgG4 (one out of five myelomas).


Urothelial carcinogenesis and portocaval anastomosis in the rat.
Portacaval anastomoses were performed in the rat to study urothelial carcinogenesis in this model and the promoting effect of dietary tryptophan. We were unable to produce any urothelial cancers or premalignant changes; some animals formed uric acid stones and developed papillary hyperplasia in the bladder. We conclude that the initiating carcinogen is likely to be exogenous and may be dietary in those experiments that have produced urothelial cancers.


Artificial Intelligence in Compound Design.
Artificial intelligence has seen an incredibly fast development in recent years. Many novel technologies for property prediction of drug molecules as well as for the design of novel molecules were introduced by different research groups. These artificial intelligence-based design methods can be applied for suggesting novel chemical motifs in lead generation or scaffold hopping as well as for optimization of desired property profiles during lead optimization. In lead generation, broad sampling of the chemical space for identification of novel motifs is required, while in the lead optimization phase, a detailed exploration of the chemical neighborhood of a current lead series is advantageous. These different requirements for successful design outcomes render different combinations of artificial intelligence technologies useful. Overall, we observe that a combination of different approaches with tailored scoring and evaluation schemes appears beneficial for efficient artificial intelligence-based compound design.


Development and validation of the system of quality of life instruments for cancer patients: lung cancer (QLICP-LU).
Using the structured group methods (nominal and focus group) and the theory in instrument development, the quality of life instrument for lung cancer (QLICP-LU) with Chinese cultural background was developed, and evaluated based on the data from a sample of 85 in-patients of lung cancer. Statistical methods used were correlation analysis, factor analysis and paired t-test, etc. The results showed that test-retest reliability of the overall scale was 0.78, and test-retest reliability and internal consistency alpha for most domains were higher than 0.70; correlation and factor analysis demonstrated good construct validity; criterion-related validity were confirmed given FACT-L and QLQ-LC43 (QLQ-C30 plus QLQ-LC13) as the criterions; statistically significant changes were found for each domain and the overall scale with standardized response mean SRM being greater than 0.80 except for the social function domain. We conclude that the QLICP-LU can be used to measure QOL for patients with lung cancer with better validity, reliability and responsiveness in China.


Use of Mixed Reality in Neurosurgery Training: A Single Centre Experience.
Mixed reality (MR) technology has opened new avenues for planning, visualization, and education in surgery. Neurosurgical pathologies require a very clear understanding of the relationships between pathology and critical neurovascular structures. The decline in cadaveric dissections and resource constraints has pushed the educators to find newer ways of rendering the same knowledge. The aim of this study was to determine the feasibility of employing a MR device in a high-volume center for neurosurgical teaching. The study also evaluated the results of the trainee experience in using the MR platform. Three neurosurgical consultants who are part of the teaching faculty were asked to facilitate the session. No prior training on utilizing the MR device was given to the trainees. HoloLens 2 was used as the MR device. Two questionnaires were used to understand the experience of the trainees. Eight active neurosurgical trainees who are currently training at our institution were recruited for the purposes of this study. Despite having no prior training on a MR platform, the learning curve was short for most of the trainees. Whether MR replace current traditional methods of teaching neuroanatomy, the response was divided across the trainees. The results of the User Experience Questionnaire were positive with the trainees finding the device as attractive, dependable, novel, and user-friendly. This study demonstrates the feasibility of using MR platform in neurosurgery training without significant preparation requirements. These data are required to justify the future investment in this technology for training institutions.


Granular cell tumors of biliary ducts. Report of two cases and review of the literature.
There were two cases of granular cell tumor of the extrahepatic biliary system; to our knowledge, 17 have previously been reported. The initial symptoms most often are those of biliary colic. The majority of the patients are black and female. The cystic duct and the common bile duct are most commonly involved. Additional biliary pathologic disorder is present in more than half of the patients.


Distinct effects of equal levels of Fusarium toxins (Deoxynivalenol) in growing pigs. Possible reasons.
The mycotoxin deoxynivalenol (DON, vomitoxin) is well documented to cause reduced feed consumption (FC) and weight gain (WG) rates in sensitive animal species (e.g. swine). In establishing feeding guidelines and diagnoses of mycotoxin related diseases it is important to establish a "dose and response" factor-but a simple relationship is not always present for DON and other Fusarium toxins. Possible reasons for these findings (e.g. experimental design and/or role of toxin source) are discussed with reference to previous research.


Effects of monocular deprivation on geniculocortical pathways in the owl monkey, Aotus trivirgatus.
As has been shown for a number of other mammals, monocular eyelid suture in the newborn owl monkey, a New World primate, is followed by a failure of neurons in the deprived layers of the lateral geniculate nucleus to reach normal size in the adult animal. This failure appears to be almost completely confined to relay cells, identified by horseradish peroxidase (HRP) injections into cortex, rather than interneurons. Similar proportions of neurons in both the deprived and non-deprived layers are labeled after HRP injections, suggesting that relay cells do not lose all cortical connections as a result of visual deprivation.


Differentiation of beta-lactamase-negative ampicillin-resistant Haemophilus influenzae from other H. influenzae strains by a disc method.
We established breakpoints for differentiating ampicillin (ABPC)-susceptible strains from resistant strains among Haemophilus influenzae isolates according to susceptibility to various beta-lactam antibiotics, using a disc method. Susceptibility testing of isolates for 13 beta-lactam agents was followed by analysis of the resistance genes, using a polymerase chain reaction (PCR) to identify the TEM-1 beta-lactamase gene ( bla) and the ftsI gene encoding penicillin-binding protein (PBP) 3, which affects beta-lactam minimum inhibitory concentrations (MICs). A total of 228 H. influenzae isolates were classified into 114 beta-lactamase-negative, ABPC-susceptible (BLNAS) strains; 29 beta-lactamase-negative, ABPC-resistant (BLNAR) strains; 53 low-BLNAR strains with a low degree of ABPC resistance; 27 TEM-1-producing strains (BLPAR); and 5 strains with ftsI gene mutations in addition to TEM-1 production (BLPACR) according to the PCR results. To identify resistant strains by disc-method susceptibility testing, the zone of inhibition was measured for ABPC (10 microg/disc), cefaclor (30 microg/disc), cefpodoxime (10 microg/disc), and cefdinir (5 microg/disc) discs. Strains were identified as BLNAS without resistant genes when the diameter was > or =27 mm for the ABPC disc and > or =21 mm for the cefaclor disc. Other strains were identified as BLNAR when the diameter was < or =22 mm for the cefpodoxime disc and < or =17 mm for the cefdinir disc. Remaining strains were identified as low-BLNAR. These criteria differentiated resistance types with high accuracy. A discrepancy was noted between genetic results and disc-testing breakpoints for differentiating resistant from susceptible H. influenzae. A disc-testing breakpoint for cefditoren (5 microg/disc) was proposed, with the susceptibility statistically defined as a diameter of > or =24 mm, which corresponds to the breakpoint (1 microg/ml) of the microdilution method recommended by the Japanese Society of Chemotherapy.


Trends and correlates of cocaine use and cocaine use disorder in the United States from 2011 to 2015.
Recent epidemiological data suggest a resurgence in cocaine use (CU) and cocaine-related problems in the United States. Demographic trends and correlates of problem CU are needed to determine potential factors that may be influencing the increased trend and to inform targeted prevention and intervention strategies. Trends in any past-year CU, weekly CU, and cocaine use disorder (CUD) were examined among persons aged ≥12 years using the National Survey on Drug Use and Health from 2011 to 2015. Logistic regression analyses were used to determine correlates of past-year and weekly CU and CUD among adolescents and adults. The prevalence of past-year CU from 2011 to 2015 increased among females, ages 18-25, ages ≥50, non-Hispanic Blacks, and persons reporting low income, past-year tobacco use, past-year alcohol use, and past-month binge and heavy alcohol use. The prevalence of weekly CU increased among persons aged ≥50 years and persons reporting past-month heavy alcohol use. A significant increase in the prevalence of CUD was only found among persons aged ≥50 years. Adjusted logistic regression showed that older age, large metropolitan residence, past-year tobacco, alcohol, cannabis, and heroin use, and major depressive episode were associated with increased odds of CU or CUD among both adolescents and adults; however, sex and race/ethnicity correlates differed among adolescents and adults. Findings have implications for increased monitoring of CU-related indicators among some high-risk groups, such as females, older adults, Blacks, and polysubstance users. Targeted screening and intervention strategies among these population subgroups may be needed.


Outreach for Annual Colorectal Cancer Screening: A Budget Impact Analysis for Community Health Centers.
Fecal immunochemical testing (FIT) is an attractive approach for colorectal cancer screening at community health centers. This budget impact analysis investigated benefits and costs of FIT outreach-with FIT kits mailed to patients, followed by reminders and phone calls-compared with point-of-care (POC) strategies. Five screening and cost outcomes were simulated over 1 year at a "base case" community health center serving 1000 screening-eligible patients: (1) FIT completion among patients due for screening; (2) proportion up-to-date on screening; (3) cost per patient due for screening; (4) cost per completed FIT; and (5) total organizational cost. Uncertainty analysis investigated potential savings from optimizing staff workflows during FIT outreach. Data were collected in 2012-2014, with analysis conducted 2014-2015. Using POC strategies, 24.0% of patients due for screening completed FIT, versus 42.4% under outreach (18.4% absolute difference). When calculations included patients up-to-date on screening from prior colonoscopy, 41.7% were up-to-date via POC, versus 55.8% for outreach (14.1% absolute difference). POC cost $4.93 per patient, versus $30.43 for outreach ($25.50 difference). Cost per patient screened was $20.60 for POC and $71.84 for outreach ($51.24 difference). Total organizational cost was $3,779 for POC distribution and $23,315 for outreach ($19,536 difference). Outreach costs decreased by approximately one fourth under optimized workflows. Outreach is an effective, practical, relatively low-cost strategy; costs could be reduced further by optimizing staff workflows. Despite its value, outreach costs more than POC distribution and may be difficult for community health centers to implement under current payment models.


Vertebral cancellous bone turn-over: microcallus and bridges in backscatter electron microscopy.
Backscatter electron microscopy (BSE) is a powerful technique for investigating cancellous bone structure. Its main function is to offer information regarding the degree of mineralization of the tissue within individual trabeculae. To illustrate the qualitative information that can be drawn from BSE imaging technique, we present a study on human vertebral cancellous bone. This tissue is continuously remodeled through osteoclastic resorption and osteoblastic new bone apposition. It is thought that osteoclastic resorption pits are especially deleterious for vertebral bone architecture since they often perforate the thin trabeculae; the osteoblasts being unable to repair the gap. In addition, excessive stress may also disrupt the architecture in case of trabecular fracture or damage accumulation. Waves of new bone formation were easy to identify in BSE. Often these waves were connecting both edges of a perforation and called bridges. Additionally, we present a few images of microcallus formations. A microcallus is described as a small mass of woven bone that generally repairs a trabecula. The microstructural aspects of different microcalluses are presented and discussed. Both bridges and microcallus should be considered as examples of the repair porcess since they obviously preserve the connectivity of the trabeculae. However, bridges were much more frequent than microcallus (396 vs 15). Both mechanisms probably illustrate the normal response to different local stimuli.


Exogenous application of transforming growth factor beta 1 stimulates arteriogenesis in the peripheral circulation.
Increased expression of transforming growth factor beta1 (TGF-beta1) during collateral artery growth, as well as its numerous effects on monocytes/macrophages and the smooth muscle cell cycle and differentiation, suggest a modulating role for this growth factor during arteriogenesis. We studied the effects of exogenously applied TGF-beta1 on arteriogenesis as well as its interactions with monocytes, endothelial cells, and smooth muscle cells. In a New Zealand White (NZW) rabbit model of femoral artery ligation, increased expression of active TGF-beta1 was found around proliferating arteries in NZW rabbits. The exogenous application of TGF-beta1 led to an increase in both the number of visible collateral arteries as well as the conductance of the collateral circulation (4.0 +/- 0.5 ml/min/100 mmHg vs. 28.9 +/- 3.7 ml/min/100 mmHg, P<0.05). Fluorescence activated cell sorting analysis showed an increase in the expression of the MAC-1 receptor in both rabbit and human monocytes after treatment with TGF-beta1 (control: 91.2 +/- 4.2/482 +/- 21.7; TGF-beta1 200 ng/ml 193.9 +/- 6.7/ 675.5 +/- 25.7, P<0.05 for all differences). TGF-beta1 treated monocytes showed an increased endothelial adhesion and transmigration in transendothelial migration assays (5.75 +/- 0.63 x 10(5) vs. 10.11 +/- 0.04 x 10(5), P<0.05). TGF-beta1 had no direct pro-angiogenic effect on human umbilical vein endothelial cells in a spheroid model of angiogenesis and inhibited the angiogenic effects of vascular endothelial growth factor.


Comparing the Effect of Adaptive Support Ventilation (ASV) and Synchronized Intermittent Mandatory Ventilation (SIMV) on Respiratory Parameters in Neurosurgical ICU Patients.
Various modes of mechanical ventilation have different effects on respiratory variables. Lack of patients' neuro-ventilatory coordination and increasing the work of breathing are major disadvantages in mechanically ventilated patients. This study is conducted to compare the respiratory parameters differences in Adaptive support ventilation (ASV) and synchronized intermittent mandatory ventilation (SIMV) modes in neurosurgical ICU patients. In a crossover study, patients under mechanical ventilation in neurosurgical ICU were enrolled. The patients alternatively experienced two types of ventilations for 30 minutes (adaptive support ventilation and synchronized intermittent mandatory ventilation). The respiratory parameters (tidal volume, respiratory rate, airway pressure, lung compliance, end-tidal carbon dioxide, peripheral oxygenation and respiratory dead space), hemodynamic variables, every 10 minutes and arterial blood gas analysis at the end of each 30 minutes were recorded. Results were compared and analyzed with SPSS v.19. Sixty patients were involved in this study. In ASV mode, values including peak airway pressure (P-peak), end-tidal carbon dioxide (EtCO2), tidal volume and respiratory dead space were significantly lower than SIMV mode. Although the mean value for dynamic compliance had no significant difference in the two types of ventilation, it was better in ASV mode. ASV mode compared with SIMV mode can lead to improve lung compliance and respiratory dead space.


The numbers of unmyelinated and myelinated axons in normal and regenerated rat saphenous nerves.
Counts have been made of the numbers of unmyelinated and myelinated axons in the proximal and distal stumps of regenerated rat saphenous nerves and from equivalent sites in normal nerves. In the proximal part of normal nerves there were averages of 1,045 myelinated axons and 4,160 unmyelinated ones. Regenerated nerves contained the same number of myelinated axons in their proximal stumps but there was a 40% reduction in the unmyelinated axon count. In the distal stumps of these nerves the myelinated axon count had increased by an average of 620; this comes about because some regenerated myelinated axons support more than one process in the distal stump. In contrast, the number of unmyelinated axons was reduced further, from a mean of 2,476 in the proximal stump to one of 2,219. The sizes of Schwann cell units in the normal and regenerated nerves were also noted. Schwann cell units in the proximal and distal stumps of the regenerated nerves were smaller than those in the normal ones. These changes associated with unmyelinated axons in regenerated nerves are likely to contribute to the sensory, vasomotor and sudomotor abnormalities that sometimes occur after peripheral nerve injury and regeneration.


Pluripotent stem cell model reveals essential roles for miR-450b-5p and miR-184 in embryonic corneal lineage specification.
Approximately 6 million people worldwide are suffering from severe visual impairments or blindness due to corneal diseases. Corneal allogeneic transplantation is often required to restore vision; however, shortage in corneal grafts and immunorejections remain major challenges. The molecular basis of corneal diseases is poorly understood largely due to lack of appropriate cellular models. Here, we described a robust differentiation of human-induced pluripotent stem cells (hiPSCs) derived from hair follicles or skin fibroblasts into corneal epithelial-like cells. We found that BMP4, coupled with corneal fibroblast-derived conditioned medium and collagen IV allowed efficient corneal epithelial commitment of hiPSCs in a manner that recapitulated corneal epithelial lineage development with high purity. Organotypic reconstitution assays suggested the ability of these cells to stratify into a corneal-like epithelium. This model allowed us identifying miR-450b-5p as a molecular switch of Pax6, a major regulator of eye development. miR-450b-5p and Pax6 were reciprocally distributed at the presumptive epidermis and ocular surface, respectively. miR-450b-5p inhibited Pax6 expression and corneal epithelial fate in vitro, altogether, suggesting that by repressing Pax6, miR-450b-5p triggers epidermal specification of the ectoderm, while its absence allows ocular epithelial development. Additionally, miR-184 was detectable in early eye development and corneal epithelial differentiation of hiPSCs. The knockdown of miR-184 resulted in a decrease in Pax6 and K3, in line with recent findings showing that a point mutation in miR-184 leads to corneal dystrophy. Altogether, these data indicate that hiPSCs are valuable for modeling corneal development and may pave the way for future cell-based therapy.


[Oncologic urology: synopsis of the 101st Congress of the French Association of Urology].
Regarding renal cell carcinoma, most recent advances concern metastatic cases in which antiangiogenic agent seem to be efficient for specific survival and outcome without progression of the disease. In prostate cancer, new urinary molecular markers, more specific than PSA, are currently under development to allow an early and non invasive diagnosis of the disease. Robot-assisted radical prostatectomy is a growing surgical approach in the treatment of localized prostate cancer, at the expense of the laparoscopic approach, with satisfactory oncologic preliminary results. Whether or not a cystectomy should be done after the resection of a pT1G3 bladder tumour, remains a moot point. However the risk of progression is far from being negligible. Regarding upper urinary tract tumors, the outcome is different from bladder tumors and several teams are working on specific molecular markers, dedicated to be useful in prognosis and specific survival.


Full genomic analysis of human rotavirus strain TB-Chen isolated in China.
A G2P[4]/NSP4[A] rotavirus strain TB-Chen was isolated from a 2-year-old patient hospitalized with acute gastroenteritis in Kunming, China. The strain TB-Chen was demonstrated having group A-specific antigenicity, a "short" (subgroup II) electropherotype. To investigate its overall genomic relatedness and to determine which group it belonged, the complete genome of strain TB-Chen was determined. Genomic comparison based on amino acid sequence identity and phylogenetic analysis revealed that all 11 gene segments of strain TB-Chen were highly identical (>91.80%) with the representative G2P[4]/NSP4[A] human strains DS-1, S2, NR1 and IS2, suggesting that this rotavirus strain was derived from human host. Besides, almost all the available representative rotavirus gene segments among group A were analyzed and identified within 15 G-types, 28 P-types, and 6 NSP4 genotypes. This is the first report of group A rotavirus genomic analyses in China and the findings have important implications for rotavirus vaccine development.


Urologic considerations in the newborn.
The management of the neonate will continue to hold a special interest for the urologic surgeon because of the many unique aspects of neonatal physiology and pathology that must be considered. I would anticipate that the number of referrals will continue to increase, from awareness of familial patterns of urologic disorders, from recognition of obstructive uropathy with prenatal ultrasound, and from improved survival of very sick neonates by our pediatric colleagues. The urologist who wishes to participate in the care of these sick newborns must become familiar with the unique aspects of care that these babies demand. The future promises to be challenging, but exciting.


Transcriptomic analysis identified up-regulation of a solute carrier transporter and UDP glucuronosyltransferases in dogs with aggressive cutaneous mast cell tumours.
Gene expression analyses have been recently used in cancer research to identify genes associated with tumorigenesis and potential prognostic markers or therapeutic targets. In the present study, the transcriptome of dogs that had died because of mast cell tumours (MCTs) was characterised to identify a fingerprint having significant influence on prognosis determination and treatment selection. A dataset (GSE50433) obtained using a commercial canine DNA microarray platform was used. The transcriptome of seven biopsies obtained from dogs with histologically confirmed, surgically removed MCTs, treated with chemotherapy, and dead for MCT-related causes, was compared with the transcriptional portrait of 40 samples obtained from dogs with histologically confirmed, surgically removed MCTs and that were still alive at the end of the follow-up period. Among the differentially expressed genes (DEGs), eight transcripts were validated by quantitative real time PCR and their mRNA levels were measured in a cohort of 22 additional MCTs. Statistical analysis identified 375 DEGs (fold change 2, false discovery rate 5%). The functional annotation analysis indicated that the DEGs were associated with drug metabolism and cell cycle pathways. Particularly, members of solute carrier transporter (SLC) and UDP glucuronosyltransferase (UGT) gene families were identified as dysregulated. Principal component analysis (PCA) of the 22 additional MCTs identified the separate cluster dogs dead for MCT-related causes. SLCs and UGTs have been recently recognised in human cancer as important key factors in tumour progression and chemo-resistance. An in-depth analysis of their roles in aggressive canine MCT is warranted in future studies.


Molecular pathogenesis of Japanese encephalitis and possible therapeutic strategies.
Japanese encephalitis virus (JEV), a single-stranded, enveloped RNA virus, is a health concern across Asian countries, associated with severe neurological disorders, especially in children. Primarily, pigs, bats, and birds are the natural hosts for JEV, but humans are infected incidentally. JEV requires a few host proteins for its entry and replication inside the mammalian host cell. The endoplasmic reticulum (ER) plays a significant role in JEV genome replication and assembly. During this process, the ER undergoes stress due to its remodelling and accumulation of viral particles and unfolded proteins, leading to an unfolded protein response (UPR). Here, we review the overall strategy used by JEV to infect the host cell and various cytopathic effects caused by JEV infection. We also highlight the role of JEV structural proteins (SPs) and non-structural proteins (NSPs) at various stages of the JEV life cycle that are involved in up- and downregulation of different host proteins and are potentially relevant for developing efficient therapeutic drugs.


Recombinant Adeno Associated Viral (AAV) vector type 9 delivery of Ex1-Q138-mutant huntingtin in the rat striatum as a short-time model for in vivo studies in drug discovery.
Huntington's disease (HD) is an inherited neurodegenerative disorder characterized by dyskinesia, cognitive impairment and emotional disturbances, presenting progressive neurodegeneration in the striatum and intracellular mutant Huntingtin (mHTT) aggregates in various areas of the brain. Recombinant Adeno Associated Viral (rAAV) vectors have been successfully used to transfer foreign genes to the brain of adult animals. In the present study we report a novel in vivo rat HD model obtained by stereotaxic injection of rAAV serotype2/9 containing Exon1-Q138 mHTT (Q138) and Exon1-Q17 wild type HTT (Q17; control), respectively in the right and in the left striatum, and expressed as C-terminal GFP fusions to facilitate detection of infected cells and aggregate production. Immunohistochemical analysis of brain slices from animals sacrificed twenty-one days after viral infection showed that Q138 injection resulted in robust formation of GFP-positive aggregates in the striatum, increased GFAP and microglial activation and neurodegeneration, with little evidence of any of these events in contralateral tissue infected with wild type (Q17) expressing construct. Differences in the relative metabolite concentrations (N-Acetyl Aspartate/Creatine and Myo-Inositol/Creatine) were observed by H1 MR Spectroscopy. By quantitative RT-PCR we also demonstrated that mHTT induced changes in the expression of genes previously shown to be altered in other rodent HD models. Importantly, administration of reference compounds previously shown to ameliorate the aggregation and neurodegeneration phenotypes in preclinical HD models was demonstrated to revert the mutant HTT-dependent effects in our model. In conclusion, the AAV2/9-Q138/Q17 exon 1 HTT stereotaxic injection represents a useful first-line in vivo preclinical model for studying the biology of mutant HTT exon 1 in the striatum and to provide early evidence of efficacy of therapeutic approaches.


The effects of prosthetic status and dementia on the chewing efficiency of seniors in nursing homes.
The oral health of seniors in nursing homes is compromised. Furthermore, reduced chewing efficiency is described to be associated with reduced cognition. However, studies investigating how prosthetic status affects the chewing efficiency of nursing home residents are not available. To evaluate associations between prosthetic status, dementia and chewing efficiency of seniors in nursing homes. This study was performed in nine nursing homes. In addition to assessing the general and medical data of the participants, a dental examination was assessed and the severity of dementia was evaluated by use of the Clinical Dementia Rating (CDR). Furthermore, chewing efficiency was assessed by use of a two-colour mixing ability test. Descriptive and bivariate statistics, as well as linear regression models with the dependent variable chewing efficiency and possible confounders, were used to analyse data at P < .05. Complete target variables of 146 participants were analysed. The mean (SD) chewing efficiency of the study sample, as expressed by the variance of hue, was .590 (.250). The type of prosthesis used (P < .001), the number of occluding tooth pairs (P < .001) and the presence of dementia (P = .002) were the main variables significantly affecting chewing efficiency. The condition of removable dentures also had an effect (P = .016). Multivariate testing predominantly confirmed these associations. The chewing efficiency of seniors in nursing homes is somewhat compromised. Beyond dental aspects, suffering from dementia seems to reduce the chewing efficiency. Further interventional/longitudinal studies are, however, encouraged to verify this outcome.


The distinct initiation sites and processing activities of TTLL4 and TTLL7 in glutamylation of brain tubulin.
Mammalian brain tubulins undergo a reversible posttranslational modification-polyglutamylation-which attaches a secondary polyglutamate chain to the primary sequence of proteins. Loss of its erasers can disrupt polyglutamylation homeostasis and cause neurodegeneration. Tubulin tyrosine ligase like 4 (TTLL4) and TTLL7 were known to modify tubulins, both with preference for the β-isoform, but differently contribute to neurodegeneration. However, differences in their biochemical properties and functions remain largely unknown. Here, using an antibody-based method, we characterized the properties of a purified recombinant TTLL4 and confirmed its sole role as an initiator, unlike TTLL7, which both initiates and elongates the side chains. Unexpectedly, TTLL4 produced stronger glutamylation immunosignals for α-isoform than β-isoform in brain tubulins. Contrarily, the recombinant TTLL7 raised comparable glutamylation immunoreactivity for two isoforms. Given the site selectivity of the glutamylation antibody, we analyzed modification sites of two enzymes. Tandem mass spectrometry analysis revealed their incompatible site selectivity on synthetic peptides mimicking carboxyl termini of α1- and β2-tubulins and a recombinant tubulin. Particularly, in the recombinant α1A-tubulin, a novel region was found glutamylated by TTLL4 and TTLL7, that again at distinct sites. These results pinpoint different site specificities between two enzymes. Moreover, TTLL7 exhibits less efficiency to elongate microtubules premodified by TTLL4, suggesting possible regulation of TTLL7 elongation activity by TTLL4-initiated sites. Finally, we showed that kinesin behaves differentially on microtubules modified by two enzymes. This study underpins the different reactivity, site selectivity, and function of TTLL4 and TTLL7 on brain tubulins and sheds light on their distinct role in vivo.


Total synthesis of a tyrosine suppressor transfer RNA gene. XIII. Synthesis of deoxyribopolynucleotide segments corresponding to the nucleotide sequence -1 to -29 in the promoter region.
Chemical syntheses of two tridecanucleotides, d(G-C-A-T-C-A-T-A-T-C-A-A-A) and d(G-C-G-T-C-A-T-T-T-G-A-T-A), and three undecanucleotides, d(G-G-A-A-G-C-G-G-G-G-C), d(T-G-A-T-G-C-G-C-C-C-C), and d(T-G-A-C-G-C-G-C-C-G-C), are described. These deoxyribo-oligonucleotide segments together represent the DNA duplex corresponding to the previously determined nucleotide sequence -1 to -29 of the promoter region of the tyrosine tRNA gene (Sekiya, T., van Ormondt, H., and Khorana, H.G. (1975) J. Biol. Chem. 250, 1087-1098). Chemical syntheses used the principles of stepwise addition of protected mono- and oligonucleotides to the 3'-hydroxyl end of growing oligonucleotide chains. The desired condensation products were isolated by solvent extraction methods in the case of di- and trincleotides and by anion exchange chromatography in the case of longer chains. All the five synthetic oligonucleotides were characterized by chromatographic and radioactive fingerprinting methods after labeling at the 5'-ends with a [32P]phosphate group.