Design and preparation of nanostructures based on Krafft point of nonionic amphiphiles for delivery of poorly water-soluble compounds.
Micellar solubilization can effectively dissolve low water-soluble compounds in aqueous environment, however, the micellar systems are not able to withstand dilution and maintain solubilization of poorly water-soluble drugs below critical micelle concentration. To overcome the drawbacks of conventional micellar solubilization, nonionic polyoxyethylated surfactants with Krafft points at or higher than body temperature were chosen to create novel micelle-based nanostructures as a delivery vehicle for poorly water-soluble compounds. A technique "thermo-spray process" was developed for the preparation of the nanostructures-containing formulation, in which the drug-containing micelle solution was first prepared and maintained at the elevated temperature above the Krafft point of the surfactant, then spray dried to solidify the obtained micelle-like nanostructure at room temperature. Lactose was used as an excipient to embed the nanostructures in the thermo-spray products. Water insoluble spherical nanoparticles with size range from 80 to 250 nm were obtained after reconstitution of the product at the temperature lower than Krafft point. When paclitaxel was used as model drug, the micelle-like nanostructures exhibited similar drug entrapment efficiency, solubility enhancement and drug release facilitation as conventional micelles, but provided lower critical micellar concentration at body temperature, and good encapsulation stability upon storage and dilution. These findings indicated that the developed thermo-spray product can serve as a promising delivery platform for drugs with low aqueous solubility.


Recent Progress in the Understanding of Obesity: Contributions of Genome-Wide Association Studies.
Since 2007, discovery of genetic variants associated with general obesity and fat distribution has advanced tremendously through genome-wide association studies (GWAS). Currently, the number of robustly associated loci is 190. Even though these loci explain <3 % of the variance, they have provided us a still emerging picture of genomic localization, frequency and effect size spectra, and hints of functional implications. The translation into biological knowledge has turned out to be an immense task. However, in silico enrichment analyses of genes involved in specific pathways or expressed in specific tissues have the power to suggest biological mechanisms underlying obesity. Inspired by this, we highlight genes in five loci potentially mechanistically linked to leptin-receptor trafficking and signaling in primary cilia. The clinical application of genetic knowledge as prediction, prevention, or treatment strategies is unfortunately still far from reality. Thus, despite major advances, further research is warranted to solve one of the greatest health problems in modern society.


Planning and surgical strategies for early management of vertebral artery and vertebrobasilar junction aneurysms.
Selection of the approach and technique for surgical repair of aneurysm of the vertebrobasilar artery system is mainly based on angiographic features. This report emphasizes that planning the surgical procedure should also include preoperative evaluation of the individual skull base configuration, as well as the relationship between aneurysm site and surrounding bony structures. These features are evaluated on thin slice CT scans using bone tissue algorithms and are particularly important for adequate exposure of distal vertebral artery (VA) or midline aneurysms, because these cases require drilling of the jugular tubercle. For the use of lateral approaches, the surgeon must be familiar with the extradural and intradural anatomy of the foramen magnum region and may rely on at least five anatomical landmarks for orientation during surgery: 1) the dural entrance of the vertebral artery; 2) the posterior condylar emissary vein; 3) the medial rim of the distal sigmoid sinus; 4) the hypoglossal canal; 5) the jugular tubercle. To increase the safety of the procedure, the authors recommend an individualized tailoring of the surgical approach according to the variable morphological situation of each patient.


Serial orthotopic transplantation of epithelial tumors in single-cell suspension.
Orthotopic transplantation of tumor tissue into recipient mice has long been established to study the role of the microenvironment in tumorigenesis and metastasis. Many of these transplantation assays involve the surgical implantation of an undissociated piece of tumor tissue. However, dissociation of tumor tissue into a single cell suspension prior to orthotopic transplantation enables the injection of fewer cell numbers, the selection of tumor-initiating populations by specific purification using antibody staining and fluorescence-activated cell sorting, and the analysis of tumor-forming efficiency.In this chapter, we provide a method to perform serial transplantation of tumor cells into their niche of origin. Visualization of the location of transplanted tumor cells is essential to confirm the success of the transplant as well as the viability of transplanted cells. We also describe an optimized immunofluorescence protocol to visualize tumor cells shortly after transplantation. This serial transplantation protocol allows for an experimental tumorigenesis assay to more closely mimic spontaneous tumor formation and is applicable to many microenvironments.


Alcoholism and correlates of mortality. Implications for epidemiology.
In 935 alcoholics, alcoholism increased the risk of death more in upper-class than in lower-class alcoholics, the middle-class alcoholics holding an intermediate position closer to that of the upper-class alcoholics.


Direct mapping of local director field of nematic liquid crystals at the nanoscale.
Liquid crystals (LCs), owing to their anisotropy in molecular ordering, are of wide interest in both the display industry and soft matter as a route to more sophisticated optical objects, to direct phase separation, and to facilitate colloidal assemblies. However, it remains challenging to directly probe the molecular-scale organization of nonglassy nematic LC molecules without altering the LC directors. We design and synthesize a new type of nematic liquid crystal monomer (LCM) system with strong dipole-dipole interactions, resulting in a stable nematic phase and strong homeotropic anchoring on silica surfaces. Upon photopolymerization, the director field can be faithfully "locked," allowing for direct visualization of the LC director field and defect structures by scanning electron microscopy (SEM) in real space with 100-nm resolution. Using this technique, we study the nematic textures in more complex LC/colloidal systems and calculate the extrapolation length of the LCM.


Viscoelastic properties and residual strain in a tensile creep test on bovine temporomandibular articular discs.
This study was designed to evaluate the creep characteristics and residual strain of bovine temporomandibular joint (TMJ) discs in tension. Twenty discs were divided into three specimens each: central, lateral and medial regions. Tension of 1.0 MPa was applied and sustained for 20 min to the specimens from 10 right-side discs, and tension of 1.5 MPa to specimens from 10 left-side discs. After the period of tension for creep, the specimens were removed from the tension devices and restoration observed for 20 min. Time-dependent creep curves showed a marked change in strain during the initial 5s. The essential time delay in strain ceased after 2 min, and strain reached an almost steady level after 3 min. At a tensile stress of 1.5 MPa, a strain of 14.5% on average was produced after 20 min creep in the central specimens; peripheral specimens showed strains of 12.4% on average. There were significant differences in strain between the central and peripheral specimens. The residual strain after 20 min restoration was 0.93% on average and there were no significant regional differences. This creep feature could be well represented by a generalized linear viscoelastic model. It was concluded that the regional differences in viscoelasticity might be caused by the complicated articulating functions of the TMJ, and that the residual strain caused by sustained stress could be an important factor in disc deformation.


Collagen multifilament spinning.
The benefits of fiber based implants and scaffolds for tissue engineering applications are their anisotropic, highly porous, and controllable macro-, micro-, and nanostructure. Collagen is one of the most commonly used material for the fabrication of scaffolds, as this biopolymer is present in the natural extracellular matrix. For textile processing and textile scaffold fabrication methods, multifilament yarns are required, however, only monofilaments can be generated by state-of-the-art collagen spinning. Hence, the research presented in here aimed at the development of a collagen multifilament wet-spinning process in reproducible quality as well as the characterization of non-crosslinked and crosslinked wet-spun multifilament yarns. Wet spun collagen yarns were comprised of 6 single filaments each having a fineness of 5 tex and a diameter of 80 μm. The tensile strength of the glutaraldehyde crosslinked yarns was 169 MPa (Young's modulus 3534 MPa) in the dry state and 40 MPa (Young's modulus 281 MPa) in the wet state. Furthermore, wet spun collagen filaments showed a characteristic fibrillar structure, which was similar the morphological structure of natural collagen fibers. The textile processing of collagen multifilament yarn was demonstrated by means of knitting technology.


Factors Associated With Unmet Supportive Care Needs and Emergency Department Visits and Hospitalizations in Ambulatory Oncology.
Patients with cancer experience multiple supportive care needs (eg, coping and financial counseling) that, if not addressed, may result in poor clinical outcomes. Limited work has assessed the factors associated with unmet needs in large and diverse samples of ambulatory oncology patients. To characterize the factors associated with unmet supportive care needs among ambulatory oncology patients and to assess whether such needs were associated with emergency department (ED) visits and hospitalizations. Between October 1, 2019, and June 30, 2022, cross-sectional retrospective analyses were performed in a large and diverse ambulatory cancer population via My Wellness Check, an electronic health record (EHR)-based supportive care needs and patient-reported outcomes (PROs) screening and referral program. Demographic characteristics, clinical characteristics, and clinical outcomes were extracted from EHRs. Data on PROs (ie, anxiety, depression, fatigue, pain, and physical function), health-related quality of life (HRQOL), and supportive care needs were also collected. Logistic regressions examined factors associated with unmet needs. Cumulative incidence of ED visits and hospitalizations were assessed by Cox proportional hazards regression models adjusting for covariates. The 5236 patients in the study had a mean (SD) age of 62.6 (13.1) years and included 2949 women (56.3%), 2506 Hispanic or Latino patients (47.9%), and 4618 White patients (88.2%); 1370 patients (26.2%) indicated Spanish as their preferred language, according to their EHR. A total of 940 patients (18.0%) reported 1 or more unmet needs. Black race (adjusted odds ratio [AOR], 1.97 [95% CI, 1.49-2.60]), Hispanic ethnicity (AOR, 1.31 [95% CI, 1.10-1.55]), 1 to 5 years after diagnosis (AOR, 0.64 [95% CI, 0.54-0.77]), more than 5 years after diagnosis (AOR, 0.60 [95% CI, 0.48-0.76]), anxiety (AOR, 2.25 [95% CI, 1.71-2.95]), depression (AOR, 2.07 [95% CI, 1.58-2.70]), poor physical function (AOR, 1.38 [95% CI, 1.07-1.79]), and low HRQOL scores (AOR, 1.89 [95% CI, 1.50-2.39]) were associated with greater unmet needs. Patients with unmet needs had a significantly higher risk of ED visits (adjusted hazard ratio [AHR], 1.45 [95% CI, 1.20-1.74]) and hospitalizations (AHR, 1.36 [95% CI, 1.13-1.63]) relative to patients without unmet needs. In this cohort study of ambulatory oncology patients, unmet supportive care needs were associated with worse clinical outcomes. Patients from racial and ethnic minority groups and those with greater emotional or physical burden were more likely to have 1 or more unmet needs. Results suggest that addressing unmet supportive care needs may be crucial for improving clinical outcomes, and targeted efforts should focus on specific populations.


De novo paracentric inversion (X)(q26q28) with features mimicking Prader-Willi syndrome.
Different genetic and non-genetic disorders, including several chromosomal abnormalities, may mimic Prader-Willi syndrome (PWS). We report on an 11-year-old girl with features reminiscent of PWS due to an unreported de novo paracentric inversion Xq26q28. Microdeletion 15q11-q13 and maternal uniparental disomy 15 were ruled out. The importance of chromosomal studies in addition to molecular analysis on patients with features suggestive of PWS is stressed.


Stress dynamically regulates behavior and glutamatergic gene expression in hippocampus by opening a window of epigenetic plasticity.
Excitatory amino acids play a key role in both adaptive and deleterious effects of stressors on the brain, and dysregulated glutamate homeostasis has been associated with psychiatric and neurological disorders. Here, we elucidate mechanisms of epigenetic plasticity in the hippocampus in the interactions between a history of chronic stress and familiar and novel acute stressors that alter expression of anxiety- and depressive-like behaviors. We demonstrate that acute restraint and acute forced swim stressors induce differential effects on these behaviors in naive mice and in mice with a history of chronic-restraint stress (CRS). They reveal a key role for epigenetic up- and down-regulation of the putative presynaptic type 2 metabotropic glutamate (mGlu2) receptors and the postsynaptic NR1/NMDA receptors in the hippocampus and particularly in the dentate gyrus (DG), a region of active neurogenesis and a target of antidepressant treatment. We show changes in DG long-term potentiation (LTP) that parallel behavioral responses, with habituation to the same acute restraint stressor and sensitization to a novel forced-swim stressor. In WT mice after CRS and in unstressed mice with a BDNF loss-of-function allele (BDNF Val66Met), we show that the epigenetic activator of histone acetylation, P300, plays a pivotal role in the dynamic up- and down-regulation of mGlu2 in hippocampus via histone-3-lysine-27-acetylation (H3K27Ac) when acute stressors are applied. These hippocampal responses reveal a window of epigenetic plasticity that may be useful for treatment of disorders in which glutamatergic transmission is dysregulated.


Map-based cloning and validation of a gall midge resistance gene, Gm8, encoding a proline-rich protein in the rice variety Aganni.
The Asian rice gall midge (ARGM), Orseolia oryzae is an important insect pest causing an annual yield loss of about US$ 80 million in India. Till now 11 R genes and seven biotypes of the pest have been characterized and reported. The indica rice variety Aganni, a landrace from the state of Kerala, is known to carry the gall midge resistance gene Gm8 with HR-type of resistance. This gene has been fine mapped within 0.43 Mb region with the flanking markers RM22685 and RM22709. We identified 63 possible candidate genes through in silico analysis in the reference Nipponbare rice genome between 7.5 and 9.5 Mb region. One of the markers targeting the proline rich protein (PRP) gene (LOC_Os08g15080) showed polymorphism between the parents and also exhibited complete co-segregation with the trait in 426 F10 RIL populations. Functional validation of this gene through RT-PCR in contrasting parents and Pre-NILs (near isogenic lines) revealed that this is an early responsive gene with rapid induction at 24 h after gall midge infestation (hai) followed by subsequent reduction in the expression levels at late hours. Validation of this gene in five gall midge resistant rice varieties carrying different resistance genes revealed that the induction was unique to Aganni rice carrying Gm8 gene. Further, cloning and sequencing of the alleles of this gene including promoter region from TN1 (susceptible parent) and Aganni (resistant parent) revealed 153 nucleotide substitution, four amino acid substitutions and three mutations at putative cis-acting elements in TN1 when compared to Aganni. In addition, we also developed a functional marker (PRP-del) for detection of the gene for use in marker-assisted introgression of Gm8.


Nemaline myopathy: comparative muscle histochemistry in the severe neonatal, moderate congenital, and adult-onset forms.
A histochemical study of biopsied muscle specimens from patients with the 3 forms of nemaline myopathy (i.e., severe neonatal, moderate congenital, and adult-onset), classified on the basis of clinical symptoms, was conducted. A close relationship could not be found between the number of rods and the severity of weakness in any form. Type 1 fiber atrophy and predominance or type 2B fiber deficiency was the usual finding in all forms. In the moderate congenital form, type 1 fiber atrophy and predominance became more apparent in patients with a protracted course, suggesting that the histochemical abnormalities are progressive. The abnormal fiber type distribution is assumed to be related to the pathogenetic mechanism of nemaline myopathy in all forms. Because acid phosphatase activity was increased in muscle fibers of patients with rapid progression, an autodegenerative process inducing lysosomal enzyme activation may be responsible for the acute clinical progression and muscle fiber loss in this disorder.


Water balance in developing leaves of four tropical savanna woody species.
Water balance was studied in developing leaves of Curatella americana, Byrsonima crassifolia, Bowdichia virgilioides and Casearia sylvestris, four widespread tropical savanna evergreen woody species that renew their leaves during the dry season. Water potential components of leaves of different ages were estimated in the laboratory by determination of pressure-volume (PV) curves. Data from PV curves were used to help interpret the significance of variations in leaf water potential (ΨL) and stomatal conductance observed in leaves of different ages in the field.Initial osmotic potential at full turgidity as well as osmotic potential at the turgor loss point changed considerably during leaf development. Values of these water potential components for mature leaves were similar to those reported for temperate mesophytic woody species. Passive osmotic adjustment defined as the change in osmotic potential between full turgidity and turgor loss averaged only 0.3 MPa, much smaller than values for temperate mesophytic and drought hardy woody species. Since environmental conditions, especially soil moisture in the rooting zone, were essentially constant during leaf development, changes in leaf water potential components were not seasonal, but rather ontogenetic responses to diurnal water stress.In all species except C. sylvestris there were differences in ΨL between adjacent leaves exposed to the same environmental conditions, with immature leaves generally maintaining higher ΨL than mature leaves. The probable causes for this were 1) lower liquid flow resistance and 2) lower transpiration rates in younger leaves. At low transpiration rates liquid flow resistance was flux-dependent in all species, but became nearly independent of transpiration flux density (E) as E increased. This and their lower flow resistance permit high E to be sustained in developing leaves without excessive ψ drop across the leaf.During two typical dry season days E was high and even though complete or nearly complete turgor loss was sustained, these water deficits were not severe enough to cause complete stomatal closure. The possible relationship between a high stomatal conductance and the maintenance of a favorable carbon balance in these species is discussed.All of the water relations features observed suggested that in spite of their scleromorphic appearance, leaves of the four species under the conditions studied were adapted only to short term diurnal water deficits.


Role of associated and covalently bound lipids in salivary mucin hydrophobicity: effect of proteolysis and disulfide bridge reduction.
The hydrophobic properties of salivary mucus glycoprotein were investigated by fluorescence spectroscopy using bis(8-anilino-1-naphthalene-sulfonate). The mucin, purified from rat submandibular salivary gland, was subjected to removal of associated and covalently bound lipids, degradation with pronase, and reduction with beta-mercaptoethanol, and titrated with the probe. Analyses of fluorescence data revealed the presence of 49 +/- 5 hydrophobic binding sites in the intact mucin molecule, a 69% increase in the number of binding sites occurred following extraction of associated lipids, while the removal of covalently bound fatty acids caused a 25% decrease in the binding sites. Proteolytic destruction of the nonglycosylated regions of the glycoprotein essentially abolished the probe binding, whereas reduction produced glycoprotein subunits whose combined number of hydrophobic binding sites was 2.4 times greater than that of mucus glycoprotein polymer. The results suggest that associated and covalently bound lipids contribute to hydrophobic characteristics of salivary mucin and that the hydrophobic binding sites reside on the nonglycosylated regions of this glycoprotein buried within its core.


Clinical neurophysiological examination of deafness associated with juvenile motor neurone disease.
Three patients with the syndrome of progressive severe deafness, associated with juvenile onset of motor neurone disease, were examined using conventional audiometry and brainstem auditory evoked response (BAER) recordings, and electrocochleographic recordings were made in two of them. Findings indicate that the deafness had resulted from loss of acoustic nerve fibers and/or sensory cells of the spiral ganglion.


Cochrane Corner: The use of anticoagulants in patients hospitalised with COVID-19.
In this Cochrane Corner, we highlight the main findings of a Cochrane Review by Flumignan et al. entitled 'Anticoagulants for people hospitalised with COVID-19' and discuss the implications of these findings for research and practice in South Africa. In particular, we underscore the need for additional, high-quality, randomised controlled trials comparing different intensities of anticoagulation in patients with COVID-19 illness. Individuals in the intensive care unit and those hospitalised with another illness who are incidentally found to be infected with SARS-CoV-2 should still only be treated with prophylactic-dose low-molecular-weight heparin. This Cochrane Corner summarises findings in a recent systematic review on the use of anticoagulation in people hospitalised with COVID-19, and provides insights on the implications of these findings for implementation by clinicians in South Africa. It highlights the need for clinicians to balance the benefits and harms of providing an anticoagulant, while considering the patients underlying risk for bleeding and thromboembolism.


Computerizing medical records in Japan.
The present study reports the current status of computerizing medical records in Japan. In 2001, the Ministry of Health, Labour and Welfare formulated the Grand Design for the Development of Information Systems in the Healthcare and Medical Fields. The Grand Design stated a numerical target for "spreading the use of electronic medical records (EMR) in at least 60% of Japan's hospitals with 400 or more beds by 2006." The objective of this study was to examine the extent to which EMR and order entry systems (OES) have been adopted as of February 2007 and to evaluate the Japanese government's policy regarding the computerization of medical records. We conducted a postal survey targeting medical institutions throughout Japan. In February 2007, we mailed self-administered questionnaires to all 1574 hospitals with 300 or more beds, and to a random selection of 1000 hospitals with less than 300 beds in addition to 4000 clinics. Responses were received from 812 (51.6%), 504 (50.5%), and 1769 (44.8%), respectively. We asked questions concerning: (i) the extent to which EMR and OES had been introduced; (ii) the reasons why certain institutions had not introduced EMR and (iii) the subjective evaluation of the efficacy and cost-effectiveness of EMR. The percentage of institutions that had introduced EMR as of February 2007 was 10.0% for hospitals and 10.1% for clinics. Even the percentage for hospitals with 400 or more beds was just 31.2%, illustrating that the government's target had not been reached. The most common reason given for not introducing EMR was: "The cost is high" which was observed in 82.0% of hospitals. It was considered that the introduction of EMR could improve 'inter-hospital networks', and 'time efficiency for physicians' by around 45% and 25% of hospitals, respectively. Healthcare information computerization in Japan is behind schedule because the introductory costs are high. For the computerization of healthcare information to be further promoted, prices of EMR systems should be lowered to a level which individual hospitals can afford. Furthermore, the communication between EMR systems should be further standardized to secure functional and semantic interoperability in Japan.


Polypeptide composition of mumps virus.
The Enders strain of mumps virus grown in ovo was purified by differential and equilibrium sucrose gradient sedimentation. Purified virus contained seven polypeptides of mol. wts 68,000, 66,000, 61,000, 54,000, 52,000, 49,000, 47,000. Nucleocapsids isolated from DOC-treated virus contained two polypeptides of mol. wts 66,000 and 61,000.


A slowly inactivating transient outward current in rat ventricular myocytes.
In rat ventricular myocytes we found two components of transient outward current, which could be discriminated time- and voltage- dependently. Besides the well known fastly inactivating transient outward current (ito,f, tau = 35 +/- 8 ms, n = 4) we investigated properties of a slowly inactivating transient outward current (ito,s, tau = 1.7 +/- 0.4 s, n = 4). Because of the slow inactivation process of ito,s tail currents were observed at -25 mV. The inactivation curve of ito,f was characterized by a half- inactivation voltage of -58.4 +/- 1.4 mV and a slope factor of 5.6 +/- 0.5 mV (n = 4). The inactivation curves of ito,s and tail currents were nearly identical but significantly different from the ito,f-curve. Half-inactivation voltages of ito,s and tail currents were -87.5 +/- 6 mV and -89.1 +/- 5 mV (n = 4), respectively. Slope factors were 10.3 +/- 2.9 mV and 9.8 +/- 1.7 mV (n = 4). The activation gate of ito,s was half-maximally opened at -11.5 +/- 2.6 mV, and the slope factor was -10.6 +/- 1.7 mV (n = 3). Ito,s tail current reversed its direction at -62 +/- 3.2 mV (n = 5). This indicates, that ito,s- current flow is carried mainly by potassium ions. Ito,s- current was not abolished by Tetrodotoxin (TTX) and Cd.


Nitric oxide-containing nerves in bowel segments of patients with Hirschsprung's disease.
To assess the role of nerves that synthesize nitric oxide (NO) in Hirschsprung's Disease (HD), the authors studied the distribution of the enzyme NADPH diaphorase (NADPHd) in normal and diseased bowel segments. In the proximal (ganglionic) segment of the colon, NADPHd-positive neurons were present in both myenteric and submucosal plexuses. In the distal involved colonic segments from HD patients, the typical pattern of the neuronal network was completely missing in the regions of the two plexuses; instead, only disorganized NADPHd-positive nerve fibers were present and NADPHd-reactive neurons were absent. Mucosal NO synthase activity was 2.76 +/- 0.38 nmol/g/min in the proximal segment and only 0.83 +/- 0.49 nmol/g/min in the distal segment (P < .05, N = 3).


Volume expansion during neonatal intensive care: do we know what we are doing?
Although volume expansion is liberally used in newborn intensive care, we know little about its effects on hemodynamics or outcomes. Given appropriately to a truly hypovolemic baby, it can be life-saving, but the clinical diagnosis of hypovolemia is probably very inaccurate. We know that volume expansion has less effect on blood pressure than dopamine, and although it seems to produce immediate increases in systemic blood flow, we do not know for how long these increases are sustained. There is evidence to show that the routine use of volume expansion in preterm babies has no effect on outcome, and there is little evidence to support its routine use during resuscitation or the treatment of metabolic acidosis. Whether crystalloids or colloids are preferable is also unclear in newborns. In situations of concern related to circulatory compromise, if possible, define the hemodynamics echocardiographically. Otherwise, if in doubt, some volume should be given, although it is probably unwise to keep expanding the volume if this is not improving physiologic (blood pressure and heart rate) or echocardiographic systemic blood flow parameters.


Quantitative analysis of trace elements in human blood and plasma by energy dispersive X-ray fluorescence.
A photon excitation, secondary target x-ray fluorescence system was used to measure concentrations of Cu, Zn, Br and Rb in whole blood and plasma of healthy adults. The samples preparation method and calculated limits of detection of the technique are reported. Correlation statistics for elemental concentrations have been determined. Verification of the results for copper was performed using atomic absorption spectrophotometry.


3D micro-CT imaging of the postmortem brain.
Magnetic resonance microscopy (microMRI) is becoming an important tool for non-destructive analysis of fixed brain tissue. However, unlike MRI, X-ray computed tomography (CT) scans show little native soft tissue contrast. In this paper, we explored the use of contrast enhanced (brains immersion stained in iodinated CT contrast media) micro-CT (microCT) for high resolution 3D imaging of fixed normal and pathological brains, compared to microMRI and standard histopathology. An optimum iodine concentration of 0.27 M resulted in excellent contrast between gray and white matter in normal brain and a wide range of anatomical structures were identified. In glioma bearing mouse brains, there was clear deliniation of tumor margin which closely matched that seen on histopathology sections. microCT tumor volume was strongly correlated with histopathology volume. Our data suggests that microCT image contrast in the immersion-stained brains is related to axonal density and myelin content. Compared to traditional histopathology, our microCT approach is relatively rapid and less labor intensive. In addition, compared to microMRI, microCT is robust and requires much lower equipment and maintenance costs. For simple measurements, such as tumor volume and non-destructive postmortem brain screening, microCT may prove to be a valuable alternative to standard histopathology or microMRI.


Tibial bone responses to 6-month calcium and vitamin D supplementation in young male jockeys: A randomised controlled trial.
Young male jockeys compromise bone health by engaging in caloric restriction and high volumes of physical activity during periods of musculoskeletal growth and development. The aim of this randomised, double-blinded, placebo-controlled trial was to establish whether calcium and vitamin D supplementation would improve bone properties of young male jockeys. We conducted a 6-month trial with two groups of weight-, height- and age-matched apprentice male jockeys (age=20.2 ± 3.2 yrs). Participants were supplemented with 800 mg of calcium and 400 IU of vitamin D (S, n=8) or a placebo (cellulose) (P, n=9) daily for 6-months. Baseline calcium intake was (669.7 ± 274.3 (S) vs 790.4 ± 423.9 (P) and vitamin D 64.6 ± 19.5 (S) vs 81.2 ± 24.4 (P) with no statistical differences. Peripheral quantitative computed tomography (pQCT) measured ultra-distal (4%) and proximal (66%) tibial bone properties at baseline and 6 months. Blood-borne markers of bone turnover, P1NP and CTX and vitamin D concentration were assessed. After co-varying for height, weight and baseline bone measurements, the supplemented group displayed greater post-intervention bone properties at the 66% proximal site with cortical content (mgmm) 6.6% greater (p<0.001), cortical area (mm(2)) 5.9% larger (p<0.001), cortical density (mgcm(2)) 1.3% greater (p=0.001), and total area (mm(2)) 4% larger (p=0.003). No other between group differences in bone variables were observed. Blood analysis indicated higher vitamin D levels (18.1%, p=0.014) and lower CTx (ng/L) (-24.8%, p=0.011) in the supplemented group with no differences observed in P1NP. This is the first randomised controlled trial to examine the efficacy of calcium and vitamin D supplementation in improving bone properties in a highly vulnerable, young athletic, weight-restricted population. Results using pQCT indicate beneficial effects of supplementation on bone properties in as little as six months. Although the study size is small, this intervention appears promising as a strategy for improving bone health in young athletes in weight-restricted sports.


Prevalence, associated factors and rifampicin resistance pattern of pulmonary tuberculosis among HIV-positive patients attending antiretroviral treatment clinic at East Gojjam Zone, Ethiopia: An institution-based cross-sectional study.
Drug-resistant tuberculosis (TB) threatens global TB care and prevention, and it remains a major public health concern in many countries particularly in sub-Saharan countries. Pulmonary TB is the most common serious opportunistic infection on HIV-positive patients and it is the leading cause of death among HIV-positive patients in developing countries. Ethiopia is one of the high TB burden countries with high morbidity and mortality. To determine the prevalence, associated factors and rifampicin resistance of pulmonary TB among HIV-positive attending antiretroviral treatment clinic at East Gojjam. Hospital-based cross-sectional study was conducted at Debre Markos Referral Hospital, from February to June 2019. A total of 112 HIV-positive TB suspected patients were included using convenient sampling techniques and a bacteriological confirmation test for tuberculosis was performed using Gene-Xpert MTB/RIF assay from a spot sputum sample. Viral load was determined by using a quantitative real-time polymerase chain reaction (RT-PCR) from the blood sample. Socio-demographic and clinical data were collected by face-to-face interview using a semi-structured questionnaire. The data were analyzed by using Statistical Package for Social Sciences (SPSS) software (version 24). Out of the 112 study participants, the prevalence of Pulmonary TB was 11.6 %. Among TB positives 23.1 % were rifampicin resistant. Rifampicin resistance was 100 % among female patients. Having family members treated for pulmonary TB (P = 0.003, [AOR = 4.5; 95 % CI = 3.59-58.8]), cigarette smoking (P = 0.039, [AOR = 2.18; 95 %CI = 1.17-40.5]), being on WHO HIV disease clinical stage II (P = 0.024, [AOR = 1.81; 95 %CI = 1.50-30.99]), and having viral load (1000-9999) RNA copies/ml (P = 0.031, [AOR = 1.54; 95 %CI = 1.32-31.41]) were found to be significantly associated with pulmonary TB. The prevalence of pulmonary TB and rifampicin resistance was high among HIV patients. Having family members treated for Pulmonary TB, history of cigarette smoking, WHO HIV clinical stage, and high viral load were associated risk factors for TB. Therefore, strengthening awareness creation on TB transmission, drug resistance, and treatment adherence are essential. Moreover, early screening and treatment are vital for preventing the transmission and occurrence of drug-resistant TB among study populations.


Spectra of photoneutrons produced by high-energy X-ray radiotherapy linacs.
Spectral fluence of photoneutrons generated in the head of the radiotherapeutic linac Varian 2100 C/D was measured by means of the Bonner spheres spectrometer whose active detector of thermal neutrons was replaced by a track detector, i.e. a sandwich of four CR-39s with the boron radiator inserted between them. Measurements with different collimator settings showed that the fluence of photoneutrons was higher for the more open collimator.


Perceived racial/ethnic bias in healthcare in Durham County, North Carolina: a comparison of community and national samples.
We sought to compare findings of a national survey of perceptions of racial/ethnic discrimination in healthcare to those of a community survey, with emphasis on the perceptions of Latinos. Responses from a national survey were compared to a telephone survey of residents of Durham County, North Carolina. Black respondents in the Durham sample were more likely than those in the national sample to feel that a healthcare provider had treated them with disrespect because of health insurance status (28% vs 14%; P < 0.001). Approximately one third of Durham Latinos and 14% of Latinos in the national sample felt they had been treated with disrespect because of their English-language ability (P < 0.01). Compared to a national sample of white participants, white respondents in Durham were more likely to believe that black persons are worse off in terms of receiving routine medical care (40% vs 27%; P < 0.01) and having health insurance (58% vs 43%; P < 0.01). As compared to their national counterparts, there was a similar trend for how white respondents in Durham perceived how Latinos fared (P < 0.001 for all comparisons). Overall the perception of bias in healthcare was greater among Durham residents, especially among newly immigrated Latinos, than among their national counterparts.


Impact of the acetate/oleic acid ratio on the performance, quorum sensing, and microbial community of sequencing batch reactor system.
This study comprehensively investigated the impact of acetate/oleic acid ratios (80%/20%, 60%/40%, 40%/60%, and 20%/80%) on sequencing batch reactor (SBR) with respect to the variations in performance, quorum sensing (QS), and microbial community. Results showed that NH+4-N removal was not affected by the acetate/oleic acid ratios, while the COD, total nitrogen (TN), and PO3-4-P removal was considerably affected. The increasing oleic acid ratios led to severe sludge bulking, which was significantly positively correlated with proteins/polysaccharides (p < 0.001). The correlation of QS with the performance and sludge properties was also observed. High-throughput sequencing demonstrated that microbial compositions considerably shifted with varying acetate/oleic acid ratios. Moreover, the potential correlation of bacterial genera with the SBR performance and QS was proposed. This study elucidated the effect of acetate/oleic acid ratios on SBR from microbial viewpoint, which provided insights into fully understanding the essential roles of carbon source on wastewater treatment.


Population pharmacokinetic-pharmacodynamic modeling of moxonidine using 24-hour ambulatory blood pressure measurements.
To develop a model for 24-hour ambulatory blood pressure measurements (ABPM) that can be applied in a pharmacokinetic-pharmacodynamic model. Four different data sets were prepared from 2 studies to accommodate different modeling strategies. In study A, a double-blind placebo-controlled study in 47 patients, 24-hour ABPM profiles (74 to 99 measurements per profile) were obtained during the placebo run-in phase and after 3, 5, and 11 weeks during the treatment. Three to 5 plasma samples were taken. Cosine and polynomial models were evaluated to describe the circadian rhythm in blood pressure based on 3 data sets (1: only run-in data; 2: only placebo data; 3: all data). In study B, a double-blind placebo-controlled study in 94 patients, two 24-hour ABPM profiles per patient (during placebo run-in and after 8 weeks) were recorded and randomly reduced to 15 measurements per profile to evaluate the robustness of the baseline model. The mean moxonidine clearance was 35 L/h, and the volume of distribution was 132 L. The final baseline model consisted of 2 cosine terms with fixed-effect parameters for rhythm-adjusted 24-hour mean blood pressure, amplitude, phase, and period; random-effect parameters for interindividual variability in rhythm-adjusted 24-hour mean, amplitude, and clock time; and interoccasion variability in rhythm-adjusted 24-hour mean and clock time. The final baseline model was combined with an Emax model for the drug effect. An effect compartment was used (kco = 0.198 h-1). The maximum decrease in diastolic blood pressure (Emax) was 16.7%, and EC50 was 0.945 microgram/L. The pharmacokinetic-pharmacodynamic model for 24-hour ABPM can be used to estimate the concentration-effect relationship of antihypertensive drugs.


Ocular trauma caused by air-gun pellets: a ten year survey.
Forty-one consecutive cases of ocular injury related to air gun pellets occurring over a ten year period have been reviewed retrospectively. The male to female ratio was 7:1 and the average age was fifteen years. Hyphaema was the most common injury (78%) while the main cause of poor final visual outcome was retinal detachment. Thirty-eight percent of the involved eyes had a final corrected visual acuity of less than 6/60 including four eyes (10%) which were enucleated following severe perforating injuries. Neither sympathetic ophthalmitis nor lead poisoning was detected in this study. The fact that these injuries are largely of an accidental nature, predominantly involve young people and appear to be occurring more frequently suggests that present legislation does not offer adequate protection. The authors suggest that the best method of prevention is through a public education campaign aimed at children and their parents.


REM sleep eye movement activity and CSF concentrations of 5-hydroxyindoleacetic acid in psychiatric patients.
The relationship between rapid-eye-movement (REM) sleep tonic and phasic activity measures and the pre- and postprobenecid cerebrospinal fluid (CSF) concentrations of the serotonin metabolite 5-hydroxyindoleacetic acid (5-HIAA) was examined in a sample of 14 unmedicated psychiatric patients. The average duration of an eye movement burst and the average number of eye movements within a burst were found to be inversely related to the probenecid corrected accumulation of 5-HIAA in the CSF. These results are interpreted as indicating an association between REM burst eye movement activity and serotonin turnover in psychiatric subjects.


Purification of a novel growth inhibitory factor for partially differentiated myeloid leukemic cells.
A novel factor termed growth inhibitory (GI) factor, which specifically inhibits the growth of mouse monocytic leukemia cells including monocytic cell lines (Mm-A and J774.1) and other partially differentiated myeloid leukemic cells, has been purified from conditioned medium of some clones of mouse myeloblastic leukemia M1 cells. The procedure for purification of the GI factor included ammonium sulfate precipitation, CM-Sepharose CL-6B and Sephadex G-200 chromatographies, reverse-phase high-performance liquid chromatography on a C18 hydrophobic support, and high-performance liquid chromatography on a gel filtration column. The purified factor gave a single band of protein with a molecular weight of 25,000 on sodium dodecyl sulfate-polyacrylamide gel. A concentration of 8 X 10(-10) M GI factor was required for 50% inhibition of growth of Mm-A cells. On chromatofocusing, the GI activity was eluted with Polybuffer 96/acetic acid at pH 8.2-8.4. The purified GI factor markedly inhibited growth of mouse bone marrow cells stimulated by macrophage colony-stimulating factor. The GI factor appeared to be a unique cytokine unrelated to known cytokines such as the tumor necrosis factor, interferons, and oncostatin M.


In vitro infectivity of cryopreserved Plasmodium berghei sporozoites to cultured cells.
Plasmodium berghei sporozoites frozen in MEM (Eagle) medium supplemented with 10% hydroxyethyl starch and 50% normal mouse serum retained 0.5% infectivity to cultured hepatoma cells, compared to 6.8% before freezing. This demonstrates that frozen-thawed sporozoites can be used in in vitro investigations of the exoerythrocytic malarial parasite and means that when large numbers of sporozoites are available, they may be frozen and preserved for later use.


Emergency department visits after hours by lung cancer patients in Japan.
The aims of this study were to clarify frequency with which Japanese lung cancer patients visited an emergency department (ED) after hours and their final outcome. This is a retrospective and single institutional study. We reviewed medical records of patients who died of lung cancer from January 2008 to June 2012 at Osaka Police Hospital who had been followed up since diagnosis of lung cancer until death. We compared patients who had visited the ED after hours on weekdays, weekends, or holidays over their lives with cancer (ED visitors) and patients who had never visited the ED (non-ED visitors). Overall, 245 patients met the inclusion criteria for analysis. There were 149 after hours ED visits by 106 lung cancer patients. Mean number of ED visits was 0.6 for all patients. Median interval from ED visit to death was 49 days. The most common chief compliant for these patients was respiratory problems (37.6%). Most patients visited the ED during chemotherapy (32.9%) or for best supportive care (42.3%). Directly after ED visits, 56.4% of ED visitors were finally hospitalized. In a multivariate analysis, performance status (PS) (odds ratio [OR]: 11.2, 95% confidence interval [CI]: 2.1-59.0, p = 0.004) and cancer stage (OR: 0.003, 95% CI: 0.0006-0.014, p < 0.001) at diagnosis were statistically associated with ED visits after hours. Japanese patients with lung cancer frequently visit ED after hours. An ED visit is itself an indicator of poor prognosis.


Chemical and genetic characterization of Phlomis species and wild hybrids in Crete.
The genus Phlomis is represented in the island of Crete (Greece, Eastern Mediterranean) by three species Phlomis cretica C. Presl., Phlomis fruticosa L., the island endemic Phlomis lanata Willd. and three hybrids Phlomis x cytherea Rech.f. (P. cretica x P. fruticosa), Phlomis x commixta Rech.f. (P. cretica x P. lanata) and Phlomis x sieberi Vierh. (P. fruticosa x P. lanata). This work describes (a) the profile of hybrids and parental species concerning their volatile compounds, (b) the suitability of ribosomal nuclear (ITS region), chloroplast (trnH-psbA), and AFLP markers to identify hybrids and (c) their competence to characterize the different chemotypes of both hybrids and their parental species. The cluster analysis and PCA constructed from chemical data (volatile oils) suggest that there are three groups of taxa. Group IA includes P. cretica and P. fruticosa, group IB includes P. x cytherea, whereas group II consists of P. x commixta, P. x sieberi and P. lanata. Volatile compounds detected only in the hybrids P. x sieberi and P. x commixta correspond to the 3% of the total compounds, value that is much higher in P. x cytherea (21%). Neighbor-joining, statistical parsimony analysis and the observations drawn from ribotypes spectrum of ITS markers divided Phlomis species in two groups, P. lanata and the complex P. cretica/P. fruticosa. In contrast to the ITS region, the plastid DNA marker follows a geographically related pattern. Neighbor-Net, PCA and Bayesian assignment analysis performed for AFLP markers separated the genotypes into three groups corresponding to populations of P. cretica, P. fruticosa, and P. lanata, respectively, while populations of P. x commixta, P. x cytherea, and P. x sieberi presented admixed ancestry. Most of the P. x cytherea samples were identified as F1 hybrids by Bayesian assignment test, while those of P. x commixta and P. x sieberi were identified as F2 hybrids. Overall, high chemical differentiation is revealed in one of the three hybrids, which is likely related with niche variation. Moreover, molecular markers show potential to identify Phlomis taxa.


Parental Distress and Stress in Association with Health-Related Quality of Life in Youth with Spina Bifida: A Longitudinal Study.
This study examined associations between 3 distinct parent factors (parent personal distress, parenting stress, and spina bifida (SB)-specific parenting stress) and youth and parent proxy reports of youth health-related quality of life (HRQOL) over time. Participants were recruited as part of a longitudinal study, and data were collected at 3 time points, spaced 2 years apart. Parents and youth completed questionnaires, and youth completed neuropsychological assessment tasks to determine youth intelligence quotient during home visits. Analyses revealed that higher levels of maternal SB-specific parenting stress were related to lower levels of youth-reported HRQOL at time 1. Other parent factors were not associated with youth report of HRQOL at the earlier time points, although higher levels of maternal SB-specific parenting stress and paternal parenting stress were associated with lower levels of youth HRQOL at time 3. For mothers and fathers, increased parent personal distress, parenting stress, and SB-specific parenting stress were associated with decreased proxy report of youth HRQOL. Of these three parent factors, SB-specific parenting stress was consistently the most strongly associated with parent proxy-report of youth HRQOL. Parenting stress and distress are important targets for interventions, and these interventions may improve youth outcomes, especially as youth age. Future research is needed to identify other factors influencing youth HRQOL over time.


Vitrectomy in uveitis associated with ankylosing spondylitis.
Chronic recurrent iridocyclitis in three eyes of two patients with ankylosing spondylitis was associated with posterior spillover of inflammatory cells into the vitreous cavity. Continued inflammation resulted in significant vitreous opacification in all three eyes. After pars plana vitrectomy (two eyes) and cataract extraction with subtotal vitrectomy (one eye), visual acuity improved and stabilized in all three instances. Ocular inflammation was not appreciably exacerbated by surgical intervention. Vitreous opacification did not recur after vitrectomy, but visual improvement was limited because of chronic cystoid macular edema.


Computer tomography navigation for the transoral anterior release of a complex craniovertebral junction deformity: A report of two cases.
The surgical correction of deformities of the craniovertebral junction (CVJ) remains a challenge due to its complex anatomy. Despite the well-known usefulness of computed tomography (CT) navigation in posterior spinal surgery, it is applied far less frequently in anterior spinal surgery, mainly due to registration difficulties. Case 1 was a 68-year-old female with rheumatoid arthritis, with a complaint of neck pain, motor weakness, and dysesthesia in the upper extremities. Case 2 was a 61-year-old male with Chiari malformation, with a complaint of neck pain and gait disturbance after a fall. Magnetic resonance imaging (MRI) showed severe atlantoaxial dislocation and multilevel cervical spinal cord compression in both patients. Continuous halo traction failed to reduce atlantoaxial dislocation, even under general anesthesia, and they were treated with combined anterior release and posterior decompression and fixation using CT navigation. Occipitocervical assimilation, which was present in both patients, enabled precise registration for navigation. The lack of anatomically characteristic landmarks on the vertebral surface makes obtaining accurate registration difficult in anterior CVJ surgery using CT navigation. The remaining mobility in the occipitocervical joint precludes the use of facial or cranial landmarks. However, occipitocervical assimilation, which is not uncommon in patients with CVJ deformities, enables accurate navigation during transoral surgery. Transoral anterior release using CT navigation is an effective treatment option for rigid complex CVJ deformities. The accurate identification of the patients' anatomical features such as occipitoatlantal assimilation, is crucial for the conducting accurate preoperative CT-based navigation during transoral surgery.


Degradation by Eco R1 endonuclease of DNA isolated from phages infecting Bacillus licheniformis and Bacillus thuringiensis.
DNA was isolated from lytic phages of two strains, Bacillus licheniformis, a producer of bacitracin, and Bacillus thuringiensis forming protein paracrystals with pronounced insecticidal effects. Its sensitivity to Eco R1 restriction endonuclease was determined. It was the aim of the work to find out whether these phages could serve as vectors in the transfer and possible amplification of genes of the two important industrial species of bacilli. Approximate values of the molecular weight of DNA of the two phages were determined after degradation of the phage DNA by Eco R1, followed by comparison of electrophoretic mobility of individual fragments with that of the Eco R1-degraded DNA of phage lambdab2.


Venipuncture in psittacine birds.
Techniques for jugular, basilic and medial metatarsal venipuncture in psittacine birds are discussed in detail. Although the number of animals used in the US for research annually is documented, birds and laboratory rats and mice are not included. It is presumed that the number of psittacine birds used for scientific purposes is relatively low. Therefore, only limited information is available about clinical techniques applicable to psittacine birds.


Very early onset multiple sclerosis.
The authors report two cases of multiple sclerosis occurring in very young children. Age at onset in one child was 13 months; he had the primary progressive form of the condition and died at nine years. Age at onset of the other child was two years; he has the classical relapsing-remitting form and is well at 15 years. These children illustrate the broad clinical spectrum of the condition even in very-early-onset disease.


Expression and production of tenascin in benign and malignant lesions of melanocyte lineage.
The distribution of the extracellular matrix (ECM) glyco-protein tenascin (TN) has been evaluated immunohistochemically in benign and malignant lesions of the melanocytic lineage. Results of our study show that TN is detectable in a variety of benign and malignant lesions. In primary melanomas, TN displays a heterogeneous distribution, and a higher degree of expression of this glycoprotein is seen in lesions of greater dermal invasiveness. TN expression does not correlate with that of other ECM glycoproteins such as fibronectin. In vivo and in vitro experiments have also demonstrated that melanoma cells are capable of producing and releasing TN in the extracellular milieu.


Conditioned fear suppresses light-induced resetting of the circadian clock.
The possibility that circadian rhythms can be modulated by emotional state is suggested by clinical evidence of altered physiological and endocrine rhythms in primary depression and related affective disorders and is supported by experiments in humans and laboratory animals showing that stress disrupts circadian rhythmicity. How emotional state might modulate circadian rhythms is not known. Here we report that induction of the emotional state of fear disrupts a process essential for stable entrainment of circadian rhythms, the resetting of the circadian clock by environmental light. A cellular correlate of light-induced clock resetting, expression of the transcription factor Fos in the suprachiasmatic nucleus of the hypothalamus (the circadian clock), and a behavioral measure of clock resetting, phase shifts in free-running activity rhythms, were found to be suppressed in rats exposed to light in a context made to induce fear by previous pairings with intermittent footshock. These findings show that fear disrupts a physiological process mediating light-induced clock resetting and suggest a mechanism through which emotional state could modulate circadian rhythms.


Fumarate reductase of Clostridium formicoaceticum. A peripheral membrane protein.
When Clostridium formicoaceticum was grown on fumarate or L-malate crude cell extracts contained a high fumarate reductase activity. Using reduced methyl viologen as electron donor the specific activity amounted to 2-3.5 U per mg of protein. Reduced benzyl viologen, FMNH2 and NADH could also serve as electron donors but the specific activities were much lower. The NADH-dependent activity was strictly membrane-bound and rather labile. Its specific activity did not exceed 0.08 U per mg of particle protein. Fumarate reductase activity was also found in cells of C. formicoaceticum grown on fructose, gluconate, glutamate and some other substrates. The methyl viologen-dependent fumarate reductase activity could almost completely be measured with intact cells whereas only about 25% of the cytoplasmic acetate kinase activity was detected with cell suspensions. The preparation of spheroplasts from cells of C. formicoaceticum in 20 mM HEPES-KOH buffer containing 0.6 M sucrose and 1 mM dithioerythritol resulted in the specific release of 88% of the fumarate reductase activity into the spheroplast medium. Only small amounts of the cytoplasmic proteins malic enzyme and acetate kinase were released during this procedure. There results indicate a peripheral location of the fumarate reductase of C. formicoaceticum on the membrane.


Cost effectiveness of nutrition support in the prevention of pressure ulcer in hospitals.
This study estimates the economic outcomes of a nutrition intervention to at-risk patients compared with standard care in the prevention of pressure ulcer. Statistical models were developed to predict 'cases of pressure ulcer avoided', 'number of bed days gained' and 'change to economic costs' in public hospitals in 2002-2003 in Queensland, Australia. Input parameters were specified and appropriate probability distributions fitted for: number of discharges per annum; incidence rate for pressure ulcer; independent effect of pressure ulcer on length of stay; cost of a bed day; change in risk in developing a pressure ulcer associated with nutrition support; annual cost of the provision of a nutrition support intervention for at-risk patients. A total of 1000 random re-samples were made and the results expressed as output probability distributions. The model predicts a mean 2896 (s.d. 632) cases of pressure ulcer avoided; 12, 397 (s.d. 4491) bed days released and corresponding mean economic cost saving of euros 2 869 526 (s.d. 2 078 715) with a nutrition support intervention, compared with standard care. Nutrition intervention is predicted to be a cost-effective approach in the prevention of pressure ulcer in at-risk patients.


An unusual foreign body in the bladder 7 years after a Stamey endoscopic bladder neck suspension.
Delayed migration of the suture and bolster after an endoscopic bladder neck suspension across tissue planes, with subsequent erosion into the bladder, is uncommon. We present a case of late migration of the suture and bolster occurring 7 years after a Stamey endoscopic bladder neck suspension. A 56-year-old woman had undergone a Stamey procedure in June 1990. In January 1997 she presented with discomfort in the left iliac fossa and the groin. A midstream sample of urine showed microscopic hematuria. Imaging and endoscopic examinations revealed a calcified lesion on the left lateral wall of the bladder, attached to the Stamey sutures. Cystolitholapaxy was attempted, but during the procedure it became obvious that there was a calcified cuff attached to the suture. This was removed endoscopically, along with its suture. Cystoscopy should be considered early in the evaluation of patients presenting with lower abdominal discomfort or irritative voiding symptoms after retropubic bladder neck suspension.


Pharmacokinetics of Ro 03-8799 in mice bearing melanosarcoma: comparison with tumors without melanin.
The pharmacokinetics of Ro 03-8799 has been studied in melanic and non-melanic tumor bearing mice after iv administration of 150 mg/kg. The peak concentration in B16 melanosarcoma tumor reached 152 micrograms/g, that is 7.6-fold higher than the plasma concentration at the same time. This concentration is 3-times greater than that obtained in the tumor of mice bearing non melanic sarcoma (DB16) or Lewis lung carcinoma (3LL). The exposure of B16 tumor (AUC) is respectively 15-times and 11-times higher than the 3LL and the DB16 ones. These experimental data confirm that this 2-nitro-imidazol compound has an important affinity for melanin and suggest that it might be used as a radiosensitizer for the treatment of malignant melanoma.


Hippocampal replays appear after a single experience and incorporate greater detail with more experience.
The hippocampus is implicated in memory formation, and neurons in the hippocampus take part in replay sequences that have been proposed to reflect memory of explored space. By recording from large ensembles of hippocampal neurons as rats explored various tracks, we show that sustained replay appears after a single experience. Further, we found that with repeated experience in a novel environment, replay slows down, taking more time to traverse the same trajectory. This effect was dependent on experience, not passage of time, and was environment specific. By investigating the slow-gamma (25-50 Hz) hover-and-jump dynamics within replays, we show that replay slows by adding more hover locations, increasing the resolution of the behavioral trajectory. We provide evidence that inhibition and cortical engagement both increase as replay slows. Thus, replays can reflect single experiences and evolve with re-exposure, in a manner consistent with the encoding of greater detail into replay memories with experience.


Limiting factors for panicle photosynthesis at the anthesis and grain filling stages in rice (Oryza sativa L.).
Panicle photosynthesis is crucial for grain yield in cereal crops; however, the limiting factors for panicle photosynthesis are poorly understood, greatly impeding improvement in this trait. In the present study, pot experiments were conducted to investigate the limiting factors for panicle photosynthesis at the anthesis stage in seven rice genotypes and to examine the temporal variations in photosynthesis during the grain filling stage in the Liangyou 287 genotype. At the anthesis stage, leaf and panicle photosynthesis was positively correlated with stomatal conductance and maximum carboxylation rate, which were in turn associated with hydraulic conductance and nitrogen content, respectively. Panicle hydraulic conductance was positively correlated with the area of bundle sheaths in the panicle neck. During grain filling, leaf and panicle photosynthesis remained constant at the early stage but dramatically decreased from 8 to 9 days after anthesis. The trends of variations in panicle photosynthesis were consistent with those in stomatal conductance but not with those in maximum carboxylation rate. At first, the maximum carboxylation rate and respiration rate in the panicle increased, through elevated panicle nitrogen content, but then drastically decreased, as a result of dehydration. The present study systematically investigated the limiting factors for panicle photosynthesis, which are vital for improving photosynthesis and crop yield.


Prevalence of atherosclerotic involvement of the internal carotid artery in hypertensive patients.
The prevalence of atherosclerotic involvement of the internal carotid arteries, as diagnosed through an echo-Doppler imaging system with pulsed Doppler spectral analysis was evaluated in 49 hypertensives who had a negative history for neurological symptoms and 49 matched controls. The prevalence was 24.5% in the hypertensive group and 10.2% in the controls with a statistically significant difference (chi-square = 6.07, P less than 0.01). Two hypertensives had severe stenosis (above 50% diameter reduction) and 7 had potentially embolic lesions (irregular surface, inhomogeneous appearance). No one of the matched controls was as severely involved. We conclude that arterial hypertension can account for enhanced prevalence of carotid artery disease in asymptomatic patients.


Embolization of a tentorial dural arterio-venous fistula presenting as atypical trigeminal neuralgia.
A patient with a tentorial dural AV fistula causing atypical trigeminal neuralgia (TN) successfully treated by embolization is reported. The patient developed persisting throbbing facial pain in the distribution of the second and third division of the right trigeminal nerve (V2, V3) after a history of typical neuralgia for one year, preceded by a two month spell of TN 6 years previously and accompanied by right-sided pulsatile tinnitus for 10 years. The patient's mother, brother and sister were also said to be affected by typical trigeminal neuralgia. A right-sided dilated vein of Rosenthal due to a dural AV fistula fed by branches of the meningeal, occipital and meningo-hypophyseal trunk of the internal carotid artery was thought to cause trigeminal nerve compression. Complete resolution of symptoms after partial intra-arterial embolization of the main feeding arteries with N-butyroacrylate is described.


Interaction of OmpR, a positive regulator, with the osmoregulated ompC and ompF genes of Escherichia coli. Studies with wild-type and mutant OmpR proteins.
The OmpR protein is a positive regulator involved in osmoregulatory expression of the ompC and ompF genes that specify the major outer membrane proteins OmpC and OmpF, respectively. We purified the OmpR protein not only from wild-type cells but also from two ompR mutants (ompR2 and ompR3) exhibiting quite different phenotypes as to osmoregulation of the ompC and ompF genes. The OmpR2 protein has an amino acid conversion in the C-terminal portion of the OmpR polypeptide, whereas the OmpR3 protein has one in the N-terminal portion. Comparative studies on these purified OmpR proteins were carried out in terms of their interaction with the ompC and ompF promoters. The nucleotide sequences involved in OmpR-binding were determined in individual promoter regions by deoxyribonuclease I footprinting. The OmpR3 protein as well as the wild-type OmpR protein appeared to bind, to similar extents, to both the ompC and ompF promoters. In contrast, the OmpR2 protein bound preferentially to the ompF promoter and failed to protect the ompC promoter against DNAse I digestion. These results support the view that the C-terminal portion of the OmpR protein is responsible for the binding of the OmpR protein to the ompC and ompF promoter DNAs. Based on these results, the structure and function of the OmpR protein are discussed in relation to the mechanism of osmoregulation.


Late Intramedullary Spinal Metastases from Esthesioneuroblastoma: Case Report and Prognostic Implications.
Esthesioneuroblastoma (ENB) is an uncommon neuroectodermal tumor that originates from the olfactory mucosa and often recurs locally. Distant metastases of ENB have been described, but there are few reports of intramedullary metastases to the spinal cord. Here we report a case of a patient presenting with a progressive paraparesis and magnetic resonance imaging findings of multiple drop metastases to thoracic and lumbar regions of the spinal cord, 17 years after diagnosis and treatment for an intracranial ENB with subsequent neck nodal metastases. The dorsal symptomatic lesion was treated with resection, radiotherapy, and adjuvant chemotherapy. The implications of spinal metastases for the clinical prognosis of this disease are discussed, with a review of the few reported cases of spinal ENB metastases in the literature. Through the presentation of this case we hope to further contribute to a better understanding of this rare disease's prognosis.


Is alexithymia related to cannabis use disorder? Results from a case-control study in outpatient adolescent cannabis abusers.
In recent decades, alexithymia has been considered a risk factor for multiple somatic and psychiatric conditions. Yet, whereas alexithymia has been extensively studied in adults with a substance misuse, only one study has reported data on cannabis abusers from the general population. Hence, our main objective was to explore alexithymia in a clinical sample of treatment-seeking young outpatients with a DSM-IV cannabis dependence or abuse diagnosis compared to controls. 120 young patients (95 males - mean age 17.9years (SD=2.8; 14 to 25)) with a cannabis dependence or abuse (DSM-IV-TR criteria evaluated with the MINI), seeking treatment in an addiction unit, and 110 healthy control subjects (77 males - mean age 18.2years (SD=3.4; 14 to 25)) participated in the study. They completed a battery of self-reports measuring alexithymia (TAS-20; BVAQ-B), depression (BDI-13) and state and trait anxiety (STAI). 35.3% of cannabis users were alexithymic, and logistic regression analysis showed that the alexithymic components of difficulties identifying and describing feelings combined with trait anxiety predicted group membership. This first study on young cannabis abusers and dependent subjects further emphasizes the importance of considering the affective style, and particularly the anxious temperament and alexithymia features, as factors associated with substance misuse during late adolescence.


The effects of ageing on the pigmented nerve cells of the human locus caeruleous and substantia nigra.
The nucleolar volume and melanin content of the human locus caeruleus and substantia nigra has been measured in 70 persons of age range new born to 91 years, all of whom were at death free from overt neurological illness. Both cell types show a reduction in nucleolar size with advancing age, which becomes disproportionately larger towards old age. At 90 years of age, the change in nucleolar volume in cells of the locus caeruleus amounts to about 5% whereas cells of the substantia nigra show a loss of 20%. The greater decrease in nucleolar volume in cells of the substantia nigra is attributed to the higher concentration of melanin pigment occurring in these cells at old age, rather than the absolute amount present. This marked decline in nucleolar volume in cells of substantia nigra indicates a reduced activity in the cell which, in turn, may be reflected in the difficulty in control and coordination in muscular activity commonly seen in normal elderly persons.


Development and validation of a Medication Adherence Universal Questionnaire: the MAUQ.
Different questionnaires assess self-reported medication adherence and others quantify aspects of patients attitudes towards medication, but not together in a single instrument. Gathering these two aspects in a single instrument could reduce patients survey burden. The aim of this study was to develop the Medication Adherence Universal Questionnaire (MAUQ) using the Maastricht Utrecht Adherence in Hypertension short version (MUAH-16) factorial structure as the hypothesized model. A multistep process started with the modification of the MUAH-16 to obtain the MAUQ. Patients using at least one antihypertensive medicine were recruited. The two questionnaires, the MUAH-16 and MAUQ, were applied. A confirmatory factor analysis (CFA) was performed using the initial MUAH-16 s-order 4-factor model. An additional bifactor model with four uncorrelated factors and an overall score was tested. The comparative fit index (CFI), root mean square error of approximation (RMSEA) with confidence intervals (CIs), and standardized root mean squared residual (SRMR) were used to assess both models. A sample of 300 hypertensive patients completed the instruments. The CFA with the second-order 4-factor solution resulted in similar results for the MUAH-16 and MAUQ: CFIs of 0.934 and 0.930, RMSEAs of 0.043 [CI 0.030-0.056] and 0.045 [CI 0.031-0.057] and SRMRs of 0.060 and 0.061, respectively. The CFA with the bifactor model showed slightly better results for both the MUAH-16 and MAUQ: CFIs of 0.974 and 0.976, RMSEAs of 0.030 [CI 0.005-0.046] and 0.028 [CI 0.001-0.044], and SRMRs of 0.043 and 0.044, respectively. CFA demonstrated that the MAUQ presented a better fit to both models than the MUAH-16, obtaining a robust universal free instrument to assess medicine-taking behaviour and four medicine beliefs components.


Current practice of rapid sequence induction in adults: A national survey among anesthesiologists in Spain.
Rapid sequence induction (RSI) in adults has undergone changes in recent years due to pharmacological and technological advances. The objective of this survey was to evaluate current practice among Spanish anesthesiologists. A 31-item questionnaire regarding RSI practice was sent to anesthesiologists working in Spanish public hospitals. Differences in responses according to the type of hospital or experience of the respondent were compared for all data using the chi-square and Fisher's exact test. Approximately 15.89% of Spanish anesthesiologists participated in the survey (1002 questionnaires). The results show considerable heterogeneity in most aspects of RSI. Less than 20% of respondents administer sodium citrate. Sixty-four percent place a nasogastric tube in patients with intestinal obstruction. Gastric residue is assessed by ultrasound in 6% of cases. Only 25% of respondents measure ETO2 to check the effectiveness of preoxygenation, and 22% use nasal oxygen insufflation with nasal prongs or THRIVE. Sixty two percent of respondents apply cricoid pressure, but only 50% release the pressure when encountering intubation difficulty. Up to 40% of respondents reported cases of aspiration despite applying cricoid pressure. Propofol was the most commonly used hypnotic (97.6%), but there was no clear preference in the choice of neuromuscular relaxant (suxamethonium versus rocuronium ratio of approximately 1:1). Only 44% of respondents calculated the dose of sugammadex that would be required for emergency reversal of rocuronium. The survey showed significant variation in RSI practice, similar to that of other countries. Quality prospective studies are needed to standardize clinical practice.


YY1-mediated overexpression of long noncoding RNA MCM3AP-AS1 accelerates angiogenesis and progression in lung cancer by targeting miR-340-5p/KPNA4 axis.
Lung cancer is famous as an aggressive malignant tumor and is the main cause of cancer-associated mortality globally. Tumor angiogenesis is a vital part in cancer, which influences cell proliferation and metastasis. Increasing studies have claimed that long noncoding RNAs (lncRNAs) were involved in the progression of several cancers. Based on previous studies, this study focused on the role and mechanism of lncRNA MCM3AP antisense RNA 1 (MCM3AP-AS1) in lung cancer. At first, MCM3AP-AS1 expression was found to be elevated in lung cancer cells. Depletion of MCM3AP-AS1 repressed cell proliferation, migration, and angiogenesis in lung cancer cells. YY1 was confirmed to mediate MCM3AP-AS1 transcription in lung cancer cells. Moreover, the molecular mechanism investigation revealed that MCM3AP-AS1 could sponge miR-340-5p and elevate KPNA4 expression. On the basis of rescue assays, we identified that the overexpression of KPNA4 partly counteracted the suppressed effect of MCM3AP-AS1 knockdown on angiogenesis and progression in lung cancer cells. Conclusively, the YY1-mediated overexpression of MCM3AP-AS1 accelerated angiogenesis and progression in lung cancer by targeting miR-340-5p/KPNA4 axis, which highlighted the possibility of MCM3AP-AS1 as a promising therapeutic target for lung cancer.


Factors increasing poly-γ-glutamic acid content of cheongguk-jang fermented by Bacillus subtilis 168.
Cheongguk-jang is a Korean traditional food produced by natural fermentation of boiled soybean. In cheongguk-jang, bacilli are dominant bacteria and produce highly viscous poly-γ-glutamic acid (γ-PGA), which improves human health functions. The purpose of this experiment was to find maximum production condition for the γ-PGA content during fermentation of cheongguk-jang with Bacillus subtilis 168. The most viscous cheongguk-jang was produced when soybean was cooked at 121 °C for 60 min in the presence of 50%(w/w) added water, followed by fermentation at 40 °C for 2 days. Additional conditions for maximum production of γ-PGA were the addition of 0.1%(w/w) FeCl3·6H2O, 3.0%(w/w) lactose and 3.0%(w/w) yeast extract as nutrients of inorganic salts, carbon source and nitrogen source, respectively. The three conditions did not show cumulative effect on the γ-PGA production and the addition of iron salt induced the most γ-PGA (0.97 ± 0.05%(w/w)), which corresponded to 2.7 times of the content in control cheongguk-jang.


In vivo therapeutic exploring for Mori folium extract against type 2 diabetes mellitus in rats.
The study was aimed to investigate the potential therapeutic effect of Mori folium aqueous extracts (MFAE) on type 2 diabetes mellitus (T2DM) in vivo. A rat model of T2DM was established with the combination of high sugar and high-fat diet (HSFD) and streptozotocin (STZ). The T2DM rats were administrated with low (2 g.kg-1) and high (5 g.kg-1) doses of MFAE for 60 consecutive days. The biochemical indices of glucose metabolism disorders, insulin resistance and oxidative stress were observed. The results indicated that MFAE significantly promoted the synthesis of hepatic glycogen, reduced the levels of fasting blood glucose and fasting blood insulin, and improved the insulin sensitivity index (ISI). MFAE administration also remarkably increased the levels of superoxide dismutase (SOD) and reduced the levels of malondialdehyde (MDA). MFAE showed a therapeutic effect on T2DM with the bioative effect of improve glucose metabolism disorders, decrease insulin resistance, and ameliorate the antioxidative ability.


Proliferative vitreoretinopathy and chemotherapeutic agents.
Proliferative vitreoretinopathy (PVR) is a disease process that occurs in eyes with rhegmatogenous retinal detachments and accounts for the majority of failures following retinal detachment surgery. PVR involves the uncontrolled proliferation of non-neoplastic cells capable of forming membranes, which may occur on either surface of the retina or along the detached surface of the vitreous gel. Contraction of these membranes creates tractional forces that can distort or detach the retina. Various surgical procedures have been used to repair retinal detachments associated with PVR. The results have not been encouraging in many instances. Recent efforts have been directed toward the chemical inhibition of cellular proliferation in PVR. The majority of drugs used in these studies have been antineoplastic agents that affect various phases in the cycle of cell growth.


[Management of a patient presenting with LAV seropositivity].
LAV-HTLV III serologic out-patient's supervision (clinical an immunological status), general practitioner and family medicine practitioners are able to follow up out-patients with a positive LAV-HTLV III serology. However the lack of IgM LAV-HTLV III serology is not enable to make secure our patients about the RETRO-VIRUS biological activity in their FUTURE.


Age-related increase in the total number of corticotropin-releasing hormone neurons in the human paraventricular nucleus in controls and Alzheimer's disease: comparison of the disector with an unfolding method.
It has been hypothesized that the corticotropin-releasing hormone (CRH) neurons of the hypothalamic paraventricular nucleus (PVN) become hyperactive with age, and even more so in Alzheimer's disease. This hyperactivity could be due to an increased production of CRH per neuron, or an increased number of PVN neurons producing CRH, or both. As a first step in elucidating which of these biological mechanisms might be operative, we have estimated the absolute number of CRH immunoreactive neurons in the PVN of 10 human control subjects between 36 and 91 years of age and 10 Alzheimer patients between 40 and 97 years of age. CRH neurons were immunocytochemically detected in 6 microns paraffin sections with the aid of a highly specific monoclonal antibody to CRH. The antibody signal was amplified by the biotin-streptavidin and alkaline phosphatase methods. The absolute number of CRH neurons in the PVN was obtained by multiplying the number of CRH neurons in a unit volume (NV) by the total volume of the PVN. Two different methods were used to estimate the NV: an unfolding method and a disector method (about three times more time-consuming). Compared to the disector, the unfolding method consistently yielded a lower cell number for all patients by 38% (+/- 2.8%; mean +/- SEM). However, both methods yielded an increase in the absolute number of CRH neurons in control and Alzheimer patients with age. No statistically significant difference in the absolute number of CRH neurons was found between control and Alzheimer patients with both methods. The age-dependent increase in the absolute number of CRH neurons within the PVN of both control and Alzheimer patients is interpreted as a sign of activation of the CRH neurons with age.


The role of microtopography in cellular mechanotransduction.
Mechanotransduction is crucial for cellular processes including cell survival, growth and differentiation. Topographically patterned surfaces offer an invaluable non-invasive means of investigating the cell response to such cues, and greater understanding of mechanotransduction at the cell-material interface has the potential to advance development of tailored topographical substrates and new generation implantable devices. This study focuses on the effects of topographical modulation of cell morphology on chromosomal positioning and gene regulation, using a microgrooved substrate as a non-invasive mechanostimulus. Intra-nuclear reorganisation of the nuclear lamina was noted, and the lamina was required for chromosomal repositioning. It appears that larger chromosomes could be predisposed to such repositioning. Microarrays and a high sensitivity proteomic approach (saturation DiGE) were utilised to identify transcripts and proteins that were subject to mechanoregulated changes in abundance, including mediators of chromatin remodelling and DNA synthesis linked to the changes in nucleolar morphology and the nucleoskeleton.


Chronic kidney disease prevention--a challenge for Asian countries: report of the Third Asian Forum of Chronic Kidney Disease Initiatives.
As an independent meeting, the third Asian Forum of Chronic Kidney Disease Initiatives was held on April 18-19, 2009, in Kaohsiung, Taiwan. Nearly 700 participants from 17 countries attended and 78 posters were presented. To begin with, status quo in Asia and in Taiwan was briefed, followed by Theme 1A "Special Epidemiology and Risk Factors", during which the interrelations between chronic kidney disease (CKD) and use of herbs, low birth weight, infections, and immunoglobulin A (IgA) nephropathy were discussed. Theme 1B dealt with both cardiovascular and renal outcomes of CKD patients. In Theme 2, five presenters from different countries shared their experiences on "Cost-effectiveness of Community-based or Nationwide CKD Prevention Programs". In between the conference themes, three international and integrative works--Kidney Disease Improving Global Outcomes, the World Kidney Day, and Kidney Early Evaluation Program--were presented and possible implications for Asia were suggested. Theme 3 was initiated with a thorough discussion on "Equation of the Estimated Glomerular Filtration Rate for Asians" and the preliminary results of a cross-country study were presented. In Theme 4, the care plan, strategies, and outcomes of timely initiation of dialysis in different countries were discussed. The final session started with a concise summary of all speeches and ended with a position statement.


Impact of labor induction, gestational age, and maternal age on cesarean delivery rates.
To quantify the impact of labor induction and maternal age on cesarean delivery rates in nulliparous and multiparous women between 36 and 42 weeks' gestation. We performed a retrospective cohort study on 14,409 women delivering at two teaching hospitals in metropolitan Boston during 1998 and 1999. Women who had contraindications to labor, including a prior cesarean delivery, were excluded. The risks for cesarean delivery by induction status, gestational age by completed week between 36 and 42 weeks, maternal age <35, 35-39, and >/=40 years, and stratified by parity, were calculated by logistic regression. In nulliparas, labor induction was associated with an increase in cesarean delivery from 13.7% to 24.7% (adjusted odds ratio [OR] 1.70; 95% confidence interval [CI] 1.48, 1.95]). In multiparas, induction was associated with an increase from 2.4% to 4.5% (OR 1.49; 95% CI 1.10, 2.00). Other variables that placed a nulliparous woman at increased risk for cesarean delivery included maternal age of at least 35 years and gestational ages over 40 weeks. For multiparas, only maternal age 40 years or older and gestational age of 41 weeks were associated with an increase in cesarean deliveries. Induction of labor, older maternal age, and gestational age over 40 weeks each independently increase the risk for cesarean delivery in both nulliparous and multiparous women. Although the relative risk from induction is similar in nulliparas and multiparas, the absolute magnitude of the increase is much greater in nulliparas (11% versus 2.1%).


Identification, developmental regulation, and response to heat shock of two antigenically related forms of a major nuclear envelope protein in Drosophila embryos: application of an improved method for affinity purification of antibodies using polypeptides immobilized on nitrocellulose blots.
An affinity-purification method has been developed for the rapid, efficient, and precise elution of antibodies specifically bound to antigens immobilized on nitrocellulose after blot transfer from SDS polyacrylamide gels. The applicability of this technology has been demonstrated using antisera raised against the nuclear matrix-pore complex-lamina fraction prepared from Drosophila melanogaster embryos. In so doing, we have established the existence in whole embryo lysates, of two nearly identical forms of the predominant 74-kilodalton polypeptide previously identified in lower resolution studies of the nuclear matrix-pore complex-lamina fraction. These species, distinguishable on the basis of a slight difference in SDS PAGE mobilities on low concentration polyacrylamide gels, are immunochemically cross-reactive and have been localized exclusively to the nuclear periphery (nuclear envelope) by indirect immunofluorescence analyses of cryosections. The steady-state levels of these two polypeptides have been examined in total embryo lysates both as a function of embryogenesis and in response to heat shock. The larger species is not detectable in early embryos but approaches levels approximately equal to that of the smaller form by about the temporal midpoint of embryonic development. In response to heat shock, this larger form appears to be converted nearly quantitatively into the lower molecular weight polypeptide. These results, as well as the general reliability of the nitrocellulose blot immunoaffinity-purification methodology, have been substantiated through the use of monoclonal antibodies.


Pharmacological and biochemical studies on a new compound, 2-(7-indeyloxymethyl)morpholine hydrochloride (YM-08054-1), and its derivatives with potential antidepressant properties.
Antidepressant properties of a new indene derivative, YM-08054-1, and its related compounds were compared with those of tricyclic antidepressants and viloxazine. The potencies of YM-08054-1 to inhibit uptake of both 14C-norepinephrine (14C-NE) and 14C-5-hydroxytryptamine (14C-5-HT) by the rat brain synaptosomes were similar to those of amitriptyline and imipramine. Other indene derivatives with an N-alkylated morpholine ring were proved to have less effect on the uptake of either 5-HT or NE than was YM-08054-1. YM-08054-1 was the most potent among all of the tested antidepressants in the inhibition of reserpine-induced facilitation of convulsions as well as in the potentiation of reserpine-induced facilitation of convulsions as well as in the potentiation of 5-hydroxytryptophan (5-HTP)-induced syndromes in mice, though the inhibitory effect of this agent on reserpine-induced hypothermia was weaker than that of either amitriptyline or desipramine, suggesting relatively selective effects of YM-08054-1 upon 5-HT rather than NE uptake in vivo. Neither viloxazine nor iprindole potentiated the responses to 5-HTP. YM-08054-1 was devoid of peripheral anticholinergic activity and exhibited weak local anesthetic effect as well as low acute toxicity when compared with amitriptyline. The results indicate that YM-08054-1 has a novel profile as an antidepressant agent which is quite different from that of either viloxazine or tricyclic compounds.


Indolequinone antitumor agents: relationship between quinone structure and rate of metabolism by recombinant human NQO1.
A series of indolequinones bearing various functional groups has been synthesized, and the effects of substituents on the metabolism of the quinones by recombinant human NAD(P)H:quinone oxidoreductase (NQO1), and on the toxicity toward nonsmall cell lung cancer cells with either high NQO1 activity (H460) or with no detectable activity (H596) were studied.


RETRACTED: Blockade of TNF-α signaling suppresses the AREG-mediated IL-6 and IL-8 cytokines secretion induced by anti-Ro/SSA autoantibodies.
The aim of this study was to analyze the Furin-TNF-α-converting enzyme (TACE)-amphiregulin (AREG)-IL-6/IL-8 secretion pathway in non-neoplastic human salivary gland epithelial cells (SGECs) stimulated with anti-Ro/SSA autoantibodies (Abs). We examined whether anti-Ro/SSA Abs-mediated TACE activation is responsible for AREG activation. As recent studies have demonstrated that AREG could induce proinflammatory cytokines secretion in epithelial cells, we discuss how TACE-mediated AREG shedding, caused by anti-Ro/SSA Abs treatment, could have a critical role in TNF-α-induced IL-6 and IL-8 secretion by SGEC. Furthermore, the effects of TNF-α blockade on AREG expression and TNF-α-AREG-mediated IL-6 and IL-8 secretion were evaluated. We have discovered that the upregulation of AREG occurs through TNF-α produced after anti-Ro/SSA Abs uptake via Fcγ receptors. Biological drug adalimumab and the gene silencing technique were used to study the AREG-IL-6/IL-8 secretion pathway, demonstrating that (i) adalimumab-mediated TNF-α blocking and TNF-α gene silencing provoke a significant decrease of proinflammatory cytokines production and AREG expression in anti-Ro/SSA Abs-treated SGEC; (ii) AREG gene silencing has a potent inhibitory effect on TNF-α-induced IL-6 and IL-8 secretion in SGEC treated with anti-Ro/SSA Abs; (iii) an inspection of the kinetics of cytokine production after exogeni TNF-α and AREG addition, and the use of cycloheximide in the presence of exogenous TNF-α as stimulant, clarified that TNF-α induces IL-6 and IL-8 secretion through AREG.Laboratory Investigation advance online publication, 20 September 2010; doi:10.1038/labinvest.2010.168.


Application of image analysis for quantitative immunoperoxidase detection of IgE containing cells in lymphoid tissues.
Methods for the quantitation of antigen or antibody specific cells within lymphoid structures rely on the laborious and often inaccurate counting of stained cells visually under the microscope. To circumvent these problems we used a combination of computer-linked image analysis, with the Quantimet 720 and specific staining for IgE by immunoperoxidase, to produce a relatively easy, accurate method for the quantitation of specifically stained cells within tonsils and lymph nodes. Frozen sections of bovine lymph nodes and tonsils were stained in an indirect immunoperoxidase technique with either rabbit anti-bovine IgE or normal rabbit serum as the primary reagent. Examination of the stained slides on the Quantimet 720 image analyzing computer revealed cells that were sufficiently dark to be considered above the 'threshold level' of background staining. These were counted as positive picture points. A quantitative evaluation of the relative number of IgE containing cells within the particular lymphoid tissue was arrived at by determining the difference between the percent positive picture in the control tissue incubated with normal rabbit serum (background) and in the sample treated with rabbit anti-IgE. Using this method, it was readily possible to quantitate the presence of IgE containing cells in the tonsils of calves; the highest percentage of positive tissue was present in the tonsils of calves exposed to ovalbumin. Image analysis is a useful technique for quantitative evaluation of any immunoperoxidase stained tissues.


Evaluation of the accuracy and intra- and interobserver reliability of three manual laxity tests for canine cranial cruciate ligament rupture-An ex vivo kinetic and kinematic study.
To investigate the accuracy and intra- and interobserver reliability of the cranial drawer test (CD), tibial compression test (TCT), and the new tibial pivot compression test (TPCT) in an experimental setting resembling acute cranial cruciate ligament rupture (CCLR) and to elucidate the ability to subjectively estimate cranial tibial translation (CTT) during testing. Experimental ex vivo study. Ten cadaveric hindlimbs of large dogs. Kinetic and 3D-kinematic data was collected while three observers performed the tests on each specimen with intact (INTACT) and transected cranial cruciate ligament (CCLD) and compared using three-way repeated-measures ANOVA. Subjectively estimated CTT (SCTT), obtained during a separate round of testing, was compared to kinematic data by Pearson correlation. CTT was significantly higher for CCLD than for INTACT for all tests, resulting in 100% sensitivity and specificity. TPCT induced the highest CTT and internal rotation. Intra- and interobserver agreement of translation was excellent. For rotation and kinetics, agreement was more variable. SCTT strongly correlated with the objectively measured values. The CD, TCT and the new TPCT were all accurate and reliable. The high translations and rotations during TPCT are promising, encouraging further development of this test. SCTT was reliable in our experimental setting. Veterinary manual laxity tests are accurate and reliable in acute CCLR. The TPCT might have potential for the assessment of subtle and rotational canine stifle instabilities. The high reliability of SCTT implies that grading schemes for stifle laxity, similar to human medicine, could be developed.


Deubiquitinating enzymes UBP12 and UBP13 regulate carbon/nitrogen-nutrient stress responses by interacting with the membrane-localized ubiquitin ligase ATL31 in Arabidopsis.
Ubiquitination is an important post-translational modification that regulates multiple cellular activities in plants including environmental stress responses. In addition to activity of ubiquitin ligases, the activity of deubiquitinating enzymes (DUBs) is critical for modulating the optimal ubiquitination status of target proteins in response to environmental stimuli. However, while several ubiquitin ligases have been isolated to date, little is known about the DUBs involved in plant stress responses. Here, we report that two DUBs, UBP12 and UBP13, function in response to disrupted carbon (C)/nitrogen (N)-nutrient stress conditions in Arabidopsis. Knockdown of UBP12 and UBP13 expression resulted in hypersensitivity to high C/low N-nutrient stress conditions, whereas overexpression of UBP13 reduced the sensitivity. Additionally, UBP13 physically interacted with and deubiquitinated the ubiquitin ligase ATL31, a key regulator of plant resistance to high C/low N-nutrient stress conditions. Genetic analysis showed that the loss of ATL31 and its homolog ATL6 suppressed the high C/low N-hyposensitivity of UBP13-overexpressing plants, suggesting that ATL31 is epistatic to UBP12 and UBP13. Taken together, our results suggest that the DUBs UBP12 and UBP13 function together with the ubiquitin ligase ATL31 to mediate C/N-nutrient stress responses in plants.