Applications of differential-display reverse transcription-PCR to molecular pathogenesis and medical mycology.
The host-fungus interaction is characterized by changes in gene expression in both host and pathogen. Differential-display reverse transcription PCR (DDRT-PCR) is a PCR-based method that allows extensive analysis of gene expression among several cell populations. Several limitations and drawbacks to this procedure have now been addressed, including the large number of false-positive results and the difficulty in confirming differential expression. Modifications that simplify the reaction time, allow the use of minute quantities of RNA, or address unusual species- or gene-specific sequences have been reported. DDRT-PCR has been used to address biological questions in mammalian systems, including cell differentiation, cell activation, cell stress, and identification of drug targets. In microbial pathogenesis and plant pathogenesis, DDRT-PCR has allowed the identification of virulence factors, genes involved in cell death, and signaling genes. In Candida albicans, DDRT-PCR studies identified TIF-2, which may play a role in the upregulation of phospholipases, and the stress-related genes, CIP1 and CIP2. In Histoplasma capsulatum and C. albicans, genes involved in the host-pathogen interaction, including a member of the 100-kDa family in Histoplasma and an ALS and 14-3-3 gene in Candida, were potentially identified by DDRT-PCR. Although very few reports have been published in medical mycology, studies in mammalian, nonfungal microbial, and plant pathogen systems are easily applied to basic questions in fungal pathogenesis and antifungal therapeutics.


Natural history of homonymous hemianopia.
To describe the characteristics of spontaneous recovery of homonymous hemianopia (HH). The authors reviewed medical records of all patients with HH confirmed by formal visual field testing and seen in follow-up in their service between 1989 and 2004. Clinical characteristics, causes, neuroradiologic definition of lesion location, final outcome, and evolution of the visual field defects were recorded. The associations among final visual field defect outcome, time from injury, and clinical features were analyzed. A total of 254 patients with 263 HH were included in this study. Spontaneous visual field defect recovery was observed in 101 HH (38.4%). The likelihood of spontaneous recovery decreased with increasing time from injury to initial visual field testing (p = 0.0003). The probability of improvement was related to the time since injury (p = 0.0003) with a 50 to 60% chance of improvement for cases tested within 1 month after injury that decreased to about 20% for cases tested at 6 months after surgery. No other factor was found to correlate with the final outcome of the visual field defects. Improvement after 6 months from injury was mild and usually related to improvement of the underlying disease. Spontaneous improvement of homonymous hemianopia is seen in at least 50% of patients first seen within 1 month of injury. In most cases, the improvement occurs within the first 3 months from injury. Spontaneous improvement after 6 months postinjury should be interpreted with caution as it is most likely related to improvement of the underlying disease or to improvement in the patient's ability to perform visual field testing reliably.


The Clinical Impact and Cost-Effectiveness of Surveillance of Incidentally Detected Gastric Intestinal Metaplasia: A Microsimulation Analysis.
Gastric intestinal metaplasia (GIM) is associated with a higher risk of noncardia intestinal gastric adenocarcinoma (GA). The aim of this study was to estimate lifetime benefits, complications, and cost-effectiveness of GIM surveillance using esophagogastroduodenoscopy (EGD). We developed a semi-Markov microsimulation model of patients with incidentally detected GIM, to compare the effectiveness of EGD surveillance with no surveillance at 10-year, 5-year, 3-year, 2-year, and 1-year intervals. We modeled a simulated cohort of 1,000,000 US individuals aged 50 with incidental GIM. Outcome measures were lifetime GA incidence, mortality, number of EGDs, complications, undiscounted life-years gained, and incremental cost-effectiveness ratio with a willingness-to-pay threshold of $100,000/quality-adjusted life-year (QALY). In the absence of surveillance, the model simulated 32.0 lifetime GA cases and 23.0 lifetime GA deaths per 1000 individuals with GIM, respectively. Among surveilled individuals, simulated lifetime GA incidence (per 1000) decreased with shorter surveillance intervals (10-year to 1-year, 11.2-6.1) as did GA mortality (7.4-3.6). Compared with no surveillance, all modeled surveillance intervals yielded greater life expectancy (87-190 undiscounted life-years gained per 1000); 5-year surveillance provided the greatest number of life-years gained per EGD performed and was the cost-effective strategy ($40,706/QALY). In individuals with risk factors of family history of GA or anatomically extensive, incomplete-type GIM intensified 3-year surveillance was cost-effective (incremental cost-effectiveness ratio $28,156/QALY and $87,020/QALY, respectively). Using microsimulation modeling, surveillance of incidentally detected GIM every 5 years is associated with reduced GA incidence/mortality and is cost-effective from a health care sector perspective. Real-world studies evaluating the impact of GIM surveillance on GA incidence and mortality in the United States are needed.


RhoA-Dependent HGF and c-Met Mediate Gas6-Induced Inhibition of Epithelial-Mesenchymal Transition, Migration, and Invasion of Lung Alveolar Epithelial Cells.
Previously, we demonstrated that growth arrest-specific protein 6 (Gas6)/Axl or Mer signaling inhibited the transforming growth factor (TGF)-β1-induced epithelial-mesenchymal transition (EMT) in lung epithelial cells. Hepatocyte growth factor (HGF) has also been shown to inhibit TGF-β1-induced changes in EMT markers. Here, we examined whether Gas6 signaling can induce the production of HGF and c-Met in lung alveolar epithelial cells to mediate the inhibition of EMT and to inhibit the migration and invasion of epithelial cells. The inhibition of the RhoA/Rho kinase pathway, using either a RhoA-targeted small interfering RNA (siRNA) or the Rho kinase pharmacologic inhibitor Y27362, prevented the inhibition of TGF-β1-induced EMT in LA-4 cells and primary alveolar type II (AT II) epithelial cells. The c-Met antagonist PHA-665752 also blocked the anti-EMT effects associated with Gas6. Moreover, treatment with Y27362 or PHA-665752 prevented the Gas6-mediated inhibition of TGF-β1-induced migration and invasion. Our data provided evidence that the RhoA-dependent production of HGF and c-Met mediated the Gas6-induced inhibition of EMT, migration and invasion in lung alveolar epithelial cells. Thus, Gas6/Axl and Mer/RhoA signaling may be necessary for the maintenance of homeostasis in the alveolar epithelium, via HGF and c-Met.


Phospholipid asymmetry in rough- and smooth-endoplasmic-reticulum membranes of untreated and phenobarbital-treated rat liver.
Phospholipase C was used as a probe for the distribution of phospholipids about the membrane of rough and smooth microsomal fractions from normal and phenobarbital-treated rat liver. All membranes exhibited an asymmetric distribution, with phosphatidylethanolamine and phosphatidylserine concentrated in the inner leaflet of the bilayer and phosphatidylcholine and sphingomyelin concentrated in the outer leaflet. The only phospholipid showing a significant difference in distribution between fractions was phosphatidylcholine, which was shifted towards the outer leaflet in the smooth microsomal fraction compared with the rough microsomal fraction, and towards the outer leaflet in both rough and smooth microsomal fractions from phenobarbital-treated liver compared with the same preparations from untreated rat liver. Apart from this small change, the asymmetric distribution of phospholipids was conserved in microsomal fractions which had proliferated in response to phenobarbital and in which the protein composition had changed.


A new framework to identify dental emergencies in the COVID-19 era.
In order to reduce the spread of COVID-19 (Coronavirus Disease 2019), it is crucial to take extraordinary prevention and safety measures in dental offices, and to defer all elective and non-urgent procedures. Dental emergencies are defined through oral symptoms but, the systemic and psychological conditions of each patient should be considered. The present short communication proposes a multilevel evaluation (oral, systemic and psychological) and risk assessment score for the management of dental emergencies following the SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2) outbreak. A comprehensive categorization and score scale assessment for dental emergencies allows a better identification of patient's treatment needs and avoids unnecessary contact between dental health care providers and patients during the SARS-CoV-2 pandemic.


Total number of individuals affected by deleterious mutant genes in a finite population.
The means and standard errors of the total numbers of heterozygotes and homozygotes affected by deleterious mutant genes and the extinction time are studied by using diffusion methods. For an overdominant mutation, the effects of an increase in population size on these quantitites are much more profound than that of an increase in initial number of mutant genes whereas for a partially recessive mutation the situation is reversed. For a completely recessive mutation, the expected total number of mutant homozygotes is independent of the population size and degree of inbreeding, though the expected total number of heterozygotes and the average extinction time are dependent on these factors, particularly the population size. The effect of inbreeding on these quantitites is very similar to that of reduction in effective population size and is usally small at the prevailing level of inbreeding, except for mutations with large degrees of overdominance in large populations. The standard errors of these quantities are large. The expected total number of sickle-cell mutant homozygotes in the U.S. population has been computed.


Parasitic plant litter input: a novel indirect mechanism influencing plant community structure.
Parasitic plants have major impacts on plant community structure through their direct negative influence on host productivity and competitive ability. However, the possibility that these parasites may also have indirect impacts on community structure (via the mechanism of nutrient-rich litter input) while long hypothesized, has remained unsupported until now. Using the hemiparasite Rhinanthus minor, we established experimental grassland mesocosms to quantify the impacts of Rhinanthus litter and parasitism across two soil fertility levels. We measured the biomass and tissue nutrient concentration of three functional groups within these communities to determine their physiological response to resource abstraction and litter input by the parasite. We show that Rhinanthus alters the biomass and nutrient status of co-occurring plants with contrasting effects on different functional groups via the mechanism of nutrient-rich litter input. Critically, in the case of grass and total community biomass, this partially negates biomass reductions caused directly by parasitism. This demonstrates that the influence of parasitic plant litter on plant community structure can be of equal importance to the much-reported direct impacts of parasitism. We must consider both positive indirect (litter) and negative direct (parasitism) impacts of parasitic plants to understand their role in structuring plant communities.


Correlation of clinical and physiological effects of cerebellar stimulation.
The value of clinical assessment of patients undergoing chronic cerebellar stimulation (CCS) is limited by lack of objective measures but neurophysiological tests can be used to "biocalibrate" the stimulator and may be used to predict effects of CCS. Eighty-seven patients undergoing CCS have been assessed clinically and neurophysiologically over the last 4 years. Somatosensory evoked responses were significantly ( p less than 0.05) reduced in amplitude in 35 patients, cortical somatosensory evoked responses in 44 patients and one or both responses were reduced in 55 patients. There were no clinical or physiological changes in 16 patients. Evoked responses showed significant changes in only 3 patients who did not show clinical improvement. The mean voltage settings were 5.2 volts and most patients were stimulated at 200 herz. These results indicate that significant changes in those somatosensory evoked potentials are a good indication of clinical benefits from CCS but clinical improvement may occur in the absence of any acute effect on evoked responses.


Surgical considerations in aortitis. Part III: Syphilitic and other forms of aortitis.
The majority of cases involving clinically significant aortitis result from Takayasu's disease, syphilis, and mycotic aneurysms, although aortitis may occur as a part of the clinical manifestation in other diseases of known and unknown etiology. Syphilitic aortitis can be treated successfully if an accurate diagnosis is made early and if appropriate antibiotic therapy is instituted. Untreated or treated inadequately, late cardiovascular complications can occur, namely, aortic aneurysm, aortic valvular insufficiency and coronary osteal stenosis. Surgical treatment should be considered when any of these late complications become manifest. Aortitis can occur with rheumatic fever and, occasionally, in rheumatoid arthritis. Giant-cell arteritis is a disease that can affect large, medium or small arteries, and usually occurs in patients over 50 years of age, chiefly women. The entire aorta may be affected. It is the only form of aortitis commonly associated with dissecting aneurysm of the aorta. Corticosteroids are effective in treating the active form of the disease and can usually prevent the development of later severe complications. Surgery is indicated when a dissecting aneurysm develops.


Influence of Context and Setting on the Mental Health and Wellbeing Outcomes of Ayahuasca Drinkers: Results of a Large International Survey.
Ayahuasca is a traditional plant decoction containing N,N-dimethyltryptamine (DMT) and various β-carbolines including harmine, harmaline, and tetrahydroharmine, which has been used ceremonially by Amazonian Indigenous groups for healing and spiritual purposes. Use of the brew has now spread far beyond its original context of consumption to North America, Europe, and Australia in neo-shamanic settings as well as Christian syncretic churches. While these groups have established their own rituals and protocols to guide use, it remains unknown the extent to which the use of traditional or non-traditional practices may affect drinkers' acute experiences, and longer term wellbeing and mental health outcomes. Hence, this study aimed to provide the first detailed assessment of associations between ceremony/ritual characteristics, additional support practices, motivations for drinking, and mental health and wellbeing outcomes. The paper uses data from a large cross-sectional study of ayahuasca drinkers in more than 40 countries who had used ayahuasca in various contexts (n = 6,877). It captured detailed information about participant demographics, patterns and history of ayahuasca drinking, the setting of consumption, and ritualistic practices employed. Current mental health status was captured via the Kessler 10 psychological distress scale and the mental health component score of the SF-12 Health Questionnaire, while reported change in prior clinically diagnosed anxiety or depression (n = 1276) was evaluated using a (PGIC) Patient Global Impression of Change tool. Various intermediate outcomes were also assessed including perceived change in psychological wellbeing, number of personal self-insights attained, and subjective spiritual experience measured via the spirituality dimension of the Persisting Effects Questionnaire (PEQ) and Short Index of Mystical Orientation. Regression models identified a range of significant associations between set and setting variables, and intermediate and final mental health and wellbeing outcomes. A generalized structural equation model (GSEM) was then used to verify relationships and associations between endogenous, mediating and final outcome variables concurrently. The present study sheds new light on the influence of ceremonial practices, additional supports and motivations on the therapeutic effects of ayahuasca for mental health and wellbeing, and ways in which such factors can be optimized in naturalistic settings and clinical studies.


SLC35B2 Acts in a Dual Role in the Host Sulfation Required for EV71 Infection.
As an important neurotropic enterovirus, enterovirus 71 (EV71) is occasionally associated with severe neurological diseases and high mortality rates in infants and young children. Understanding the interaction between host factors and EV71 will play a vital role in developing antivirals and optimizing vaccines. Here, we performed a genome-wide CRISPR-Cas9 knockout screen and revealed that scavenger receptor class B member 2 (SCARB2), solute carrier family 35 member B2 (SLC35B2), and beta-1,3-glucuronyltransferase 3 (B3GAT3) are essential in facilitating EV71 replication. Subsequently, the exploration of molecular mechanisms suggested that the knockout of SLC35B2 or B3GAT3, not SCARB2, led to a remarkable decrease in the binding of EV71 to cells and internalization into cells. Furthermore, we found that the infection efficiency for EV71 was positively correlated with the level of host cell sulfation, not simply with the amount of heparan sulfate, suggesting that an unidentified sulfated protein(s) must contribute to EV71 infection. In support of this idea, we screened possible sulfated proteins among the proteinous receptors for EV71 and confirmed that SCARB2 could uniquely interact with both tyrosyl protein sulfotransferases in humans. We then performed mass spectrometric analysis of SCARB2, identifying five sites with tyrosine sulfation. The function verification test indicated that there were more than five tyrosine-sulfated sites on SCARB2. Finally, we constructed a model for EV71 entry in which both heparan sulfate and SCARB2 are regulated by SLC35B2 and act cooperatively to support viral binding, internalization, and uncoating. Taken together, this is the first time that we performed the pooled CRISPR-Cas9 genetic screening to investigate the interplay of host cells and EV71. Furthermore, we found that a novel host factor, SLC35B2, played a dual role in regulating the overall sulfation comprising heparan sulfate sulfation and protein tyrosine sulfation, which are critical for EV71 entry. IMPORTANCE As the most important nonpolio neurotropic enterovirus lacking specific treatments, EV71 can transmit to the central nervous system, leading to severe and fatal neurological complications in infants and young children. The identification of new factors that facilitate or inhibit EV71 replication is crucial to uncover the mechanisms of viral infection and pathogenesis. To date, only a few host factors involved in EV71 infection have been characterized. Herein, we conducted a genome-wide CRISPR-Cas9 functional knockout (GeCKO) screen for the first time to study EV71 in HeLa cells. The screening results are presented as a ranked list of candidates, including 518 hits in the positive selection that facilitate EV71 replication and 1,044 hits in the negative selection that may be essential for cell growth and survival or for suppressing EV71 infection. We subsequently concentrated on the top three hits in the positive selection: SCARB2, SLC35B2, and B3GAT3. The knockout of any of these three genes confers strong resistance against EV71 infection. We confirmed that EV71 infection is codependent on two receptors, heparan sulfate and SCARB2. We also identified a host entry factor, SLC35B2, indirectly facilitating EV71 infection through regulation of the host cell sulfation, and determined a novel posttranslational modification, protein tyrosine sulfation existing in SCARB2. This study revealed that EV71 infectivity exhibits a significant positive correlation with the level of cellular sulfation regulated by SLC35B2. Due to the sulfation pathway being required for many distinct viruses, including but not limited to EV71 and respiratory syncytial virus (RSV), which were tested in this study, SLC35B2 represents a target of broad-spectrum antiviral therapy.


Conditional Control of CRISPR/Cas9 Function.
The recently discovered CRISPR/Cas9 endonuclease system, comprised of a guide RNA for the recognition of a DNA target and the Cas9 nuclease protein for binding and processing the target, has been extensively studied and has been widely applied in genome editing, synthetic biology, and transcriptional modulation in cells and animals. Toward more precise genomic modification and further expansion of the CRISPR/Cas9 system as a spatiotemporally controlled gene regulatory system, several approaches of conditional activation of Cas9 function using small molecules and light have recently been developed. These methods have led to improvements in the genome editing specificity of the CRISPR/Cas9 system and enabled its activation with temporal and spatial precision.


Carotid chemoreceptor control of muscle sympathetic nerve activity in hypobaric hypoxia.
What is the central question of this study? High-altitude hypoxia increases muscle sympathetic nerve activity (MSNA), but whether intravenous infusion of dopamine, to blunt the responsiveness of the carotid chemoreceptors, reduces MSNA at high altitude is not known. What is the main finding and its importance? Muscle sympathetic nerve activity was elevated after 15-17 days of high-altitude hypoxia (3454 m) compared with values at 'sea level' (432 m). However, intravenous dopamine infusion to blunt the responsiveness of the carotid chemoreceptors did not significantly decrease MSNA either at sea level or at high altitude, suggesting that high-altitude sympathoexcitation arises via a different mechanism. High-altitude hypoxia causes pronounced sympathoexcitation, but the underlying mechanisms remain unclear. We tested the hypothesis that i.v. infusion of dopamine to attenuate carotid chemoreceptor responsiveness would reduce muscle sympathetic nerve activity (MSNA) at high altitude. Nine healthy individuals [mean (SD); 26 (4) years of age] were studied at 'sea level' (SL; Zurich) and at high altitude (ALT; 3454 m; 15-17 days after arrival), both while breathing the ambient air and during an acute incremental hypoxia test (eight 3 min stages; partial pressure of end-tidal O2 90-45 mmHg). Intravenous infusions of dopamine (3 μg kg-1 min-1 ) and placebo (saline) were administered on both study days, according to a single-blind randomized cross-over design. Sojourn to high altitude decreased the partial pressure of end-tidal O2 (to ∼60 mmHg) and increased minute ventilation [V̇E; mean ± SEM, SL versus ALT: saline, 8.6 ± 0.5 versus 11.3 ± 0.6 l min-1 ; dopamine, 8.2 ± 0.5 versus 10.6 ± 0.8 l min-1 ; P < 0.05] and MSNA burst frequency by ∼80% [SL versus ALT: saline, 16 ± 3 versus 28 ± 4 bursts min-1 ; dopamine, 16 ± 4 versus 31 ± 4 bursts min-1 ; P < 0.05) when breathing the ambient air, but were not different with dopamine. Increases in MSNA burst frequency and V̇E during the acute incremental hypoxia test were greater at ALT than SL (P < 0.05). Dopamine did not affect the magnitude of the MSNA burst frequency response to acute incremental hypoxia at either SL or ALT. However, V̇E was lower with dopamine than saline administration throughout the acute incremental hypoxia test at ALT. These data indicate that i.v. infusion of low-dose dopamine to blunt the responsiveness of the carotid chemoreceptors does not significantly decrease MSNA at high altitude.


Impact of a pressure-responsive flow-limiting valve on bag-valve-mask ventilation in an airway model.
Using a simulated airway model, we compared ventilation performance by emergency medical services (EMS) providers using a traditional bag-valve-mask (Easy Grip) resuscitator to their performance when using a new device, the SMART BAG resuscitator, which has a pressure-responsive flow-limiting valve. We recruited EMS providers at an EMS educational forum and performed a randomized, non-blinded, prospective crossover comparison of ventilation with 2 devices on a non-intubated simulated airway model. Subjects were instructed to ventilate a Mini Ventilation Training Analyzer as they would an 85-kg adult patient in respiratory arrest. After being randomized to order of device use, they performed ventilation for 1 minute with each device. Primary outcomes were ventilation rates and peak airway pressures. We also measured average tidal volume, gastric inflation volume, minute ventilation and inspiratory:expiratory (I:E) ratio, and compared our results to the American Heart Association standards (2005 edition). We observed statistically significant differences between the SMART BAG and the traditional bag-valve-mask for respiratory rate (12 v. 14 breaths/min), peak airway pressure (15.6 v. 18.9 cm H(2)O), gastric inflation (239.6 v. 1598.4 mL), minute ventilation (7980 v. 8775 mL), and I:E ratio (1.3 v. 1.1). Average tidal volume was similar with both devices (679.6 v. 672.2 mL). The SMART BAG(R) provided ventilation performance that was more consistent with American Heart Association guidelines and delivered similar tidal volumes when compared with ventilation with a traditional bag-valve-mask resuscitator.


Metabolic profiling of ligands for the chemokine receptor CXCR3 by liquid chromatography-mass spectrometry coupled to bioaffinity assessment.
Chemokine receptors belong to the class of G protein-coupled receptors and are important in the host defense against infections and inflammation. However, aberrant chemokine signaling is linked to different disorders such as cancer, central nervous system and immune disorders, and viral infections [Scholten DJ et al. (2012) Br J Pharmacol 165(6):1617-1643]. Modulating the chemokine receptor function provides new ways of targeting specific diseases. Therefore, discovery and development of drugs targeting chemokine receptors have received considerable attention from the pharmaceutical industry in the past decade. Along with that, the determination of bioactivities of individual metabolites derived from lead compounds towards chemokine receptors is crucial for drug selectivity, pharmacodynamics, and potential toxicity issues. Therefore, advanced analytical methodologies are in high demand. This study is aimed at the optimization of a new analytical method for metabolic profiling with parallel bioaffinity assessment of CXCR3 ligands of the azaquinazolinone and piperazinyl-piperidine class and their metabolites. The method is based on mass spectrometric (MS) identification after liquid chromatographic (LC) separation of metabolic mixtures. The bioaffinity assessment is performed "at-line" via high-resolution nanofractionation onto 96-well plates allowing direct integration of radioligand binding assays. This new method enables identification of metabolites from lead compounds with associated estimation of their individual bioaffinity. Moreover, the identification of the metabolite structures via accurate mass measurements and MS(2) allows the identification of liable metabolic "hotspots" for further lead optimization. The efficient combination of chemokine receptor ligand binding assays with analytical techniques, involving nanofractionation as linking technology, allows implementation of comprehensive metabolic profiling in an early phase of the drug discovery process.


Effects of deuterium oxide on mechanics and energetics of skeletal muscle contraction.
Mechanics and energetics of contraction were studied in isolated frog semitendinosus muscles (n = 15) in deuterium oxide (D2O) at 5 degrees C. Twitch force and energy liberation (heat + work) in 99.8% D2O-Ringer at lo were 4 +/- 0.8 and 22 +/- 1%, respectively, of control values. Tetanus force was slowed in development, and developed tetanus force was reduced to 89 +/- 7% of control values. With repeated tetanic contractions, the rate of force development increased, and peak force increased above control values in some muscles. The twitch was potentiated 10-fold (10 +/- 2) after repeated tetanic contractions. Energy liberation increased but not in proportion to force. Ca2+ cycling energy liberation or activation heat, estimated from stretched muscles, was 60 +/- 3% of control values. With maximum posttetanic twitch potentiation, the activation heat increased by 25 +/- 3%. Peak twitch force and energy liberation occurred at longer muscle lengths than in normal Ringer. After correcting for small differences in force development at lo and 1.38 +/- 0.01 lo, it was possible to estimate the effect of length per se on energy liberation. Under these conditions, there was an 8 +/- 2% decrease in energy liberation at 1.38 lo.


[Electron paramagnetic resonance study of the structural characteristics of the envelope of the Venezuelan equine encephalitis virus].
The method of electron paramagnetic resonance and spin labels was used to study the structural characteristics of Venezuelan equine encephalomyelitis virus envelope and cell membranes of chick embryo fibroblasts. The virus envelope lipids form a bilayer structural typical of biological membranes. The lipid bilayer of virions is more rigid than the cytoplasmic membrane of chick embryo fibroblasts. Investigation of interaction of a small hydrophilic molecule with Venezuelan equine encephalomyelitis virus particles showed free water to be absent in the virions. A structural translation was detected at 25 degrees-30 degrees C in the virion membrane. The role of membrane structures in virus reproduction is discussed.


Clinical and epidemiological features of keratoconus genetic and external factors in the pathogenesis of the disease.
Clinical and epidemiological features of keratoconus (KC) were studied in a series comprising all the 212 KC patients treated at Oulu University Central Hospital from 1964 to 1984. Altogether 294 keratoconus patients and relatives were examined ophthalmologically by the author. The prevalence rate of KC needing ophthalmic care was estimated to be 0.03% (75/260,000). The annual incidence was 0.0015% (75/260,000 per 20 years) and remained the same throughout the period studied. 62.7% (133) of the patients were male and 37.3% (79) female. 73% were aged 24 years or younger at the onset of symptoms, the mean age of the males at the first examination being 26.5 +/- 8.2 years, and that of the females 30.6 +/- 13.7 years. Corneal transplantation was carried out on 65 of the 144 patients coming from the area served by Oulu University Central Hospital. Familial KC was found in 19 of the 101 families investigated in the north of Finland (19%) and in 5 of the 58 from the south (9%). The higher frequency of familial KC in the north is probably due to the more pronounced effect of gene pooling in the larger families (mean family size 4.9 persons as compared with 3.5 in the south). The inheritance was found to be attributable to a dominant autosomal mode in 24 out of 28 multiple-case families (85%), the disease being inherited from the mother in 15 cases and the father in 9. Data on the order of birth of keratoconic children were obtained from 159 families. 169 out of a total of 688 children were affected (25%). If families with only one child were excluded, then 47 of the 149 first children (32%) and 44 of the 149 second children (30%) had KC. Thus the disease is characterized by incomplete penetrance and variable expressivity. 122 HLA-A,B,C antigen typings were performed in 18 multiple-case families and the HLA genotypes expressed as haplotypes. In 15 families with more than one child affected, 27 keratoconic children were noted to share the mutual haplotype with the affected parent, whereas 3 had inherited the mutual haplotype from the healthy parent (p less than 0.001). The HLA haplotype could thus serve as a marker for KC inside the family. Connective tissue symptoms and abnormalities were seen in 31 out of 46 KC patients (67%) and in 60 out of 122 first-degree relatives from the town of Oulu and its surroundings (49%).(ABSTRACT TRUNCATED AT 400 WORDS)


Usability and Accuracy of a Smartwatch for the Assessment of Physical Activity in the Elderly Population: Observational Study.
Regular physical activity (PA) contributes to the primary and secondary prevention of several chronic diseases and reduces the risk of premature death. Physical inactivity is a modifiable risk factor for cardiovascular disease and a variety of chronic disorders such as diabetes, obesity, hypertension, bone and joint diseases (eg, osteoporosis and osteoarthritis), depression, and colon and breast cancer. Population aging and the related increase in chronic diseases have a major impact on the health care systems of most Western countries and will produce an even more significant effect in the future. Monitoring PA is a valuable method of determining whether people are performing enough PA so as to prevent chronic diseases or are showing early symptoms of those diseases. The aim of this study was to estimate the accuracy of wearable devices in quantifying the PA of elderly people in a real-life setting. Participants aged 70 to 90 years with the ability to walk safely without any walking aid for at least 300 meters, who had no walking disabilities or episodes of falling while walking in the last 12 months, were asked to walk 150 meters at their preferred pace wearing a vívoactive HR device (Garmin Ltd) and actual steps were monitored and tallied by a researcher using a hand-tally counter to assess the performance of the device at a natural speed. A Bland-Altman plot was used to analyze the difference between manually counted steps and wearable device-measured steps. The intraclass correlation coefficient (ICC) was computed (with a 95% confidence interval) between step measurements. The generalized linear mixed-model (GLMM) ICCs were estimated, providing a random effect term (random intercept) for the individual measurements (gold standard and device). Both adjusted and conditional ICCs were computed for the GLMM models considering separately the effect of age, sex, BMI, and obesity. Analyses were performed using R software (R Foundation for Statistical Computing) with the rms package. A total of 23 females and 26 males were enrolled in the study. The median age of the participants was 75 years. The Bland-Altman plot revealed that, excluding one observation, all differences across measurements were in the confidence bounds, demonstrating the substantial agreement between the step count measurements. The results were confirmed by an ICC equal to .98 (.96-.99), demonstrating excellent agreement between the two sets of measurements. The level of accuracy of wearable devices in quantifying the PA of elderly people in a real-life setting that was found in this study supports the idea of considering wrist-wearable nonmedical devices (widely available in nonspecialized stores) as reliable tools. Both health care professionals and informal caregivers could monitor the level of PA of their patients.


Navigating Molecular Pathways: An Update on Drugs in Colorectal Cancer Treatment.
Colorectal cancer (CRC) is a multifaceted and heterogeneous ailment that affects the colon or rectum of the digestive system. It is the second most commonly occurring form of cancer and ranks third in terms of mortality rate. The progression of CRC does not occur due to a single mutational event; rather, it is the result of the sequential and cumulative accumulation of mutations in key driver genes of signaling pathways. The most significant signaling pathways, which have oncogenic potential due to their deregulation, include Wnt/β-catenin, Notch, TGF-β, EGFR/MAPK, and PI3K/AKT pathways. Numerous drug target therapies have been developed to treat CRC using small molecule inhibitors, antibodies, or peptides. Although drug-targeted therapy is effective in most cases, the development of resistance mechanisms in CRC has raised questions about their efficacy. To overcome this issue, a novel approach of to drug repurposing has come to light, which utilizes already FDA-approved drugs to treat CRC. This approach has shown some promising experimental results, making it a crucial avenue of research in the treatment of CRC.


Mitochondrial DNA control region analysis of three ethnic groups in the Republic of Macedonia.
A total of 444 individuals representing three ethnic groups (Albanians, Turks and Romanies) in the Republic of Macedonia were sequenced in the mitochondrial control region. The mtDNA haplogroup composition differed between the three groups. Our results showed relatively high frequencies of haplogroup H12 in Albanians (8.8%) and less in Turks (3.3%), while haplogroups M5a1 and H7a1a were dominant in Romanies (13.7% and 10.3%, respectively) but rare in the former two. This highlights the importance of regional sampling for forensic mtDNA databasing purposes. These population data will be available on EMPOP under accession numbers EMP00644 (Albanians), EMP00645 (Romanies) and EMP00646 (Turks).


Reactivation of multiple viruses in patients with sepsis.
A current controversy is whether patients with sepsis progress to an immunosuppressed state. We hypothesized that reactivation of latent viruses occurred with prolonged sepsis thereby providing evidence of clinically-relevant immunosuppression and potentially providing a means to serially-monitor patients' immune status. Secondly, if viral loads are markedly elevated, they may contribute to morbidity and mortality. This study determined if reactivation of herpesviruses, polyomaviruses, and the anellovirus TTV occurred in sepsis and correlated with severity. Serial whole blood and plasma samples from 560 critically-ill septic, 161 critically-ill non-septic, and 164 healthy age-matched patients were analyzed by quantitative-polymerase-chain-reaction for cytomegalovirus (CMV), Epstein-Barr (EBV), herpes-simplex (HSV), human herpes virus-6 (HHV-6), and TTV. Polyomaviruses BK and JC were quantitated in urine. Detectable virus was analyzed with respect to secondary fungal and opportunistic bacterial infections, ICU duration, severity of illness, and survival. Patients with protracted sepsis had markedly increased frequency of detectable virus. Cumulative viral DNA detection rates in blood were: CMV (24.2%), EBV (53.2%), HSV (14.1%), HHV-6 (10.4%), and TTV (77.5%). 42.7% of septic patients had presence of two or more viruses. The 50% detection rate for herpesviruses was 5-8 days after sepsis onset. A small subgroup of septic patients had markedly elevated viral loads (>104-106 DNA copies/ml blood) for CMV, EBV, and HSV. Excluding TTV, DNAemia was uncommon in critically-ill non-septic patients and in age-matched healthy controls. Compared to septic patients without DNAemia, septic patients with viremia had increased fungal and opportunistic bacterial infections. Patients with detectable CMV in plasma had higher 90-day mortality compared to CMV-negative patients; p<0.05. Reactivation of latent viruses is common with prolonged sepsis, with frequencies similar to those occurring in transplant patients on immunosuppressive therapy and consistent with development of an immunosuppressive state. Whether reactivated latent viruses contribute to morbidity and mortality in sepsis remains unknown.


Effects of NG-nitro-L-arginine methyl ester on renal function and blood pressure.
The dose-dependent effects of intravenous infusions of nitric oxide (NO) synthesis inhibitor, NG-nitro-L-arginine methyl ester (L-NAME; 0.1, 1, 10, and 50 micrograms.kg-1.min-1), were studied in anesthetized rats to determine whether the inhibitory actions of L-NAME are manifested primarily in alterations of renal function or whether they are the consequences of the increase in systemic blood pressure. Mean arterial pressure (MAP) was not altered by the intravenous L-NAME infusions of 0.1 and 1.0 microgram.kg-1.min-1. However, 0.1 microgram.kg-1.min-1 L-NAME induced a 30% decrease in urine flow rate (UV). The administration of 1.0 microgram.kg-1.min-1 L-NAME, in addition to decreasing UV, also decreased urinary sodium excretion (UNaV) and renal plasma flow (RPF). The intravenous L-NAME infusions of 10.0 and 50.0 microgram.kg-1.min-1 intravenous L-NAME infusions of 10.0 and 50.0 microgram.kg-1.min-1 produced significant increases in MAP that reversed the initial fall in UV and UNaV, despite decreasing RPF and glomerular filtration rate (GFR). The administration of L-arginine alone (10 micrograms.kg-1.min-1) did not modify any of the parameters measured, but it effectively prevented all the hemodynamic and renal changes induced by the infusion of 50 micrograms.kg-1.min-1 L-NAME. These results suggest that the decrease in nitric oxide production induced by the intravenous infusion of L-NAME affects renal excretion of sodium and water in the absence of any significant change in blood pressure. At larger doses, L-NAME also produces hypertension that overrides the initial antinatriuretic effect.


Racial differences in the anatomical distribution of colorectal cancer: a study of differences between American and Chinese patients.
To compare the racial differences of anatomical distribution of colorectal cancer (CRC) and determine the association of age, gender and time with anatomical distribution between patients from America (white) and China (oriental). Data was collected from 690 consecutive patients in Cleveland Clinic Florida, U.S.A. and 870 consecutive patients in Nan Fang Hospital affiliated to the First Military Medical University, China over the past 11 years from 1990 to 2000. All patients had colorectal adenocarcinoma diagnosed by histology and underwent surgery. The anatomical subsite distribution of tumor, age and gender were significantly different between white and oriental patients. Lesions in the proximal colon (P<0.001) were found in 36.3 % of white vs 26.0 % of oriental patients and cancers located in the distal colon and rectum in 63.7 % of white and 74 % of oriental patients (P<0.001). There was a trend towards the redistribution from distal colon and rectum to proximal colon in white males over time, especially in older patients (>80 years). No significant change of anatomical distribution occurred in white women and Oriental patients. The mean age at diagnosis was 69.0 years in white patients and 48.3 years in Oriental patients (P<0.001). This is the first study comparing the anatomical distribution of colorectal cancers in whites and Chinese patients. White Americans have a higher risk of proximal CRC and this risk increased with time. The proportion of white males with CRC also increased with time. Chinese patients were more likely to have distal CRC and developed the disease at a significantly earlier age than white patients. These findings have enhanced our understanding of the disease process of colorectal cancer in these two races.


Spin-1 neutron resonance peak cannot account for electronic anomalies in the cuprate superconductors.
In certain cuprates, a spin-1 resonance mode is prominent in the magnetic structure measured by neutron scattering. It has been proposed that this mode is responsible for significant features seen in other spectroscopies, such as photoemission and optical absorption, which are sensitive to the charge dynamics, and even that this mode is the boson responsible for "mediating" the superconducting pairing. We show that its small (measured) intensity and weak coupling to electron-hole pairs (as deduced from the measured lifetime) disqualifies the resonant mode from either proposed role.


Continuing professional development in public health medicine.
A survey of public health doctors was undertaken in the South Thames region to support coordination of continuing professional development (CPD) and to guide appropriate provision of educational activities for those working in the specialty. A postal survey of 130 public health doctors in service, academic and military posts in the South Thames region of the UK. The response rate was 79% with good representation of grades and geographical areas. Public health doctors preferred personal and small group learning particularly focused on problem solving. Epidemiology as a basic discipline was rated highly by respondents, but senior doctors also showed preferences for topics beyond the textbooks. Respondents demonstrated that they had acquired such knowledge and skills, which could be shared in CPD sessions with colleagues. There was support for learning with and from non-medical colleagues. In the service posts structural change was ongoing, expectations of performance by management were high, and appropriate CPD was an expressed need, although time was seriously limited for this activity. Appropriate CPD could support the effectiveness of public health physicians in challenging circumstances. However, a critical evaluation of the relevance of CPD programmes to public health practice is essential if CPD is to support doctors in delivering the public health agenda in the changing structures within which they work.


Geographic and temporal analysis of folate-sensitive fetal malformations.
To identify potential geographic and temporal clustering of folate-sensitive fetal malformations as a prelude to a targeted preconception curriculum in folic acid supplementation. Our comprehensive prenatal anomaly database was queried to select fetal malformations presumed to be sensitive to preconception folate insufficiency. Evidence of geographic clustering was evaluated by distribution of individual cases using zip codes of maternal residence. Potential temporal clustering of anomalies was sought by tabulating the frequency of each anomaly category during 5 consecutive 2-year intervals between 1992 and 2001. Over a 10-year period, approximately 2000 fetal anomalies were identified, of which 400 (20%) were considered potentially folate sensitive. We found geographic clustering of ventral wall defects as well as obstructive uropathy by zip code analysis. Significant increases in the frequencies of cardiac defects (P <.001) and obstructive uropathy (P <.001) were noted during the epoch of this study. A moderate increase in anomaly frequency was also seen in the diagnostic subcategory of gastroschisis, in which 15 of 27 total gastroschisis cases occurred in 2000-2001. Geographic clustering and temporal trends in anomaly rates were noted in certain folate-sensitive malformation categories. Identification of specific, high-incidence regions may provide an opportunity for targeted interventions designed to supplement the national folic acid campaign.


Femoral impaction bone grafting with the Elite Plus stem.
This study evaluates the results of impaction grafting of the femoral stem with the Elite Plus system (DePuy, Leeds, UK) in revision hip arthroplasty with irradiated allograft. Nineteen hips in 19 patients (12 men and 7 women), at an average age of 59 years were followed for an average of 33 months. Endo-Klinik grading of bone stock loss was 5 grade 1, 6 grade 2, 8 grade 3 and no grade 4. Radiographic analysis revealed evidence of graft incorporation in 32% but no evidence of trabecular remodelling or cortical repair. Four patients have undergone revision of the femoral stem, one patient died while awaiting revision and one patient is unfit for revision. Most complications occurred in patients classified as Endo-Klinik grade 3. We found that impaction grafting with the Elite Plus system was associated with a high failure rate, especially in those cases with more severe bone stock loss. (Hip International 2002; 1: 11-6).


The QQS orphan gene of Arabidopsis modulates carbon and nitrogen allocation in soybean.
The genome of each species contains as high as 8% of genes that are uniquely present in that species. Little is known about the functional significance of these so-called species specific or orphan genes. The Arabidopsis thaliana gene Qua-Quine Starch (QQS) is species specific. Here, we show that altering QQS expression in Arabidopsis affects carbon partitioning to both starch and protein. We hypothesized QQS may be conserved in a feature other than primary sequence, and as such could function to impact composition in another species. To test the potential of QQS in affecting composition in an ectopic species, we introduced QQS into soybean. Soybean T1 lines expressing QQS have up to 80% decreased leaf starch and up to 60% increased leaf protein; T4 generation seeds from field-grown plants contain up to 13% less oil, while protein is increased by up to 18%. These data broaden the concept of QQS as a modulator of carbon and nitrogen allocation, and demonstrate that this species-specific gene can affect the seed composition of an agronomic species thought to have diverged from Arabidopsis 100 million years ago.


Topical bovine thrombin and adverse events: a review of the literature.
To review published evidence suggesting a link between topical bovine thrombin (TBT) and important adverse events (AEs). English language articles and abstracts were obtained from MEDLINE using combinations of text and MeSH terms for thrombin, bovine thrombin and their trade names. References from summary articles were also retrieved. Published case reports, review articles, and retrospective, prospective or observational studies involving either immunogenicity or AEs were selected for further assessment. Retrieved articles were evaluated separately as AE case reports, quantitative studies of antibodies, or quantitative studies of AEs. Presence of case causal information, temporal pattern of case report publication, reproducibility of aggregate data findings, and study design features. The major limitations of reviewed publications were insufficient information regarding TBT and other exposures, and designs in which linkage between laboratory immune phenomena and AEs could not be evaluated. While immunogenicity studies did support an increased risk for post-TBT antibodies, there was no consistent evidence that this led to an increased AE risk or severity. Common evidentiary deficiencies included case reports from high incidence environments, studies of combination or mixture products, biased study designs, lack of patient-level exposure data, inadequate control groups and insufficient sample sizes. The best designed study (a randomized, controlled comparison of TBT to a recombinant bovine product) documented post-TBT antibody production, but no important efficacy or AE differences. An examination of publication dates for case reports showed a peak between 1992 and 1994 followed by a substantial drop. Since 1997 the number of published AE case reports has continued to decline. TBT increases the risk for antibody elevations in patients. A careful review of published evidence does not show that either TBT itself or any associated elevations in anti-bovine antibodies are risk factors for clinically important AEs.


What do relatives value most in end-of-life care for people with dementia?
End-of-life care for older people with dementia is often sub-optimal. Understanding the experiences of the relatives involved in the care of the person with dementia may help to improve care practice. To investigate relatives' experiences with end-of-life care for people with dementia, comparing the nursing home and home setting. In-depth interviews were conducted with 32 individuals who were bereaved of someone with dementia. Thematic analysis was performed to identify main themes from the data. Experiences translated into four themes: acknowledging human dignity; being recognised as an important caregiver; (not) talking about death and dying and making decisions together. A lack of person-centered care was mainly evident in nursing homes. Relatives took on a more prominent role in the care of the older person with dementia when the relative was cared for in a home setting and this involvement in the care of the older person was something that the relatives valued. Surrogate decision-making induced similar challenges in both settings. It is important that healthcare professionals inform and support relatives to help them make decision about end-of-life care and preferences. Nursing homes should learn to offer the same standard of person-centred care as a home care setting, and ensure that relatives are still involved in the care of the person with dementia.


Visualizing the knowledge outburst in global research on COVID-19.
The scholarly output of the new coronavirus research has been proliferating. During five months, an amount of 14,588 scientific publications about nCoV-2 and COVID-19 has been generated intensively (as indexed in Scopus on 31 May 2020). Such a knowledge outburst has created ample interest in understanding the research landscape of this newly configured area. This paper demonstrates on scientometric dimensions of the novel coronavirus (2019-nCov) research using quantifiable characteristics of the publication dataset. Findings reveal that the rate of publication growth (1600%) is very significant to a synergic response of the researchers to combat with the most extended sequence of an RNA virus. Indeed their response has geared up to an average of 100 articles per day. Many scholarly publishers have disclosed their preprint servers to make the publications available immediately, even by enabling Open Access. The scientific contents have published in more than 500 journals from 240 academic publishers. While the top-ten publishers occupied almost 70% of the articles, then about 25% of the studies were sponsored by 300 funding agencies. Among the notable journals Lancet, Nature, BMJ, JAMA, JMV, and NEJM are prominent. Findings also reveal that majority of the contributions have occurred in Medical Science, focusing on virology, immunology, epidemiology, pharmacology, public health, critical care, and emergency medicine. However, the closely associated terms are virus transmission, infection control, asymptomatic, quarantine, pneumonia, human, disease severity, clinical trials, viral pathogenesis, pandemic, risk, and mortality. The study suggests that academic hubs are located mostly in the USA, China, Italy, and the UK. Among the productive institutions; Huazhong Univ (China), Tongji Med. College (China), Harvard Med. School (USA), Univ of Milan (Italy), INSERM (France), UCL (UK) are outstanding. The G7 countries together produced 50% of the global research output on nCov-2. It also noted an encouraging trend of collaborative research across many countries and disciplines, where the values of CI (6.46), DC (0.79), and CC (0.59) are very significant. It examines the geographical diversity of the collaborating authors, thereby visualized their linkages via co-authorship occurrences. Finally, it analyzed the publications' impact to showcase the most influential contributions of the new coronavirus research.


Dentate granule cells in reeler mutants and VLDLR and ApoER2 knockout mice.
We have studied the organization and cellular differentiation of dentate granule cells and their axons, the mossy fibers, in reeler mutant mice lacking reelin and in mutants lacking the reelin receptors very low density lipoprotein receptor (VLDLR) and apolipoprotein E receptor 2 (ApoER2). We show that granule cells in reeler mice do not form a densely packed granular layer, but are loosely distributed throughout the hilar region. Immunolabeling for calbindin and calretinin revealed that the sharp border between dentate granule cells and hilar mossy cells is completely lost in reeler mice. ApoER2/VLDLR double-knockout mice copy the reeler phenotype. Mice deficient only in VLDLR showed minor alterations of dentate organization; migration defects were more prominent in ApoER2 knockout mice. Tracing of the mossy fibers with Phaseolus vulgaris leukoagglutinin and calbindin immunolabeling revealed an irregular broad projection in reeler mice and ApoER2/VLDLR double knockouts, likely caused by the irregular wide distribution of granule cell somata. Mutants lacking only one of the lipoprotein receptors showed only minor changes in the mossy fiber projection. In all mutants, mossy fibers respected the CA3-CA1 border. Retrograde labeling with DiI showed that malpositioned granule cells also projected as normal to the CA3 region. These results indicate that ( 1 ) reelin signaling via ApoER2 and VLDLR is required for the normal positioning of dentate granule cells and (2) the reelin signaling pathway is not involved in pathfinding and target recognition of granule cell axons.


Predictors of perception of pregnancy risk among nulliparous women.
To determine factors associated with perception of pregnancy risk using a conceptual framework based on a review of the relevant literature and the psychometric model of risk perception. A correlational study. Ambulatory care and antepartum units of two tertiary hospitals and selected obstetricians' offices and prenatal classes in Winnipeg, Canada. A convenience sample of nulliparous women in their third trimester with a singleton pregnancy. Between December 2009 and January 2011, the following questionnaires were completed by 159 nulliparous women: the Perception of Pregnancy Risk Questionnaire, the Pregnancy-related Anxiety scale, Knowledge of Maternal Age-related Risks of Childbearing Questionnaire, the SF-12v2 Health Status Survey, the Multidimensional Health Locus of Control, and the Prenatal Scoring Form. Pearson's r correlations and stepwise multivariable linear regression analyses were conducted to achieve the research objectives. Of the eight proposed factors in the conceptual framework, five factors were significant predictors of perception of pregnancy risk, including pregnancy-related anxiety, maternal age, medical risk, perceived internal control, and gestational age, accounting for 47% to 49% of the variance in risk perception. An interaction between the pregnancy-related anxiety score and maternal age was found. These results contribute to the literature on perception of pregnancy risk by identifying a new predictor (gestational age), supporting the role of previously known factors in the state of pregnancy, and proposing pregnancy-related anxiety as a pregnancy dread factor in risk perception theories. This knowledge may have implications for developing more effective risk communication models.


Nrf2-dysregulation correlates with reduced synthesis and low glutathione levels in experimental autoimmune encephalomyelitis.
This study investigates the possible mechanism(s) underlying glutathione (GSH) deficiency in the mouse spinal cord during the course of myelin oligodendrocyte glycoprotein35-55 peptide-induced experimental autoimmune encephalomyelitis (EAE), a commonly used animal model of multiple sclerosis. Using the classical enzymatic recycling method and a newly developed immunodot assay, we first demonstrated that total GSH levels (i.e. free GSH plus all its adducts) are reduced in EAE, suggesting an impaired synthesis. The decline in the levels of this essential antioxidant tripeptide in EAE coincides temporally and in magnitude with a reduction in the amount of γ-glutamylcysteine ligase, the rate-limiting enzyme in GSH synthesis. Other enzymes involved in GSH biosynthesis, whose genes also contain antioxidant-response elements, including glutathione synthetase, cystine/glutamate antiporter, and γ-glutamyl transpeptidase (γ-GT) are diminished in EAE as well. Low levels of γ-glutamylcysteine ligase, glutathione synthetase, and γ-GT are the consequence of reduced mRNA expression, which correlates with diminished expression of the nuclear factor (erythroid-derived 2)-like 2 (Nrf2) in both the cytosol and nucleus. Interestingly, the low Nrf2 expression does not seem to be caused by increased degradation via Kelch-like ECH-associated protein 1-dependent or Kelch-like ECH-associated protein 1-independent mechanisms (such as glycogen synthetase kinase-3β activation), or by reduced levels of Nrf2 mRNA. This suggests that translation of this important transcription factor and/or other still unidentified post-translational processes are altered in EAE. These novel findings are central toward understanding how critical antioxidant and protective responses are lost in inflammatory demyelinating disorders.


Mechanical properties, phenolic composition and extractability indices of Barbera grapes of different soluble solids contents from several growing areas.
Phenolic compounds, extractable from grape skins and seeds, have a notable influence on the quality of red wines. Many studies have clearly demonstrated the relationship between the phenolic composition of the grape at harvest time and its influence on the phenolic composition of the red wine produced. In many previous works the evolution of phenolic composition and relative extractability was normally studied on grapes sampled at different times during ripening, but at the same date the physiological characteristics of grape berries in a vineyard are often very heterogeneous. Therefore, the main goal of the study is to investigate the differences among mechanical properties, phenolic composition and relative extractability of Vitis vinifera L. cv Barbera grape berries, harvested at the same date from several vineyards, and calibrated according to their density at three levels of soluble solids (A=235+/-8, B=252+/-8 and C=269+/-8 g L(-1) sugar) with the aim of studying the influence of ripeness stages and growing locations on these parameters. Results on mechanical properties showed that the thickness of the berry skin (Sp(sk)) was the parameter most affected by the different level of sugars in the pulp, while different skin hardnesses, evaluated by the break skin force (F(sk)), were related to the cultivation sites. The latter were also observed to influence the mechanical characteristics of seeds. Generally, the anthocyanin content increased with the level of soluble solids, while the increase in the tannin content of the berry skin and seeds was less marked. However, significant changes in flavanols reactive to vanillin in the seeds were found. The cellular maturity index (EA%) was little influenced by the soluble solids content of grapes.


Host plant use among closely related Anaea butterfly species (Lepidoptera, Nymphalidae, Charaxinae).
There is a great number of Charaxinae (Lepidoptera: Nymphalidae) species in the tropics whose larvae feed on several plant families. However the genus Anaea is almost always associated with Croton species (Euphorbiaceae). This work describes patterns of host plant use by immature and adult abundance on different vertical strata of sympatric Anaea species in a forest of Southeastern Brazil. Quantitative samples of leaves were taken in April/1999 and May/2000 to collect eggs and larvae of four Anaea species on C. alchorneicarpus, C. floribundus and C. salutaris in a semideciduous forest. Sampled leaves were divided into three classes of plant phenological stage: saplings, shrubs and trees. The results showed that the butterfly species are segregating in host plant use on two scales: host plant species and plant phenological stages. C. alchorneicarpus was used by only one Anaea species, whereas C. floribundus was used by three species and C. salutaris by four Anaea species. There was one Anaea species concentrated on sapling, another on sapling/shrub and two others on shrub/tree leaves. Adults of Anaea were more frequent at canopy traps but there were no differences among species caught in traps at different vertical positions. This work supplements early studies on host plant use among Charaxinae species and it describes how a guild of closely related butterfly species may be organized in a complex tropical habitat.


Towards minimally invasive monitoring for gastroenterology - An external Squamocolumnar Junction Locator.
Transient lower oesophageal sphincter relaxations (TLOSRs) occur frequently and are the main mechanism of acid reflux. The only means of currently detecting TLOSRs is intra-luminal manometry and the probes themselves may stimulate TLOSRs. The squamo-columnar junction moves 4-5 centimeters proximally during TLOSRs and this provides a means of detecting such episodes. The objective of this work is to develop a sensor system capable of detecting the movement of a miniature magnet attached to the squamo-columnar junction from outside the body and thus allow detection of TLOSRs without the artifact associated with intraluminal detection probes. A GaAs Hall effect sensor was selected and an alternating current supply was developed with a combination of filters and a Phase Sensitive Detector, to detect the magnet. The oscillation frequency of the current was chosen in order to reduce electronic noise, and filtering outside this frequency means the signal to noise ratio was greatly improved. The phase sensitive detector was employed to accurately convert the amplitude of the sensor's output to a DC signal. With the addition of paired Flux Concentrators increases the range up to 10.2 centimetres, an improvement of 580% over commercial Hall effect sensors. The AC circuit and flux concentrator device far exceeds the sensitivity of the current Hall effect sensors supplied in the market, by rejecting noise and providing accurate measurement over significantly larger distances. The development of this sensor has applications beyond this specific medical device.


Association of physical fitness with health-related quality of life in early postmenopause.
To assess the association of different components of physical fitness with HRQoL in early postmenopause and to test which physical fitness components are independently associated with the physical and mental components of HRQoL. The final sample comprised 67 early postmenopausal women. Physical fitness was assessed with the Senior Fitness Test battery (additionally including handgrip strength test), and HRQoL was evaluated with the Short-Form Health Survey-36 (SF-36). We also analyzed plasma gonadotropic hormones and estradiol. Overall, most of the fitness components were positively associated with HRQoL. Lower-body flexibility, upper-body muscle strength and cardiorespiratory fitness were the fitness components more strongly associated with HRQoL (r range from 0.28 to 0.56). Static balance was especially associated with mental health (r = -0.46, P < 0.001). Lower-body flexibility (assessed with the chair sit-and-reach test) and upper-body muscle strength (assessed with handgrip dynamometry) were independently associated with the SF-36 Physical Component Summary (both, P < 0.001). Upper-body muscle strength (P < 0.01) and cardiorespiratory fitness (assessed with the 6-min walk test, P < 0.05) were independently associated with the SF-36 Mental Component Summary. Higher physical fitness is associated with better HRQoL in early postmenopause. Lower-body flexibility and upper-body muscle strength were the most important independent fitness indicators, explaining ~30 % of HRQoL.


Oligonucleotide Therapies in the Treatment of Arthritis: A Narrative Review.
Osteoarthritis (OA) and rheumatoid arthritis (RA) are two of the most common chronic inflammatory joint diseases, for which there remains a great clinical need to develop safer and more efficacious pharmacological treatments. The pathology of both OA and RA involves multiple tissues within the joint, including the synovial joint lining and the bone, as well as the articular cartilage in OA. In this review, we discuss the potential for the development of oligonucleotide therapies for these disorders by examining the evidence that oligonucleotides can modulate the key cellular pathways that drive the pathology of the inflammatory diseased joint pathology, as well as evidence in preclinical in vivo models that oligonucleotides can modify disease progression.


Molecular Dynamics Simulation of a Feather-Boa Model of a Bacterial Chromosome.
The chromosome of a bacterium consists of a mega-base pair long circular DNA, which self-organizes within the micron-sized bacterial cell volume, compacting itself by three orders of magnitude. Unlike in eukaryotes, it lacks a nuclear membrane, and freely floats in the cytosol confined by the cell membrane. It is believed that strong confinement, cross-linking by associated proteins, and molecular crowding all contribute to determine chromosome size and morphology. Modeling the chromosome simply as a circular polymer decorated with closed side-loops in a cylindrical confining volume, has been shown to already recapture some of the salient properties observed experimentally. Here, we describe how a computer simulation can be set up to study structure and dynamics of bacterial chromosomes using this model.


IRT1, an Arabidopsis transporter essential for iron uptake from the soil and for plant growth.
Plants are the principal source of iron in most diets, yet iron availability often limits plant growth. In response to iron deficiency, Arabidopsis roots induce the expression of the divalent cation transporter IRT1. Here, we present genetic evidence that IRT1 is essential for the uptake of iron from the soil. An Arabidopsis knockout mutant in IRT1 is chlorotic and has a severe growth defect in soil, leading to death. This defect is rescued by the exogenous application of iron. The mutant plants do not take up iron and fail to accumulate other divalent cations in low-iron conditions. IRT1-green fluorescent protein fusion, transiently expressed in culture cells, localized to the plasma membrane. We also show, through promoter::beta-glucuronidase analysis and in situ hybridization, that IRT1 is expressed in the external cell layers of the root, specifically in response to iron starvation. These results clearly demonstrate that IRT1 is the major transporter responsible for high-affinity metal uptake under iron deficiency.


Estimating global mortality from potentially foodborne diseases: an analysis using vital registration data.
Foodborne diseases (FBD) comprise a large part of the global mortality burden, yet the true extent of their impact remains unknown. The present study utilizes multiple regression with the first attempt to use nonhealth variables to predict potentially FBD mortality at the country level. Vital registration (VR) data were used to build a multiple regression model incorporating nonhealth variables in addition to traditionally used health indicators. This model was subsequently used to predict FBD mortality rates for all countries of the World Health Organization classifications AmrA, AmrB, EurA, and EurB. Statistical modeling strongly supported the inclusion of nonhealth variables in a multiple regression model as predictors of potentially FBD mortality. Six variables were included in the final model: percent irrigated land, average calorie supply from animal products, meat production in metric tons, adult literacy rate, adult HIV/AIDS prevalence, and percent of deaths under age 5 caused by diarrheal disease. Interestingly, nonhealth variables were not only more robust predictors of mortality than health variables but also remained significant when adding additional health variables into the analysis. Mortality rate predictions from our model ranged from 0.26 deaths per 100,000 (Netherlands) to 15.65 deaths per 100,000 (Honduras). Reported mortality rates of potentially FBD from VR data lie within the 95% prediction interval for the majority of countries (37/39) where comparison was possible. Nonhealth variables appear to be strong predictors of potentially FBD mortality at the country level and may be a powerful tool in the effort to estimate the global mortality burden of FBD. The views expressed in this document are solely those of the authors and do not represent the views of the World Health Organization.


Use of animals in dental research: Trends and ethical reflections.
This study aimed to survey and analyze the profile of experimental dental research in animals, verifying its trends. We evaluated studies developed with the use of animals in vivo, published in 10 dental journals with high impact factors, from 2015 to 2020. From 1652 studies retrieved, 594 involved in vivo experimentation on animals and were analyzed further. Rats were the species most used and with the highest mean of animals per study. Ferrets, although presenting the lowest rate of publications, had the second highest mean of animals per study. Periodontics was the dental specialty with the highest number of publications, while oral rehabilitation had the lowest number. The data on the institution responsible for reviewing animal research protocols approval, sample size, anesthesia and analgesia were provided in 93.10%, 83%, 70.54% and 23.74% of studies, respectively. In 53% of studies, euthanasia was specified and anesthetic overdose was the method most used. Over the period analyzed, there was a reduction in animal studies in vivo, and periodontics was the specialty that most used this experimental model. Although most studies mentioned approval by an ethics committee, some publications neglected to mention sample size, anesthesia and euthanasia. The omission of essential information may raise scientific and ethical concerns.


Age-related changes in the chemical composition of gallstones.
The association between pigment cholelithiasis and advancing age has been previously described but little is known about the time-course of these changes. To determine the specific changes that occur in the chemical composition of gallstones with increasing age. Gallstones were collected from 387 non-cirrhotic patients and visually classified as either cholesterol or pigment. All stones were quantitatively analyzed by Fourier transform infrared spectroscopy for cholesterol, bilirubin, carbonate and phosphate and the results correlated with stone type and patient age. Forty-five patients had pigment stones (12 %) and 342 had cholesterol stones (88 %). No patient had both types. There was a reciprocal relationship between the mean cholesterol and bilirubin contents of stones over time with cholesterol accounting for 54% of the weight of gallstones before age 30 and only 17% after age 70. Similarly, the mean content (by weight) of bilirubin was 35% before age 30 but 61% after age 70. In addition, the fraction of gallstones containing carbonate or phosphate salts increased sequentially with age (6% at age 30 to 57% at age 70). (1). The ratio of pigment to cholesterol gallstones increases directly with age. (2) The cholesterol content of stones steadily decreases after age 50 while the content of bilirubin, phosphate and carbonate gradually increases. (3) These data suggest that, during aging, cholesterol may become solubilized and may be replaced by calcium salts of carbonate, phosphate or bilirubinate.


Efficacy of vagus nerve stimulation as a treatment for medically intractable epilepsy in brain tumor patients. A case-controlled study using the VNS therapy Patient Outcome Registry.
Vagus nerve stimulation (VNS) therapy is a procedure to control seizure frequency in patients with medically intractable epilepsy. However, there is no data on efficacy in the subset of these patients with brain tumors. The purpose of this study is to evaluate the efficacy of VNS therapy in patients with brain tumor-associated medically intractable epilepsy. Data from the VNS therapy Patient Outcome Registry, maintained by the manufacturer of the device, Cyberonics Inc. (Houston, TX, USA), was queried to characterize the response of patients in whom a brain tumor was listed as the etiology of epilepsy. A case-control analysis was implemented and patient outcome was measured by Engel classification, median seizure response and responder rate (≥50% seizure reduction) using t-tests and chi-squared tests. In 107 patients with an epilepsy etiology related to a brain tumor, seizure reduction was 45% at 3 months and 79% at 24 months with a responder rate of 48% at 3 months and 79% at 24 months. There was no statistical difference in seizure reduction compared with 326 case-control patients from the registry without brain tumors. There was no significant difference in anti-epileptic drug (AED) usage from baseline to 24 months post implant in either group. VNS therapy is equally effective in patients who suffer seizures secondary to brain tumors as in patients without history of a brain tumor. VNS therapy is a viable treatment option for patients with brain tumor associated medically intractable epilepsy, assuming cytoreductive and other adjuvant therapies have been fully explored.


PGRMC1 effects on metabolism, genomic mutation and CpG methylation imply crucial roles in animal biology and disease.
Progesterone receptor membrane component 1 (PGRMC1) is often elevated in cancers, and exists in alternative states of phosphorylation. A motif centered on PGRMC1 Y180 was evolutionarily acquired concurrently with the embryological gastrulation organizer that orchestrates vertebrate tissue differentiation. Here, we show that mutagenic manipulation of PGRMC1 phosphorylation alters cell metabolism, genomic stability, and CpG methylation. Each of several mutants elicited distinct patterns of genomic CpG methylation. Mutation of S57A/Y180/S181A led to increased net hypermethylation, reminiscent of embryonic stem cells. Pathways enrichment analysis suggested modulation of processes related to animal cell differentiation status and tissue identity, as well as cell cycle control and ATM/ATR DNA damage repair regulation. We detected different genomic mutation rates in culture. A companion manuscript shows that these cell states dramatically affect protein abundances, cell and mitochondrial morphology, and glycolytic metabolism. We propose that PGRMC1 phosphorylation status modulates cellular plasticity mechanisms relevant to early embryological tissue differentiation.


A retrospective case series of ultrasound-guided suprascapular nerve pulsed radiofrequency treatment for hemiplegic shoulder pain in patients with chronic stroke.
Hemiplegic shoulder pain (HSP) is the most common pain condition after stroke. Pulsed radiofrequency (PRF) treatment of the suprascapular nerve (SSN) effectively relieves shoulder pain conditions. To date, there is no study about the effects of PRF treatment for HSP. Thus, our aim was to report on a case series about its use in chronic stroke. Six chronic stroke patients with HSP (visual analog scale [VAS] score for pain ≥30 mm) underwent ultrasound-guided SSN PRF treatment. All were evaluated before treatment and at 4 and 16 weeks of follow-up. The main outcome was VAS score. Secondary outcomes were Modified Ashworth Scale, shoulder passive range of motion (PROM), Disability Assessment Scale (DAS), Fugl-Meyer Assessment, and EuroQol-5 dimension questionnaire (EuroQol-5D) scores. As compared with baseline, improvement was observed in the following parameters: VAS for pain (at 4 weeks, P=0.023; at 16 weeks, P=0.023); shoulder PROM for abduction (at 4 weeks, P=0.023; at 16 weeks, P=0.024), flexion (at 4 and 16 weeks, P=0.024), extension (at 4 and 16 weeks, P=0.02), and external rotation (4 and 16 weeks, P=0.02); DAS for hygiene (at 4 and 16 weeks, P=0.024), dressing (at 4 weeks, P=0.02; at 16 weeks, P=0.024), and pain (at 4 weeks, P=0.024; at 16 weeks, P=0.023); and EuroQol-5D (at 4 and 16 weeks, P=0.024). Our observations support the use of ultrasound-guided SSN PRF treatment for HSP in chronic stroke patients.


Heterogeneous solution deposition of high-performance adhesive hybrid films.
Interfaces between organic and inorganic materials are of critical importance to the lifetime of devices found in microelectronic chips, organic electronics, photovoltaics, and high-performance laminates. Hybrid organic/inorganic materials synthesized through sol-gel processing are best suited to address these challenges because of the intimate mixing of both components. We demonstrate that deposition from heterogeneous sol-gel solutions leads to the unique nanolength-scale control of the through-thickness film composition and therefore the independent optimization of both the bulk and interfacial film properties. Consequently, an outstanding 3-fold improvement in the adhesive/cohesive properties of these hybrid films can be obtained from otherwise identical precursors.


Non-invasive sudden death risk stratification.
Most sudden cardiac deaths are caused by fatal ventricular arrhythmias (ventricular tachycardia [VT] and fibrillation) in patients with and without known structural heart diseases. Given the large number of patients potentially at risk for developing ventricular arrhythmias, any strategy for treating them prophylactically requires efficient and effective risk stratification. Both non-invasive and invasive testing may be used for prognostic evaluation of patients with heart diseases. The optimal way to use them in the risk stratification for sudden cardiac death will depend in part on the goals of screening. At present risk markers perform better at identifying low-risk patients who may not need an implantable cardioverter-defibrillator (ICD), because all tests have a high negative predictive accuracy. In our opinion an electrophysiological test should not be performed and an ICD should not be implanted in post-myocardial infarction patients with moderate left ventricular dysfunction (left ventricular ejection fraction 30-40%) with a preserved autonomic balance and without non-sustained VT. In MADIT II-like patients electrophysiological testing does not seem necessary and an ICD could not be implanted only in patients with a negative T-wave alternans test. Most of the data available refer to patients with ischemic cardiomyopathy but the preliminary data on T-wave alternans suggest its usefulness in patients with non-ischemic cardiomyopathy too, although a large definitive study has not yet been completed in this important population.


[Bipolar I disorder in France: prevalence of manic episodes and hospitalisation-related costs].
Bipolar disorder is a chronic, highly disabling illness. However, few studies have evaluated the economic impact of this illness. The objective of this study was to estimate: 1) the annual number of manic episodes in patients with bipolar I disorder, and 2) the costs of hospitalisations related to manic episodes in France. We only included data on bipolar I disorder, as there is greater consensus and better documentation for this subgroup of patients with bipolar disorder. The prevalence of manic episodes was estimated using published epidemiological data. A computerised literature search was performed using the traditional scientific and medical databases. Additional epidemiological references were identified from published studies and textbooks. For hospitalisation data, we used the statistics of the Medical Information Department of a large psychiatric hospital in Paris for the year 1999. We estimated the annual number of manic episodes in France based on: 1) the lifetime prevalence of bipolar I disorder, 2) the average cycle duration, 3) the proportion of rapid cycling patients, and 4) the proportion of depressive vs. manic episodes for patients with bipolar I disorder. In order to estimate the prevalence of bipolar I disorder, we conducted a random effects meta-analysis using published international data. Results of the meta-analysis, which was based on a total of 62 736 patients, showed the lifetime prevalence of bipolar I disorder to be 0.82% [95% CI: 0.42, 1.21]. Applied to the adult population in France, this prevalence implies that the number of persons who have ever experienced a bipolar I -disorder is approximately 390,000 [95% CI: 200,000, 575,000]. Few studies provide information on the duration of cycles in patients with bipolar I disorder. Available estimates suggest the cycle duration to be approximately 12 months. Regarding the proportion of rapid cyclers, data from the meta-analysis by Tondo et al. show that 18% of patients with bipolar disorder experience at least four episodes of mood disorder per year. Finally, based on findings provided by cohort studies, the number of depressive episodes appears to be roughly equal to the number of manic episodes during the course of bipolar disorder. A rapid cycling rate of 18% and a cycle duration of 12 months imply that, on average, among 100 bipolar patients, 18 will have a 3-month cycle duration and 82 a 14-month cycle duration. Given an equal proportion of manic and depressive episodes, the annual number of manic episodes would then be 68 for a cohort of 100 bipolar patients (0.68 episode per patient per year). Applying this figure to the estimate of the total number of patients with bipolar I disorder in France suggests that the annual number of manic episodes in France is 265,000 [95% CI: 136,000, 391,000]. Based on data from a psychiatric hospital in Paris, the proportion of manic episodes that require hospitalisation was estimated to be around 63% with an average length of stay of 32.4 days. Hence the annual number of hospitalisations for manic episodes in France is estimated to be 167 000 [95% CI: 86 000, 246 000] and the hospitalisation-related costs 1,3 billion euros approximately. Our review of literature highlights the lack of medical and economic data at the national level on the frequency and hospitalisation-related costs of manic episodes in patients with bipolar I disorder in France. Given the lifetime prevalence of bipolar I disorder which may be as high as 3% among adults, further studies are required in order to provide representative national data and to allow economic evaluations of costs related to bipolar disorder in France.


Depression Anxiety Stress Scale: is it valid for children and adolescents?
The Depression Anxiety Stress Scale (Lovibond & Lovibond, 1995) is used to assess the severity of symptoms in child and adolescent samples although its validity in these populations has not been demonstrated. The authors assessed the latent structure of the 21-item version of the scale in samples of 425 and 285 children and adolescents on two occasions, one year apart. On each occasion, parallel analyses suggested that only one component should be extracted, indicating that the test does not differentiate depression, anxiety, and stress in children and adolescents. The results provide additional evidence that adult models of depression do not describe the experience of depression in children and adolescents.


Elevation of proteasomal substrate levels sensitizes cells to apoptosis induced by inhibition of proteasomal deubiquitinases.
Inhibitors of the catalytic activity of the 20S proteasome are cytotoxic to tumor cells and are currently in clinical use for treatment of multiple myeloma, whilst the deubiquitinase activity associated with the 19S regulatory subunit of the proteasome is also a valid target for anti-cancer drugs. The mechanisms underlying the therapeutic efficacy of these drugs and their selective toxicity towards cancer cells are not known. Here, we show that increasing the cellular levels of proteasome substrates using an inhibitor of Sec61-mediated protein translocation significantly increases the extent of apoptosis that is induced by inhibition of proteasomal deubiquitinase activity in both cancer derived and non-transformed cell lines. Our results suggest that increased generation of misfolded proteasome substrates may contribute to the mechanism(s) underlying the increased sensitivity of tumor cells to inhibitors of the ubiquitin-proteasome system.


Neural-network methods for boundary value problems with irregular boundaries.
Partial differential equations (PDEs) with boundary conditions (Dirichlet or Neumann) defined on boundaries with simple geometry have been successfully treated using sigmoidal multilayer perceptrons in previous works. This article deals with the case of complex boundary geometry, where the boundary is determined by a number of points that belong to it and are closely located, so as to offer a reasonable representation. Two networks are employed: a multilayer perceptron and a radial basis function network. The later is used to account for the exact satisfaction of the boundary conditions. The method has been successfully tested on two-dimensional and three-dimensional PDEs and has yielded accurate results.


Positionally-dependent chondrogenesis induced by BMP4 is co-regulated by Sox9 and Msx2.
Cranial neural crest cells emigrate from the posterior midbrain and anterior hindbrain to populate the first branchial arch and eventually differentiate into multiple cell lineages in the maxilla and mandible during craniofacial morphogenesis. In the developing mouse mandibular process, the expression profiles of BMP4, Msx2, Sox9, and type II collagen demonstrate temporally and spatially restrictive localization patterns suggestive of their functions in the patterning and differentiation of cartilage. Under serumless culture conditions, beads soaked in BMP4 and implanted into embryonic day 10 (E10) mouse mandibular explants induced ectopic cartilage formation in the proximal position of the explant. However, BMP4-soaked beads implanted at the rostral position did not have an inductive effect. Ectopic chondrogenesis was associated with the up-regulation of Sox9 and Msx2 expression in the immediate vicinity of the BMP4 beads 24 hours after implantation. Control beads had no effect on cartilage induction or Msx2 and Sox9 expression. Sox9 was induced at all sites of BMP4 bead implantation. In contrast, Msx2 expression was induced more intensely at the rostral position when compared with the proximal position, and suggested that Msx2 expression was inhibitory to chondrogenesis. To test the hypothesis that over-expression of Msx2 inhibits chondrogenesis, we ectopically expressed Msx2 in the mandibular process organ culture system using adenovirus gene delivery strategy. Microinjection of the Msx2-adenovirus to the proximal position inhibited BMP4-induced chondrogenesis. Over-expression of Msx2 also resulted in the abrogation of endogenous cartilage and the down-regulation of type II collagen expression. Taken together, these results suggest that BMP4 induces chondrogenesis, the pattern of which is positively regulated by Sox9 and negatively by Msx2. Chondrogenesis only occurs at sites where Sox9 expression is high relative to that of Msx2. The combinatorial action of these transcription factors appear to establish a threshold for Sox9 function and thereby restricts the position of chondrogenesis.


Influence of various beverages on urine acid output.
High dose methotrexate therapy requires a high alkaline urine output. In order to evaluate the effect of various beverages on the urine acid output, healthy volunteers (n = 6) took 1.5-2 liters of a test drink during a 2-h period. On the test day urine pH and urine acid excretion (titratable acid plus ammonia minus bicarbonate) were measured. Controls received water and tea as test drink. Orange juice (pH 3.64) and tube feeding (pH 6.78) both led to alkaline urine pH and significantly decreased urine acid output compared to the control group (n = 4, P less than 0.01 and n = 3, P less than 0.001, respectively). Yoghurt (pH 4.1), buttermilk (pH 4.58), and Coca-Cola (pH 2.54), on the other hand, all induced a higher acid output than the control group (n = 6) and a urine pH less than 7.0 during the whole test day (n = 6, NS; n = 6, P less than 0.02; n = 4, P less than 0.05, respectively). If high urine output with an alkaline pH is required, fruit juices or well balanced tube feeding, both with low cation and low sulfur-bound amino acid content, can accomplish this. Drinks with high inorganic acid content (such as Coca-Cola) or high sulfur-bound amino acid content such as yoghurt and buttermilk will result in acidification of the urine.


Activation of c-Jun N-terminal kinase in the absence of NFkappaB function prior to induction of NK cell death triggered by a combination of anti-class I and anti-CD16 antibodies.
Addition of antibodies to major histocompatibility complex class I (MHC class I) and F(c) gamma RIII (CD16) antigens resulted in the synergistic augmentation of natural killer (NK) cell death, and the loss of NK cell cytotoxic function. The binding of anti-CD16 and anti-class I antibodies to the same population of NK cells is required for the synergistic augmentation of NK cell death. Moreover, the addition of antibodies to leukocyte function antigen-1 (LFA-1), which significantly inhibited the phorbol 12-myristate 13-acetate (PMA) and ionomycin mediated NK cell death, had no effect on NK cell death mediated by anti-CD16 and anti-class I antibodies. The increase in NK cell death was associated with an increase in tumor necrosis factor-alpha (TNF-alpha) secretion, and concomitant inhibition of nuclear factor kappa B (NFkappaB) activation and the induction of c-jun N-terminal kinase (JNK) activity in NK cells treated with the combination of anti-class I and anti-CD16 antibodies. Furthermore, the inhibition of NFkappaB activation in anti-CD16 and anti-class I antibody treated NK cells was paralleled with a significant increase in inhibitor of kappa B (IkappaB) protein expression. Overexpression of IkappaB super-repressor in YT, a NK cell line, caused significant up-regulation of TNF-alpha, PMA and ionomycin and Fusobacterium nucleatum mediated NK cell death. Overall, our studies suggest an important regulatory role for NFkappaB and JNK activities in MHC class I mediated NK cell death.


Pupillary Response as Assessment of Effective Seizure Induction by Electroconvulsive Therapy.
Electroconvulsive therapy (ECT) is reported to be effective for severe neuropsychiatric disorders. In ECT, electrical stimulation is applied to the brain, inducing seizure activity. Adequate seizure induction with ECT is associated with seizure duration, symmetrical high amplitude waveforms during slow-wave activity, postictal suppression, and activation of the sympathetic nervous system. Sympathetic nervous system activation is influenced by anesthetic agents or cardiovascular drugs during ECT. Pupillary responses can reflect sympathetic nervous activity or the degree of brain damage. Pupillary response measurement can be conducted in a simple, precise, and objective way using an automated infrared pupillometer, enabling the measurement of pupil diameter (mm) to two decimal places. The white light used for measuring light reflexes is not overly bright, and patients do not typically report discomfort. Pupillary light reflexes were measured before anesthesia induction and immediately after electrical stimulation using this equipment. Pupil diameter is typically enlarged after brain damage or sympathetic nervous activation. Adequate seizure induction using ECT could induce pupillary enlargement immediately after electrical stimulation. In the current method, the constriction ratio of pupil size was calculated automatically and compared with seizure quality. Pupillary responses immediately after electrical stimulation may provide a useful assessment of the efficacy of seizure induction with ECT.


Student assessment in community settings: a comprehensive approach.
Student assessment in community settings presents problems for medical teachers, e.g., difficulties in assessing the contribution of individual members to group work, and lack of test standardization due to varying field conditions. The Faculty of Medicine, University of Gezira, Sudan is a community-oriented, community-based medical school which has adopted a comprehensive approach to student assessment in community settings using various methods, including peer assessment, a supervisory checklist, community feedback, reports from students, short essay questions (SEQs) and multiple choice questions (MCQs). Each method focuses on a specific aspect of the objectives of the community-based programme and is weighted in the final grade according to the extent to which objectives were covered. This assessment programme contrasts with the conventional teacher-centred approach, and is continuously monitored and improved using a variety of sources of information. A total of 105 students participated in a study designed to measure the reliability and validity of this approach. The reliability of the methods was tested by computing the alpha coefficient and was found to range between 0.77 and 0.92. This was considered acceptable. The validity of the instruments was examined using confirmatory factor analysis, and their content validity was reviewed. The results show that the comprehensive approach used is fairly valid. It is suggested that the University's approach is successful in solving some of the problems of student assessment in community settings.


Resting energy expenditure based on equation estimation can predict renal outcomes in patients with type 2 diabetes mellitus and biopsy-proven diabetic kidney disease.
The aim of this study was to investigate the relationship between resting energy expenditure (REE) based on equation estimation and renal outcomes in patients with diabetes kidney disease (DKD). A total of 124 patients were enrolled from a retrospective cohort of Type 2 Diabetes mellitus (T2DM) patients with biopsy-proven DKD. Renal outcome defined as End-Stage Renal Disease (ESRD). To compare the predictive ability of different REE estimation equations on ESRD. Patients' REE was assessed according to the estimating equation with the best predictive power, and then the relationship between REE and ESRD risk was fitted using a restricted cubic spline curve (RCS) plot and REE cutoff values were obtained. Grouping using cutoff values, and ultimately evaluate the relationship between REE and the risk of ESRD using a Multivariate Cox regression model. The strongest predictive validity for renal outcomes was the NDCKD-equation. The patients were divided into the higher-REE group (n = 78) and the lower-REE group (n = 46), based on the cutoff value. During the follow-up, 30 of 124 patients (24.2%) proceeded to ESRD. Multivariate Cox regression models showed that the risk of ESRD in patients with lower REE was 6.08 times increased compared with that in those with higher REE (HR = 6.08; 95% CI, 1.28-28.80, p = 0.023). These findings suggested that the lower REE was an independent risk factor for unfavorable renal outcomes in patients with DKD.