Tools for diagnosis, monitoring and screening of Schistosoma infections utilizing lateral-flow based assays and upconverting phosphor labels.
The potential of various quantitative lateral flow (LF) based assays utilizing up-converting phosphor (UCP) reporters for the diagnosis of schistosomiasis is reviewed including recent developments. Active infections are demonstrated by screening for the presence of regurgitated worm antigens (genus specific polysaccharides), whereas anti-Schistosoma antibodies may indicate ongoing as well as past infections. The circulating anodic antigen (CAA) in serum or urine (and potentially also saliva) is identified as the marker that may allow detection of single-worm infections. Quantitation of antigen levels is a reliable method to study effects of drug administration, worm burden and anti-fecundity mechanisms. Moreover, the ratio of CAA and circulating cathodic antigen (CCA) is postulated to facilitate identification of either Schistosoma mansoni or Schistosoma haematobium infections. The UCP-LF assays allow simultaneous detection of multiple targets on a single strip, a valuable feature for antibody detection assays. Although antibody detection in endemic regions is not a useful tool to diagnose active infections, it gains potential when the ratio of different classes of antibody specific for the parasite/disease can be determined. The UCP-LF antibody assay format allows this type of multiplexing, including testing a linear array of up to 20 different targets. Multiple test spots would allow detection of specific antibodies, e.g. against different Schistosoma species or other pathogens as soil-transmitted helminths. Concluding, the different UCP-LF based assays for diagnosis of schistosomiasis provide a collection of tests with relatively low complexity and high sensitivity, covering the full range of diagnostics needed in control programmes for mapping, screening and monitoring.


Controlled dual delivery of low doses of BMP-2 and VEGF in a silk fibroin-nanohydroxyapatite scaffold for vascularized bone regeneration.
The controlled co-release of osteoinductive and angiogenic factors is an efficient approach to promote vascularized bone regeneration, and a suitable controlled release system can largely reduce the usage of these factors to avoid cost and safety problems. In this study, a cell-free vascularized bone tissue engineering system based on a silk fibroin (SF)/nanohydroxyapatite (nHAp) scaffold was developed, in which very low doses of osteoinductive and angiogenic factors, bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF), were embedded and released in a controlled manner to facilitate bone formation and vascularization, respectively. BMP-2 and VEGF were adsorbed onto SF microspheres (diameter of 1.5 ± 0.3 μm) that were prepared using a co-flow capillary device, and these microspheres were subsequently incorporated within the SF/nHAp scaffolds to provide controlled release. BMP-2 and VEGF were incorporated into SF microspheres via chemical covalent bonding and physical adsorption, respectively, leading to their controlled and sustained release from the SF/nHAp scaffolds. The rapid initial release of VEGF mimicked its expression at the early bone healing stage and promoted angiogenesis, and the relatively slow and sustained release of BMP-2 facilitated osteogenic differentiation both in vitro and in vivo, and the bone completely bridged the rat calvarial defects after 12 weeks of implantation. Overall, our findings suggest that the controlled dual release of very low doses of BMP-2 (300 ng per scaffold) and VEGF (20 ng per scaffold) from SF/nHAp scaffolds results in a synergistic effect on vascularized bone regeneration; this controlled release system can largely reduce the usage of BMP-2 as compared to other systems.


High-sensitivity C-reactive protein is an independent marker of abnormal coronary vasoreactivity in patients with non-obstructive coronary artery disease.
Coronary endothelial dysfunction (CED) is an early stage of atherosclerosis and is associated with adverse cardiovascular events. Inflammation may play a role in the development of endothelial dysfunction. To date no study has evaluated the relationship between C-reactive protein and CED. We aimed to determine if C-reactive protein is associated with CED. In 1016 patients (mean age 50.7±12.3 years, 34% male) presenting to the catheterization laboratory with chest pain and non-obstructive coronary artery disease, coronary vasoreactivity was assessed by measuring the percent change in coronary blood flow (%ΔCBF) and coronary artery diameter (%ΔCAD) in response to intracoronary acetylcholine. Plasma high sensitivity C-reactive protein (hs-CRP) was measured and patients were divided into 2 groups: hs-CRP≤3.0 mg/L (low-intermediate cardiovascular risk n=169) and 3 mg/L<hs-CRP≤10 mg/L (high cardiovascular risk n=847). Patients with a high risk hs-CRP had a significantly lower %ΔCBF and %ΔCAD in response to acetylcholine vs low risk hs-CRP (43.8±6.1 vs 65.8±4.5, P=.004 and -17.2±1.5 vs -13.1±0.8, P=.02 respectively). Low risk hs-CRP was associated with significantly higher %ΔCBF and %ΔCAD vs high risk hs-CRP (27.1±11.0, P=.01 and 4.5±1.9, P=.02 respectively). CED was associated with significantly higher hs-CRP levels and high risk hs-CRP was independently associated with abnormal coronary vasoreactivity, OR 1.82 (95% CI 1.25-2.69). Hs-CRP is independently associated with and a strong predictor of abnormal coronary vasoreactivity in patients with non-obstructive coronary artery disease.


A histologic and cytologic method for the spatial definition of gliomas.
Because of the increasing application of stereotactic neurosurgical techniques not only in the diagnosis but also in the treatment of brain tumors, a demand for data regarding the spatial delimitation of gliomas is being created. Traditional histologic criteria were not established for detecting isolated tumor cells located at the boundaries of gliomas. Standard histologic techniques used for processing surgical specimens result in preparations of insufficient quality to allow the detection of minimal tumor infiltrates within normal brain parenchyma. Consequently, histologic examination has been considered an unreliable method of determining the boundaries of gliomas. In this study, we describe in detail an improved technique of biopsy specimen preparation that involves the use of glutaraldehyde-fixed and hemalum-phloxine-stained paraffin sections as well as alcohol-fixed and hemalum-phloxine-stained smears. In concert, these methods allow visualization of delicate cytologic details and are complementary in assessing the presence of isolated tumor cells at the periphery of gliomas. We define morphologic criteria for the accurate identification of isolated neoplastic cells and for their distinction from normal or reactive glial elements. These morphologic criteria include classic cytologic features of malignant lesions (such as nuclear pleomorphism and hyperchromasia) as well as the assessment of tumor architecture and cellular spatial relationships.


Bioactive phenylpropanoids from Daucus crinitus Desf. from Algeria.
The volatile constituents of Daucus crinitus Desf. from Algeria were analyzed by GC and GC-MS, The main constituent was isochavicol isobutyrate (39.0%), an uncommon phenylpropanoid. By synthesis of a series of homologous esters, it was also possible to determine the presence of small amounts of isochavicol propionate, which has never been described previously as a natural product. The antibacterial and antifungal activities of the whole essential oil, of these two esters, and of isochavicol itself were investigated against a wide range of bacteria and fungi. Additionally, their antimalarial and antiradical properties were also evaluated, showing an interesting antiplasmodial activity of isochavicol.


Optimized techniques for the extraction of grape allergens appropriate for in vivo and in vitro testing and diagnosis.
Standardized allergen extracts are needed for diagnosis and therapy purposes. For grapes, standardization is hampered by low protein and high tannin and pectin concentrations. The aim of the current study was to develop an optimized method for the extraction of grape proteins and possibly extend this to other fruits. Several existing or modified extraction methods were compared by means of protein concentration determination, SDS-PAGE, immunoblotting and radioallergosorbent test (RAST). An optimized extraction protocol was obtained in which we combined a high concentration of plant tissue, a concentrated, enriched and neutral buffer able to remove sugars and keep proteins soluble and a bivalent buffer for pectin removal. Both the quantitative (protein concentration) and qualitative parameters (SDS-PAGE protein patterns and IgE reactivity) were compared to standard protocols and commercial extracts used as diagnostic tools in the clinical practice. This method proved to be the most efficient mainly compared to the standard Björksten protocol in extracting the low molecular weight proteins, including the major grape allergen (lipid transfer protein, Vit v 1). It proved to be an easy, low cost and reproducible method proposed to prepare grape extracts that could replace the commercially available ones, used for diagnosis and possibly extend the method to other fruits especially in extracting LTPs.


[Long term effects o gastric banding for weight reduction].
Obesity is defined as body weight exceeding normal weight by 30%, or a body mass index (BMI) > 30. Various surgical procedures have been introduced to treat obesity. Gastric banding is a variant of the gastroplastic method. Our aim was to study the long term effect of this procedure. Over the years 1982 to 1988, gastric banding was performed at the county hospital of Haugesund in 26 patients with a mean age of 35.7 years and a mean preoperative BMI of 41.2. Data were obtained from patient files and from questionnaires returned by 24 patients, on average 14 years after the operation. Mean BMI for 16 patients with an intact banding device at follow-up was 35.2. Mean weight reduction was 15 kilograms. 58% of the patients regretted the procedure. Only a few were completely satisfied. Long term results of gastric banding are not satisfactory.


MicroRNA-561 promotes acetaminophen-induced hepatotoxicity in HepG2 cells and primary human hepatocytes through downregulation of the nuclear receptor corepressor dosage-sensitive sex-reversal adrenal hypoplasia congenital critical region on the X chromosome, gene 1 (DAX-1).
One of the major mechanisms involved in acetaminophen (APAP)-induced hepatotoxicity is hepatocyte nuclear factor 4α (HNF4α)-mediated activation of pregnane X receptor (PXR) and constitutive androstane receptor (CAR). In the present study, we investigated the role of miR-561 and its target gene DAX-1 encoding a corepressor of HNF4α in the process of APAP-induced hepatotoxicity. We used both human hepatocellular liver carcinoma cell line (HepG2) cells and primary human hepatocytes in this study and monitored the levels of reactive oxygen species, lactate dehydrogenase, and glutathione. Our bioinformatics study suggests an association between miR-561 and DAX-1, but not HNF4α. Treatment of HepG2 cells with APAP significantly reduced the expression of DAX-1 in a concentration-dependent manner. miR-561 was induced by APAP treatment in HepG2 cells. Transfection of HepG2 cells with an miR-561 mimic exacerbated APAP-induced hepatotoxicity. HNF4α is physically associated with DAX-1 in HepG2 cells. A decreased protein level of DAX-1 by APAP treatment was also enhanced by miR-561 mimic transfection in HepG2 cells and primary human hepatocytes. The basal and APAP-induced expression of PXR and CAR was enhanced by miR-561 mimic transfection; however, transfection of HepG2 cells or primary human hepatocytes with a miR-561 inhibitor or DAX-1 small interfering RNA reversed these effects. Additionally, the chromatin immunoprecipitation assay revealed that recruitment of DAX-1 onto the PXR promoter was inversely correlated with the recruitment of peroxisome proliferator-activated receptor-α coactivator-1α and HNF4α on APAP treatment. These results indicate that miR-561 worsens APAP-induced hepatotoxicity via inhibition of DAX-1 and consequent transactivation of nuclear receptors.


Seroprevalence survey of rubella infection in pregnancy at the University of Benin Teaching Hospital, Benin City, Nigeria.
To determine the serosusceptibility of rubella infection in pregnancy and the feasibility of establishing an organized prevention program in a tertiary hospital in Nigeria. This prospective, cross-sectional, laboratory-based study involved 300 consecutive pregnant women who gave informed consent and were screened for rubella immunoglobulins G (IgG) and M (IgM), using the ELISA-based quantitative assay at the University of Benin Teaching Hospital, Nigeria. Of the cohort, 30 women later withdrew. IgG seropositive samples were screened for IgM antibodies. The mean age and parity of the women were 30.0 ± 4.8 years, 95% CI 29.727-30.873 and 2.0 ± 1.4; 95% CI 1.317-1.661, respectively. IgG seroprevalence was 53%, while 10.0% of all IgG seropositive women were IgM seropositive. Most infections were acquired before the age of 35. None of the women ever had previous rubella vaccination. Rubella vaccine is scarce in Nigeria. Prevalence of rubella seromarkers for previous and current infection is high. Facilities for routine diagnosis and vaccination are lacking. Initiation of organized screening and vaccination programs is limited by lack of vaccine. We recommend immunization of children and women of child-bearing age as a cost-effective public health intervention strategy for managing the sequelae of the congenital rubella syndrome.


Effects of alcohol on exogenous precueing of attention.
Transient visual events cause a rapid allocation of attention to the event location. We studied the effect of alcohol on this aspect of attention. Subjects responded to targets presented either left or right of fixation and the time between target presentation and response (reaction time: RT) was measured both before and after ingestion of either alcohol or a placebo. Transient cues were presented prior to each target presentation. Cues were, variously, a brightening of either the box in which the target was to occur ("valid"), the box on the opposite side of the display from where the target was to occur ("invalid") or the fixation stimulus ("neutral"). The interval between cue and target presentation (stimulus onset asynchrony: SOA) was variously 50, 100, 150 or 800 msecs. At short SOAs, RT was faster to targets presented following valid cues relative to the other cues. At 800 msecs the opposite pattern was found. Alcohol selectively slowed RT only following neutral or invalid cues at short SOAs. Alcohol does not disrupt the normal attentional allocation to visual transients. It may delay the subsequent response to events at other locations, however.


Allium-Derived Compound Propyl Propane Thiosulfonate (PTSO) Attenuates Metabolic Alterations in Mice Fed a High-Fat Diet through Its Anti-Inflammatory and Prebiotic Properties.
Propyl propane thiosulfonate (PTSO) is an organosulfur compound from Allium spp. that has shown interesting antimicrobial properties and immunomodulatory effects in different experimental models. In this sense, our aim was to evaluate its effect on an experimental model of obesity, focusing on inflammatory and metabolic markers and the gut microbiota. Mice were fed a high-fat diet and orally treated with different doses of PTSO (0.1, 0.5 and 1 mg/kg/day) for 5 weeks. PTSO lessened the weight gain and improved the plasma markers associated with glucose and lipid metabolisms. PTSO also attenuated obesity-associated systemic inflammation, reducing the immune cell infiltration and, thus, the expression of pro-inflammatory cytokines in adipose and hepatic tissues (Il-1ẞ, Il-6, Tnf-α, Mcp-1, Jnk-1, Jnk-2, Leptin, Leptin R, Adiponectin, Ampk, Ppar-α, Ppar-γ, Glut-4 and Tlr-4) and improving the expression of different key elements for gut barrier integrity (Muc-2, Muc-3, Occludin, Zo-1 and Tff-3). Additionally, these effects were connected to a regulation of the gut microbiome, which was altered by the high-fat diet. Allium-derived PTSO can be considered a potential new tool for the treatment of metabolic syndrome.


Managing the innovation supply chain to maximize personalized medicine.
Personalized medicine epitomizes an evolving model of care tailored to the individual patient. This emerging paradigm harnesses radical technological advances to define each patient's molecular characteristics and decipher his or her unique pathophysiological processes. Translated into individualized algorithms, personalized medicine aims to predict, prevent, and cure disease without producing therapeutic adverse events. Although the transformative power of personalized medicine is generally recognized by physicians, patients, and payers, the complexity of translating discoveries into new modalities that transform health care is less appreciated. We often consider the flow of innovation and technology along a continuum of discovery, development, regulation, and application bridging the bench with the bedside. However, this process also can be viewed through a complementary prism, as a necessary supply chain of services and providers, each making essential contributions to the development of the final product to maximize value to consumers. Considering personalized medicine in this context of supply chain management highlights essential points of vulnerability and/or scalability that can ultimately constrain translation of the biological revolution or potentiate it into individualized diagnostics and therapeutics for optimized value creation and delivery.


Study of the Structure and Antimicrobial Activity of Ca-Deficient Ceramics on Chlorhexidine Nanoclay Substrate.
Novel biomedical composites, based on organically modified vermiculite and montmorillonite with deposited Ca-deficient hydroxyapatite (CDH), were prepared. The monoionic sodium forms of vermiculite and montmorillonite were intercalated with chlorhexidine diacetate (CA). The surfaces of organoclays were used for the precipitation of Ca-deficient hydroxyapatite. The composites with Ca-deficient hydroxyapatite showed very good antibacterial effects, similar to the antimicrobial activity of pure organoclay samples. Better antibacterial activity was shown in the organically modified montmorillonite sample with Ca-deficient hydroxyapatite compared with the vermiculite composite, but, in the case of Staphylococcus aureus, both composites showed the same minimum inhibitory concentration (MIC) value. The antimicrobial effect of composites against bacteria and fungi increased with the time of exposure. The structural characterization of all the prepared materials, performed using X-ray diffraction and FT infrared spectroscopy analysis, detected no changes in the original clay or CDH during the intercalation or precipitation process, therefore we expect the strength of the compounds to be in the original power.


Digital artifacts reveal development and diffusion of climate research.
Research for development organizations generate tremendous amount of accessible knowledge, but given their scale, time and resource constraints, the impact of outputs is not systematically analyzed. This is because traditional bibliometric analyses present limitations to synthesize accumulated knowledge and retrofitting indicators to historical outputs. To address these shortcomings, this study proposes an integrated, web-based approach to systematically analyze the production and diffusion of knowledge from large-scale research programs, using climate research of the International Maize and Wheat Improvement Center (CIMMYT) as a case study. Our analytical framework employs text mining, social network analysis and hyperlink analysis to an unstructured mass of publicly available digital artifacts such as institutional repositories, citation databases, and social media to uncover narratives, dynamics, and relationships. Findings show CIMMYT's climate research is strongly incorporated into a holistic systems approach and that the institution is actively engaged in knowledge exchanges with key actors from the scientific, development and public policy communities. The proposed analytical framework establishes an effective approach for research for development organizations to leverage existing online data sources to assess the extent of their knowledge production, dissemination, and reach.


The shape-memory effect in ionic elastomers: fixation through ionic interactions.
Shape-memory elastomers based on a commercial rubber cross-linked by both ionic and covalent bonds have been developed. The elastomeric matrix was a carboxylated nitrile rubber (XNBR) vulcanized with magnesium oxide (MgO) providing ionic interactions that form hierarchical structures. The so-named ionic transition is used as the unique thermal transition responsible for the shape-memory effect (SME) in these elastomers. These ionic interactions fix the temporary shape due to their behavior as dynamic cross-links with temperature changes. Covalent cross-links were incorporated with the addition of different proportions of dicumyl peroxide (DCP) to the ionic elastomer to establish and recover the permanent shape. In this article, the SME was modulated by modifying the degree of covalent cross-linking, while keeping the ionic contribution constant. In addition, different programming parameters, such as deformation temperature, heating/cooling rate, loading/unloading rate and percentage of tensile strain, were evaluated for their effects on shape-memory behavior.


The ubiquitin ligase TRIM25 targets ERG for degradation in prostate cancer.
Ets related gene (ERG) is a transcription factor that is overexpressed in 40% of prostate tumors due to a gene fusion between ERG and TMPRSS2. Because ERG functions as a driver of prostate carcinogenesis, understanding the mechanisms that influence its turnover may provide new molecular handles to target the protein. Previously, we found that ERG undergoes ubiquitination and then is deubiquitinated by USP9X in prostate cancer cells to prevent its proteasomal degradation. Here, we identify Tripartite motif-containing protein 25 (TRIM25) as the E3 ubiquitin ligase that ubiquitinates the protein prior to its degradation. TRIM25 binds full-length ERG, and it also binds the N-terminally truncated variants of ERG that are expressed in tumors with TMPRSS2-ERG fusions. We demonstrate that TRIM25 polyubiquitinates ERG in vitro and that inactivation of TRIM25 resulted in reduced polyubiquitination and stabilization of ERG. TRIM25 mRNA and protein expression was increased in ERG rearrangement-positive prostate cancer specimens, and we provide evidence that ERG upregulates TRIM25 expression. Thus, overexpression of ERG in prostate cancer may cause an increase in TRIM25 activity, which is mitigated by the expression of the deubiquitinase USP9X, which is required to stabilize ERG.


Intranasal therapeutic strategies for management of Alzheimer's disease.
Alzheimer's disease (AD) is a chronic and progressive age-related irreversible neurodegenerative disorder that represents 70% of all dementia with 35 million cases worldwide. Successful treatment strategies for AD have so far been limited, and present therapy is based on cholinergic replacement therapy and inhibiting glutamate excitotoxicity. In this context, role of neuroprotective drugs has generated considerable interest in management of AD. Recently, direct intranasal (IN) delivery of drug moieties to the central nervous system (CNS) has emerged as a therapeutically viable alternative to oral and parenteral routes. IN delivery bypasses the blood-brain barrier by delivering and targeting drugs to the CNS along the olfactory and trigeminal neural pathways which are in direct contact with both the environment and the CNS. In an attempt to understand how neurotherapeutics/nanoparticulate delivery systems can be transported from the nose to the CNS, the present review sets out to discuss the mechanism of transport from nose to brain. The aim of this review is to discuss and summarize the latest findings of some of the major studies on IN drug delivery in AD models, with a focus on the potential efficacy of neuroprotective treatments.


Comprehensive genomic profiling of relapsed and metastatic adenoid cystic carcinomas by next-generation sequencing reveals potential new routes to targeted therapies.
We hypothesized that next-generation sequencing could reveal actionable genomic alterations (GAs) and potentially expand treatment options for patients with advanced adenoid cystic carcinoma (ACC). Genomic profiling using next-generation sequencing was performed on hybridization-captured, adapter ligation libraries derived from 28 relapsed and metastatic formalin-fixed paraffin-embedded ACC. The 3230 exons of 182 cancer-related genes and 37 introns of 14 genes frequently rearranged in cancer were fully sequenced using the Illumina HiSeq 2000. All classes of GAs were evaluated. Actionable GAs were defined as those impacting targeted anticancer therapies on the market or in registered clinical trials. A total of 44 GAs were identified in the 28 ACC tumors, with 12 of 28 (42.9%) of tumors harboring at least 1 potentially actionable GA. The most common nonactionable GAs were identified in KD6MA (5 cases; 18%), ARID1A (4 cases; 14%), RUNX1 (2 cases; 7%), and MYC (2 cases; 7%). Actionable GAs included NOTCH1 (3 cases; 11%), MDM2 (2 cases; 7%), PDGFRA (2 cases; 7%), and CDKN2A/B (p16) (2 cases; 7%). Other potentially actionable GAs identified in a single case included: mutations in AKT1, BAP1, EGFR, and PIK3CA, homozygous deletion of FBXW7, and amplifications of CDK4, FGFR1, IGF1R, KDR, KIT, and MCL1. The frequency of GA in ACC is lower than that seen in the more common solid tumors. Comprehensive genomic profiling of ACC can identify actionable GAs in a subset of patients that could influence therapy for these difficult-to-treat progressive neoplasms.


Improving the reliability of visual acuity measures in young children.
Whilst the methodology of adult letter acuity measurement has been substantially refined over the last two decades, relatively little development has occurred in methods for quantifying letter acuity in young children. This study compares a recently developed visual acuity test (Glasgow Acuity Cards), which incorporates several key design features used in adult test charts to improve the sensitivity and reliability of visual acuity measurements. The equivalence of acuity measurements made with Glasgow Acuity Cards were compared with the Bailey-Lovie logMAR chart and Snellen chart in adults, and with traditional Single Letter Acuity and a modified Single Letter Acuity test in children. The test-retest reliability of acuity measurements made with Glasgow Acuity Cards and the Single Letter Acuity tests were also assessed in a large group of visually normal children. In addition, the ability of the pre-school letter acuity tests to detect differences in acuity between the two eyes, and to detect amblyopia were examined. Ninety-five percent of vision measurements made with the Bailey-Lovie chart and Glasgow Acuity Cards differ by less than 0.07 log unit. Furthermore, the sensitivity of Glasgow Acuity Cards to detecting changes in acuity longitudinally and inter-ocular differences in acuity is considerably greater as compared with traditional Single Letter Acuity tests. Improvements in paediatric acuity chart design are important for the effective detection and management of children with amblyopia.


Angular dependence of the response of the nanoDot OSLD system for measurements at depth in clinical megavoltage beams.
The purpose of this investigation was to assess the angular dependence of a commercial optically stimulated luminescence dosimeter (OSLD) dosimetry system in MV x-ray beams at depths beyond d(max) and to find ways to mitigate this dependence for measurements in phantoms. Two special holders were designed which allow a dosimeter to be rotated around the center of its sensitive volume. The dosimeter's sensitive volume is a disk, 5 mm in diameter and 0.2 mm thick. The first holder rotates the disk in the traditional way. It positions the disk perpendicular to the beam (gantry pointing to the floor) in the initial position (0°). When the holder is rotated the angle of the disk towards the beam increases until the disk is parallel with the beam ("edge on," 90°). This is referred to as Setup 1. The second holder offers a new, alternative measurement position. It positions the disk parallel to the beam for all angles while rotating around its center (Setup 2). Measurements with five to ten dosimeters per point were carried out for 6 MV at 3 and 10 cm depth. Monte Carlo simulations using GEANT4 were performed to simulate the response of the active detector material for several angles. Detector and housing were simulated in detail based on microCT data and communications with the manufacturer. Various material compositions and an all-water geometry were considered. For the traditional Setup 1 the response of the OSLD dropped on average by 1.4% ± 0.7% (measurement) and 2.1% ± 0.3% (Monte Carlo simulation) for the 90° orientation compared to 0°. Monte Carlo simulations also showed a strong dependence of the effect on the composition of the sensitive layer. Assuming the layer to completely consist of the active material (Al2O3) results in a 7% drop in response for 90° compared to 0°. Assuming the layer to be completely water, results in a flat response within the simulation uncertainty of about 1%. For the new Setup 2, measurements and Monte Carlo simulations found the angular dependence of the dosimeter to be below 1% and within the measurement uncertainty. The dosimeter system exhibits a small angular dependence of approximately 2% which needs to be considered for measurements involving other than normal incident beams angles. This applies in particular to clinical in vivo measurements where the orientation of the dosimeter is dictated by clinical circumstances and cannot be optimized as otherwise suggested here. When measuring in a phantom, the proposed new setup should be considered. It changes the orientation of the dosimeter so that a coplanar beam arrangement always hits the disk shaped detector material from the thin side and thereby reduces the angular dependence of the response to within the measurement uncertainty of about 1%. This improvement makes the dosimeter more attractive for clinical measurements with multiple coplanar beams in phantoms, as the overall measurement uncertainty is reduced. Similarly, phantom based postal audits can transition from the traditional TLD to the more accurate and convenient OSLD.


Patient satisfaction and survival of maxillary overdentures supported by four or six splinted implants: a systematic review with meta-analysis.
Implant-supported overdentures offer enhanced mechanical properties, which lead to better patient satisfaction and survival rates than conventional dentures. However, it is unclear whether these satisfaction levels and survival rates depend on the number of implants supporting the overdenture. Therefore, this systematic review aimed to compare maxillary overdentures supported by four or six splinted implants in terms of patient satisfaction, implant survival, overdenture survival, and prosthodontic complications. Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE (PubMed), and EMBASE databases were systematically searched and complemented by hand searching from 2000 to 2019, employing a combination of specific keywords. Studies comparing the use of four versus six implants for supporting overdentures with at least one-year of follow-up after prosthesis installation and including ten fully edentulous patients were included. The risk of bias (RoB) was analyzed with Cochrane's RoB 2 and Newcastle-Ottawa tools. Implants and prosthesis survival rates were analyzed by random-effects meta-analysis and expressed as risk ratios or risk differences, respectively, and by the non-parametric unpaired Fisher's test. A total of 15 from 1865 articles were included, and reported follow-up times after implant placement ranged from 1 to 10 years. Irrespective of the number of implants used, high scores were reported by all studies investigating patient satisfaction. Meta-analysis and non-parametric Fisher's test showed no statistical differences regarding the survival rate of implants (P = 0.34, P = 0.3) or overdentures (P = 0.74, P = 0.9) when using 4 versus 6 splinted implants to support overdentures, and no significant differences regarding prosthodontic complications were found between groups. Randomized studies presented high RoB and non-randomized studies presented acceptable quality. Within the limits of this systematic review, we can conclude that the bar-supported overdenture on four implants is not inferior to the overdenture supported by six implants for rehabilitating the edentulous maxilla, in terms of patient satisfaction, survival rates of implants and overdentures, and prosthodontic complications.


How Does Prior Experience Pay Off in Large-Scale Quality Improvement Initiatives?
To examine the association of prior investment on the effectiveness of organizations delivering large-scale external support to improve primary care. Mixed-methods study of 7 EvidenceNOW grantees (henceforth, Cooperatives) and their recruited practices (n = 1720). Independent Variable: Cooperatives's experience level prior to EvidenceNOW, defined as a sustained track record in delivering large-scale quality improvement (QI) to primary care practices (high, medium, or low). Dependent Variables: Implementation of external support, measured as facilitation dose; effectiveness at improving (1) clinical quality, measured as practices' performance on Aspirin, Blood Pressure, Cholesterol, and Smoking (ABCS); and (2) practice capacity, measured using the Adaptive Reserve (AR) score and Change Process Capacity Questionnaire (CPCQ). Data were analyzed using multivariable linear regressions and a qualitative inductive approach. Cooperatives with High (vs low) levels of prior experience with and investment in large-scale QI before EvidenceNOW recruited more geographically dispersed and diverse practices, with lower baseline ABCS performance (differences ranging from 2.8% for blood pressure to 41.5% for smoking), delivered more facilitation (mean=+20.3 hours, P = .04), and made greater improvements in practices' QI capacity (CPCQ: +2.04, P < .001) and smoking performance (+6.43%, P = .003). These Cooperatives had established networks of facilitators at the start of EvidenceNOW and leadership experienced in supporting this workforce, which explained their better recruitment, delivery of facilitation, and improvement in outcomes. Long-term investment that establishes regionwide organizations with infrastructure and experience to support primary care practices in QI is associated with more consistent delivery of facilitation support, and greater improvement in practice capacity and some clinical outcomes.


Stability indicating LC method for simultaneous determination of irbesartan and hydrochlorothiazide in pharmaceutical preparations.
A simple and precise stability-indicating liquid chromatography method is developed and validated for the quantitative simultaneous estimation of irbesartan (IRB) and hydrochlorothiazide (HCTZ) in combined pharmaceutical dosage form. A chromatographic separation of the two drugs was achieved with an Ace5 C(18) 25-cm analytical column using buffer-acetonitrile (70:30 v/v). The buffer used in mobile phase contains 50 mM ammonium acetate pH adjusted 5.5 with acetic acid. The instrumental settings are flow rate of 1.5 mL/min, column temperature at 30 degrees C, and detector wavelength of 235 nm using a photodiode array detector. IRB, HCTZ, and their combination drug products were exposed to thermal, photolytic, hydrolytic, and oxidative stress conditions, and the stressed samples were analyzed by the proposed method. Peak homogeneity data of IRB and HCTZ is obtained using photodiode array detector. In the stressed sample chromatograms, it demonstrated the specificity of the assay method for their estimation in presence of degradation products. The described method shows excellent linearity over a range of 10-200 microg/mL for IRB and 5-100 microg/mL for HCTZ. Methylparaben was used as internal standard. The correlation coefficient for IRB and HCTZ are 0.998 and 0.999. The mean recovery values for IRB and HCTZ ranged from 100.45% to 101.25%. The limit of detection for IRB and HCTZ were 0.019 and 0.023 microg/mL, respectively, and the limit of quantification were 0.053 and 0.070 microg/mL, respectively. The proposed method was suitable for quantitative determination and stability study of IRB and HCTZ in pharmaceutical preparations and also can be used in the quality control of bulk manufacturing and pharmaceutical dosage forms.


A French nationwide study compared various conditions and healthcare use of individuals < 65 years with a Down's syndrome to those without.
Few regular national clinical data are available for individuals with Down's syndrome (IDS) bearing in mind that they are subject to countries variations in medical termination of pregnancy and screening. Individuals < 65 in 2019 were selected in view of the low number of older IDS. Thus, 98% of 52.4 million people with correct data were included from the national health data system. IDS (35,342) were identified on the basis of the International Classification of Diseases 10th revision code (Q90). Risk ratios (RR) were calculated to compare the frequencies in 2019 between IDS and individual without Down's syndrome (IWDS) of use of health care. The prevalence of IDS was 0.07% (48% women), comorbidities were more frequent, especially in younger patients (24% < 1 year had another comorbidity, RR = 20), as was the percentage of deaths (4.6%, RR = 10). Overall, tumours were less frequent in IDS compared with IWDS (1.2%, RR = 0.7) except for certain leukaemias and testicular tumours (0.3%, RR = 4). Cardiac malformations (5.2%, RR = 52), dementia (1.2%, RR = 29), mental retardation (5%, RR = 21) and epilepsy (4%, RR = 9) were also more frequent in IDS. The most frequent hospital diagnoses for IDS were: aspiration pneumonia (0.7%, RR = 89), respiratory failure (0.4%, RR = 17), sleep apnoea (1.1%, RR = 8), cryptorchidism (0.3%, RR = 5.9), protein-energy malnutrition (0.1%, RR = 7), type 1 diabetes (0.2%, RR = 2.8) and hypothyroidism (0.1%, RR = 72). IDS were more likely to use emergency services (9%, RR = 2.4), short hospital stay (24%, RR = 1.6) or hospitalisation at home (0.6%, RR = 6). They consulted certain specialists two to three times more frequently than IWDS, for example cardiologists (17%, RR = 2.6). This study is the first detailed national study comparing IDS and non-IDS by age group. These results could help to optimize prenatal healthcare, medical and social support.


Monascus-fermented yellow pigments monascin and ankaflavin showed antiobesity effect via the suppression of differentiation and lipogenesis in obese rats fed a high-fat diet.
Monascus-fermented monascin and ankaflavin are found to strongly inhibit differentiation and lipogenesis and stimulate lipolysis effects in a 3T3-L1 preadipocyte model, but the in vivo regulation mechanism is unclear. This study uses obese rats caused by a high-fat diet to examine the effects of daily monascin and ankaflavin feeding (8 weeks) on antiobesity effects and modulation of differentiation, lipogenesis, and lipid absorption. The results show that monascin and ankaflavin had a significant antiobesity effect, which should result from the modulation of monascin and ankaflavin on the inhibition of differentiation by inhibiting CCAT/enhancer-binding protein β (C/EBPβ) expression (36.4% and 48.3%) and its downstream peroxisome proliferator-activated receptor γ (PPARγ) (55.6% and 64.5%) and CCAT/enhancer-binding protein α (C/EBPα) expressions (25.2% and 33.2%) and the inhibition of lipogenesis by increasing lipase activity (14.0% and 10.7%) and decreasing heparin releasable lipoprotein lipase (HR-LPL) activity (34.8% and 30.5%). Furthermore, monascin and ankaflavin are the first agents found to suppress Niemann-Pick C1 Like 1 (NPC1L1) protein expression (73.6% and 26.1%) associated with small intestine tissue lipid absorption. Importantly, monascin and ankaflavin are not like monacolin K, which increases creatine phosphokinase (CPK) activity, known as a rhabdomyolysis indicator.


Growth charts of human development.
This article reviews and compares two types of growth charts for tracking human development over age. Both charts assume the existence of a continuous latent variable, but relate to the observed data in different ways. The D-score diagram summarizes developmental indicators into a single aggregate score measuring global development. The relations between the indicators should be consistent with the Rasch model. If true, the D-score is a measure with interval scale properties, and allows for the calculation of meaningful differences both within and across age. The stage line diagram describes the natural development of ordinal indicators. The method models the transition probabilities between successive stages of the indicator as smoothly varying functions of age. The location of each stage is quantified by the mid-P-value. Both types of diagrams assist in identifying early and delayed development, as well as finding differences in tempo. The relevant techniques are illustrated to track global development during infancy and early childhood (0-2 years) and Tanner pubertal stages (8-21 years). New reference values for both applications are provided.


Advanced MRI of the Optic Nerve.
Clinical orbital MRI protocols are routinely used to study the optic nerves and exclude compressive lesions, infarctions, or inflammation. However, the small caliber and divergent oblique orientations of the optic nerves make it challenging to characterize them well with conventional MRI, especially since adjacent air-filled bony structures distort the MRI signal and motion is a problem even in cooperative, healthy volunteers. Over the past 3 years we have experimented with multiple novel MRI approaches and sequences to better characterize the optic nerves. The perfect MRI protocol would be quantitative and sensitive to subtle optic nerve pathologic changes, provide high spatial resolution, be rapidly acquired, and resistant to motion degradation. This review provides an update of recent MRI sequence innovations for the optic nerves being currently translated into clinical practice. Methods discussed include rapid MRI with compressed sensing or simultaneous multislice approaches, postprocessing techniques for quantitative T2 mapping or track density imaging, and multiple MRI sequences for measuring diffusion in the optic nerves. Recently-developed orbit-specific MRI coils, quantitative sequences, and rapid acquisition techniques can improve our future ability to study optic nerve pathologies noninvasively. As advanced MRI becomes more proficient at characterizing the optic nerves, its role in the clinical management of patients should increase.


Simpler Certified Radius Maximization by Propagating Covariances.
One strategy for adversarially training a robust model is to maximize its certified radius - the neighborhood around a given training sample for which the model's prediction remains unchanged. The scheme typically involves analyzing a "smoothed" classifier where one estimates the prediction corresponding to Gaussian samples in the neighborhood of each sample in the mini-batch, accomplished in practice by Monte Carlo sampling. In this paper, we investigate the hypothesis that this sampling bottleneck can potentially be mitigated by identifying ways to directly propagate the covariance matrix of the smoothed distribution through the network. To this end, we find that other than certain adjustments to the network, propagating the covariances must also be accompanied by additional accounting that keeps track of how the distributional moments transform and interact at each stage in the network. We show how satisfying these criteria yields an algorithm for maximizing the certified radius on datasets including Cifar-10, ImageNet, and Places365 while offering runtime savings on networks with moderate depth, with a small compromise in overall accuracy. We describe the details of the key modifications that enable practical use. Via various experiments, we evaluate when our simplifications are sensible, and what the key benefits and limitations are.


Direct effects of muscarinic agents on the outflow pathways in human eyes.
Recent studies demonstrating the presence of muscarinic receptors and contractile-like cells in the trabecular meshwork tissue and/or cell cultures from human eyes suggest the possibility that there may be a direct effect of muscarinic agonists on outflow facility. The present studies were conducted to determine whether muscarinic agonists could change outflow facility in perfused human ocular anterior segments, which lack an intact ciliary muscle. Human eyes were dissected and perfused according to previously described methods. A steady state baseline facility was established for 90 minutes, after which up to four sequential concentrations ranging from 10(-9) to 10(-3) M of pilocarpine, aceclidine, or carbachol were added to the perfusion medium. In other studies, 10(-6) M atropine was perfused alone followed by 10(-7) M carbachol with 10(-6) M atropine, whereas fellow control eyes received carbachol alone. Outflow facility was measured for 60 minutes after each drug addition. The outflow facility measurement in each eye after drug administration was compared with the baseline measurement. Outflow facility increased from baseline facility in eyes treated with pilocarpine, aceclidine, or carbachol at lower concentrations (10(-9) to 10(-6) M) but remained unchanged at higher concentrations (10(-4) to 10(-2) M). The effects of carbachol at 10(-7) M were completely blocked by atropine. Muscarinic agonists increase outflow facility in human eyes by a direct stimulation of the outflow tissues in the absence of an intact ciliary muscle. This effect is biphasic, occurring at concentrations of 10(-6) M and lower with no effect at higher concentrations.


An epigenetic hypothesis for the genomic memory of pain.
Chronic pain is accompanied with long-term sensory, affective and cognitive disturbances. What are the mechanisms that mediate the long-term consequences of painful experiences and embed them in the genome? We hypothesize that alterations in DNA methylation, an enzymatic covalent modification of cytosine bases in DNA, serve as a "genomic" memory of pain in the adult cortex. DNA methylation is an epigenetic mechanism for long-term regulation of gene expression. Neuronal plasticity at the neuroanatomical, functional, morphological, physiological and molecular levels has been demonstrated throughout the neuroaxis in response to persistent pain, including in the adult prefrontal cortex (PFC). We have previously reported widespread changes in gene expression and DNA methylation in the PFC many months following peripheral nerve injury. In support of this hypothesis, we show here that up-regulation of a gene involved with synaptic function, Synaptotagmin II (syt2), in the PFC in a chronic pain model is associated with long-term changes in DNA methylation. The challenges of understanding the contributions of epigenetic mechanisms such as DNA methylation within the PFC to pain chronicity and their therapeutic implications are discussed.


Surfing the internet for health information: an italian survey on use and population choices.
Recent international sources have described how the rapid expansion of the Internet has precipitated an increase in its use by the general population to search for medical information. Most studies on e-health use investigated either through the prevalence of such use and the social and income patterns of users in selected populations, or the psychological consequences and satisfaction experienced by patients with particular diseases. Few studies have been carried out in Europe that have tried to identify the behavioral consequences of Internet use for health-related purposes in the general population.The aims of this study are to provide information about the prevalence of Internet use for health-related purposes in Italy according to demographic and socio-cultural features, to investigate the impact of the information found on health-related behaviors and choices and to analyze any differences based on health condition, self-rated health and relationships with health professionals and facilities. A multicenter survey was designed within six representative Italian cities. Data were collected through a validated questionnaire administered in hospital laboratories by physicians. Respondents were questioned about their generic condition, their use of the Internet and their health behaviors and choices related to Internet use. Data were analyzed using descriptive statistics and logistic regression to assess any differences by socio-demographic and health-related variables. The sample included 3018 individuals between the ages of 18 and 65 years. Approximately 65% of respondents reported using the Internet, and 57% of them reported using it to search for health-related information. The main reasons for search on the Internet were faster access and a greater amount of information. People using the Internet more for health-related purposes were younger, female and affected by chronic diseases. A large number of Internet users search for health information and subsequently modify their health behaviors and relationships with their medical providers. This may suggest a strong public health impact with consequences in all European countries, and it would be prudent to plan educational and prevention programs. However, it could be important to investigate the quality of health-related websites to protect and inform users.


Interaction of methotrexate polyglutamates and dihydrofolate during leucovorin rescue in a human breast cancer cell line (MCF-7).
Previous investigations have suggested that high-dose methotrexate with leucovorin rescue is a potentially useful strategy for overcoming antifolate resistance. Interactions between methotrexate (MTX) and leucovorin and their respective metabolites appear to occur at multiple intracellular sites, including dihydrofolate reductase (MTX/MTX polyglutamates versus dihydrofolate) and other folate-dependent enzymes (MTX polyglutamates versus reduced folate substrates). The present studies were designed to test the ability of dihydrofolate to compete with methotrexate and methotrexate polyglutamates for dihydrofolate reductase activity using an intact human breast carcinoma cell line (MCF-7) as the model system. Exposure of the breast cells to methotrexate for 24 h resulted in a concentration-dependent formation of methotrexate polyglutamates that markedly exceeded the dihydrofolate reductase-binding capacity for up to 24 h after the removal of drug from the growth media. Under these conditions of dihydrofolate reductase inhibition, we found that tritium-labeled dihydrofolate was capable of competing with methotrexate and its metabolites for dihydrofolate reductase activity as evidenced by the appearance of tritium-labeled reduced folates in the treated cells. We found the interaction between dihydrofolate and methotrexate to be dependent on the exposure concentrations of both methotrexate and dihydrofolate. These studies provide direct evidence that competition during leucovorin rescue occurs at the level of dihydrofolate reductase between methotrexate polyglutamates and dihydrofolate polyglutamates in intact human cells.


Research design and statistical methods in Indian medical journals: a retrospective survey.
Good quality medical research generally requires not only an expertise in the chosen medical field of interest but also a sound knowledge of statistical methodology. The number of medical research articles which have been published in Indian medical journals has increased quite substantially in the past decade. The aim of this study was to collate all evidence on study design quality and statistical analyses used in selected leading Indian medical journals. Ten (10) leading Indian medical journals were selected based on impact factors and all original research articles published in 2003 (N = 588) and 2013 (N = 774) were categorized and reviewed. A validated checklist on study design, statistical analyses, results presentation, and interpretation was used for review and evaluation of the articles. Main outcomes considered in the present study were - study design types and their frequencies, error/defects proportion in study design, statistical analyses, and implementation of CONSORT checklist in RCT (randomized clinical trials). From 2003 to 2013: The proportion of erroneous statistical analyses did not decrease (χ2=0.592, Φ=0.027, p=0.4418), 25% (80/320) in 2003 compared to 22.6% (111/490) in 2013. Compared with 2003, significant improvement was seen in 2013; the proportion of papers using statistical tests increased significantly (χ2=26.96, Φ=0.16, p<0.0001) from 42.5% (250/588) to 56.7 % (439/774). The overall proportion of errors in study design decreased significantly (χ2=16.783, Φ=0.12 p<0.0001), 41.3% (243/588) compared to 30.6% (237/774). In 2013, randomized clinical trials designs has remained very low (7.3%, 43/588) with majority showing some errors (41 papers, 95.3%). Majority of the published studies were retrospective in nature both in 2003 [79.1% (465/588)] and in 2013 [78.2% (605/774)]. Major decreases in error proportions were observed in both results presentation (χ2=24.477, Φ=0.17, p<0.0001), 82.2% (263/320) compared to 66.3% (325/490) and interpretation (χ2=25.616, Φ=0.173, p<0.0001), 32.5% (104/320) compared to 17.1% (84/490), though some serious ones were still present. Indian medical research seems to have made no major progress regarding using correct statistical analyses, but error/defects in study designs have decreased significantly. Randomized clinical trials are quite rarely published and have high proportion of methodological problems.


[Accident prevention and emergency treatment of traumatic hand injuries in the metropolitan area of São Paulo].
The workman's hand is a "tool"; it is his most valuable asset. Worker selection and training, associated with a preventional approach, by employee and employer, humanises work and makes it more effective. The rate of hand injuries in the city of São Paulo is very high (of the order of 500 cases per day) and its aggregate cost is extremely high. The primary care of lesions is so poor that it well deserves the label of "second accident". The CIPAM-SUDS/SP system will provide a simpler and proper treatment by trained surgeons in full time duty in first-aid decentralized stations, located inside the industrial areas.


Paper persistence, workarounds, and communication breakdowns in computerized consultation management.
The consultation request process between primary care and specialty services often contains paper-based components that may be inefficient and difficult to track. Other barriers may include workarounds and communication breakdowns with the potential to adversely impact delivery of quality medical care. We investigated current challenges to the electronic outpatient consult management process in the United States Veterans Health Administration (VHA). We conducted ethnographic observation and semi-structured interviews in nine different specialty clinics and three primary care clinics in a large, tertiary Veterans Affairs Medical Center (VAMC). We also performed a national-level query of 'electronic error and enhancement requests' (E3Rs) related to the consult package in the VA's electronic health record (EHR) submitted over a 5-year period (2005-2009). Two researchers recorded the observable interactions and interview responses of 16 healthcare workers related to their work with consultations. Two separate coding schemes were applied to both the observational and the interview data. E3Rs from the national query were reviewed and categorized based on the nature of the enhancement requests. We identified several examples of paper persistence, as well as workarounds, communication breakdowns, and redundancies in computerized consult management. An analysis of enhancement requests for the consults also revealed three broad needs related to reporting, configuration or customization, and user interface enhancements. Understanding these challenges to the current consult management process is important to help design enhanced informatics tools integrated into workflow to support coordination of care and tracking of consults requests.


Brain tumors with ipsilateral cerebral hemiatrophy.
Brain tumors with secondary ipsilateral cerebral hemiatrophy are so rare that only seven cases have been reported in the literature. Three new cases are presented and the clinical findings in all 10 cases are reviewed. The diagnostic value of computed tomography (CT) is emphasized. The authors conclude that nodular high-density lesions located deep in the cerebrum and showing slight contrast enhancement on CT may be associated with ipsilateral cerebral hemiatrophy in young male patients with slowly progressive hemiparesis, dementia, and personality change. In such cases, early diagnosis of germinoma and subsequent radiotherapy may prevent unnecessary surgery.


The effect of bracing on patella alignment and patellofemoral joint contact area.
To examine the influence of two patellofemoral braces on pain response, patellar alignment, and patellofemoral joint contact area in persons with patellofemoral pain. Fifteen women between the ages of 18 and 45 yr with a diagnosis of patellofemoral pain participated. After the assessment of pain response using a visual analog scale, subjects underwent axial plane magnetic resonance imaging of patellofemoral joint at 0 degrees, 20 degrees, 40 degrees, and 60 degrees of knee flexion. Imaging was done with the knee extensors contracted (25% body weight) under three conditions: 1) no brace, 2) On-Track brace, and 3) Patellar Tracking Orthosis (PTO). Measures of mediolateral patellar displacement and tilt and medial and lateral facet contact area were obtained from the magnetic resonance images. On average, the On-Track brace reduced symptoms by 50%, whereas the PTO reduced pain by 44%. When averaged across all knee flexion angles, the PTO and the On-Track brace significantly increased total patellofemoral joint contact area by 52.0 mm (21%) and 59.3 mm (24%), respectively, when compared with the no-brace condition. Bracing had no influence on lateral patellar tilt; however, small but significant changes in lateral patellar displacement were observed. Large changes in pain and contact area occurred without sizable changes in patellar alignment. The results of this study suggest that changes in patellar alignment by itself may not be responsible for pain alleviation after patellar bracing.


Concise review: chromatin and genome organization in reprogramming.
The ability to reprogram somatic cells to pluripotency is continuingly attracting increasing amounts of attention, providing both potential opportunities for regenerative medicine, as well as an intriguing model to study basic mechanisms of developmental reversal and epigenetic erasure. Currently, nuclear reprogramming is an inefficient process and a better understanding of its components and the underlying mechanisms will no doubt enable us to increase its robustness and to gain a deeper understanding of its regulation. Here we focus on the reprogramming process from the chromatin and genome organization perspective, describing the chromatin changes that occur both globally and locally. At the global level, chromatin decondenses toward the characteristic 'open' state, while locally, chromatin reorganization supports the silencing of lineage-specific genes and the activation of pluripotency-related genes. Importantly, the proteins that regulate this process are being identified, revealing different layers of chromatin regulation, including histone modifications, histone variants, chromatin remodeling and genomic DNA methylation. The emerging theme is that chromatin and genome organization are not only altered during the transition from a somatic to a pluripotent state, but also play active, regulatory roles during the reprogramming process.


Hyperthermia: the immune response and tumor metastasis.
Evidence is circumstantial that in animals and, to some extent, in man, an antitumor immune response may be generated after curative hyperthermia that leads to disappearance of metastases (abscopal response) and the acquisition of host immunity. In rodents, tumor regression after heating does not occur in immunosuppressed hosts, and the tumor cure rate is significantly reduced by inhibition of macrophage activity with silica; cured immune rats succumb to tumor inoculation when immunosuppressed. Quantitative data for cellular and humoral immunocompetence in these situations exist for only a few tumor types, and the bulk of evidence indicates that host response following tumor heating is nonspecific in type, with a major macrophage component. Little is distinctive about the regression of heated tumors, i.e., an abscopal response can follow treatment of tumors by excision or hypothermia. In man, an immune response is seldom evoked by the heating of the common solid tumors. Whole-body heating in animals can cause immunosuppression, probably from a direct damaging effect on lymphoid tissue, and enhanced metastatic spread may follow in the tumor-bearing host, but this has not been proved in man. The differences in response of tumors to heat in animals and man may be due to variations in tumor immunogenicity and host tolerance to heat.


Cobalamin deficiency: clinical picture and radiological findings.
Vitamin B12 deficiency causes a wide range of hematological, gastrointestinal, psychiatric and neurological disorders. Hematological presentation of cobalamin deficiency ranges from the incidental increase of mean corpuscular volume and neutrophil hypersegmentation to symptoms due to severe anemia, such as angor, dyspnea on exertion, fatigue or symptoms related to congestive heart failure, such as ankle edema, orthopnea and nocturia. Neuropsychiatric symptoms may precede hematologic signs and are represented by myelopathy, neuropathy, dementia and, less often, optic nerve atrophy. The spinal cord manifestation, subacute combined degeneration (SCD), is characterized by symmetric dysesthesia, disturbance of position sense and spastic paraparesis or tetraparesis. The most consistent MRI finding is a symmetrical abnormally increased T2 signal intensity confined to posterior or posterior and lateral columns in the cervical and thoracic spinal cord. Isolated peripheral neuropathy is less frequent, but likely overlooked. Vitamin B12 deficiency has been correlated negatively with cognitive functioning in healthy elderly subjects. Symptoms include slow mentation, memory impairment, attention deficits and dementia. Optic neuropathy occurs occasionally in adult patient. It is characterized by symmetric, painless and progressive visual loss. Parenteral replacement therapy should be started soon after the vitamin deficiency has been established.


Human phenotype caused by biallelic KDM4B frameshift variant.
KDM4B (MIM*609765, NM_015015.3, formerly JMJD2B) encodes a histone demethylase and regulates gene expression via demethylation, mainly of H3K9 tri-methylation. Heterozygous KDM4B loss-of-function variants cause autosomal dominant intellectual developmental disorder 65 (MIM#619320), which is characterized by global developmental delay, intellectual disability, language and gross motor delays, structural brain anomalies, characteristic facial features, and clinodactyly. Although the majority of reported patients have de novo pathogenic variants, some patients inherit pathogenic variants from affected parents. To our knowledge, only 23 patients with heterozygous KDM4B variants have been reported to date, and there are no reports of patients with biallelic KDM4B pathogenic variants. Herein, we report a female patient with a biallelic KDM4B frameshift variant (NM_015015.3: c.1384_1394delinsGGG, p.(Leu462Glyfs*43)) located at exon 12 of 23 protein-coding exons, which is thought to be subject to nonsense-mediated mRNA decay and no protein production. She presented developmental and language delays and a hypotonic and characteristic face. The patient's phenotype was more obvious than that of her mother, who is heterozygous for the same variant. Although declining birth rate (embryonic lethality in male mice) in homozygous knockout mice has been demonstrated, our report suggests that homozygous KDM4B frameshift variants can be viable in humans at least female.


Acceptable donor-specific antibody levels allowing for successful deceased and living donor kidney transplantation after desensitization therapy.
The aims of this study were to determine the level of donor-specific antibody (DSA) that allows for successful transplantation after desensitization with IVIG and rituximab and to identify patients at risk for antibody-mediated rejection (AMR). Pre- and posttransplant sera from 16 patients with DSA before desensitization were tested. Strength of DSA was determined by single antigen Luminex bead assay and results expressed as standard fluorescence intensity (SFI). T-cell flow crossmatch results were expressed as mean channel shifts (MCS). AMR was determined by biopsy and C4d deposition. Six had negative pretransplant flow crossmatches with a mean DSA of 8,805 SFI. Five had positive flow crossmatches (78-192 MCS) with mean DSA of 55,869 SFI. No patients in either group had AMR. Five had positive flow crossmatches (222-266 MCS) with mean DSA of 118,063 SFI. Three experienced AMR. The MCS and DSA levels for patients with AMR were significantly higher than patients without (P < or = 0.001). For patients without complications (n=7), DSA remained less than 10(5) SFI and usually decreased to approximately 10(4) SFI posttransplant for both class I and II. For patients with AMR (n=3), predominant increases in class II DSA more than 10(5) SFI were observed. All three patients continue to have DSA approximately 10(5) SFI with stable creatinine after treatment for AMR. Approximately 63% of patients were transplanted with a positive flow crossmatch. The results show that patients with DSA more than 10(5) and FCM more than 200 MCS are at higher risk for AMR. Treatment of AMR improves renal function without significant changes in DSA.


Cell death and long-term maintenance of neuron-like state after differentiation of rat bone marrow stromal cells: a comparison of protocols.
Recent literature suggests that bone marrow stromal cells (BMSCs) may be differentiated into neuron-like and/or glia-like cells, implying that differentiated BMSCs may have potential use in cell replacement therapy for central nervous system disorders. However, many questions remain concerning the nature of BMSCs differentiated to express CNS antigens. For example, how long after differentiation do cells express CNS markers, and do differentiation procedures alter cell viability? This study compared neuronal differentiation methods in sister cell preparations, evaluating cell death and maintenance of the CNS antigen positivity after the Deng or Woodbury methods. Rat BMSCs were harvested, passaged, differentiated, placed in growth or maintenance media, and processed for cell viability or immunocytochemistry for NeuN and GFAP post-differentiation. We report that the Woodbury differentiation protocol produced maximally 51% neuron-like cells, yet also produced significant cell death. The Deng differentiation method produced 13% neuron-like cells and without marked cell death. No significant increases in GFAP immunoreactivity (IR) were seen after differentiation by either protocol. Following both protocols, removal of cells from the maintenance media significantly decreased expression of NeuN. Thus, differentiation procedures may be substantially affected BMSC potential, and maintenance of immunoreactivity to neuronal antigens was dependent on specific, nonphysiological environmental conditions.


Trophoblast PR-SET7 dysfunction induces viral mimicry response and necroptosis associated with recurrent miscarriage.
Recurrent miscarriage (RM) is a distressing pregnancy complication. While the etiology of RM remains unclear, growing evidence has indicated the relevance of trophoblast impairment to the pathogenesis of RM. PR-SET7 is the sole enzyme catalyzing monomethylation of H4K20 (H4K20me1) and has been implicated in many pathophysiological processes. However, how PR-SET7 functions in trophoblasts and its relevance to RM remain unknown. Here, we found that trophoblast-specific loss of Pr-set7 in mice led to defective trophoblasts, resulting in early embryonic loss. Mechanistic analysis revealed that PR-SET7 deficiency in trophoblasts derepressed endogenous retroviruses (ERVs), leading to double-stranded RNA stress and subsequent viral mimicry, which drove overwhelming interferon response and necroptosis. Further examination discovered that H4K20me1 and H4K20me3 mediated the inhibition of cell-intrinsic expression of ERVs. Importantly, dysregulation of PR-SET7 expression and the corresponding aberrant epigenetic modifications were observed in the placentas of RM. Collectively, our results demonstrate that PR-SET7 acts as an epigenetic transcriptional modulator essential for repressing ERVs in trophoblasts, ensuring normal pregnancy and fetal survival, which sheds new light on potential epigenetic causes contributing to RM.


Dose fractionation schedules--biologic aspects and applications to high LET radiotherapy.
The main differences between biologic effects of high and low LET irradiation are reviewed with special reference to multiple dose fractionation schedules. Some new results relating to RBE values of normal tissues (skin) at low doses per fraction are presented. Recent results of experiments to investigate optimum x-ray or fast neutron fractionation schedules are summarized. It is concluded that, until much more is known about the processes occurring inside tumors during treatment, high LET radiation is likely to be more reliable for eradicating tumors.


Homoacetogenesis and microbial community composition are shaped by pH and total sulfide concentration.
Biological CO2 sequestration through acetogenesis with H2 as electron donor is a promising technology to reduce greenhouse gas emissions. Today, a major issue is the presence of impurities such as hydrogen sulfide (H2 S) in CO2 containing gases, as they are known to inhibit acetogenesis in CO2 -based fermentations. However, exact values of toxicity and inhibition are not well-defined. To tackle this uncertainty, a series of toxicity experiments were conducted, with a mixed homoacetogenic culture, total dissolved sulfide concentrations ([TDS]) varied between 0 and 5 mM and pH between 5 and 7. The extent of inhibition was evaluated based on acetate production rates and microbial growth. Maximum acetate production rates of 0.12, 0.09 and 0.04 mM h-1 were achieved in the controls without sulfide at pH 7, pH 6 and pH 5. The half-maximal inhibitory concentration (IC50 qAc ) was 0.86, 1.16 and 1.36 mM [TDS] for pH 7, pH 6 and pH 5. At [TDS] above 3.33 mM, acetate production and microbial growth were completely inhibited at all pHs. 16S rRNA gene amplicon sequencing revealed major community composition transitions that could be attributed to both pH and [TDS]. Based on the observed toxicity levels, treatment approaches for incoming industrial CO2 streams can be determined.


Effects of Polyphenol-Rich Fruit Extracts on Diet-Induced Obesity in Rodents: Systematic Review and Meta-Analysis.
Obesity is a complex condition of high prevalence and cost to the public health system. Recent research has demonstrated the potential of natural products, such as polyphenol-rich fruit extracts, for use in the treatment of obesity. The goal of this systematic review and meta-analysis is to determine the metabolic effects of polyphenol-rich fruit extracts on diet-induced obesity (DIO) in rodents. We searched MEDLINE, EMBASE, and Web of Science databases to identify preclinical studies that assessed polyphenol-rich fruit extracts compared to placebo on DIO in rodents in December 2018. Two researchers selected the studies, extracted the data, and assessed the quality of studies. Meta-analyses of standardized mean difference (SMD) of outcomes were calculated in Stata 11, and causes of heterogeneity were assessed by meta-regression. We included 14 studies in the systematic review and 13 studies with 21 matched groups in the metaanalysis. Polyphenol-rich fruit extracts reduced the total body weight gain (SMD = -1.48; confidence interval: - 1.95, -1.01), energy intake (SMD = -0.42; -0.67, -0.17), visceral adipose tissue (SMD = -0.96; -1.25, -0.66), triglycerides (SMD = -1.00; -1.39, -0.62), cholesterol (SMD = -1.18, -1.66, -0.69), LDL- c (SMD = -1.15; -1.65, - 0.65), fasting glucose (SMD = -1.05; -1.65, -0.46), and fasting insulin (SMD = -1.40; -1.80, -1.00) when compared to vehicle. Polyphenol-rich fruit extract had positive effects on weight gain, dyslipidaemia, insulin resistance at different doses, and fruit source in male mice.


Cysteine protease activities during maturation stages of Paragonimus westermani.
In mature Paragonimus westermani, specific activity of parasitic cysteine protease declines. To clarify which of the known 17-, 27-, and 28-kDa enzyme activities is decreased, the cysteine proteases were purified from the crude extracts of metacercariae, 4- and 7-wk juveniles, and 16-wk adults by gel filtration, ion-exchange, and affinity matrix chromatographies; the enzyme activity was monitored with the fluorogenic substrate, Cbz-phe-arg-AMC. In addition to 3 known enzymes, 2 other cysteine proteases at 15 and 53 kDa were identified in juveniles and adults and were purified. The 2 novel enzymes were most active in 0.1 M ionic strength and pH 5-6 and were inhibited by N-(N-[L-3-transcarboxyrane-2-carbonyl]-L-leucyl)agamatine, iodoacetamide, and leupeptin. Of the 5 enzymes, specific activities of metacercarial 27- and 28-kDa enzymes were lowered from metacercaria to 16 wk. Between 4 and 16 wk, activities of 3 cysteine proteases of juveniles and adults were additionally exhibited. The activity changes of 5 different cysteine proteases may be associated with migration and immune evasion during the maturation stage of P. westermani when the parasite environment is changing.


Telomere-targeted retrotransposons in the rice blast fungus Magnaporthe oryzae: agents of telomere instability.
The fungus Magnaporthe oryzae is a serious pathogen of rice and other grasses. Telomeric restriction fragments in Magnaporthe isolates that infect perennial ryegrass (prg) are hotspots for genomic rearrangement and undergo frequent, spontaneous alterations during fungal culture. The telomeres of rice-infecting isolates are very stable by comparison. Sequencing of chromosome ends from a number of prg-infecting isolates revealed two related non-LTR retrotransposons (M. oryzae Telomeric Retrotransposons or MoTeRs) inserted in the telomere repeats. This contrasts with rice pathogen telomeres that are uninterrupted by other sequences. Genetic evidence indicates that the MoTeR elements are responsible for the observed instability. MoTeRs represent a new family of telomere-targeted transposons whose members are found exclusively in fungi.


Unique structural characteristics and evolution of a cluster of venom phospholipase A2 isozyme genes of Protobothrops flavoviridis snake.
Protobothrops flavoviridis (Crotalinae) venom gland phospholipase A(2) (PLA(2)) isozyme genes have evolved in an accelerated manner to acquire diverse physiological activities in their products. For elucidation of the multiplication mechanism of PLA(2) genes, a 25,026 bp genome segment harboring five PLA(2) isozyme genes was obtained from Amami-Oshima P. flavoviridis liver and sequenced. The gene PfPLA 2 encoded [Lys(49)]PLA(2) called BPII, the gene PfPLA 4 neurotoxic [Asp(49)]PLA(2) called PLA-N, the gene PfPLA 5 basic [Asp(49)]PLA(2) called PLA-B, and PfPLA 1(psi) and PfPLA 3(psi) were the inactivated genes. The 5' truncated reverse transcriptase (RT) elements, whose intact forms constitute long interspersed nuclear elements (LINEs), were found in close proximity to the 3' end of PLA(2) genes and named PLA(2) gene-coupled RT fragments (PcRTFs). The facts that PcRTFs have the stem-loop and repetitive sequence in the 3' untranslated region (UTR) which is characteristic of CR1 LINEs suggest that PcRTFs are the debris of P. flavoviridis ancestral CR1 LINEs, denoted as PfCR1s. Since the associated pairs of PLA(2) genes and PcRTFs are arranged in tandem in the 25,026 bp segment, it is thought that an ancestral PLA(2) gene-PfCR1 unit (PfPLA-PfCR1) which was produced by retrotransposition of PfCR1 by itself to the 3' end of PLA(2) gene duplicated several times to form a multimer of PfPLA-PfCR1, a cluster of PLA(2) genes, in the period after Crotalinae and Viperinae snakes branched off. Recombinational hot spot of a 37bp segment, named Scomb, was found in the region 548 bp upstream from the TATA box of PLA(2) genes. Thus, it could be assumed that multiplication of PfPLA-PfCR1 occurred by unequal crossing over of the segment, -Scomb-PfPLA-PfCR1-Scomb-. The PfCR1 moieties were afterward disrupted in the 5' portion to PcRTFs. The detection of two types of PcRTFs different in length which were produced by elimination of two definitive sequences in PfCR1 moiety possibly by gene conversion clearly supports such process but not multiplication of the PLA(2) gene-PcRTF unit.


[Sources for the history of health sciences in Brazil (1808-1930)].
The article presents a panorama of documented sources and of the most representative archives, libraries, and documentation centers pertinent to the history of health sciences in Brazil (1808-1930). Historians, especially those investigating the biomedical sciences, will find here a broad and diversified set of sources. It is precisely from such a convergence of different witnesses that historical investigation is viable.


Improved therapeutic index of cis-diammine dichloro platinum by diethyldithiocarbamate in rodents.
cis-Diammine Dichloro Platinum (DDP) is an effective antitumor agent. However, nephrotoxicity remains a major dose limiting factor in its clinical application. The present experiments with Fischer 344 rats showed that diethyldithiocarbamate, an alkyl dithiocarbamate, prevents DDP induced acute nephrotoxicity, and increases the LD50 from 8.3 to 10.8 mg/kg. The subacute DDP toxicity was also prevented by the DTC in B6D2F1/J mice. Further, studies with L1210 Leukemic mice indicate that DTC in dosages up to 250 mg/kg did not interfere with the antitumor effects of DDP. The present experiments, therefore, add evidence to the thesis that thiol compounds may "selectively" protect normal tissues against agents which form interstrand or intrastrand cross links with DNA. Thiol compounds, therefore, may have a wider clinical application in patients receiving cancer chemotherapy.


Impact of the community healthcare plan and the free maternity services programme on maternal and child healthcare utilisation in rural Kenya: a dairy farmer population-based study.
Access to and utilisation of quality maternal and child healthcare services is generally recognized as the best way to reduce maternal and child mortality. We evaluated whether the introduction of a voluntary family health insurance programme, combined with quality improvement of healthcare facilities [The Community Health Plan (TCHP)], and the introduction of free access to delivery services in all public facilities [Free Maternity Services programme (FMS)] increased antenatal care utilisation and use of facility deliveries among pregnant women in rural Kenya. TCHP was introduced in 2011, whilst the FMS programme was launched in 2013. To measure the impact of TCHP, percentage points (PP) changes in antenatal care utilisation and facility deliveries from the pre-TCHP to the post-TCHP period between the TCHP programme area and a control area were compared in multivariable difference-in-differences analysis. To measure the impact of the FMS programme, PP changes in antenatal care utilisation and facility deliveries from the pre-FMS to the post-FMS period in the pooled TCHP programme and control areas was assessed in multivariable logistic regression analysis. Data was collected through household surveys in 2011 and 2104. Households (n=549) were randomly selected from the member lists of 2 dairy companies, and all full-term pregnancies in the 3.5 years preceding the baseline and follow-up survey among women aged 15-49 at the time of pregnancy were eligible for this study (n=295). Because only 4.1% of eligible women were insured through TCHP during pregnancy, any increase in utilisation attributable to the TCHP programme could only have come about as a result of the quality improvements in TCHP facilities. Antenatal care utilisation significantly increased after TCHP was introduced (14.4 PP; 95% CI: 4.5-24.3; P=0.004), whereas no effect was observed of the programme on facility deliveries (8.8 PP; 95% CI: -14.1 to +31.7; P=0.450). Facility deliveries significantly increased after the introduction of the FMS programme (27.9 PP; 95% CI: 11.8-44.1; P=0.001), but antenatal care utilisation did not change significantly (4.0 PP; 95% CI: -0.6 to +8.5; P=0.088). Access to the FMS programme increased facility deliveries substantially and may contribute to improved maternal and new-born health and survival if the quality of delivery services is sustained or further improved. Despite low up-take, TCHP had a positive effect on antenatal care utilisation among uninsured women by improving the quality of existing healthcare facilities. An alignment of the two programmes could potentially lead to optimal results. The study was funded by the Health Insurance Fund (http://www.hifund.org/), through a grant from the Dutch Ministry of Foreign Affairs.


Mapping the Hsp90 genetic interaction network in Candida albicans reveals environmental contingency and rewired circuitry.
The molecular chaperone Hsp90 regulates the folding of diverse signal transducers in all eukaryotes, profoundly affecting cellular circuitry. In fungi, Hsp90 influences development, drug resistance, and evolution. Hsp90 interacts with -10% of the proteome in the model yeast Saccharomyces cerevisiae, while only two interactions have been identified in Candida albicans, the leading fungal pathogen of humans. Utilizing a chemical genomic approach, we mapped the C. albicans Hsp90 interaction network under diverse stress conditions. The chaperone network is environmentally contingent, and most of the 226 genetic interactors are important for growth only under specific conditions, suggesting that they operate downstream of Hsp90, as with the MAPK Hog1. Few interactors are important for growth in many environments, and these are poised to operate upstream of Hsp90, as with the protein kinase CK2 and the transcription factor Ahr1. We establish environmental contingency in the first chaperone network of a fungal pathogen, novel effectors upstream and downstream of Hsp90, and network rewiring over evolutionary time.


Influence of single nucleotide polymorphisms of cytokine genes on anti-HBs antibody production after hepatitis B vaccination in a Japanese young adult population.
Hepatitis B (HB) vaccination is one of the most efficient tools to prevent the transmission of the virus. Considerable variability exists in HB vaccine responses, with 5-10% of healthy Japanese adults demonstrating no response following a standard vaccination. Recently, polymorphisms of immune-regulatory genes, such as cytokine genes, have been reported to influence the immune response to HB vaccine. The aim of this study was to investigate the underlying mechanisms of the genetic association between several cytokine gene polymorphisms and the immune response to HB vaccination in a Japanese population. One hundred and twenty three vaccinated young adults were classified according to the level of antibody-titer (anti-HBs). Single nucleotide polymorphism typing for IFN-γ (+874, 3'-UTR), IL-10 (-591, -819, -1082), and TNF-α (-308, -857), was accomplished using the PCR-RFLP or SSP-PCR method. The TNF-α (-857) CC type and the IL-10 (-1082) AG type were present more frequently in the low titer group than in the high titer group. The TNF-α (-857) CC type was found to be significantly associated with low response of serum anti-HBs. The anti-HBs antibody was not readily produced in the IL-10 (-1082) AG and TNF-α (-857) CC haplotype. Conversely, the antibody was readily produced in the IL-10 (-1082) AA and TNF-α (-857) CC haplotype, and the IL-10 (-1082) AA and TNF-α (-857) CT haplotype, suggesting a high likelihood of the IL-10 (-1082) AG type to be included in the low anti-HBs group, and high anti-HBs antibody production in those with the TNF-α (-857) CT type. These SNPs may produce ethnically-specific differences in the immune response to HB vaccine in the Japanese population. J. Med. Invest. 63: 256-261, August, 2016.


Resident aerobic microbiota of the adult human nasal cavity.
Recent evidence strongly suggests that the microbiota of the nasal cavity plays a crucial role in determining the reaction patterns of the mucosal and systemic immune system. However, little is known about the normal microbiota of the nasal cavity. The purpose of this study was to determine the microbiota in different parts of the nasal cavity and to develop and evaluate methods for this purpose. Samples were collected from 10 healthy adults by nasal washes and by swabbing of the mucosa through a sterile introduction device. Both methods gave results that were quantitatively and qualitatively reproducible, and revealed significant differences in the density of the nasal microbiota between individuals. The study revealed absence of gram-negative bacteria that are regular members of the commensal microbiota of the pharynx. Likewise, viridans type streptococci were sparsely represented. The nasal microbiota was dominated by species of the genera Corynebacterium, Aureobacterium, Rhodococcus, and Staphylococcus, including S. epidermis, S. capitis, S. hominis, S. haemolyticus, S. lugdunensis and S. warneri. These studies show that the microbiota of the nasal cavity of adults is strikingly different from that of the pharynx, and that the nasal cavity is a primary habitat for several species of diphtheroids recognized as opportunistic pathogens. Under special circumstances, single species, including IgA1 protease-producing bacteria, may become predominant in a restricted area of the nasal mucosa.


Assaying Circuit Specific Regulation of Adult Hippocampal Neural Precursor Cells.
Adult neurogenesis is a dynamic process by which newly activated neural stem cells (NSCs) in the subgranular zone (SGZ) of the dentate gyrus (DG) generate new neurons, which integrate into an existing neural circuit and contribute to specific hippocampal functions. Importantly, adult neurogenesis is highly susceptible to environmental stimuli, which allows for activity-dependent regulation of various cognitive functions. A vast range of neural circuits from various brain regions orchestrates these complex cognitive functions. It is therefore important to understand how specific neural circuits regulate adult neurogenesis. Here, we describe a protocol to manipulate neural circuit activity using designer receptor exclusively activated by designer drugs (DREADDs) technology that regulates NSCs and newborn progeny in rodents. This comprehensive protocol includes stereotaxic injection of viral particles, chemogenetic stimulation of specific neural circuits, thymidine analog administration, tissue processing, immunofluorescence labeling, confocal imaging, and imaging analysis of various stages of neural precursor cells. This protocol provides detailed instructions on antigen retrieval techniques used to visualize NSCs and their progeny and describes a simple, yet effective way to modulate brain circuits using clozapine N-oxide (CNO) or CNO-containing drinking water and DREADDs-expressing viruses. The strength of this protocol lies in its adaptability to study a diverse range of neural circuits that influence adult neurogenesis derived from NSCs.


[Transcapillary filtration of electrolytes in the brain after craniocerebral injuries].
A new method was used to determine the value and direction of electrolyte filtration through the walls of capillaries in the brain in patients with craniocerebral injury. It was found that the period of the development of brain edema coincided in time with the prevalence of the processes of sodium, calcium, and fluid escape from the cerebral vessels into the cerebral tissue. The movement of potassium through the capillary wall was counter-current to that of sodium filtration in most cases. Transcapillary filtration of electrolytes in patients with craniocerebral injury during the posttraumatic period was characterized by recurrent changes in the direction of filtration. Electrolytes (sodium, potassium, and calcium) possess their own, often independent of one another, mechanisms of transfer through the membranes of the histohematic barriers.


Vortex Keratopathy as a Presenting Feature of Ocular Mucous Membrane Pemphigoid.
The aim of this study was to describe the presenting feature of vortex keratopathy in 3 patients with biopsy-proven ocular mucous membrane pemphigoid (OMMP). The first patient was a 52-year-old woman with chronic redness and foreign body sensation for 3 years who presented with unilateral vortex keratopathy. Seven months later, in the same eye, she developed conjunctival signs suggestive of OMMP. The second patient was a 33-year-old woman with similar chronic symptoms that were exacerbated by pterygium surgery. Clinical examination revealed vortex keratopathy in the right eye with subtle conjunctival signs suggestive of OMMP. The third patient was a 70-year-old woman with complaints of repeated episodes of redness and foreign body sensation for 18 months who presented with vortex keratopathy in the right eye and conjunctival signs suggestive of OMMP in the same eye. To confirm the clinical diagnosis, bilateral conjunctival biopsy was performed in all patients. OMMP was diagnosed based on conjunctival signs and confirmed on direct immunofluorescence positivity, demonstrating antibodies characteristic and diagnostic of OMMP in the basement membrane zone. A unique feature that preceded or coexisted with the conjunctival signs was unilateral vortex keratopathy seen in all 3 patients, independent of the stage of the disease at which they presented. Vortex keratopathy can be a presenting feature in patients with OMMP. A complete ocular surface examination, especially checking the medial canthus for keratin and the inferior fornix for foreshortening, is necessary. Conjunctival biopsy should be performed in all cases to confirm the clinical diagnosis wherever required.


Role of prostaglandin D2 in mast cell activation-induced sensitization of esophageal vagal afferents.
Sensitization of esophageal afferents plays an important role in esophageal nociception, but the mechanism is less clear. Our previous studies demonstrated that mast cell (MC) activation releases the preformed mediators histamine and tryptase, which play important roles in sensitization of esophageal vagal nociceptive C fibers. PGD2 is a lipid mediator released by activated MCs. Whether PGD2 plays a role in this sensitization process has yet to be determined. Expression of the PGD2 DP1 and DP2 receptors in nodose ganglion neurons was determined by immunofluorescence staining, Western blotting, and RT-PCR. Extracellular recordings were performed in ex vivo esophageal-vagal preparations. Action potentials evoked by esophageal distension were compared before and after perfusion of PGD2, DP1 and DP2 receptor agonists, and MC activation, with or without pretreatment with antagonists. The effect of PGD2 on 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI)-labeled esophageal nodose neurons was determined by patch-clamp recording. Our results demonstrate that DP1 and DP2 receptor mRNA and protein were expressed mainly in small- and medium-diameter neurons in nodose ganglia. PGD2 significantly increased esophageal distension-evoked action potential discharges in esophageal nodose C fibers. The DP1 receptor agonist BW 245C mimicked this effect. PGD2 directly sensitized DiI-labeled esophageal nodose neurons by decreasing the action potential threshold. Pretreatment with the DP1 receptor antagonist BW A868C significantly inhibited PGD2 perfusion- or MC activation-induced increases in esophageal distension-evoked action potential discharges in esophageal nodose C fibers. In conclusion, PGD2 plays an important role in MC activation-induced sensitization of esophageal nodose C fibers. This adds a novel mechanism of visceral afferent sensitization.


Gap Junction Dependent Cell Communication Is Modulated During Transdifferentiation of Mesenchymal Stem/Stromal Cells Towards Neuron-Like Cells.
In vitro transdifferentiation of patient-derived mesenchymal stem/stromal cells (MSCs) into neurons is of special interest for treatment of neurodegenerative diseases. Although there are encouraging studies, little is known about physiological modulations during this transdifferentiation process. Here, we focus on the analysis of gap junction dependent cell-cell communication and the expression pattern of gap junction-building connexins during small molecule-induced neuronal transdifferentiation of human bone marrow-derived MSCs. During this process, the MSC markers CD73, CD90, CD105, and CD166 were downregulated while the neuronal marker Tuj1 was upregulated. Moreover, the differentiation protocol used in the present study changed the cellular morphology and physiology. The MSCs evolved from a fibroblastoid morphology towards a neuronal shape with round cell bodies and neurite-like processes. Moreover, depolarization evoked action potentials in the transdifferentiated cells. MSCs expressed mRNAs encoding Cx43 and Cx45 as well as trace levels of Cx26, Cx37- and Cx40 and allowed transfer of microinjected Lucifer yellow. The differentiation protocol increased levels of Cx26 (mRNA and protein) and decreased Cx43 (mRNA and protein) while reducing the dye transfer. Cx36 mRNA was nearly undetectable in all cells regardless of treatment. Treatment of the cells with the gap junction coupling inhibitor carbenoxolone (CBX) only modestly altered connexin mRNA levels and had little effect on neuronal differentiation. Our study indicates that the small molecule-based differentiation protocol generates immature neuron-like cells from MSCs. This might be potentially interesting for elucidating physiological modifications and mechanisms in MSCs during the initial steps of differentiation towards a neuronal lineage.


Breast cancer patients' presentation for oncological treatment: a single centre study.
Breast cancer patients are presenting at advanced stages for oncological treatment in Nigeria and World Health Organization predicted developing countries' breast cancer incidence and mortality to increase by year 2020. Prospective observational hospital based study that enrolled breast cancer patients from catchment area of an oncology service hospital in Nigeria between 2007 and 2013. Patients' demographics, breast cancer burden and health care giver presentation variables were analysed for causal factors of seeking medical help and what determines commencement of effective oncological treatment. Forty-six patients were enrolled, 19.6% of them presented primarily to oncologist while 80.4% presented secondarily for oncological treatment. There is a significant difference in presentation time for oncological treatment (t = -3.56, df = 42.90, p = 0.001) between primary (M =11.56 ± 5.21 weeks) and secondary presentation (M= 52.56 ± 10.27weeks). Tumor burden of those that presented secondarily were significantly more advanced (U = 78.5, p = 0.011) and, univariate analysis reveals that: patients' matrimonial setting, breast cancer awareness and mode of discovery of breast symptoms are patient related factors that determines their choice of health care providers and, determinant of effective oncological treatment is patient first contact health care provider. Patients' bio-characteristics that determine their choice of health care provider should be incorporated into community breast cancer sensitization drives. Additionally, there is a need for a government agency assign the task of accrediting and defining scope of enterprise of health care institutions and their health care providers in our pluralist health system.


Detecting and characterizing gene conversions between multigene family members.
We used a variety of methods to detect known gene conversions in the actin gene families of five angiosperm species, the beta-globin gene families of two primate species, and the Zfx/Zfy gene families of seven mammalian species. Our goal was to devise a working strategy which would allow the analysis of the members of a multigene family in order to determine whether there had been gene conversions between its members, identify the genes involved in the gene conversions, establish the lengths of the converted regions, and determine the polarities of the gene conversions. We show that three phylogenetic methods and the homoplasy test of Maynard Smith and Smith perform relatively poorly on our data sets because the sequences we analyzed had large levels of multiple substitutions. The method of Sawyer, the compatibility method of Jakobsen and Easteal, the partition matrix method of Jakobsen, Wilson, and Easteal, and the co-double method of Balding, Nichols, and Hunt can be used to identify the genes which have been involved in gene conversions. The co-double method is more powerful than other methods but requires orthologous sequences from related species. Compatibility, phylogenetic, and nucleotide substitution distribution statistics methods can be used to identify the location of the converted region(s). Site-by-site compatibility analyses can also be used to identify the direction of the conversion event(s). Combinations of these methods can therefore be used to establish the presence, locations, and polarities of gene conversions between multigene family members.