Allogenic Bone Graft in Dentistry: A Review of Current Trends and Developments.
In an effort to prepare non-autologous bone graft or biomaterial that would possess characteristics comparable to autologous bone, many different allogenic bone derivatives have been created. Although different existing processing methods aim to achieve the very same results, the specific parameters applied during different stages material preparation can result in significant differences in the material's mechanical and biological properties The properties, including osteoconductive, osteoinductive, and even osteogenic potential, can differ vastly depending on particular preparation and storage techniques used. Osteogenic properties, which have long been thought to be characteristic to autogenic bone grafts only, now seem to also be achievable in allogenic materials due to the possibility to seed the host's stem cells on a graft before its implantation. In this article, we aim to review the available literature on allogenic bone and its derivatives as well as the influence of different preparation methods on its performance.


[Role of genetic features of Helicobacter pylori in development of inflammation of the gastric mucosa in chronic gastritis and ulcer disease].
This review, which is devoted to the analysis of current data on genetic properties of Helicobacteria, is the first work in the Russian literature summarizing the research results published before and after the sequencing of the genome of two Helicobacter pylori pathogenic strains--26695 and J99. It discloses information about the genetic control of pathogenecity factors and their localization on the chromosome. There is a review of genes participating in the colonization of Helicobacteria, lipopolysaccharide biosynthesis, urease synthesis, and pH control. Special attention is focused on the problem of so-called "viable" but non-cultured forms of Helicobacter pylori.


Strategies for faster detachment of corneal cell sheet using micropatterned thermoresponsive matrices.
The development of transplantable cell sheets of functional keratocytes embedded within an aligned collagen type I matrix is a viable approach for constructing a bioequivalent of corneal stroma. Thermoresponsive materials based on poly(N-isopropylacrylamide) (PolyNIPA) have been utilized to recover carrier-free corneal cell sheets by inducing temperature changes. In this study, we employed direct-write assembly (DWA) to develop microperiodic parallel patterns of silk-PolyNIPA and gelatin-PolyNIPA. Semi-interpenetrating networks of PolyNIPA hybrids (with silk/gelatin) exhibited temperature-responsive nature and thereby have potential use in cell sheet engineering. Silk-PolyNIPA and gelatin-PolyNIPA hybrids demonstrated a hydrophobic surface at 37 °C (i.e. above their lower critical solution temperature) with a contact angle of 59°± 0.3° and 55°± 3°, respectively, whereas the surface roughness of silk-PolyNIPA was double that of gelatin-PolyNIPA. The reduction of temperature to 20 °C resulted in a decrease in the value of surface roughness and water contact angle for both hybrids. All four parallel patterned substrates guided corneal cell alignment along the direction of the patterns. Collagen type-I and aggrecan gene expression was higher when the cells were grown over the gelatin-PolyNIPA matrix after 3 weeks of culture when compared to silk-PolyNIPA. In addition, a significantly higher metabolic activity as well as enhanced vinculin expression of keratocytes on the gelatin-PolyNIPA matrix indicated the improved cytocompability compared to the silk, gelatin and silk-PolyNIPA matrices. Interestingly, the detachment of keratocytes cell sheet was achieved from the silk-PolyNIPA and gelatin-PolyNIPA planar films only within 10 min and 30 min, respectively, but the patterns could not yield intact sheet recovery. Hence, we conclude that while gelatin-PolyNIPA hybrids with parallel patterns fabricated using DWA will benefit from the application of cellular alignment, some optimization in the pattern parameters may be required for rapid sheet recovery from such substrates. Understanding the keratocytes responses to such hybrid biomaterials suggests viable options to develop a corneal stromal bioequivalent.


Rapid plate method for the isolation of lysogenic bacteriophages.
We have devised a rapid plate method for the detection and isolation of lysogenic phages and have used the procedure to screen 15 strains of Staphylococcus aureus. This method should prove applicable to a wide variety of bacterial species and should be especially valuable for those pathogens for which no phage-typing system is available.


Care for perinatal illness in rural Nepal: a descriptive study with cross-sectional and qualitative components.
BACKGROUND: Maternal, perinatal and neonatal mortality rates remain high in rural areas of developing countries. Most deliveries take place at home and care-seeking behaviour is often delayed. We report on a combined quantitative and qualitative study of care seeking obstacles and practices relating to perinatal illness in rural Makwanpur district, Nepal, with particular emphasis on consultation strategies. METHODS: The analysis included a survey of 8798 women who reported a birth in the previous two years [of whom 3557 reported illness in their pregnancy], on 30 case studies of perinatal morbidity and mortality, and on 43 focus group discussions with mothers, other family members and health workers. RESULTS: Early pregnancy was often concealed, preparation for birth was minimal and trained attendance at birth was uncommon. Family members were favoured attendants, particularly mothers-in-law. The most common recalled maternal complications were prolonged labour, postpartum haemorrhage and retained placenta. Neonatal death, though less definable, was often associated with cessation of suckling and shortness of breath. Many home-based care practices for maternal and neonatal illness were described. Self-medication was common.There were delays in recognising and acting on danger signs, and in seeking care beyond the household, in which the cultural requirement for maternal seclusion, and the perceived expense of care, played a part. Of the 760 women who sought care at a government facility, 70% took more than 12 hours from the decision to seek help to actual consultation. Consultation was primarily with traditional healers, who were key actors in the ascription of causation. Use of the government primary health care system was limited: the most common source of allopathic care was the district hospital. CONCLUSIONS: Major obstacles to seeking care were: a limited capacity to recognise danger signs; the need to watch and wait; and an overwhelming preference to treat illness within the community. Safer motherhood and newborn care programmes in rural communities, must address both community and health facility care to have an impact on morbidity and mortality. The roles of community actors such as mothers-in-law, husbands, local healers and pharmacies, and increased access to properly trained birth attendants need to be addressed if delays in reaching health facilities are to be shortened.


Comparison of different bonding techniques for efficient strain transfer using piezoelectric actuators.
In this paper, strain transfer efficiencies from a single crystalline piezoelectric lead magnesium niobate-lead titanate substrate to a GaAs semiconductor membrane bonded on top are investigated using state-of-the-art x-ray diffraction (XRD) techniques and finite-element-method (FEM) simulations. Two different bonding techniques are studied, namely, gold-thermo-compression and polymer-based SU8 bonding. Our results show a much higher strain-transfer for the "soft" SU8 bonding in comparison to the "hard" bonding via gold-thermo-compression. A comparison between the XRD results and FEM simulations allows us to explain this unexpected result with the presence of complex interface structures between the different layers.


Scalp potentials evoked by amplitude-modulated tones in dyslexia.
We recorded the far-field EEG potential evoked by amplitude modulation of acoustic stimuli (the amplitude modulation following response, AMFR) in adults with developmental dyslexia and in a matched control group of adults with no history of reading problems. The mean AMFR recorded from participants with dyslexia was significantly smaller than that recorded from members of the control group. In contrast, the amplitude of the click-evoked auditory brainstem response (ABR) was not significantly different between participant groups. Also, there was no difference between participant groups in the latency of the AMFR or ABR. The reduced AMFR in listeners with dyslexia may reflect impaired ability of the auditory system to follow rapid changes in stimulus energy, a cue believed to be important in the perception of speech.


The serum pentraxin-3 is elevated in patients with cardiac syndrome X.
Cardiac syndrome X (CSX) is a clinical entity that is defined as normal coronary arteries with angina pectoris and objective sins of ischemia. The correlation between CSX and inflammatory markers such as high-sensitivity C-reactive protein (hs-CRP) is well established, however an association with pentraxin-3 (PTX-3) has not been examined. The aim of this study was to investigate the association between PTX-3 and CSX. A total of 122 patients (58 female, 64 male, mean age 49.6±5.8 years) with suspected of coronary artery disease (CAD) were included in the study. Those with evidence of ischemia (50 patients with positive treadmill tests, 32 patients with positive myocardial perfusion scintography) underwent coronary angiography (82 patients). Patients with a normal angiogram were considered the CSX group (n=41) and patients with coronary lesions were referred to as the CAD group (n=41). Patients without signs of ischemia served as the control group. Serum PTX-3 and hs-CRP levels were measured in all patients. The CSX group had significantly increased PTX-3 levels relative to the control group (0.46±0.16 vs. 0.23±0.09 ng/ml, p<0.001). However there were no differences in levels of PTX-3 and hs-CRP between the CSX and the CAD groups (PTX-3: 0.46±0.16 vs. 0.51±0.13 ng/ml, p=0.21; hs-CRP: 1.04±0.45 vs. 1.16±0.64 mg/dl, p=0.62). The control group had significantly lower hs-CRP levels (0.73±0.51 mg/dl) when compared to the both CSX and CAD groups (p=0.03 and p=0.002, respectively). Serum PTX-3 levels were weakly correlated with hs-CRP levels (r=0.30, p=0.001). PTX-3, a novel inflammatory marker, is elevated in patients with CSX, similar to the well known inflammatory marker hs-CRP, and may be a promising biomarker reflecting inflammatory status in these patients.


Timing of development of measles-specific immunoglobulin M and G after primary measles vaccination.
A standard method for diagnosing measles is to detect measles-specific immunoglobulin M (IgM) in the serum of infected persons. Interpreting a positive IgM result from a person with suspected measles can be difficult if the person has recently received a measles vaccine. We have previously demonstrated that measles-specific IgM may persist for at least 8 weeks after primary vaccination, but it is unknown how quickly IgM appears. This study determined the timing of the rise of measles-specific IgM and IgG after primary measles vaccination with Schwartz vaccine. Two hundred eighty 9-month-old children from Ethiopia presenting for routine measles vaccination were enrolled. Sera were collected before and either 1, 2, 3, or 4 weeks after vaccination and tested for measles-specific antibodies by an IgM capture enzyme immunoassay (EIA) and by an indirect IgG EIA. A total of 209 of the 224 children who returned for the second visit had prevaccination sera that were both IgM and IgG negative. The postvaccination IgM positivity rates for these 209 children were 2% at 1 week, 61% at 2 weeks, 79% at 3 weeks, and 60% at 4 weeks. The postvaccination IgG positivity rates were 0% at 1 week, 14% at 2 weeks, 81% at 3 weeks, and 85% at 4 weeks. We conclude that an IgM-positive result obtained by this antibody capture EIA is difficult to interpret if serum is collected between 8 days and 8 weeks after vaccination; in this situation, the diagnosis of measles should be based on an epidemiologic linkage to a confirmed case or on the detection of wild-type measles virus.


Downregulation of the alpha5 subunit of the GABA(A) receptor in the pilocarpine model of temporal lobe epilepsy.
Specific subunits of gamma-aminobutyric acid (GABA)A receptors may be regulated differentially in animal models of temporal lobe epilepsy during the chronic stage. Although several subunits may be upregulated, other subunits may be downregulated in the hippocampal formation. The alpha5 subunit is of particular interest because of its relatively selective localization in the hippocampus and its potential role in tonic inhibition. In normal rats, immunolabeling of the alpha5 subunit was high in the dendritic layers of CA1 and CA2 and moderate in these regions of CA3. In chronic pilocarpine-treated rats displaying recurrent seizures, alpha5 subunit-labeling was substantially decreased in CA1 and nearly absent in CA2. Only slight decreases in immunolabeling were evident in CA3. In situ hybridization studies demonstrated that the alpha5 subunit mRNA was also strongly decreased in stratum pyramidale of CA1 and CA2. Thus, the alterations in localization of the alpha5 subunit peptide and its mRNA were highly correlated. The large decreases in labeling of the alpha5 subunit did not appear to be related to loss of pyramidal neurons in CA1 or CA2 since these neurons were generally preserved in pilocarpine-treated animals. No comparable decreases in labeling of the alpha2 subunit of the GABA(A) receptor were detected. These findings indicate that the alpha5 subunit of the GABA(A) receptor is capable of substantial and prolonged downregulation in remaining pyramidal neurons in a model of temporal lobe epilepsy. The results raise the possibility that presumptive extrasynaptic GABA(A) receptor subunits, such as the alpha5 subunit, may be regulated differently than synaptically located subunits, such as the alpha2 subunit, within the same brain regions in some pathological conditions.


Pathophysiological analysis of the progression of hepatic lesions in STAM mice.
Nonalcoholic steatohepatitis (NASH) is a current health issue since the disease often leads to hepatocellular carcinoma; however, the pathogenesis of the disease has still not been fully elucidated. In this study, we investigated the pathophysiological changes observed in hepatic lesions in STAM mice, a novel NASH model. STAM mice, high fat-diet (HFD) fed mice, and streptozotocin (STZ) treated mice were prepared, and changes over time, such as biological parameters, mRNA expression, and histopathological findings, were evaluated once animal reached 5, 7, and 10 weeks of age. STZ mice presented with hyperglycemia and an increase in oxidative stress in immunohistochemical analyses of Hexanoyl-lysine: HEL from 5 weeks, with fibrosis in the liver also being observed from 5 weeks. HFD mice presented with hyperinsulinemia from 7 weeks and the slight hepatosteatosis was observed at 5 weeks, with changes significantly increasing until 10 weeks. STAM mice at 10 weeks showed significant hepatic changes, including hepatosteatosis, hypertrophic hepatocytes, and fibrosis, indicating pathological changes associated with NASH. These results suggested that the increase in oxidative stress with hyperglycemia triggered hepatic lesions in STAM mice, and insulin resistance promoted lesion formation with hepatic lipid accumulation. STAM mice may be a useful model for elucidating the pathogenesis of NASH with diabetes.


[Infected common iliac aneurysm due to Salmonella: a case report].
A 62-year-old male, presenting with chills and fever and RLQ abdominal pain, was found to have an infected aneurysm of the right common iliac artery infected by Salmonella group C1. CT scan of abdomen and arteriography definitely identified the lesions. Antibiotic treatment and surgical intervention with aneurysmectomy and suprapubic crossover femoro-femoral artery graft resulted in a good recovery and outcome.


Distortion of the three-dimensional structure of the vnd/NK-2 homeodomain bound to DNA induced by an embryonically lethal A35T point mutation.
The three-dimensional solution structure obtained by NMR of the A35T mutant vnd/NK-2 homeodomain bound to the vnd/NK-2 consensus 16 bp DNA sequence was determined. This mutation to threonine from alanine in position 35 in helix II of the vnd/NK-2 homeodomain is associated with early embryonic lethality in Drosophila melanogaster. Although the unbound mutant protein is not structured, in the DNA-bound state it adopts the three-helix fold characteristic of all known homeodomains, but with alterations relative to the structure of the wild-type analogue. These structural modifications occur, and are accompanied by a 50-fold reduction in the DNA binding affinity, even though most of the protein-DNA interactions originally seen for the wild-type homeodomain are found likewise in the threonine analogue. Alterations include torsional angle changes in the loop between helix I and helix II, and in the turn between helix II and helix III, as well as in a distortion of the usual antiparallel orientation of helix I with respect to helix II. The alteration of the position of leucine 40 in the A35T mutant is proposed to explain the observed 1.27 ppm upfield shift of the corresponding amide proton resonance relative to the value observed for the wild-type analogue. A detailed comparison of the structures of the mutant A35T and wild-type vnd/NK-2 homeodomains bound to the cognate DNA is presented. The consequences of the structural alteration of the DNA-bound A35T mutant vnd/NK-2 protein may constitute the basis of the observed early embryonic lethality.


Identification of regulatory and catalytic domains in the apoptosis nuclease DFF40/CAD.
The DNA fragmentation factor (DFF) is composed of two subunits, the 40-kDa caspase-3-activated nuclease (DFF40/CAD) and its 45-kDa inhibitor (DFF45/ICAD). During apoptosis, DFF-40/CAD is activated by caspase-3-mediated cleavage of DFF45/ICAD. Mutational analysis of DFF40/CAD revealed that DFF40/CAD is composed of a C-terminal catalytic domain and an N-terminal regulatory domain. Deletion of the catalytic domain (residues 290-345) abrogated the caspase-3-induced nuclease activity of DFF40/CAD but not its ability to interact with DFF45/ICAD. Conversely, removal of the regulatory domain (residues 1-83) yielded a constitutively active DFF40/CAD nuclease that neither bound to its inhibitor nor required caspase-3 for activation. Amino acid alignment revealed that the regulatory domain of DFF40/CAD has homology to the N-terminal region of mammalian and Drosophila DFF45/ICAD and CIDE-N, a regulatory domain previously identified in pro-apoptotic CIDE proteins. Mutational analysis of the N-terminal region revealed mutants with diminished nuclease activity but with intact ability to bind DFF45/ICAD. Thus, CIDE-N represents a new type of domain that is associated with the regulation of the apoptosis/DNA fragmentation pathway.


Reduced gaze following and attention to heads when viewing a "live" social scene.
Social stimuli are known to both attract and direct our attention, but most research on social attention has been conducted in highly controlled laboratory settings lacking in social context. This study examined the role of social context on viewing behaviour of participants whilst they watched a dynamic social scene, under three different conditions. In two social groups, participants believed they were watching a live webcam of other participants. The socially-engaged group believed they would later complete a group task with the people in the video, whilst the non-engaged group believed they would not meet the people in the scene. In a third condition, participants simply free-viewed the same video with the knowledge that it was pre-recorded, with no suggestion of a later interaction. Results demonstrated that the social context in which the stimulus was viewed significantly influenced viewing behaviour. Specifically, participants in the social conditions allocated less visual attention towards the heads of the actors in the scene and followed their gaze less than those in the free-viewing group. These findings suggest that by underestimating the impact of social context in social attention, researchers risk coming to inaccurate conclusions about how we attend to others in the real world.


Spatiotemporal variation and risk assessment of pesticides in water of the lower catchment basin of Acheloos River, Western Greece.
A three-year monitoring survey (March 2005-February 2008) was conducted to investigate, on monthly basis, the presence of thirty pesticides belonging to various categories and metabolites, in Acheloos River (Western Greece), one of the most important water resources in Greece. Six sampling stations along the river were established. Water analyses were performed using solid-phase extraction combined with gas chromatography with flame thermionic detector and mass spectrometry. Statistical analysis using one-way ANOVA and Duncan's multiple range test (P < 0.05) was used to compare annual mean concentrations of pesticides, seasonal and spatial distribution. In general, the highest mean concentrations of the pesticides were recorded at the three stations downstream. The greatest average concentrations were determined during spring and summer in agreement with the pesticide application period. The observed lower concentrations after 2006 reflect the land-use change because of the elimination of tobacco, the main cultivation of the area for many decades. The compounds most frequently detected were diazinon (78.6%), DEA (69.3%), and fenthion (52.6%). Environmental risk assessment using risk quotient (RQ) approach showed high risk for six insecticides in 2005 and one in 2007. A compliance with the European Environmental Quality Standards (EQS) was observed for the priority pesticides.


Carcinogenesis by hepatic peroxisome proliferators: evaluation of the risk of hypolipidemic drugs and industrial plasticizers to humans.
In this critical review, I would like to provide a brief outline of the morphology, biochemical composition, distribution, and functions of peroxisomes. The induction of peroxisome proliferation and peroxisome-associated enzymes in the rodent liver by two classes of chemicals (hypolipidemic drugs and the industrial plasticizers) will be considered. The role of peroxisomes in lipid metabolism will be discussed. Carcinogenicity studies in rats and mice with these peroxisome proliferators will be evaluated critically. Careful consideration will be given to the hypothesis that "potent hepatic peroxisome proliferators as a class are carcinogenic." The possible mechanism(s) by which peroxisome proliferators induce liver tumors will be outlined. Particular attention will be paid to the possible role of peroxisome proliferation-mediated radical toxicity and generation of endogenous initiators of carcinogenesis.


[Adaptive potential of the secretory system of the stomach in different phases of peptic ulcer in children].
A conjugated shift in the levels of total acidity and lysozyme in the fasting portion of gastric juice within the system of coordinates enables the diagnosis of the stages of the adaptation syndrome and the respective levels of the functioning of the secretory gastric apparatus, which in turn specify differentiation of peptic ulcer phases. The levels of total acidity and lysozyme, the indirect parameter of the level of the secretory apparatus functioning have been measured in accordance with gastric juice fractions in healthy children.


An Integrative Review of Measuring Caregiver Burden in Substance Use Disorder.
Family caregivers contribute to engagement in treatment and adherence, reduced substance misuse and relapse, and increased well-being of recipients with substance use disorder. However, providing care has also been associated with negative emotional and physical health outcomes for caregivers. The purpose of this integrative review was to determine what instruments are used to measure caregiver burden in informal caregivers of individuals with substance use disorder. An integrative review framework was applied to examine empirical and theoretical literature to answer the guiding research question, "How is caregiver burden measured in caregivers of individuals with substance use disorder?" PubMed, CINAHL, and APA PsychINFO were searched using a combination of search terms. The initial 1,198 articles were narrowed to 32 that fit the search criteria and purpose of the review. A variety of scales have been used to measure caregiver burden. Caregiver burden is operationalized as objective or subjective burden. Objective burden refers to changes in the home, finances, employment, social life, and leisure, whereas subjective burden refers to the emotional reaction of the caregiver in coping with providing care. Caregiver burden was most often reported as moderate to severe in populations with substance use disorder. Attributes measured included anxiety, depression, stress, worry, displeasure, care recipient behavioral problems and substance abuse, stigma, relationship strain, financial expenses, social support, family disruption, and the effect on caregiver physical and emotional health. Specific instruments that can accurately evaluate objective and subjective caregiver burden are needed to measure the quality of caregiver health. More research is necessary to better understand the physical and emotional health of caregivers of persons with substance use disorder and the factors that contribute to increased quality of life. Understanding the relationship between outcomes and protective factors could help nurses to develop prevention strategies and treatment interventions aimed at decreasing the psychosocial trauma and stress associated with caregiver burden.


Relationship between Corneal Sensation, Blinking, and Tear Film Quality.
To examine the possible role of corneal sensitivity and tear film quality in triggering a blink by investigating the relationship between blink rate, central corneal sensitivity threshold (CST), ocular surface temperature (OST), tear meniscus height (TMH), tear film quality (noninvasive tear break-up time [NIBUT]), and tear film lipid pattern under normal conditions. Forty-two volunteers (average age, 27.76 ± 5.36 years; 11 males) with good ocular health (Ocular Surface Disease Index, <15.0) were recruited for this cross-sectional cohort study. Blink rate, CST (noncontact corneal air gas aesthesiometry, NCCA), minimum and maximum OST in the central and inferior cornea between blinks (thermal infrared camera), TMH, NIBUT, and lipid pattern of the tear film (Keeler Tearscope Plus) were recorded on the right eye only. Median blink rate was 11 blinks/min (interquartile range [IR], 6.95 to 17.05), CST was 0.35 mbars (IR, 0.30 to 0.40), minimum OST in the central cornea was 35.15°C (IR, 34.58 to 35.50), and NIBUT was 34.55 s (IR, 12.45 to 53.80). Moderate but statistically significant correlations were observed between CST and NIBUT (r = 0.535, p < 0.001), CST and blink rate (r = -0.398, p < 0.001), lipid pattern and OST (r = 0.556, p < 0.001), and between CST and OST (r = 0.371, p = 0.008). The correlations between blink rate and NIBUT (r = -0.696, p < 0.001) and between OST and NIBUT (r = 0.639, p < 0.001; Spearman test) achieved higher significance; this was highlighted by the linear regression model where NIBUT and minimum central and inferior OST were identified as significant predictor variables. There is strong evidence for significant interactions between corneal sensitivity, NIBUT, OST, and blink frequency, emphasizing that ocular surface conditions represent a possible important trigger for the initiation of a blink. However, the mechanisms involved in the initiation of a blink are complex, with local ocular sensory input as only one trigger, along with other external influences and internal factors under cortical control.


Fgf3 is crucial for the generation of monoaminergic cerebrospinal fluid contacting cells in zebrafish.
In most vertebrates, including zebrafish, the hypothalamic serotonergic cerebrospinal fluid-contacting (CSF-c) cells constitute a prominent population. In contrast to the hindbrain serotonergic neurons, little is known about the development and function of these cells. Here, we identify fibroblast growth factor (Fgf)3 as the main Fgf ligand controlling the ontogeny of serotonergic CSF-c cells. We show that fgf3 positively regulates the number of serotonergic CSF-c cells, as well as a subset of dopaminergic and neuroendocrine cells in the posterior hypothalamus via control of proliferation and cell survival. Further, expression of the ETS-domain transcription factor etv5b is downregulated after fgf3 impairment. Previous findings identified etv5b as critical for the proliferation of serotonergic progenitors in the hypothalamus, and therefore we now suggest that Fgf3 acts via etv5b during early development to ultimately control the number of mature serotonergic CSF-c cells. Moreover, our analysis of the developing hypothalamic transcriptome shows that the expression of fgf3 is upregulated upon fgf3 loss-of-function, suggesting activation of a self-compensatory mechanism. Together, these results highlight Fgf3 in a novel context as part of a signalling pathway of critical importance for hypothalamic development.


Role of the acidic hirudin-like COOH-terminal amino acid region of factor Va heavy chain in the enhanced function of prothrombinase.
Prothrombinase activates prothrombin through initial cleavage at Arg(320) followed by cleavage at Arg(271). This pathway is characterized by the generation of an enzymatically active, transient intermediate, meizothrombin, that has increased chromogenic substrate activity but poor clotting activity. The heavy chain of factor Va contains an acidic region at the COOH terminus (residues 680-709). We have shown that a pentapeptide from this region (DYDYQ) inhibits prothrombin activation by prothrombinase by inhibiting meizothrombin generation. To ascertain the function of these regions, we have created a mutant recombinant factor V molecule that is missing the last 30 amino acids from the heavy chain (factor V(Delta680-709)) and a mutant molecule with the (695)DYDY (698) --> AAAA substitutions (factor V(4A)). The clotting activities of both recombinant mutant factor Va molecules were impaired compared to the clotting activity of wild-type factor Va (factor Va (Wt)). Using an assay employing purified reagents, we found that prothrombinase assembled with factor Va(Delta680-709) displayed an approximately 39% increase in k cat, while prothrombinase assembled with factor Va(4A) exhibited an approximately 20% increase in k cat for the activation of prothrombin as compared to prothrombinase assembled with factor Va(Wt). Gel electrophoresis analyzing prothrombin activation by prothrombinase assembled with the mutant molecules revealed a delay in prothrombin activation with persistence of meizothrombin. Our data demonstrate that the COOH-terminal region of factor Va heavy chain is indeed crucial for coordinated prothrombin activation by prothrombinase because it regulates meizothrombin cleavage at Arg(271) and suggest that this portion of factor Va is partially responsible for the enhanced procoagulant function of prothrombinase.


The genome and transcriptome of a newly described psychrophilic archaeon, Methanolobus psychrophilus R15, reveal its cold adaptive characteristics.
We analysed the cold-responsive gene repertoire for a psychrophilic methanogen, Methanolobus psychrophilus R15 through genomic and RNA-seq assayed transcriptomic comparisons for cultures at 18°C (optimal temperature) versus 4°C. The differences found by RNA-seq analysis were verified using quantitative real time-PCR assay. The results showed that as in the Antarctic methanogen, Methanococcoides burtonii, genes for methanogenesis, biosynthesis and protein synthesis were all downregulated by the cold in R15. However, the RNA polymerase complex was upregulated at cold, as well as a gene cluster for a putative exosome complex, suggesting that exosome-mediated RNA decay may be cold-accelerated. Unexpectedly, the chaperonin genes for both thermosome and GroES/EL were all upregulated at 4°C. Strain R15 possessed eight protein families for oxygen detoxification, including both anaerobe-specific superoxide reductase (SOR) and the aerobe-typical superoxide dismutase (SOD)-catalase oxidant-removing system, implying the higher oxidative tolerance. Compared with a mesophilic methanogen, R15 survived in higher paraquat, a redox-cycling drug. Moreover, 71 one-component systems and 50 two-component systems for signal transduction ranked strain R15, together with M. burtonii, as being highly adaptive among archaea. Most of them exhibited cold-enhanced expression, indicating their involvement in cold adaptation. This study has added new perspectives on the cold adaptation of methanogenic archaea.


Oxidative stress abolishes leptomycin B-sensitive nuclear export of transcription repressor Bach2 that counteracts activation of Maf recognition element.
The mammalian transcription activator Nrf2 plays critical roles in executing oxidative stress response by binding to the regulatory DNA sequence Maf recognition element. Bach2 is an Nrf2-related transcription repressor and a tissue-specific partner of the Maf oncoprotein family. We show here how Bach2 is regulated by an oxidative stress-sensitive conditional nuclear export. In cultured cells, Bach2 was localized in cytoplasm through its C-terminal evolutionarily conserved cytoplasmic localization signal (CLS). The CLS directed leptomycin B-sensitive nuclear export of reporter proteins, suggesting its dependence on the nuclear exporter Crm1/exportin 1. However, the CLS sequence does not bear a resemblance to the leucine-rich class of nuclear export signal, and mutagenesis analysis indicated that a stretch of nonhydrophobic amino acids is essential for its activity. Oxidative stressors aborted the CLS activity and induced nuclear accumulation of Bach2. Whereas oxidative stress is known to activate MARE-dependent transcription, overexpression of Bach2 in cultured cells silenced the inducibility of MARE. The results suggest that Bach2 mediates nucleocytoplasmic communication to couple oxidative stress and transcription repression in mammalian cells.


Overview of current and future chemotherapeutic agents in non-small cell lung cancer.
The two-drug regimen consisting of a platinum compound (cisplatin or carboplatin) combined with either a vinca alkaloid or a podophyllotoxin has been considered by many to be the standard chemotherapy treatment for non-small cell lung cancer (NSCLC). Randomized trials with these regimens have demonstrated modest but statistically significant increases in survival for patients with stage IV disease compared to treatment with best supportive care, and especially for selected patients with stage III disease when combined with radiotherapy or surgery compared to these treatments alone. Recently, several new compounds with promising efficacy and acceptable toxicity profiles have been investigated for the treatment of NSCLC, including the taxanes paclitaxel and docetaxel, the novel pyrimidine analog gemcitabine, and the topoisomerase inhibitors irinotecan and topotecan. Small but significant improvements in response rates and survival have been achieved with two-drug combinations, which include several of these new agents combined with a platinum-based compound, in patients with advanced NSCLC. Modifications of dosing schedules and the use of premedication regimens have resulted in better efficacy and more manageable side effects with such combinations. These encouraging gains in patients with advanced NSCLC suggest a potentially greater impact in patients with early stage disease. Given the manageable toxicity profiles of many of these newer agents, various three-drug regimens may be feasible in the future.


Use of hyperbaric oxygen therapy in quinine-associated visual disturbances.
Hyperbaric oxygen (HBO2 ) therapy is infrequently reported as a treatment for poison-induced retinal damage. We describe a case in which HBO2 therapy was used to treat suspected retinal toxicity induced by quinine. We present a case in which HBO2 was used to treat visual disturbances thought to be caused by quinine-induced retinal damage. The patient intentionally ingested undisclosed amounts of citalopram and quinine. Following a complicated hospital course, including profound shock requiring treatment with four vasopressors and a peripheral left-ventricular assist device, the patient, once extubated, reported visual abnormalities consistent with those described from quinine-induced retinal toxicity. Visual disturbances seemed to show improvement following HBO2 treatment. Several months following hospitalization visual defects continued to be present on examination. However, with corrective lenses the patient's visual acuity was normal. No adverse events were attributed to the use of HBO2. HBO2 for treatment of quinine-induced retinal damage is infrequently reported or studied. In the reported case, use of HBO2 appeared to be associated with substantial improvement in visual disturbances occurring in the setting of an overdose of quinine. The patient's improvement is remarkable, given her retinas were also jeopardized by her profound shock. Additional data are needed to understand the risks and benefits of this procedure, but due to limited treatment options for poison-induced retinal toxicity and the low likelihood for implementation of a controlled randomized trial of HBO2 in this population, the procedure may be considered in quinine-induced retinal toxicity.


Variations in fluconazole susceptibility and DNA subtyping of multiple Candida albicans colonies from patients with AIDS and oral candidiasis suffering one or more episodes of infection.
Five Candida albicans colonies from each infection in AIDS patients receiving fluconazole therapy for oropharyngeal candidiasis over a 2-year period were evaluated by antifungal susceptibility testing and DNA subtyping, and the results were correlated with clinical response to determine the occurrence of clinically significant selection of more-resistant C. albicans over multiple infections. A total of 534 C. albicans isolates were obtained from 38 patients who exhibited 84 episodes of infection. Antifungal susceptibility testing revealed that the MICs for 93% of the isolates were < or = 8.0 microg/ml and the MICs for 7% of the isolates were > or = 64 microg/ml. DNA subtyping revealed 70 different subtypes, with 78% of patients with one infection exhibiting one DNA subtype and 80% of patients with more than one infection exhibiting multiple DNA subtypes. Also, patients who had multiple infections had lower CD4 counts than those with single infections. Differences between the single-infection group and the multiple-infection group regarding the number of DNA subtypes and CD4 counts were both statistically significant. Of the 74 evaluable infections all were successfully treated with regular-dose (100-mg/day) fluconazole, except for three patients who ultimately responded to higher-dose fluconazole. Only one patient may have shown clinically significant selection of a more-resistant C. albicans strain over multiple courses of treatment. Interestingly, MICs reached only 8.0 microg/ml, even though doses of 400 mg of fluconazole were necessary for clinical cure.


Microscopic characters of the leaf and stem of Lavandula dentata L. (Lamiaceae).
Lavandula dentata L. is an aromatic plant used in folk medicine for different purposes and, for this reason, phytochemical surveys have been carried out in the search for bioactive substances aiming to support its uses. Since there is little knowledge on the structural aspects of L. dentata, this work has studied the anatomical characters of the leaf and stem using light and scanning electron microscopy, in order to assist the species identification. As a result, there are different types of trichomes: capitate glandular with uni- or bicellular head, peltate glandular with multicellular head, and branched non-glandular. The leaf is hypostomatic showing diacytic stomata. The epidermis is uniseriate and coated with striate cuticle. The mesophyll is dorsiventral and the midrib is concave-convex and traversed by a single collateral vascular bundle. The stem is quadrangular and has alternating strands of collenchyma and cortical parenchyma as well as a typical endodermis in the cortex. The phloem and xylem cylinders are traversed by narrow rays and there is an incomplete sclerenchymatic sheath adjoining the phloem. These results are a novelty for the species and contribute to distinguish it from other lavenders.


Self-compassion, sleep quality and psychological well-being in type 2 diabetes: a cross-sectional study.
Low self-compassion and poor sleep quality have been identified as potential key predictors of distress in type 2 diabetes (T2D). This study investigated relationships between sleep behaviors (sleep duration, social jetlag and daytime sleepiness), diabetes-related distress (DRD) and self-compassion in people with T2D. This cross-sectional study used data from 467 people with T2D derived from self-report questionnaires, accelerometer-assessed sleep measures and demographic information (clinicaltrials.gov registration: NCT02973412). All participants had a diagnosis of T2D and no comorbid sleep disorder (excluding obstructive sleep apnea). Hierarchical multiple regression and mediation analysis were used to quantify relationships between self-compassion, sleep variables and DRD. Significant predictors of DRD included two negative subscales of the Self-Compassion Scale (SCS), and daytime sleepiness. The 'overidentified' and 'isolation' SCS subscales were particularly important in predicting distress. Daytime sleepiness also partially mediated the influence of self-compassion on DRD, potentially through self-care around sleep. Daytime sleepiness and negative self-compassion have clear associations with DRD for people with T2D. The specific negative subscale outcomes suggest that strengthening individuals' ability to mindfully notice thoughts and experiences without becoming enmeshed in them, and reducing a sense of separateness and difference, might be key therapeutic targets for improving well-being in T2D. Psychological interventions should include approaches focused on reducing negative self-compassion and improving sleep behavior. Equally, reducing DRD may carry beneficial outcomes for sleep and self-compassion. Further work is however crucial to establish causation and long-term impact, and for development of relevant clinical resources.


Enhanced enzyme-linked immunosorbent assay for detection of antibodies to virus-like particles of human papillomavirus.
Measurement of antibodies to human papillomavirus (HPV) is complicated by many factors. Although enzyme-linked immunosorbent assays (ELISAs) that use virus-like particles (VLPs) have proved useful, the assays have, in general, had moderate sensitivities and low signal-to-noise ratios. To enhance the performance of the assay, a systematic investigation was undertaken to examine key variables used in ELISAs for the detection of antibodies to VLPs of HPV. Incorporation of two vinyl polymers, polyvinyl alcohol (molecular weight, 50,000) (PVA-50) and polyvinylpyrrolidone (molecular weight, 360,000) (PVP-360), was found to increase the sensitivity as well as the specificity of the assay for the detection of antibodies to VLPs of HPV. In particular, the addition of PVA-50 to the blocking solution reduced the amount of nonspecific binding of antibodies to VLPs and the microplate surface, whereas the addition of PVP-360 increased the sensitivity of antibody detection. The new ELISA demonstrated increased sensitivity and specificity for the detection of cervical HPV type 16 infection compared to those of a prototype assay with coded clinical serum samples from women with known cervicovaginal HPV infection status. It is anticipated that the enhanced ELISA conditions will have wide application to a large number of clinical diagnostic assays.


Delayed cord clamping is associated with improved dynamic cerebral autoregulation and decreased incidence of intraventricular hemorrhage in preterm infants.
Delayed cord clamping (DCC) improves neurologic outcomes in preterm infants through a reduction in intraventricular hemorrhage (IVH) incidence. The mechanism behind this neuroprotective effect is not known. Infants born <28 wk gestation were recruited for longitudinal monitoring. All infants underwent 72 h of synchronized near-infrared spectroscopy (NIRS) and mean arterial blood pressure (MABP) recording within 24 h of birth. Infants with DCC were compared with control infants with immediate cord clamping (ICC), controlling for severity of illness [clinical risk index for babies (CRIB-II) score], chorioamnionitis, antenatal steroids, sedation, inotropes, and delivery mode. Autoregulatory dampening was calculated as the transfer function gain coefficient between the MABP and NIRS signals. Forty-five infants were included (DCC; n = 15, paired 2:1 with ICC controls n = 30). ICC and DCC groups were similar including gestational age (25.5 vs. 25.2 wk, P = 0.48), birth weight (852.3 vs. 816.6 g, P = 0.73), percent female (40 vs. 40%, P = 0.75), and dopamine usage (27 vs. 23%, P = 1.00). There was a significant difference in IVH incidence between the DCC and ICC groups (20 vs. 50%, P = 0.04). Mean MABP was not different (35.9 vs. 35.1 mmHg, P = 0.44). Compared with the DCC group, the ICC group had diminished autoregulatory dampening capacity (-12.96 vs. -15.06 dB, P = 0.01), which remained significant when controlling for confounders. Dampening capacity was, in turn, strongly associated with decreased risk of IVH (odds ratio = 0.14, P < 0.01). The results of this pilot study demonstrate that DCC is associated with improved dynamic cerebral autoregulatory function and may be the mechanism behind the decreased incidence of IVH. NEW & NOTEWORTHY The neuroprotective mechanism of delayed cord clamping in premature infants is unclear. Delayed cord clamping was associated with improved cerebral autoregulatory function and a marked decrease in intraventricular hemorrhage (IVH). Improved dynamic cerebral autoregulation may decrease arterial baroreceptor sensitivity, thereby reducing the risk of IVH.


Complaints of heroin-maintained patients: A survey of symptoms ascribed to diacetylmorphine.
Prescribing of injectable diacetylmorphine (DAM) for heroin dependence has raised concerns about its safety. In light of various reports by heroin-maintained patients of DAM-related adverse events, and previously established unwanted effects of opioids in pain management, we undertook a survey in February 2001 of a random sample of 132 (127 participated) of 1061 patients prescribed DAM in Switzerland at that time. The purpose was to document the prevalence rates of a list of unintended symptoms experienced and attributed to DAM by patients. To assess symptom complaints and other data, staff administered a six-page self-report questionnaire. The patients ascribed numerous symptoms to DAM, with the best-known being the most frequently reported (e.g. skin itching, sweating, constipation). Among potentially more problematic complaints ranged irregular menses, cognitive deficits, muscle twitches, labored breathing, pains in the cardiac region, and temporary paralysis of limbs. In the absence of a control group, however, these may also be due to other factors, such as expectation, co-medication, concomitant substance use or co-morbidity. This pilot study emphasizes the necessity of rigorous assessment of the true rates, types, severity and preventability of such complications, especially given the current efforts to establish heroin maintenance as an optional treatment for heroin dependence.


How Men with Prostate Cancer Choose Specialists: A Qualitative Study.
The specific specialist that a patient sees can have a large influence on the type of care they receive. We administered semistructured interviews with 47 men diagnosed with prostate adenocarcinoma between 2012 and 2014. Telephone interviews were recorded, transcribed, and analyzed using a systematic thematic approach. Three profiles of patients emerged for choosing specialists: active (21.3%), partially active (53.2%), and passive (25.5%). Active patients conducted substantial research when choosing a diagnosing urologist and a treating specialist: they searched online, consulted other men with prostate cancer, and/or visited multiple specialists for opinions. Partially active patients took only 1 additional step to find a treating specialist on their own after receiving a referral from their diagnosing urologist. Passive patients relied exclusively on referrals from their primary care physicians (PCPs) and diagnosing urologists. The majority of patients relied on their PCPs for referrals to diagnosing urologists and on their diagnosing urologists to choose the treating specialist. Given these findings and the significance of specialist choice in determining treatment, it is important that PCPs recognize their indirect but potentially important effect on treatment choice when making referrals for prostate cancer. PCPs should consider counseling patients about seeking second opinions from providers with different treatment perspectives and participating in treatment decisions.


[Osteo-articular manifestations in dialysis of patients over 70 years].
A clinical, biological and radiological prospective study was carried out in 21 patients over 70 years of age and treated by hemodialysis or chronic ambulatory peritoneal dialysis (CAPD) to evaluate the frequency and specificity of rheumatic diseases observed in aged chronic renal failure patients. Some are caused or favored by old age and are not in any way related to renal failure and its replacement therapy. Such was the case with arthrosis which was present in 85% of patients, ankylosing vertebral hyperostosis (14%), Paget's disease (5%) and gouty arthritis (10%). Elsewhere there is an implication between abnormalities due to aging and those linked to renal failure and/or dialysis, some of which can worsen or accelerate others. Secondary hyperparathyroidism seems less frequent in the elderly than in the young patients. Common vitamin dependent osteomalacia should not be neglected because it can be either prevented or efficiently treated. Osteoporosis is another important factor in osteopenia. Extra-skeletal calcifications are frequent: periarticular calcifications (38%), chondrocalcinosis (14%) and disc calcifications (24%). Dialysis arthropathy comprising: carpal tunnel syndrome, erosive lesions of large and intermediate limb joint articulations and destructive spondylarthropathy is observed in 43% of patients after an average dialysis period of 44 months. The advent of this complication seems to be quite early in the elderly, as compared to the young population, which confirms the role played by age as a favoring factor.


Transgastric approach to posterior juxta-esophageal gastric ulcer.
Three cases are reported to illustrate the transgastric techniques for approach to posterior wall juxta-esophageal gastric ulcers. The author emphasizes the importance of making the incision or suture line parallel to the esophagus to avoid compromise of esophageal entry. The advantages of this technique include speed, good direct exposure for a mucosal view, and minimal interruption of gastric blood supply in the exploratory phase before planning a definitive procedure. It is particularly suitable for occasions when surgical risks are increased by obesity or critical clinical status, or when the pathology is strictly localized.


Regional recurrence after negative sentinel lymph node biopsy for melanoma.
Sentinel lymph node (SLN) biopsy has shown great utility in the management of melanoma. An analysis of regional recurrence in previously mapped negative SLN basins as the first site of relapse is performed. A retrospective query of a prospective melanoma database from 1994 to 2006 identified 1287 patients who underwent successful SLN biopsy. One thousand sixty patients (82.4%) were SLN negative and 227 (17.6%) patients SLN positive. Clinical variables were examined for the impact on regional recurrence by multivariate analysis. Mean follow-up was 44.3 months (range 3-155 months). Thirty-five patients (3.3%) presented with false-negative (FN) SLN biopsy. Pathologic review of the SLNs harvested from these basins found 7 (20.0%) samples positive for metastatic melanoma. Multivariate analysis found head and neck site [hazard ratio 3.67; 95% confidence interval (CI), 1.77-7.60, P < 0.001] and tumor thickness (hazard ratio 1.16; 95% CI, 1.04-1.30, P = 0.01) to be predictive of FN SLN biopsy. The 5-year melanoma specific survival calculated from the date of the SLN biopsy was 57.6% (95%CI, 35.7-41.9) in the FN group, which was not statistically different than the SLN positive group 60.0% (95% CI, 29.6-40.1; P = 0.14). Head and neck tumor site and tumor thickness are predictors of a FN SLN biopsy. Mechanisms other than pathologic SLN sampling error may contribute to the failure of the SLN biopsy in some patients. Patients with regional recurrence after negative SLN biopsy have a similar 5-year survival compared with patients with positive SLNs.


Lanthanide-doped bismuth oxobromide nanosheets for self-activated photodynamic therapy.
Low tissue penetration depth of the excited light and complicated synthetic procedures greatly hinder the clinical application of photodynamic therapy (PDT). Here we present a facile and mass production route to fabricate Yb3+/Tm3+ co-doped BiOBr nanosheets. In contrast to the complicated combination of photosensitizers (PSs) with up-conversion nanoparticles (UCNPs), which generates a PDT effect by a fluorescence resonance energy transfer process from UCNPs to PSs upon near-infrared light excitation, this as-synthesized material can be self-activated by deep-penetrating 980 nm laser light to produce a large amount of reactive oxygen species, giving rise to a high PDT efficiency which has been proven by in vitro and in vivo therapeutic assays. Surface modification of the BiOBr:Yb,Tm nanosheets with polyethylene glycol endows the system with improved biocompatibility. Through the combination of inherent fluorescence and CT imaging properties, an imaging-monitored therapeutic system has been realized. The system overcomes the problems of low tissue penetration depth, complicated structure-induced low efficiency, and potential safety concerns. Our finding presents the first demonstration of a self-activated nanoplatform for targeted and noninvasive deep-cancer therapy.


Lens capsule abnormalities in Alport's syndrome.
The ultrastructure of the glomerular basement membrane in Alport's syndrome is well known and characteristic of this disease, but the anterior lenticonus that frequently occurs in this syndrome has not been similarly studied. An anterior lens capsule from a 30-year-old patient with lenticonus who had Alport's syndrome was obtained at capsulectomy and found to be one third the normal thickness centrally and to be more fibrillar than usual. There were large numbers of partial capsular dehiscences containing fibrillar material and vacuoles. Cellular debris and more recent ruptures of lens epithelial cell membranes underlaid the breaks. The anterior capsule is clearly fragile in this disease, forming the basis for the progressive lenticonus and anterior polar cataract. These abnormalities correlate well with a defect in the type IV collagen molecule, as has been recently reported in Alport's syndrome.


PKR inhibitors suppress endoplasmic reticulum stress and subdue glucolipotoxicity-mediated impairment of insulin secretion in pancreatic beta cells.
Type 2 diabetes mellitus is characterized by insulin resistance and hypersecretion of insulin from the pancreas to compensate for decreased insulin sensitivity in the peripheral tissues. In later stages of the disease insulin-secreting beta cell degeneration commences and patients require insulin replacement therapy in order to accomplish proper regulation of their blood glucose. Endoplasmic reticulum (ER) stress in the beta cells is one of the factors contributing to this detrimental effect. Protein kinase R (PKR) is a cellular stress kinase activated by ER stress and contributing to degeneration of pancreatic islets. In order to determine whether inhibition of PKR activation by specific small molecule inhibitors of PKR ameliorates pancreatic insulin secretion capacity, we treated beta cells with two imidazole/oxindole-derived inhibitors of PKR kinase, imoxin (C16) and 2-aminopurine (2-AP), in the presence of ER stress. Our results demonstrate that PKR inhibition suppresses tunicamycin-mediated ER stress without altering the insulin production capacity of the cells. Palmitic acid-mediated suppression of insulin secretion, however, was subdued significantly by PKR inhibitor treatment through an ER stress-related mechanism. We suggest that PKR inhibitor treatment may be used to increase the insulin secretion capacity of the pancreas in later stages of diabetes.


Plasma amino acid and metabolite signatures tracking diabetes progression in the UCD-T2DM rat model.
Elevations of plasma concentrations of branched-chain amino acids (BCAAs) are observed in human insulin resistance and type 2 diabetes mellitus (T2DM); however, there has been some controversy with respect to the passive or causative nature of the BCAA phenotype. Using untargeted metabolomics, plasma BCAA and other metabolites were assessed in lean control Sprague-Dawley rats (LC) and temporally during diabetes development in the UCD-T2DM rat model, i.e., prediabetic (PD) and 2 wk (D2W), 3 mo (D3M), and 6 mo (D6M) post-onset of diabetes. Plasma leucine, isoleucine, and valine concentrations were elevated only in D6M rats compared with D2W rats (by 28, 29, and 30%, respectively). This was in contrast to decreased plasma concentrations of several other amino acids in D3M and/or D6M relative to LC rats (Ala, Arg, Glu, Gln, Met, Ser, Thr, and Trp). BCAAs were positively correlated with fasting glucose and negatively correlated with plasma insulin, total body weight, total adipose tissue weight, and gastrocnemius muscle weight in the D3M and D6M groups. Multivariate analysis revealed that D3M and D6M UCD-T2DM rats had lower concentrations of amino acids, amino acid derivatives, 1,5-anhydroglucitol, and conduritol-β-opoxide and higher concentrations of uronic acids, pantothenic acids, aconitate, benzoic acid, lactate, and monopalmitin-2-glyceride relative to PD and D2W UCD-T2DM rats. The UCD-T2DM rat does not display elevated plasma BCAA concentrations until 6 mo post-onset of diabetes. With the acknowledgement that this is a rodent model of T2DM, the results indicate that elevated plasma BCAA concentrations are not necessary or sufficient to elicit an insulin resistance or T2DM onset.


Detection, localization, and action of the INSL3 receptor, LGR8, in rat kidney.
Recent molecular and pharmacologic studies have identified LGR8, a member of the leucine-rich repeat-containing G-protein-coupled receptor family, as a cognate receptor for insulin-like peptide-3 (INSL3). LGR8 mRNA has been detected in various tissues, but the precise roles of these INSL3-LGR8 systems are unknown. In this study we first investigated the presence and cellular localization of LGR8 mRNA in both adult and developing rat kidney and subsequently examined the possible role of INSL3-LGR8 signaling in cultured mesangial cells. LGR8 mRNA was detected in the kidney by polymerase chain reaction and localized by in situ hybridization in mature glomerular mesangial cells within the renal cortex, with highest levels detected at embryonic day 18 and lowest levels in adult kidney. Synthetic INSL3 inhibited the proliferation of mesangial cells in primary culture, indicating the presence of functional LGR8 on these cells. These findings suggest that INSL3/LGR8 signaling may be involved in the genesis and/or developmental maturation of renal glomeruli and in the regulation of mesangial cell density in the adult kidney.


[Influence of carbidine on the noradrenaline level in the adrenergic nerve fibers in the peripheral tissues].
By employing fluorescent histochemistry and spectrofluorimetry the influence of an original psychotropic substance -- carbidine on the level, localization and accumulation of the adrenergic mediator in Vas deferens, dura mater and the kidney capsule was studied "in vivo" and "in vitro". Carbidine was shown to capable of liberating the adrenergic mediator from the sympathetic nerve fibres of the peripheral tissues without affecting the capture and accumulation of exogenous norepinephrine by adrenergic nerve fibres.


A molecular and pathological study of peste des petits ruminants virus (PPRV) from field outbreaks in Palestine, 2017-2019.
Peste des petits ruminants (PPR) is a severe, highly contagious and fatal viral disease of small ruminants causing high economic losses. Therefore, the disease was targeted for eradication by 2030. The aim of this study was to investigate for the first time the molecular and pathological characterization of the circulating PPR virus (PPRV) in sheep and goat in Palestine. Samples were collected from suspected necropsy cases of sheep and goats during recent outbreaks in two provinces in Palestine between 2017 and 2019. In this study, severe PPR outbreaks occurred in sheep and goats causing typical lesions which include erosive and ulcerative stomatitis, bronchointerstitial pneumonia and severe enteritis. For the molecular investigation of PPRV, suspected animals were examined for the presence of PPRV by RT-PCR. PPRV genome was detected in all samples. Subsequently, two samples were used for N gene sequencing and phylogenetic analysis of PPRV isolates. The nucleotide sequence and phylogenetic analysis indicated that the Palestinian PPRV isolates were genetically clustered within the lineage IV isolates of the virus among populations of sheep and goats which most prevalent in Asia, the Middle East and recently Africa. Further analysis showed that the Palestinian isolates were closely related to those described in Turkey and Iraq, suggesting a common origin of PPRV isolates in the region. This information is critical to understand the molecular epidemiology of this disease in the region and helps to develop appropriate control measures for eradication of this disease.


A comparison of the home-care and healthcare service use and costs of older Australians randomised to receive a restorative or a conventional home-care service.
Restorative home-care services, or re-ablement home-care services as they are now known in the UK, aim to assist older individuals who are experiencing difficulties in everyday living to optimise their functioning and reduce their need for ongoing home care. Until recently, the effectiveness of restorative home-care services had only been investigated in terms of singular outcomes such as length of home-care episode, admission to hospital and quality of life. This paper reports on a more complex and perhaps more significant measure--the use and cost of the home-care and healthcare services received over the 2-year period following service commencement. Seven hundred and fifty older individuals referred for government-funded home care were randomly assigned to a restorative or standard service between June 2005 and August 2007. Health and aged care service data were sourced and linked via the Western Australian Data Linkage System. Restorative clients used fewer home-care hours (mean [SD], 117.3 [129.4] vs. 191.2 [230.4]), had lower total home-care costs (AU$5570 vs. AU$8541) and were less likely to be approved for a higher level of aged care (N [%], 171 [55.2] vs. 249 [63.0]) during follow-up. They were also less likely to have presented at an emergency department (OR = 0.69, 95% CI = 0.50-0.94) or have had an unplanned hospital admission [OR (95% CI), 0.69 (0.50-0.95)]. Additionally, the aggregated health and home-care costs of the restorative clients were lower by a factor of 0.83 (95% CI 0.72-0.96) over the 2-year follow-up (AU$19,090 vs. AU$23,428). These results indicate that at a time when Australia is facing the challenges of population ageing and an expected increase in demand for health and aged care services, the provision of a restorative service when an older person is referred for home care is potentially a more cost-effective option than providing conventional home care.


Adverse effects of high glucose levels on somite and limb development in avian embryos.
Gestational diabetes has an adverse impact on fetal musculoskeletal development, but the mechanism involved is still not completely understood. In this study, we investigated the effects of high glucose on the developing somites and their derivate using the chick embryo as a model. We demonstrated that under high glucose, the number of generated somites was reduced and their morphology altered in 2-day old chick embryos. In addition, high glucose repressed the development of the limb buds in 5.5-day old chick embryos. We also demonstrated that high glucose abridged the development of the sclerotome and the cartilage in the developing limb bud. The sonic hedgehog (Shh) gene has been reported to play a crucial role in the development and differentiation of sclerotome. Hence, we examined how Shh expression in the sclerotome was affected under high glucose. We found that high glucose treatment significantly inhibited Shh expression. The high glucose also impaired myotome formation at trunk level - as revealed by immunofluorescent staining with MF20 antibodies. In the neural tube, we established that Wnt3a expression was also significantly repressed. In summary, our study demonstrates that high glucose concentrations impair somite and limb bud development in chick embryos, and suggests that Shh and Wnt genes may play a role in the underlying mechanism.


Recent progress in covalent organic framework thin films: fabrications, applications and perspectives.
As a newly emerging class of porous materials, covalent organic frameworks (COFs) have attracted much attention due to their intriguing structural merits (e.g., total organic backbone, tunable porosity and predictable structure). However, the insoluble and unprocessable features of bulk COF powder limit their applications. To overcome these limitations, considerable efforts have been devoted to exploring the fabrication of COF thin films with controllable architectures, which open the door for their novel applications. In this critical review, we aim to provide the recent advances in the fabrication of COF thin films not only supported on substrates but also as free-standing nanosheets via both bottom-up and top-down strategies. The bottom-up strategy involves solvothermal synthesis, interfacial polymerization, room temperature vapor-assisted conversion, and synthesis under continuous flow conditions; whereas, the top-down strategy involves solvent-assisted exfoliation, self-exfoliation, mechanical delamination, and chemical exfoliation. In addition, the applications of COF thin films including energy storage, semiconductor devices, membrane-separation, sensors, and drug delivery are summarized. Finally, to accelerate further research, a personal perspective covering their synthetic strategies, mechanisms and applications is presented.


Parent-of-origin effect in multiple sclerosis: observations from interracial matings.
Multiple sclerosis (MS) is a complex neurologic disease with a striking geographical distribution. In Canada, prevalence is high in Caucasians of Northern European ancestry and uncommon in North American Aboriginals, many of whom now have Caucasian admixture. The population-based Canadian Collaborative Project on the Genetic Susceptibility to MS provided the characteristics of 58 individuals with 1 Caucasian and 1 North American Aboriginal parent from a database of 30,000 MS index cases. We found that MS index cases with a Caucasian mother and a North American Aboriginal father had a higher sib recurrence risk and greater F:M sex ratio (p = 0.043) than patients with a North American Aboriginal mother and Caucasian father. Maternal parent-of-origin effects in multiple sclerosis disease etiology previously seen in studies of half-siblings and avuncular pairs are also seen in Caucasian-North American Aboriginal admixture matings and warrant further investigation. A differential influence of maternal risk transmission on the sex ratio of affected offspring is implied. The method of analysis used may have broader implications for detection of parent-of-origin effects in admixture cohorts.


Identification and characterization of two distinct alpha-(1-3)-L-fucosyltransferase activities in human colon carcinoma.
Two distinct alpha-(1-3)-fucosyltransferase activities have been identified in the colon carcinoma cell lines HT-29 and COLO-205. While both enzymatic activities exhibit similar affinities for a synthetic alpha-(1-3) acceptor and GDP-fucose, they differ with respect to divalent cation requirements, N-ethylmaleimide inhibition, and glycoprotein substrate specificity. The COLO-205 alpha-(1-3) activity exhibits maximal enzymatic activity in the presence of 20 mM Mn2+ but retains less than 10% activity in the absence of divalent cations. In contrast, the optimal Mn2+ concentration for the HT-29 enzyme is 1 mM, although this activity is relatively insensitive to divalent cation stimulation. In addition, the HT-29 alpha-(1-3)-fucosyltransferase activity is resistant to inhibition by 30 mM N-ethylmaleimide and relatively inactive toward the glycoprotein substrate fetuin as compared to its desialylated derivative, asialofetuin. The COLO-205 activity is inhibited approximately 90% by N-ethylmaleimide and is equally active with either glycoprotein acceptor. Although the alpha-(1-3) specific activities are similar in both cell lines, N-ethylmaleimide-sensitive alpha-(1-4) fucosyltransferase activity is 40-fold higher in COLO-205 as compared to HT-29, suggesting that the COLO-205 fucosyltransferase activity may be an alpha-(1-3/4) enzyme, while the HT-29 activity appears to be an alpha-(1-3) specific form. Further examination of a panel of cell lines, tumor biopsies, and xenografts, based on the effect of metal ions and N-ethylmaleimide, indicated that both enzyme activities are similarly expressed in human colon carcinoma tissue.


Does Self-Efficacy Mediate the Link Between Impulse Control and Diabetes Adherence?
The goal of this study was to test the hypothesis that diabetes self-efficacy mediates the relationship between impulse control and type 1 diabetes (T1D) management from ages 8 to 18 years, using multilevel modeling. Participants included 117 youth with T1D and their parents. Youth (aged 8-16 years at baseline) and parents were assessed 5 times over 2 years. Using a cohort sequential design, we first estimated the growth trajectory of adherence from age 8 to 18 years, then specified a multilevel mediation model using impulse control as the main predictor, diabetes self-efficacy as the mediator, and changes in adherence (both within- and between-individuals) as the outcome. According to youth-reported adherence only, self-efficacy partially mediated the within-person effect of impulse control on adherence. On occasions when youth reported increases in impulse control, they tended to report higher adherence, and this was, in part, due to increases in youths' perceived self-efficacy. Self-efficacy accounted for approximately 21% of the within-person relationship between impulse control and youth-reported adherence. There was no association between impulse control and adherence between-individuals. Impulse control and self-efficacy were not related to parent-reported adherence. Environments that enrich youth with confidence in their own diabetes-related abilities may benefit self-care behaviors in youth with T1D, but such increases in youths' perceived competence do not fully account for, or override, the behavioral benefits of impulse control. Efforts to improve adherence in youth with T1D will benefit from consideration of both impulse control and self-efficacy.


A Middle-aged Man With Progressive Ophthalmoparesis, Ataxia, and Spastic Paraparesis.
A 50-year old man presented for evaluation of progressive gait ataxia with a superimposed spastic paraparesis. During his clinic visit, he was also observed to have slow and limited eye movements. In this article, we discuss the clinical approach to this triad of symptoms and guide the reader to discover the patient's ultimate genetic diagnosis.


Enfortumab Vedotin, a fully human monoclonal antibody against Nectin 4 conjugated to monomethyl auristatin E for metastatic urothelial Carcinoma.
Introduction: The conventional management of most patients with metastatic urothelial carcinoma (UC) is platinum-based chemotherapy followed by immunotherapy. Erdafitinib is an option in post-platinum patients with activating mutations in fibroblast growth factor receptor (FGFR)-3 and -2. Salvage therapy with taxanes or vinflunine has demonstrated minimal efficacy. Enfortumab Vedotin (EV), a monoclonal antibody-drug conjugate (ADC) targeting nectin-4 is under investigation in patients with advanced UC. Areas covered: This review describes the epidemiology and unmet needs of patients with metastatic UC and is focused specifically on heavily treated patients. We explore the rationale for targeting nectin 4 and the clinical development of EV; efficacy and safety data from the completed phase I and II studies are examined. Ongoing trials to definitively assess clinical outcomes in comparison to current therapy and trials exploring EV in combination are also highlighted. Expert opinion: There is an unmet need for new therapies in most patients with advanced UC and who progress after platinum and immunotherapy. EV has shown promising efficacy and safety in this population in phase 1 and 2 trials including those with poor prognostic factors such as liver metastases. Ongoing trials exploring this agent in combination will continue to advance the treatment of UC.


Towards a pan marsupial sero-immunological tool in the demanding field of wildlife serology: Marsupial immunoglobulin-binding capability with protein A/G, protein L and anti-kangaroo antibody.
Detection of infections in wildlife species is increasingly important to reduce the risk of spreading zoonotic and economically important parasites, understand disease epidemiology and promote the conservation of wildlife species. Serological tests are key in disease diagnosis and surveillance by detecting immunoglobulins against infectious agents. However, the need for species-specific reagents has limited the application of serological tests in wildlife species. This study evaluated the serum immunoglobulin-binding capability of polyclonal anti-kangaroo antibody and two non-species-specific reagents, including protein A/G and protein L, with the largest range of Australian marsupial species so far, including 32 species representing three major marsupial orders. Immunoglobulin-binding capability was assessed using immunoblotting, enzyme-linked immunosorbent assay and Western blot techniques. Variation in immunoglobulin-binding capability was observed between the three reagents and across the species tested, both across but also within taxonomic groups. Taxonomic distance was thus not always a good predictor of immunoglobulin-binding affinity, emphasizing the need to validate these reagents for each species separately. However, all three reagents bound with the serum immunoglobulins of most marsupial species tested. The findings of this study provide a valuable reference for species differences in affinity to protein A/G, protein L and anti-kangaroo antibody, assisting in the selection of appropriate reagents and the development of sero-immunological assays in Australian marsupials.


Determining chemotactic responses by two subsurface microaerophiles using a simplified capillary assay method.
A simplified capillary chemotaxis assay utilizing a hypodermic needle, syringe, and disposable pipette tip was developed to measure bacterial tactic responses. The method was applied to two strains of subsurface microaerophilic bacteria. This method was more convenient than the Adler method and required less practice. Isolate VT10 was a strain of Pseudomonas syringae, which was isolated from the shallow subsurface. It was chemotactically attracted toward dextrose, glycerol, and phenol, which could be used as sole carbon sources, and toward maltose, which could not be used. Isolate MR100 was phylogenetically related to Pseudomonas mendocina and was isolated from the deep subsurface. It showed no tactic response to these compounds, although, it could use dextrose, maltose, and glycerol as carbon sources. The chemotaxis results obtained by the new method were verified by using the swarm plate assay technique. The simplified technique may be useful for routine chemotactic testing.


Synthesis and biological evaluation of the 1-arylpyrazole class of σ(1) receptor antagonists: identification of 4-{2-[5-methyl-1-(naphthalen-2-yl)-1H-pyrazol-3-yloxy]ethyl}morpholine (S1RA, E-52862).
The synthesis and pharmacological activity of a new series of 1-arylpyrazoles as potent σ(1) receptor (σ(1)R) antagonists are reported. The new compounds were evaluated in vitro in human σ(1)R and guinea pig σ(2) receptor (σ(2)R) binding assays. The nature of the pyrazole substituents was crucial for activity, and a basic amine was shown to be necessary, in accordance with known receptor pharmacophores. A wide variety of amines and spacer lengths between the amino and pyrazole groups were tolerated, but only the ethylenoxy spacer and small cyclic amines provided compounds with sufficient selectivity for σ(1)R vs σ(2)R. The most selective compounds were further profiled, and compound 28, 4-{2-[5-methyl-1-(naphthalen-2-yl)-1H-pyrazol-3-yloxy]ethyl}morpholine (S1RA, E-52862), which showed high activity in the mouse capsaicin model of neurogenic pain, emerged as the most interesting candidate. In addition, compound 28 exerted dose-dependent antinociceptive effects in several neuropathic pain models. This, together with its good physicochemical, safety, and ADME properties, led compound 28 to be selected as clinical candidate.


PICALM Variation Moderates the Relationships of APOE ɛ4 with Alzheimer's Disease Cerebrospinal Biomarkers and Memory Function Among Non-Demented Population.
APOE ɛ4 and PICALM are established genes associated with risk of late-onset Alzheimer's disease (AD). Previous study indicated interaction of PICALM with APOE ɛ4 in AD patients. To explore whether PICALM variation could moderate the influences of APOE ɛ4 on AD pathology biomarkers and cognition in pre-dementia stage. A total of 1,034 non-demented participants (mean age 74 years, 56% females, 40% APOE ɛ4 carriers) were genotyped for PICALM rs3851179 and APOE ɛ4 at baseline and were followed for influences on changes of cognition and cerebrospinal fluid (CSF) AD markers in six years. The interaction effects were examined via regression models adjusting for age, gender, education, and cognitive diagnosis. The interaction term of rs3851179×APOE ɛ4 accounted for a significant amount of variance in baseline general cognition (p = 0.039) and memory function (p = 0.002). The relationships of APOE ɛ4 with trajectory of CSF Aβ42 (p = 0.007), CSF P-tau181 (p = 0.003), CSF T-tau (p = 0.001), and memory function (p = 0.017) were also moderated by rs3851179 variation. APOE ɛ4 carriers experienced slower clinical and pathological progression when they had more protective A alleles of PICALM rs3851179. These findings firstly revealed the gene-gene interactive effects of PICALM with APOE ɛ4 in pre-dementia stage.


Work volume and strength training responses to resistive exercise improve with periodic heat extraction from the palm.
Body core cooling via the palm of a hand increases work volume during resistive exercise. We asked: (a) "Is there a correlation between elevated core temperatures and fatigue onset during resistive exercise?" and (b) "Does palm cooling between sets of resistive exercise affect strength and work volume training responses?" Core temperature was manipulated by 30-45 minutes of fixed load and duration treadmill exercise in the heat with or without palm cooling. Work volume was then assessed by 4 sets of fixed load bench press exercises. Core temperatures were reduced and work volumes increased after palm cooling (Control: Tes = 39.0 ± 0.1° C, 36 ± 7 reps vs. Cooling: Tes = 38.4 ± 0.2° C, 42 ± 7 reps, mean ± SD, n = 8, p < 0.001). In separate experiments, the impact of palm cooling on work volume and strength training responses were assessed. The participants completed biweekly bench press or pull-up exercises for multiple successive weeks. Palm cooling was applied for 3 minutes between sets of exercise. Over 3 weeks of bench press training, palm cooling increased work volume by 40% (vs. 13% with no treatment; n = 8, p < 0.05). Over 6 weeks of pull-up training, palm cooling increased work volume by 144% in pull-up experienced subjects (vs. 5% over 2 weeks with no treatment; n = 7, p < 0.001) and by 80% in pull-up naïve subjects (vs. 20% with no treatment; n = 11, p < 0.01). Strength (1 repetition maximum) increased 22% over 10 weeks of pyramid bench press training (4 weeks with no treatment followed by 6 weeks with palm cooling; n = 10, p < 0.001). These results verify previous observations about the effects of palm cooling on work volume, demonstrate a link between core temperature and fatigue onset during resistive exercise, and suggest a novel means for improving strength and work volume training responses.


QUANTITATIVE STUDIES ON GLYCOLYTIC ENZYMES IN LACTOBACILLUS PLANTARUM. II. INTRACELLULAR CONCENTRATIONS OF GLYCOLYTIC INTERMEDIATES IN GLUCOSE-METABOLIZING WASHED CELLS.
Mizushima, Shôji (University of Tokyo, Tokyo, Japan), and Kakuo Kitahara. Quantitative studies on glycolytic enzymes in Lactobacillus plantarum. II. Intracellular concentrations of glycolytic intermediates in glucose-metabolizing washed cells. J. Bacteriol. 87:1429-1435. 1964.-The intracellular concentrations of glycolytic intermediates in glucose-metabolizing cells could be estimated by using C(14)-glucose of high specific activity. Separation of each intermediate was accomplished by column chromatography with Dowex 1-X8 (formate) ion-exchange resin. If the intermediates were inseparable, one was converted into another substance separable by chromatography. Almost all the glycolytic intermediates were found to be radioactive, and all of the radioactive substances appearing on the chromatograms were limited to glycolytic intermediates or their related substances. The results clearly show the role of the glycolytic system for lactic acid fermentation in the organism. Among the intermediates, fructose-1,6-diphosphate and 3-phosphoglycerate were found to be present in relatively high concentrations. The reason for the high concentrations of these compounds is discussed.


LRRC52 (leucine-rich-repeat-containing protein 52), a testis-specific auxiliary subunit of the alkalization-activated Slo3 channel.
KSper, a pH-dependent K(+) current in mouse spermatozoa that is critical for fertility, is activated by alkalization in the range of pH 6.4-7.2 at membrane potentials between -50 and 0 mV. Although the KSper pore-forming subunit is encoded by the Slo3 gene, heterologously expressed Slo3 channels are largely closed at potentials negative to 0 mV at physiological pH. Here we identify a Slo3-associating protein, LRRC52 (leucine-rich repeat-containing 52), that shifts Slo3 gating into a range of voltages and pH values similar to that producing KSper current activation. Message for LRRC52, a homolog of the Slo1-modifying LRRC26 protein, is enriched in testis relative to other homologous LRRC subunits and is developmentally regulated in concert with that for Slo3. LRRC52 protein is detected only in testis. It is markedly diminished from Slo3(-/-) testis and completely absent from Slo3(-/-) sperm, indicating that LRRC52 expression is critically dependent on the presence of Slo3. We also examined the ability of other LRRC subunits homologous to LRRC26 and LRRC52 to modify Slo3 currents. Although both LRRC26 and LRRC52 are able to modify Slo3 function, LRRC52 is the stronger modifier of Slo3 function. Effects of other related subunits were weaker or absent. We propose that LRRC52 is a testis-enriched Slo3 auxiliary subunit that helps define the specific alkalization dependence of KSper activation. Together, LRRC52 and LRRC26 define a new family of auxiliary subunits capable of critically modifying the gating behavior of Slo family channels.


Hematopoietic colony-stimulating factors in head and neck cancers: Recent advances and therapeutic challenges.
Colony-stimulating factors (CSFs) are key cytokines responsible for the production, maturation, and mobilization of the granulocytic and macrophage lineages from the bone marrow, which have been gaining attention for playing pro- and/or anti-tumorigenic roles in cancer. Head and neck cancers (HNCs) represent a group of heterogeneous neoplasms with high morbidity and mortality worldwide. Treatment for HNCs is still limited even with the advancements in cancer immunotherapy. Novel treatments for patients with recurrent and metastatic HNCs are urgently needed. This article provides an in-depth review of the role of hematopoietic cytokines such as granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), and interleukin-3 (IL-3; also known as multi-CSF) in the HNCs tumor microenvironment. We have reviewed current results from clinical trials using CSFs as adjuvant therapy to treat HNCs patients, and also clinical findings reported to date on the therapeutic application of CSFs toxicities arising from chemoradiotherapy.


Musical empathy (Einfuhling).
In this paper an "alpha-effect" is defined as a plethysmograph amplitude increase to the hearing of music concurrently with strong affinity for music, pleasurable feeling and reported empathy. Based on this definition a hypothetical model of empathy to music is formulated and the psychophysiological alpha-effect mechanism is related to the processes of empathy and cognitive attribution. In order to understand the psychophysiological mechanism of the alpha-effect, a network is proposed made up of several physiological processes with feedback circuits and feedback circuits at both the sensory and physiological level, and at the cognitive and psychological level. It was aimed also to give a theoretical account of the efficacy of music and to explicate the underlying psychophysiological mechanisms of that efficacy. The model may also help clinical psychologists who want to use music as a therapeutic tool in arranging optimal programs suited to the individual needs, experience, and background of a patient.


Developmental dyslexia: A new look at clinical features and brain mechanisms.
Developmental dyslexia is the commonest "specific learning disorder" (DSM-5) or "developmental learning disorder with impairment in reading" (ICD-11). This impairment in reading acquisition is related to a defect in the installation of cognitive precursors necessary to master the grapheme-phoneme conversion. Its origin is largely genetic, but many environmental factors seem capable of modulating symptom intensity. Three types of presentation, roughly equal in occurrence, are useful to distinguish according to the associated disorders (language, attentional, and/or motor coordination), thus suggesting, at least in part, potentially different mechanisms at their origin. In adolescence and adulthood the clinical presentation tends to bear a more uniform pattern, covering a large range of severity depending on each person's ability to compensate for their deficit. Research has demonstrated dysfunction of specific brain areas during reading-related tasks (using fMRI), essentially in the left cerebral hemisphere, but also atypical patterns of connectivity (using diffusion imaging), further supplemented by functional connectivity studies at rest. The current therapeutic recommendations emphasize the need for multidisciplinary care, giving priority, depending on the clinical form, to the language, psychomotor, or neuropsychologic aspects of rehabilitation. Various training methods whose effectiveness has been scientifically tested are reviewed, emphasizing those exploiting the hypothesis of a lack of intermodal connectivity between separate cognitive systems.


MRI in T staging of rectal cancer: How effective is it?
Rectal cancer constitutes about one-third of all gastrointestinal (GI) tract tumors. Because of the high recurrence rates (30%) in rectal cancer, it is vitally important to accurately stage these tumours preoperatively so that appropriate surgical resection can be undertaken. MRI is the ideal technique for the preoperative staging of these tumours. To determine the accuracy of local T staging of rectal cancer with MRI, using histopathological staging as the gold. Forty consecutive patients admitted with rectal cancer over a period of 18 months were included in this retrospective study. MRI scans were performed prior to surgery in all patients, on 1.5T scanners. Two radiologists, with a special interest in gastrointestinal imaging reported all images. Two dedicated histopathologists reported the histology slides. The accuracy of preoperative local MRI T staging was assessed by comparison with postoperative histopathological staging. There was agreement between MRI and histopathology (TNM) staging in 12 patients (30%). The sensitivity and specificity of MRI for T staging was 89% and 67% respectively. The circumferential resection margin (CRM) status was accurately staged in 94.1% of the patients. Preoperative staging with MRI is sensitive in identifying CRM involvement, which is the main factor affecting the outcome of surgery.


A case report of anti-GAD65 antibody-positive autoimmune encephalitis in children associated with autoimmune polyendocrine syndrome type-II and literature review.
Glutamic acid decarboxylase (GAD) is the rate-limiting enzyme for the synthesis of gamma-aminobutyric acid (GABA), the major inhibitory neurotransmitter in the central nervous system. Antibodies against glutamic acid decarboxylase (GAD) are associated with various neurologic conditions described in patients, including stiff person syndrome, cerebellar ataxia, refractory epilepsy, and limbic and extra limbic encephalitis. While there are few case reports and research on anti-GAD65 antibody-associated encephalitis in adults, such cases are extremely rare in pediatric cases. For the first time, we report a case of anti-GAD65-positive autoimmune encephalitis associated with autoimmune polyendocrine syndrome (APS) type II. We reviewed previously published pediatric cases of anti-GAD65 autoimmune encephalitis to discuss their clinical features, laboratory tests, imaging findings, EEG patterns, and prognosis. An 8-year-old, male child presented to the outpatient department after experiencing generalized convulsions for twenty days. The child was admitted for epilepsy and had received oral sodium valproate (500 mg/day) in another center, where investigations such as USG abdomen and MRI brain revealed no abnormalities, however, had abnormal EEG with diffuse mixed activity in the left anterior middle prefrontal temporal region. On the follow-up day, a repeat blood test showed a very low serum drug concentration of sodium valproate hence the dose was increased to 750 mg/day. Then, the child experienced adverse effects including increased sleep, thirst, and poor appetite, prompting the parents to discontinue the medication. A repeat MRI showed increased signals on FLAIR sequences in the right hippocampus hence admitted for further management. The child's past history included a diagnosis of hypothyroidism at the age of 4, and receiving levothyroxine 75 mcg once daily. His parents are healthy with no history of any similar neurological, autoimmune, or genetic diseases, but his uncle had a history of epilepsy. At presentation, he had uncontrolled blood glucose levels with elevated HbA1c levels. Additionally, the serum and CSF autoantibodies were positive against the anti-GAD65 antibody with the titer of 1:100 and 1:32 respectively. The patient was managed with a mixed type of insulin regimen and received first-line immunotherapy (intravenous immunoglobulin, IVIG) for five consecutive days, followed by oral prednisone and sodium valproate as an antiepileptic drug. Upon achieving a favorable clinical outcome, the patient was discharged with oral medications. Among the 15 pediatric patients reported in this literature, nine presented with limbic encephalitis (LE), three with extralimbic encephalitis (ELE), and three with a combination of limbic and extralimbic encephalitis. Most of these cases exhibited T2-W FLAIR hyperintensities primarily localized to the temporal lobes in the early phase, progressing to hippocampal sclerosis/atrophy in the later phase on MRI. EEG commonly showed slow or spike waves on frontotemporal lobes with epileptic discharges. Prognostic factors varied among patients, with some experiencing persistent refractory seizures, type-1 diabetes mellitus (T1DM), persistent memory impairment, persistent disability requiring full assistance, and, in severe cases, death. Our findings suggest that anti-GAD65 antibody-positive autoimmune encephalitis patients may concurrently present with other APS. Our unique case presented with multiple endocrine syndromes and represents the first reported occurrence in children. Early diagnosis and timely initiation of immunotherapy are crucial for improving clinical symptoms and reducing the likelihood of relapses or permanent disabilities. Therefore, emphasis should be placed on prompt diagnosis and appropriate treatment implementation to achieve better patient outcomes.


Oncogenic splicing abnormalities induced by DEAD-Box Helicase 56 amplification in colorectal cancer.
Alternative splicing, regulated by DEAD-Box Helicase (DDX) families, plays an important role in cancer. However, the relationship between the DDX family and cancer has not been fully elucidated. In the present study, we identified a candidate oncogene DDX56 on Ch.7p by a bioinformatics approach using The Cancer Genome Atlas (TCGA) dataset of colorectal cancer (CRC). DDX56 expression was measured by RT-qPCR and immunochemical staining in 108 CRC patients. Clinicopathological and survival analyses were carried out using three CRC datasets. Biological roles of DDX56 were explored by gene set enrichment analysis (GSEA), and cell proliferation in vitro and in vivo, cell cycle assays, and using DDX56-knockdown or overexpressed CRC cells. RNA sequencing was carried out to elucidate the effect of DDX56 on mRNA splicing. We found that DDX56 expression was positively correlated with the amplification of DDX56 and was upregulated in CRC cells. High DDX56 expression was associated with lymphatic invasion and distant metastasis and was an independent poor prognostic factor. In vitro analysis, in vivo analysis and GSEA showed that DDX56 promoted proliferation ability through regulating the cell cycle. DDX56 knockdown reduced intron retention and tumor suppressor WEE1 expression, which functions as a G2-M DNA damage checkpoint. We have identified DDX56 as a novel oncogene and prognostic biomarker of CRC that promotes alternative splicing of WEE1.


Intravenous Lipid Emulsion Therapy for Severe Diphenhydramine Toxicity: A Randomized, Controlled Pilot Study in a Swine Model.
Diphenhydramine is a moderately lipophilic antihistamine with sodium channel blockade properties. It is consumed recreationally for mild hallucinogenic and hypnotic effects and causes dysrhythmias, seizures, and death with overdose. Intravenous lipid emulsion is a novel agent used to treat lipophilic drug overdose. Two case reports describe clinical improvement with intravenous lipid emulsion after diphenhydramine toxicity, but no prospective studies have been reported. Our objective is to determine whether intravenous lipid emulsion improved hypotension compared with sodium bicarbonate for severe diphenhydramine toxicity in a model of critically ill swine. Twenty-four swine weighing 45 to 55 kg were infused with diphenhydramine at 1 mg/kg per minute until the mean arterial pressure reached 60% of baseline. Subjects were randomized to receive intravenous lipid emulsion (bolus of 7 mL/kg and then 0.25 mL/kg per minute) or sodium bicarbonate (2 mEq/kg plus an equal volume of normal saline solution). We measured pulse rate, systolic blood pressure, mean arterial pressure, cardiac output, QRS interval, and serum diphenhydramine level. Twelve animals per group provided a power of 0.8 and α of .05 to detect a 50% difference in mean arterial pressure. We assessed differences between groups with a repeated-measures linear model (MIXED) and Kaplan-Meier estimation methods. We compared systolic blood pressure, mean arterial pressure, and cardiac output with repeated measures ANOVA. Baseline weight, hemodynamic parameters, QRS interval, time to hypotension, and diphenhydramine dose required to achieve hypotension were similar between groups. After hypotension was reached, there was no overall difference between intravenous lipid emulsion and sodium bicarbonate groups for cardiac output or QRS intervals; however, there were transient differences in mean arterial pressure and systolic blood pressure, favoring intravenous lipid emulsion (difference: mean arterial pressure, sodium bicarbonate versus intravenous lipid emulsion -20.7 [95% confidence interval -31.6 to -9.8]; systolic blood pressure, sodium bicarbonate versus intravenous lipid emulsion -24.8 [95% confidence interval -37.6 to -12.1]). Time to death was similar. One intravenous lipid emulsion and 2 sodium bicarbonate pigs survived. End-of-study mean total serum diphenhydramine levels were similar. The mean lipid layer diphenhydramine level was 6.8 μg/mL (SD 3.1 μg/mL) and mean aqueous layer level 8.6 μg/mL (SD 5.5 μg/mL). In our study of diphenhydramine-induced hypotensive swine, we found no difference in hypotension, QRS widening, or diphenhydramine levels in aqueous layers between intravenous lipid emulsion and sodium bicarbonate.