Editing of the OsACS locus alters phosphate deficiency-induced adaptive responses in rice seedlings.
Phosphate (Pi) deficiency severely influences the growth and reproduction of plants. To cope with Pi deficiency, plants initiate morphological and biochemical adaptive responses upon sensing low Pi in the soil, and the plant hormone ethylene plays a crucial role during this process. However, how regulation of ethylene biosynthesis influences the Pi-induced adaptive responses remains unclear. Here, we determine the roles of rice 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS), the rate-limiting enzymes in ethylene biosynthesis, in response to Pi deficiency. Through analysis of tissue-specific expression of OsACS in response to Pi deficiency and OsACS mutants generated by CRISPR/Cas9 [clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9] genome editing, we found that two members of the OsACS family, i.e. OsACS1 and OsACS2, are involved but differed in their importance in controlling the remodeling of root system architecture, transcriptional regulation of Pi starvation-induced genes, and cellular phosphorus homeostasis. Interestingly, in contrast to the known inhibitory role of ethylene on root elongation, both OsACS mutants, especially OsACS1, almost fail to promote lateral root growth in response to Pi deficiency, demonstrating a stimulatory role for ethylene in lateral root development under Pi-deficient conditions. Together, this study provides new insights into the roles of ethylene in Pi deficiency response in rice seedlings and the isoform-specific function of OsACS genes in this process.


Human immunodeficiency virus-1 Nef protein interacts with Tat and enhances HIV-1 gene expression.
The human immunodeficiency virus (HIV-1) Nef protein is now regarded as a regulatory protein responsible not only for establishment of infection and increased pathogenesis but also for enhancement of viral replication. However, the mechanism of Nef-induced activation of viral replication remains to be clearly understood. Using transient transfection assay, co-immunoprecipitation and pull-down analysis, we demonstrate in this report that the HIV-1 Nef protein physically interacts with Tat, the principal transactivating protein of HIV-1. Our observations with single cycle replication experiments further indicate that this interaction results not only in enhancement of Tat-induced HIV-1 long terminal repeat-mediated gene expression but also in virus production.


Incremental layers bonding of silorane composite: the initial bonding properties.
Lack of oxygen inhibition layer of silorane composite with cationic polymerization raises the question of the bonding of incremental layers of the composite. This study aimed to evaluate the bond strength of the silorane composite layers. Fresh, 20 s, 5 min aged silorane composite (Silorane, 3M-ESPE) was used as substrate to adhere new silorane composite. For a comparison, dimethacrylate-based composite resin (Z250, 3M-ESPE) was adhered to the silorane composite with or without intermediate adhesive resin. As a control, dimethacrylate composite with oxygen inhibition layer was attached to fresh dimethacrylate composite. The bonded specimens (n=12/group) were water stored for 24 h. The shear bond strengths (SBS) were measured with a crosshead speed of 1.0 mm/min. Failure modes were assessed. Data were analysed by ANOVA, Tukey's post hoc tests and Chi-square tests (p=0.05). Dimethacrylate-dimethacrylate composite resin combination showed the highest mean SBS (33.0 MPa) values with no adhesive failures. Fresh silorane-silorane SBS was slightly lower (26.7 MPa) and was further decreased by aging the substrate for 20s (25.4 MPa) and 5 min (22.4 MPa). The percent of adhesive failures increased from 25% to 75%, respectively. The failure modes were significantly different (Chi-square, p<0.001). Silorane-dimethacrylate composite showed the lowest (4.0 MPa) SBS among the groups, which was increased significantly by use of phosphate-methacrylate-based intermediate resin (p<0.05). In order to bond dimethacrylate composite to silorane composite, a phosphate-methacrylate-based intermediate resin is required. The silorane composite showed slightly lower incremental bonding properties than conventional dimethacrylate composites.


[Neuromuscular relaxation and CCMDP. The Zilgrei and Feldenkrais methods 2].
The Authors show two neuromuscular release methods employed in the treatment of cranio-cervico-mandibular syndrome; these methods work at the place of origin of the pathology resolving the symptoms in different districts of the body.


Adult derived mononuclear bone marrow cells improve survival in a model of acetaminophen-induced acute liver failure in rats.
Acute liver failure (ALF) is characterized by a rapid loss of hepatic function, with high mortality. Acetaminophen (APAP) intoxication and viral hepatitis are common causes of ALF. Several studies have shown the capacity of adult bone marrow cells to differentiate in hepatocytes, suggesting their use for treating ALF. In the present study, we tested the use of adult derived mononuclear bone marrow fraction to improve the survival of Wistar rats with APAP-induced ALF. Forty-eight female Wistar rats pre-induced with phenobarbital were given APAP in a single dose of 1g/kg via intraperitoneal injection. Bone marrow mononuclear cells were purified from male rats using FICOLL gradient and injected through the portal vein in a volume of 0.2mL containing 1x10(6) cells stained with DAPI. Treatment was administered 24h after APAP injection. The sham group (n=24), received 0.2mL of saline through the portal vein 24h after APAP administration. Survival, liver histology and ALT levels were observed. Survival 72h post-APAP administration was 33% in the sham group and 70.8% in the group receiving bone marrow cells. Liver histology in treated animals showed less intense necrosis and the presence of DAPI-positive cells. We have shown that bone marrow derived cells are capable of significantly increasing the survival rate of APAP-induced ALF in 37.5% (95% CI, 27.8-40.3%).


Novel topical skin hydration agent containing Anadenanthera colubrina polysaccharide-standardized herbal preparation.
Hydration is an important factor to promote skin barrier function, metabolism, and appearance. In this process, the presence of aquaglyceroporins, envelope and lipid synthesis, and metabolism proteins are essential to provide greater corneocyte cohesion and to form a barrier avoiding transepidermal water loss. We evaluated the effects of a new topical pigment-free agent containing an Anadenanthera colubrina polysaccharide-rich dermocosmetic preparation (ACP) on the aquaporin-3 (AQP-3), filaggrin (FLG), involucrin (INV), glucocerebrosidase (GBA), and elongation of very-long-chain fatty acid (ELOVL) proteins production in skin human fragments, as well as on the transepidermal water loss in a double-blind placebo-controlled clinical trial. AQP3, FLG, INV, GBA, and ELOVL3 levels were measured by immunofluorescence analysis in human skin explants. Clinical trial was conducted to evaluate the effects of ACP 1% and ACP 3% on the transepidermal water loss (TEWL). Image and statistical analysis showed that ACP 3% significantly increased at 90% the expression of AQP3. Similarly, ACP 3% was able to promote a significant increase of 68% and 51% in FLG and INV, respectively. ACP 3% produced no effects on the GBA and ELOVL3 proteins. Transepidermal water loss was significantly reduced in human volunteers under treatment with ACP 1% and ACP 3%. ACP reduced transepidermal water loss in a clinical trial, promoting human skin hydration. These effects were related to modulation of the AQP3, FLG, and INV as evidenced by immunofluorescence assay. This way, A colubrina polysaccharide-rich phytopharmaceutical preparation is an effective additive product to skin hydration.


Obstetric float nurse role redesign in a small rural community hospital.
The purpose of this process improvement project in a rural community hospital with a small volume perinatal service was to foster a cooperative working environment between the obstetric and medical-surgical nursing teams and promote quality nursing care by evaluating, revising, and clearly defining the obstetric float nurse role. Members of the obstetric and medical-surgical nursing team were convened to evaluate the obstetric float nurse role and develop plans for improvement. Nurses from both units participated in a written survey before and after the project was completed. Based on issues identified from review of the surveys and group discussion, a guideline was developed that defined the expectations and responsibilities of the obstetric float nurse on the medical-surgical unit. The redesigned obstetric float nurse role helped nurses of both units understand the expectations and responsibilities of the obstetric float nurse and promoted better working relationships. An additional benefit was improved communication regarding care and needs of the medical-surgical patients. Clearly defined obstetric float nurse guidelines were successful in our small rural community hospital and may be of benefit to other similar volume perinatal units.


Methionine Supplementation Affects Fecal Bacterial Community and Production Performance in Sika Deer (Cervus nippon).
Amino acid balance is central to improving the efficiency of feed protein utilization and for reducing environmental pollution caused by intensive farming. In previous studies, supplementation with limiting amino acids has been shown to be an effective means of improving animal nutrient utilization and performance. In this experiment, the effects of methionine on the apparent digestibility of nutrients, antler nutrient composition, rumen fluid amino acid composition, fecal volatile fatty acids and intestinal bacteria in antler-growing sika deer were investigated by randomly adding different levels of methionine to the diets of three groups of four deer at 0 g/day (CON), 4 g/day (LMet) and 6 g/day (HMet). Methionine supplementation significantly increased the apparent digestibility of organic matter, neutral detergent fiber (NDF) and acid detergent fiber (ADF) in the LMet group (p < 0.05). The crude protein and collagen protein of antlers were significantly higher in the LMet and HMet groups compared to the CON group and also significantly higher in the HMet group compared to the LMet group, while the calcium content of antlers was significantly lower in the HMet group (p < 0.05). Ruminal fluid free amino acid composition was altered in the three groups of sika deer, with significant changes in aspartic acid, citrulline, valine, cysteine, methionine, histidine and proline. At the phylum level, Firmicutes and Bacteroidetes were highest in the rectal microflora. Unidentified bacterial abundance was significantly decreased in the HMet group compared to the CON group. Based on the results of principal coordinate analysis (PCoA) and Adonis analysis, there was a significant difference in the composition of the intestinal flora between the CON and HMet groups (p < 0.05). At the genus level, compared with the CON group, the abundance of Rikenellaceae_RC9_gut_group and Lachnospiraceae_UCG-010 in the LMet group increased significantly (p < 0.05), the abundance of dgA-11_gut_group in the HMet group decreased significantly (p < 0.05) and the abundance of Lachnospiraceae_UCG-010, Saccharofermentans and Lachnospiraceae_NK3A20_group increased significantly. Taken together, the results showed that methionine supplementation was beneficial in increasing the feed utilization efficiency and improving antler quality in sika deer, while affecting the composition of fecal bacteria.


Automated rare sperm identification from low-magnification microscopy images of dissociated microsurgical testicular sperm extraction samples using deep learning.
To develop a machine learning algorithm to detect rare human sperm in semen and microsurgical testicular sperm extraction (microTESE) samples using bright-field (BF) microscopy for nonobstructive azoospermia patients. Spermatozoa were collected from fertile men. Testis biopsies were collected from microTESE samples determined to be clinically negative for sperm. A convolutional neural network based on the U-Net architecture was trained using 35,761 BF image patches with fluorescent ground truth image pairs to segment sperm. The algorithm was validated using 7,663 image patches. The algorithm was tested using 7,663 image patches containing abundant sperm, as well as 7,985 image patches containing rare sperm. In vitro fertilization center and university laboratories. Normospermic and nonobstructive azoospermia patients. None. Precision (positive predictive value [PPV]), recall (sensitivity), and F1-score of detected sperm locations. For sperm-only samples, our algorithm achieved 91% PPV, 95.8% sensitivity, and 93.3% F1-score at ×10 magnification. For dissociated microTESE samples doped with an abundant quantity of sperm, our algorithm achieved 84.0% PPV, 72.7% sensitivity, and 77.9% F1-score. For dissociated microTESE samples doped with rare sperm, our algorithm achieved 84.4% PPV, 86.1% sensitivity, and 85.2% F1-score. Rare sperm can be detected in patients' testis biopsy samples for potential subsequent use in in vitro fertilization-intracytoplasmic sperm injection. A machine learning algorithm can use BF images at ×10 magnification to accurately detect sperm locations using automated imaging.


Role of stromal and Thy 1,2+ cells in the mechanisms of action of immobilized granulocyte colony-stimulating factor during cytostatic-induced myelosuppression.
The effects of immobilized granulocyte colony-stimulating factor (mediated by cells of the hemopoiesis-inducing microenvironment) on hemopoietic precursors of various classes were studied on the model of cytostatic-induced myelosuppression (administration of cyclophosphamide). The action of this preparation was compared with that of the standard preparation of granulocyte colony-stimulating factor. Thy 1,2(+)cells potentiated the effects of immobilized and standard granulocyte colony-stimulating factors on granulocyte-erythroid-macrophage-megakaryocyte precursors. Stromal cells were shown to potentiate the influence of these agents on granulocyte precursors. Induction of proliferation of precursor cells by the immobilized factor mediated by cells of the hemopoiesis-inducing microenvironment persisted for a longer period compared to that induced by the standard product.


What is the role of the supplementary motor area in movement initiation?
The hierarchical position of the supplementary motor area (SMA) relative to the primary motor cortex is discussed on the basis of neurological observations and of animal experiments. In the last 10 years evidence has accumulated, especially from studies on the human brain, that the supplementary motor area is a hierarchically superior structure involved in the processes of movement initiation. Single unit studies in subhuman primates also revealed neuronal populations related to aspects of movement preparation rather than to the movement per se. However, we report that a surprisingly large subpopulation of SMA neurones has features classically found in the primary motor cortex (MI). These MI-like neurones precede movement onset by a relatively short interval. The occurrence of such "short-lead neurones" was somewhat higher in MI, but the histograms of lead-times were completely overlapping in the two areas. Taken together with the fact that the SMA is microexcitable and is part of the origin of the pyramidal tract, these findings suggest that the SMA functions also in parallel with MI as concluded by Woolsey and coworkers (1952). Finally, the SMA and MI are reciprocally interconnected, a situation which is not unlike that of the cortical visual areas.


Zinc finger protein 852 is essential for the proliferation, drug sensitivity, and self-renewal of gastric cancer cells.
Exploring cellular and molecular mechanisms responsible for gastric cancer growth, survival, self-renewal, and metastasis helps develop efficacious therapeutic strategies. In this study, the expression and function of zinc finger protein 852 (ZNF852) in human gastric cancer cell lines were characterized. ZNF852 was upregulated in gastric cancer cell lines relative to normal gastric epithelial cell line GES-1. When the ZNF852 gene was ablated in gastric cancer cell line MGC-803 using the CRISPR/Cas9-encoding lentivirus, the proliferation of MGC-803 was suppressed. ZNF852 deficiency also resulted in the inhibition of MGC-803 sphere formation, along with decreases in SRY-box 2 (SOX2), octamer-binding transcription factor 4 (OCT4), and Nanog homeobox (NANOG), suggesting that ZNF852 sustains self-renewal of MGC-803 cells. Furthermore, ZNF852 deficiency increased oxaliplatin-induced MGC-803 cell death, implying the role of ZNF852 in drug sensitivity. Subcutaneous infusion of MGC-803 cells into nude mice illustrated the same effects of ZNF852 on the proliferation and self-renewal of gastric cancer cells. Similar effects of ANF852 were also seen in gastric cancer cell line SNU-1. Interestingly, ZNF852 deficiency caused downregulation of epidermal growth factor receptor (EGFR) on gastric cancer cells. In summary, this study uncovers the positive regulatory role of ZNF852 in gastric cancer growth and maintenance. ZNF852 could be a potential therapeutic target for inhibiting gastric cancer initiation or progression.


The development of monoclonal antibodies to respiratory syncytial virus and their use in diagnosis by indirect immunofluorescence.
Twelve clones of murine hybridoma cells secreting antibody specific for respiratory syncytial (RS) virus were classified into four groups on the basis of their pattern of staining of unfixed RS virus-infected HEp-2 cells in an indirect immunofluorescence test. Three of the groups reacted with virus antigens present on the membrane of the cells, whilst the fourth group failed to stain most live cells, suggesting specificity for an antigen expressed internally. Representative monoclonals from the membrane antigen staining groups immunoprecipitated the 86K glycoprotein (G), 50K plus 19K glycoprotein (F1,2) and a 23K non-glycosylated protein (VP23). A representative monoclonal from the fourth group that appeared to stain an internally expressed protein immunoprecipitated the virion 34K phospho-protein (P). All four monoclonals stained acetone-fixed tissue culture cells infected with either the Long strain of RS virus or with strains isolated in Newcastle during the 1965, 1972, and 1983 winter epidemics. The anti-fusion protein antibody stained acetone-fixed cells from all of 26 nasopharyngeal secretions from infants with RS virus infection. The anti-G glycoprotein antibody and the anti-VP23 antibody stained cells from secretions poorly or not at all, whilst the anti-P protein antibody stained cells in half the secretions tested but reacted with only a small proportion of cells in comparison with the anti-F or polyclonal antibodies. A pool of all four monoclonals produced more intense staining than the anti-F monoclonal alone and gave a more clearly defined staining reaction than the polyclonal antiserum used for routine diagnosis in over half the secretions. These results indicate that monoclonal antibodies will be of value in the diagnosis of RS virus by indirect immunofluorescence if care is taken in the selection of a suitable pool.


Growth and nutrition after discharge.
Preterm infants are frequently discharged from hospital growth retarded and show reduced growth during childhood. However, nutrition during the post-discharge period has been neglected. Randomized trials show growth benefits, particularly in boys, when nutrient-enriched diets (preterm formula or post-discharge formula) rather than term formula are fed after discharge, with effects persisting beyond the intervention period. These findings suggest that post-discharge period may be a critical time for influencing the growth trajectory, but further research is required to investigate longer-term effects on growth, health and development. Preterm infants who are breast-fed after discharge show slower weight and length gain and lower bone mass than formula-fed infants. Further studies should establish whether differences persist or have long-term consequences, and clarify whether such infants would benefit from post-discharge nutritional supplementation. The introduction of solids may have a significant effect on growth and nutritional status, but insufficient data are currently available for evidence-based guidelines.


Improving surgical ward care: development and psychometric properties of a global assessment toolkit.
To develop a toolkit that covers the clinical, nontechnical, and empathic skills required for effective, safe surgical ward care. Despite the explosion of interest in patient safety, little attention has been placed on the skill set required for safe and effective surgical ward care. Currently, there is a lack of a systematic approach to improving ward care via assessing and improving residents' ward care skills. A comprehensive evidence-based and expert-derived toolkit was developed, including a novel clinical checklist for ward care (Clinical Skills Assessment for Ward Care: C-SAW-C); a novel team assessment scale for wards rounds (Teamwork Skills Assessment for Ward Care: T-SAW-C); and a revised version of a physician-patient interaction scale (Physician-Patient Interaction Global Rating Scale: PP-GIS). Interassessor reliability (κ, intraclass correlation), internal consistency (Cronbach α), and convergent validity (Pearson r correlations) were evaluated statistically in 38 simulated scenarios (during which a patient rapidly deteriorated) involving 185 residents. Excellent interassessor reliability was obtained for C-SAW-C [median κ = 0.89; median intraclass correlation coefficient (ICC) = 0.94], T-SAW-C (median ICC = 0.99), and the revised PP-GIs (κ = 1.00; ICC = 0.98 or higher). Internal consistency was also very high for all team skills assessed by T-SAW-C (Cronbach α range 0.87-0.94 across 6 skills) and the revised PP-GIS (Cronbach α = 0.96)-all P's < 0.001. Significant positive correlations were obtained between the 3 assessments (r = 0.73-0.92, P < 0.001) thus showing evidence for convergent validity. We developed a toolkit that captures comprehensively the skills that are required for safe and effective ward care, including the high-risk situation where a patient decompensates. The toolkit offers a systematic evaluation of the quality and safety of surgical ward care and can be used to train and debrief residents' skills and performance.


Indexation criteria of ventricular mass and predictive role of blood pressure and body composition.
There is no definite consensus on which indexation of left ventricular mass (LVM) should be used to better identify left ventricular hypertrophy (LVH). Left ventricular mass has been adjusted to height, to height2.7(h2.7) and to body surface area (BSA). The aims of the present study were to evaluate the prevalence of LVH according to different indexations and different cut-offs and to identify the most useful indexation of LVM to detect hypertension-related LVH. Echocardiographic LVH was defined as LVM to h2.7, LVM to BSA, LVM to height, LVM values in the upper 5th percentile of our gender-related LVM distribution, using different partition values suggested in previous population-based studies. Prevalence of LVH in the general population was 32% using the less restrictive criterion (LVH 49.2/46.7 g/m2.7), 15% with the criterion of LVH 116/104 g/m2, and 3.8% with the most restrictive one (LVH 134/110 g/m2). Prevalence of LVH in hypertensive subjects was almost twice than in normotensive subjects with all criteria. Only 20 subjects out of the 707 evaluated were found to have LVH with all six criteria. In multiple regression analysis SBP was independently associated with nonindexed LVM and was indexed to both BSA and h2.7. On the other hand, fat-free mass was a powerful predictor of nonindexed LVM or of LVM to BSA, whereas body mass index was the strongest predictor of LVM to h2.7. The indexation of LVM to BSA, possibly with the cut-off of LVH 116/104, is probably the best criterion for identifying blood pressure-related LVH.


Automated Raman feed-back control of multiple supplemental feeds to enable an intensified high inoculation density fed-batch platform process.
Biologics manufacturing is increasingly moving toward intensified processes that require novel control strategies in order to achieve higher titers in shorter periods of time compared to traditional fed-batch cultures. In order to implement these strategies for intensified processes, continuous process monitoring is often required. To this end, inline Raman spectroscopy was used to develop partial least squares models to monitor changes in residual concentrations of glucose, phenylalanine and methionine during the culture of five different glutamine synthetase piggyBac® Chinese hamster ovary clones cultured using an intensified high inoculation density fed-batch platform process. Continuous monitoring of residual metabolite concentrations facilitated automated feed-rate adjustment of three supplemental feeds to maintain glucose, phenylalanine, and methionine at desired setpoints, while maintaining other nutrient concentrations at acceptable levels across all clones cultured on the high inoculation density platform process. Furthermore, all clones cultured on this process achieved high viable cell concentrations over the course of culture, indicating no detrimental impacts from the proposed feeding strategy. Finally, the automated control strategy sustained cultures inoculated at high cell densities to achieve product concentrations between 5 and 8.3 g/L over the course of 12 days of culture.


The active synthesis of phosphatidylcholine is required for very low density lipoprotein secretion from rat hepatocytes.
Hepatocytes obtained from rats fed a choline-deficient diet for 3 days were cultured in a medium +/- choline (100 microM) or methionine (200 microM). We investigated how choline deficiency affected hepatic lipogenesis, apolipoprotein synthesis, and lipoprotein secretion. The mass of triacylglycerol and phosphatidylcholine secreted was increased about 3-fold and 2-fold, respectively, by the addition of either choline or methionine to the cultured cells. Similarly, a 3-fold stimulation in the secretion of [3H]triacylglycerol and [3H]phosphatidylcholine derived from [3H]oleate was observed after the addition of choline or methionine. Fractionation of secreted lipoproteins by ultracentrifugation revealed that the reduced secretion of triacylglycerol and phosphatidylcholine from choline-deficient cells was mainly due to impaired secretion of very low density lipoproteins (VLDL) (but not high density lipoproteins (HDL)). Fluorography of L-[4,5-3H]leucine-labeled lipoproteins showed a remarkable inhibition of VLDL secretion by choline deficiency. The addition of choline or methionine stimulated the synthesis of phosphatidylcholine and increased the cellular phosphatidylcholine levels to that in normal cells. While there was little effect of choline on the synthesis and amount of cellular phosphatidylethanolamine, the addition of methionine diminished cellular phosphatidylethanolamine levels. Choline deficiency did not change the rate of incorporation of L-[4,5-3H]leucine into cellular VLDL apolipoproteins, nor the rate of disappearance of radioactivity from L-[4,5-3H]leucine-labeled cellular apoB, apoE, and apoC. These results suggest that hepatic secretion of VLDL, but not HDL, requires active phosphatidylcholine biosynthesis. Secondly, the inhibitory effect of choline deficiency on VLDL secretion can be compensated by the methylation of phosphatidylethanolamine.


Phase 1B ventricular arrhythmia in the dog: localized reentry within the mid-myocardium.
Intramural and epicardial composite electrograms, signal-averaged orthogonal bipolar electrograms across the ischemic zone, and closely-spaced bipolar electrograms from subendocardium, mid-myocardium, and subepicardium were utilized to determine if phase 1B reentry resulted from localized reentry within ischemic mid-myocardium. During the first 10 minutes following coronary ligation, activation delays were largest in ischemic subepicardium, with continuous electrical activity in ischemic epicardium linking a ventricular extrasystole to the preceding beat. During the 15-30 minute period, activation delay observed in ischemic mid-myocardium exceeded activation delay on the epicardial surface. Ventricular extrasystoles were associated with mid-myocardial delays > 130 msec. With short-coupled extrasystoles (< 300 msec), electrical activity in ischemic mid-myocardium linked an extrasystole with the preceding beat. Although single extrasystoles with coupling intervals > 300 msec were also associated with mid-myocardial delays > 130 msec, most extrasystoles (68%) demonstrated an isoelectric gap > 20 msec. The data demonstrate an association between delayed activation within ischemic mid-myocardium and phase 1B arrhythmia resulting from (1) localized reentry in ischemic mid-myocardium and (2) a delay-dependent "non-reentrant" mechanism.


Evolving biosynthetic tangos negotiate mechanistic landscapes.
The dependence of polyketide synthase and terpene cyclase mechanistic adaptation on the chemistry of their oligomeric substrates illuminates a convergent evolutionary strategy for shaping cyclization in these otherwise disparate reactions. Evolution of these enzyme families relies on rhythmic tangos, in which the enzymes and substrates together determine product outcome by negotiating decision networks governing intrinsic and induced chemical reactivities.


[Basic principles in rheumatologic patient education. Theoretical principles and didactic aspects].
Nowadays, the handling of a chronic disease is seen as a process, influenced by a variety of factors. Case studies, taken from health behavior, help to illustrate the conduct of patients suffering from chronic diseases and develop strategies of how to increase a patient's skills in coping with his illness. The concept of self-efficacy beliefs is of great importance and will be discussed in detail. In addition, possible effects of patient education in the comprehensive treatment of rheumatic diseases will be presented. In teaching methods, equal emphasis is placed on the transmission of information as on the development of practical know-how. They are on a par with emotional support, motivation, assistance in accepting the disease and coping with fear. The special methods and media required will be discussed, along with the framework for training methods and qualification required of the staff in charge of training.


Production of recombinant butyrylcholinesterase from transgenic rice cell suspension cultures in a pilot-scale bioreactor.
Producing recombinant proteins in transgenic plant cell suspension cultures in bioreactors provides controllability, reproducibility, scalability, and low-cost production, although low yields remain the major challenge. The studies on scaling-up to pilot-scale bioreactors, especially in conventional stainless-steel stirred tank bioreactors (STB), to produce recombinant proteins in plant cell suspension cultures are very limited. In this study, we scaled-up the production of rice recombinant butyrylcholinesterase (rrBChE), a complex hydrolase enzyme that can be used to prophylactically and therapeutically treat against organophosphorus nerve agents and pesticide exposure, from metabolically regulated transgenic rice cell suspension cultures in a 40-L pilot-scale STB. Employing cyclical operation together with a simplified-process operation (controlling gas sparging rate rather than dissolved oxygen and allowing natural sugar depletion) identified in lab-scale (5 L) bioreactor studies, we found a consistent maximum total active rrBChE production level of 46-58 µg/g fresh weight in four cycles over 82 days of semicontinuous operation. Additionally, maintaining the overall volumetric oxygen mass transfer coefficient (kL a) in the pilot-scale STB to be equivalent to the lab-scale STB improves the maximum total active rrBChE production level and the maximum volumetric productivity to 85 µg/g fresh weight and 387 µg L-1 day-1 , respectively, which are comparable to the lab-scale culture. Here, we demonstrate pilot-scale bioreactor performance using a metabolically regulated transgenic rice cell culture for long-term, reproducible, and sustained production of rrBChE.


Claudin-2 promotes colorectal cancer growth and metastasis by suppressing NDRG1 transcription.
Colorectal cancer (CRC) is one of the most common malignant tumours, with multiple driving factors and biological transitions involved in its development. Claudin-2 (CLDN2), a well-defined component of cellular tight junction, has been indicated to associate with CRC progression. However, the function of CLDN2 and the underlying mechanism whereby the downstream signalling transduction is regulated in CRC remains largely unclear. In this study, we demonstrated that CLDN2 is upregulated in CRC samples and associated with poor survival. And CLDN2 depletion significantly promotes N-myc downstream-regulated gene 1 (NDRG1) transcription, leading to termination of the CRC growth and metastasis in vitro and in vivo. Mechanistically, this process promotes CLDN2/ZO1/ZONAB complex dissociation and ZONAB shuttle into nucleus to enrich in the promoter of NDRG1. Thus, this study reveals a novel CLDN2/ZO1/ZONAB-NDRG1 axis in CRC by regulating the expression of EMT-related genes and CDKIs, suggesting CLDN2 may serve as a promising target for CRC treatment.


Approach to anaerobic bio-degradation of natural and synthetic fabrics: Physico-chemical study of the alteration processes.
In this paper, the mesophilic Biochemical Methane Potential of several fabrics was assessed at different Total Solid concentrations (1-4%TS). Physico-chemical techniques were applied to explore the arising structural changes on fibers during the anaerobic digestion process. Additionally, the modified Gompertz model was used to assess and compare the AD performance of the fabrics. In cellulose-based fibers the production of biogas was enhanced thanks to the easy solubilization of acetate, which is generated upon partial breakage of cellulose bonds. The crystallinity of vegetal fibers decreased significantly from day 19. The highest methane yields were attained for silk and wool fabrics at the lowest TS concentrations. Conformational changes in fibroin and keratin were detected. The highest degrees of degradation were observed in solid samples with lower solid concentrations. Accordingly, the maximum methane yields were reported in the reactors operating with lower TS.


Particulate matter and polycyclic aromatic hydrocarbon concentrations in indoor and outdoor microenvironments in Boston, Massachusetts.
Estimating personal exposures to air pollution is a crucial component in identifying high-risk populations and determining efficient control strategies. Because of the difficulty of comprehensively measuring personal exposure, data on air pollution patterns in homogenous microenvironments linked with activity data are often used as surrogates. In this study, we focus on strengthening the available information about nonresidential microenvironmental exposures to particulate matter and other combustion pollutants. During the summer of 2000, we measured ultrafine particles, fine particulate matter (PM2.5), and particle-bound polycyclic aromatic hydrocarbons (PAHs) outdoors and in indoor microenvironments in Boston, Massachusetts. In indoor microenvironments averaged across sample days, mean ultrafine particle concentrations ranged from 3800 to 140,000 particles/cm(3), with 7-200 microg/m(3) of PM2.5 and 5-12 ng/m(3) of particle-bound PAH. PM2.5 indoor-outdoor ratios generally exceeded 1 in settings with high levels of human activity, with lower ratios for ultrafine particles. Cooking activities contributed significantly to elevated levels of all three pollutants. Using Linear Mixed Effects models with AR-1 autoregressive correlation structures, 10-min average outdoor concentrations were generally weak predictors of indoor levels, with stronger relationships in an apartment without mechanical ventilation than in air-conditioned nonresidential settings. Although further study would be needed to determine whether these patterns could be generalized beyond the monitored sites, these data support previous findings and enhance our knowledge about nonresidential exposure patterns.


Analysis of porphyrins and enzymes in porphyrin synthesis in Taenia solium cysticercus from man and pig.
Porphyrins and activities of heme biosynthetic enzymes in Taenia solium cysticerci from porcine and human hosts, were examined in order to clarify the possible step where heme synthesis is interrupted. Porphyrins in the vesicular fluid of the parasite were predominantly coproporphyrin, followed by penta-carboxylated porphyrin, which together accounted for 90% of the accumulated porphyrins. Coproporphyrin and penta-carboxylated porphyrin were both type I and III isomers. Small amounts of protoporphyrin and uroporphyrin, and trace amounts of tri-, hexa- and hepta-carboxylated porphyrins were also detected. Fluorescence and phosphorescence spectra and lifetime studies revealed that at least 75% of the porphyrins were bound to metal, probably Zn, while the rest was free. Reverse phase high performance liquid chromatography monitored at an excitation wavelength of 417 nm and at an emission wavelength of 585 nm demonstrated that approximately 90% of these porphyrins were Zn-coproporphyrin. A fluorescence excitation peak at 283 nm with an emission peak at 585 nm and 625 nm indicated that some of the porphyrins were associated with proteins in the vesicular fluid of the parasite. Low levels of delta-aminolevulinic acid dehydratase, porphobilinogen deaminase and uroporphyrinogen decarboxylase activities, and heme concentrations were found in the extract of the parasite walls and scolex, but not in the vesicular fluid. The porphyrin accumulation pattern in this parasite can best be explained by postulating a deficiency of coproporphyrinogen oxidase activity, similar to that in human patients with hereditary coproporphyria. A parasite dissected from a human host was considerably less porphyric than those from pigs, but the pattern of accumulated porphyrins was quite similar in both. In view of their porphyrin contents, T. solium cysticerci could be light sensitive.


Suicidal ideation, mental health problems, and social impairment are increased in adolescents with acne: a population-based study.
We performed a cross-sectional, questionnaire-based study to explore the relationship of suicidal ideation, mental health problems, and social functioning to acne severity among adolescents aged 18-19 years. A total of 4,744 youth were invited and 3,775 (80%) participated. In all, 14% reported having substantial acne (a lot and very much). Among those with very much acne, as compared those with no/little acne, suicidal ideation was twice as frequently reported among girls (25.5 vs. 11.9%) and three times more frequently reported among boys (22.6 vs. 6.3%). Suicidal ideation remained significantly associated with substantial acne (odds ratio 1.80, 95% confidence interval 1.30-2.50) in a multivariate model including adjustments of symptoms of depression, ethnicity, and family income. Mental health problems, as assessed by the Strengths and Difficulties Questionnaire (2.25, 1.69-3.00), low attachment to friends (1.52, 1.21-1.91), not thriving at school (1.41, 1.12-1.78), never having had a romantic relationship (1.35, 1.05-1.70), and never having had sexual intercourse (1.51, 1.21-1.89) were all associated with substantial acne in a multivariate model. Acne is frequently found in late adolescence and is associated with social and psychological problems. Adverse events including suicidal ideation and depression that have been associated with therapies for acne may reflect the burden of substantial acne rather than the effects of medication.


Fatty Acid Composition of Proximal Femur Bone Marrow Adipose Tissue in Subjects With Systemic Lupus Erythematous Using 3 T Magnetic Resonance Spectroscopy.
Systemic lupus erythematosus (SLE) is a chronic, inflammatory disease with common musculoskeletal manifestations, notably reductions in bone quality. Bone marrow adipose tissue composition and quantity has been previously linked to bone quality and may play a role in SLE pathophysiology but has not been thoroughly studied. To use magnetic resonance spectroscopy (MRS) to investigate bone marrow adipose tissue quantity and composition in proximal femur subregions of untreated SLE patients compared to controls and treated patients. Prospective. A total of 64 female subjects: 28 SLE, 15 glucocorticoid (GC)-treated SLE and 21 matched controls. Stimulated echo acquisition mode (STEAM) sequence at 3 T. MRS was performed at multiple echo times in the femoral neck and trochanter regions and fatty acids (FA) composition was computed. Intergroup comparisons were carried out using ANOVA. A P value < 0.05 was considered statistically significant. SLE patients had significantly higher saturated FA compared to controls in both the femoral neck (+0.12) and trochanter (+0.11), significantly lower monounsaturated FA in the trochanter compared to controls (-0.05), and significantly lower polyunsaturated FA in the femoral neck compared to both controls (-0.07) and SLE patients on GC therapy (-0.05). SLE patients have altered proximal femur marrow fat metabolism, which may reflect a manifestation of, or play a role in, the altered inflammatory response of these patients. 2 TECHNICAL EFFICACY: Stage 2.


Types and arrangements of neurons in the crayfish optic lamina.
The neural arrangements in the optic lamina of the crayfish Pacifastacus leniusculus Dana have been studied by light microscopy by means of silver impregnation techniques. The lamina is composed of columnar synaptic compartments (cartridges). Each cartridge is composed of seven receptor terminals distributed in two layers and second-order monopolar neurons connecting the lamina with the second synaptic region, the medulla externa. The neurons found in the lamina consist of five classes: monopolar neurons, centrifugal small-field neurons, tangential neurons, multipolar cells (possibly of a glial nature) and photoreceptor axons (fig. 13). Among the monopolar cells, five types are classified (M1-M5) according to their lamina arborizations. Two types are stratified (M3 and M5) corresponding to the photoreceptor terminal strata. On this basis, the lamina plexiform layer is subdivided into two layers (epl1 and epl2). The remaining monopolar neurons have lateral processes in both layers, two of them within one cartridge (M1 and M2) and one over several cartridges (M5). There is one type of small-field centrifugal neuron (C1) and two types of tangential medulla to lamina neurons (Tan1 and Tan2), both having processes covering a large number of cartridges. Multipolar cells with cell bodies distal (MP1) or proximal (MP2) to the plexiform layer send processes to several cartridges. The receptor axons consit of three types. One has terminals in epl1 or epl2, the second has its terminal in epl1 and a thin process to epl2, and the third (corresponding to the 8th retinular cell) bypasses the lamina and has a terminal in the medulla externa. A brief comparison is made with the neural arrangements in the lamina of the Norway lobster Nephrops norvegicus L.


Regulatory functions of T- and accessory-cells for hepatitis B surface antigen induced specific antibody production and proliferation of human peripheral blood lymphocytes, in vitro.
In vitro assays for lymphocyte proliferation and production of specific antibodies to hepatitis B surface antigen (HBsAg) are described. Peripheral B-cells from healthy donors with acquired immunity after infection with hepatitis B virus (HBV) were activated by purified HBsAg at low concentrations (approximately 2.5 ng/ml), into plasma cell differentiation and maturation to specific immunoglobulin (anti-HBs) secretion. This process was dependent upon and strictly regulated by T-cells and anti-HBs secretion of IgG class was registered at T/B ratios less than or equal to 4.0. Higher T/B ratios provoked secretion of large amounts of total IgM but no specific antibodies. Maximal DNA-synthesis was induced in purified T-cells or the total peripheral blood lymphocyte population from HBV-immune donors at higher concentrations of HBsAg (7-15 ng/ml and 60-125 ng/ml respectively). Proliferation was dependent upon and regulated by monocytic cells. Ten percent of autologous monocytic cells within the cell cultures was optimal, while excessive amounts effectively suppressed the proliferative response. DNA-synthesis was low and extremely shortlived and the induction phase to maximal proliferation for different HBV immune donors varied between 5-8 days in culture.


Amputation as a First Treatment is Highly Associated With Late Presentation: An Underestimated Modifiable Major Risk Factor for Diabetic Foot Ulcer.
Several risk factors for lower extremity amputation in diabetic patients have been identified; however, late presentation has been poorly investigated. Very few studies looked at the impact of such risk factor. As none has investigated the link between admission to amputation and late presentation in this population, we investigated such association. The study is a retrospective comparative cohort of continuous series of patients admitted for amputation related to diabetic foot ulcer (DFU) as a first treatment. A late presentation was defined as a period of 3 weeks or more from the onset of the ulcer. Forty-six patients admitted for amputation and all wounds were infected and located on the plantar forefoot in 32 cases (69.5%), on the toes in 10 cases (21.7%), and in the heel area in 4 cases (8.8%). The mean duration to admission was 5.2 ±1.8 weeks (range 4-10 weeks). Late presentation was recorded in 42 (91.3%) patients. This study demonstrates that amputation due to DFU is highly associated with late presentation. Educational campaigns targeting patient and first-line health care providers and highlighting the urgency of diabetic ulcers are needed to impact this major modifiable risk factor.Levels of Evidence: Therapeutic, Level III: Retrospective Comparative.


Infection cases in infants and small children with atopic dermatitis--own observations.
The purpose of the work is to estimate the frequency of occurrence of various infections in infants and small children, aged between 0 and 36 months with atopic dermatitis in the practice of a family doctor. The study has been conducted on the basis of the retrospective analysis of medical documentation of disease among children born in 2005-2008 treated in Outpatient Clinic of the Polish Mother's Memorial Hospital in Lodz. Children suffering from atopic dermatitis are more prone to develop infectious diseases. Infectious diseases in children with atopic dermatitis are of more chronic nature and more often require antibiotic therapy. Such children often manifest chronic diarrhea and their stool contains Staphylococcus aureus. They are more often hospitalized because of difficulty in setting a diagnosis and applying treatment in outpatient clinics.


Analysis of Mutations in Epidermal Growth Factor Receptor Gene in Korean Patients with Non-small Cell Lung Cancer: Summary of a Nationwide Survey.
Analysis of mutations in the epidermal growth factor receptor gene (EGFR) is important for predicting response to EGFR tyrosine kinase inhibitors. The overall rate of EGFR mutations in Korean patients is variable. To obtain comprehensive data on the status of EGFR mutations in Korean patients with lung cancer, the Cardiopulmonary Pathology Study Group of the Korean Society of Pathologists initiated a nationwide survey. We obtained 1,753 reports on EGFR mutations in patients with lung cancer from 15 hospitals between January and December 2009. We compared EGFR mutations with patient age, sex, history of smoking, histologic diagnosis, specimen type, procurement site, tumor cell dissection, and laboratory status. The overall EGFR mutation rate was 34.3% in patients with non-small cell lung cancer (NSCLC) and 43.3% in patients with adenocarcinoma. EGFR mutation rate was significantly higher in women, never smokers, patients with adenocarcinoma, and patients who had undergone excisional biopsy. EGFR mutation rates did not differ with respect to patient age or procurement site among patients with NSCLC. EGFR mutation rates and statuses were similar to those in published data from other East Asian countries.


Poisoning in ostriches following ingestion of toxic plants--field observations.
Data from post-mortem and field studies were obtained that discussed poisoning in ostriches following ingestion of toxic plants. From the notes, all plants studied caused death in ostriches and there was systemic organ damage. Poisoning from Sarcostemma viminale (Melktou) resulted in beak patting, muscular tremors and head flopping, followed by collapse and violent kicking before death. Ingestion of Combretum oatesii (Red wings) seeds from plants in free grazing pastures resulted in vomiting, restlessness, eyelid flicking, collapse and kicking movements. Dichapetalum cymosum (Gifblaar) killed an ostrich after episodes of shaking legs, rapid respiration and bradycardia and hyperaemia of the lungs, liver and kidneys. Poisoning from Senecio sceleratus (Ragwort) caused skin haemorrhages and bleeding in tracheal mucous membranes, the pericardium, diaphragm and interperitoneal membrane. Consumption of drupes from Melia azedarach (Syringa berry) caused muscle tremors, kicking movements and respiratory distress. Lantana camara (Cherry pie) poisoning resulted in extremely inflamed eyes with copious yellow exudates extending down their beaks and onto their necks. Bentonite was administered by gavage at a dose of 5 g/kg. Poisoning in these cases is usually associated with the farmer allowing his/her birds to roam free-range in paddocks in which toxic plants are growing. Toxic plants should be removed from grass cut for hay.


Effects of sex and sexual orientation on self-reported attraction and viewing times to images of men and women: testing for category specificity.
In a paradigm that asked participants to rate the sexual attractiveness of male and female swimsuit models, Lippa, Patterson, and Marelich (2010) showed that heterosexual men's category specificity exceeded heterosexual women's in two ways: (1) Heterosexual men showed much larger differences in their attraction and viewing times to male versus female photo models than heterosexual women, and (2) heterosexual men's attractions to female but not male models increased with model attractiveness whereas heterosexual women's attractions to both sexes increased with model attractiveness. The current study used the same paradigm to study category specificity in homosexual and heterosexual participants. In addition to replicating previous findings for heterosexual men and women, the results showed that homosexual men were high on category specificity, like heterosexual men, whereas lesbians showed lower levels of category specificity than men, but sometimes higher levels than heterosexual women.


Genetic variation, gene-expression and circulating levels of matrix metalloproteinase-9 in patients with stable coronary artery disease.
Mediators involved in atherosclerosis and plaque rupture may have importance as risk markers for coronary artery disease (CAD). We have investigated the influence of matrix metalloproteinase (MMP)-9 genetic variations on gene- and protein expression in stable CAD patients. The promoter -1562C/T and exon 6 R279Q A/G polymorphisms were determined in 1001 patients with angiographically verified stable CAD and in 204 healthy controls. Genotype and gene-expression were determined by real-time PCR. Serum levels of MMP-9 and its inhibitor TIMP-1were measured immunologically and by zymography (MMP-9 activity). None of the polymorphisms associated with the presence of CAD, myocardial infarction or type 2 diabetes, whereas the variant allele of the R279Q polymorphism associated with hypertension (adjusted p=0.015). The T- and G alleles associated with lower and higher mRNA levels, respectively (p<0.005 both), also shown in an experimental ex-vivo LPS stimulated model. T-allele carriers had higher concentrations of MMP-9 (adjusted p=0.032) and the GG genotype induced lower MMP-9 gelatinolytic activity (p=0.01). Higher MMP-9 gene-expression and TIMP-1 levels were observed in patients with previous myocardial infarction, the latter also was elevated in diabetics (<0.05, all). The investigated MMP-9 polymorphisms influenced gene- and protein expression differently and the R279Q polymorphism associated significantly with hypertension.


Regulation of AmtR-controlled gene expression in Corynebacterium glutamicum: mechanism and characterization of the AmtR regulon.
AmtR, the master regulator of nitrogen control in Corynebacterium glutamicum, represses transcription of a number of genes during nitrogen surplus. Repression is released by an interaction of AmtR with signal transduction protein GlnK. As shown by pull-down assays and gel retardation experiments, only adenylylated GlnK, which is present in the cells during nitrogen limitation, is able to bind to AmtR. The AmtR regulon was characterized in this study by a combination of bioinformatics, transcriptome and proteome analyses. At least 33 genes are directly controlled by the repressor protein including those encoding transporters and enzymes for ammonium assimilation (amtA, amtB, glnA, gltBD), urea and creatinine metabolism (urtABCDE, ureABCEFGD, crnT, codA), a number of biochemically uncharacterized enzymes and transport systems (NCgl1099, NCgl1100, NCgl 1915-1918) as well as signal transduction proteins (glnD, glnK). For the AmtR regulon, an AmtR box has been defined which comprises the sequence tttCTATN6AtAGat/aA. Furthermore, the transcriptional organization of AmtR-regulated genes and operons was characterized.


Primary multifocal tuberculous osteomyelitis with involvement of the ribs.
Two cases of primary multifocal tuberculous osteomyelitis with involvement of the rib cage are presented. The lungs were normal and the appearance of the skeletal lesions did not suggest tuberculosis. These lesions were predominantly lytic, with minimal soft tissue involvement. Tuberculosis should be high in the differential diagnosis of multiple destructive bone lesions, especially in patients from regions where tuberculosis is endemic.


Diagnostic procedures for Paget's disease. Radiologic, pathologic, and laboratory testing.
Paget's disease is a relatively common skeletal disorder that affects older individuals. Although its causes are unknown, it is characterized by abnormal remodeling and a high rate of bone turnover that determines its pathologic, radiologic, and biochemical findings. The findings in the three phases of Paget's disease are described.


Assessing child abuse: "We need to talk!"
This discussion paper adds to the recent critical debate concerning retrospective measurement of childhood abuse and adverse experiences. A series of recent articles found only modest overlap of prospective informant-based records with retrospective self-report questionnaires, with biases evident in the latter. However, this literature has omitted the use of investigator-based interviews as an alternative retrospective tool for triangulation of measurement. Validated interview approaches can ascertain accurate data using expert scoring that can be used to test both dose-effect and specificity analyses for further research and treatment purposes. Arguments for the retention and further promotion of intensive interview measures for retrospective assessment of childhood neglect and abuse in relation to lifetime clinical outcomes are presented, with illustrative analyses. A network analytic approach is outlined, with six types of childhood abuse or neglect experiences scored via a well-validated interview (the Childhood Experiences of Care and Abuse). This indicates distinct pathways between types of neglect and abuse, but also from more common emotional abuse (antipathy, critical parenting) through to more pernicious psychological abuse (coercive, sadistic control) involving physical abuse or sexual abuse pathways. This is supplemented by a case vignette to illustrate the different pathways indicated. The interview approach gives victims a voice with their narrative (chance to talk) needed for better understanding of the specific dynamics of child abuse and neglect and for an entry into psychotherapeutic work. We need to ensure that such methods are retained in the research and practitioner portfolio of measurement tools.


Virus Propagation and Plaque Assay for Dengue Virus.
This chapter describes the methods of propagation and titration for DENV-1 to -4, which are required for most of the experiments using live viruses. DENV does not reach titers as high as those of other viruses or as high as desired for their use in biological assays. Although DENV grows in many different cell lines derived from both vertebrate and invertebrate cells, the most common cell lines used for virus isolation and propagation are mosquito cells C6/36 from Aedes albopictus. Amongst several methods for virus quantification, plaque assay stands out as being very practical and inexpensive. This technique is carried out essentially to estimate virion density in a particular material, being extremely important when evaluating the effect of an antiviral treatment or antibody neutralization capacity, for example. In this assay, viral particles are serially diluted and added onto confluent cell monolayers immersed in a semisolid medium, which is responsible for limiting virus spread throughout the culture. Therefore, regarding the medium consistency, once a virion successfully infects a cell, the newly produced particles can only infect neighboring cells, eventually leading them to death. This phenomenon can be observed as round gaps or plaques in the culture after staining live cells with a crystal violet solution. Then, plaques are counted and used to determine plaque-forming units per milliliter. Here, we describe a protocol established by our group for dengue virus (DENV) titration with porcine kidney (PS) cells.


The effect of needle-insertion depth on the irrigant flow in the root canal: evaluation using an unsteady computational fluid dynamics model.
The aim of this study was to evaluate the effect of needle-insertion depth on the irrigant flow inside a prepared root canal during final irrigation with a syringe and two different needle types using a Computational Fluid Dynamics (CFD) model. A validated CFD model was used to simulate irrigant flow from either a side-vented or an open-ended flat 30-G needle positioned inside a prepared root canal (45 .06) at 1, 2, 3, 4, or 5 mm short of the working length (WL). Velocity, pressure, and shear stress in the root canal were evaluated. The flow pattern in the apical part of the root canal was similar among different needle positions. Major differences were observed between the two needle types. The side-vented needle achieved irrigant replacement to the WL only at the 1-mm position, whereas the open-ended flat needle was able to achieve complete replacement even when positioned at 2 mm short of the WL. The maximum shear stress decreased as needles moved away from the WL. The flat needle led to higher mean pressure at the apical foramen. Both needles showed a similar gradual decrease in apical pressure as the distance from the WL increased. Needle-insertion depth was found to affect the extent of irrigant replacement, the shear stress on the canal wall, and the pressure at the apical foramen for both needle types.


Acute isolated sphenoid sinusitis in children: A case series and systematic review of the literature.
This study aims to present a case series and systematic review of acute isolated sphenoid sinusitis (AISS) in children in order to better characterize clinical presentation, diagnosis, treatment, and outcomes of this condition. Ovid MEDLINE, Pubmed, Embase, Cochrane Library, and Google Scholar. Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Full-text, peer-reviewed journal publications from 1994 to 2020 in English; focus on acute sphenoid sinusitis; pediatric patients (<18 years of age); series with two or more children. Studies were assessed for data including demographics, presenting symptoms and signs, radiological investigations, treatment, outcomes and complications. Ten studies identifying 71 patients were included. Average age at presentation was 12.0 years (range 5-17 years). M:F ratio 1:1. The most common presenting symptoms were headache (98.6%), fever (50.7%), nasal symptoms (22.5%) ocular symptoms (19.7%) and decreased level of consciousness (12.7%). Twenty patients (28.1%) had neurological signs. Twenty-three patients (32.4%) presented with headache in isolation. Unsuspected diagnosis at presentation was noted in 54.0%. Average time to initial presentation was 14.0 days (median = 5.5 days, range 1-90 days). The majority of children were treated with antibiotics (98.6%) with 31.0%, 2.8% and 2.8% also undergoing sinus surgery, revision sinus surgery and neurosurgery, respectively. Intracranial complications occurred in 16.9% of patients. Significant long term sequelae occurred in 2 children (2.8%) and one death (1.4%) was also reported. All studies were retrospective case note reviews. Acute sphenoid sinusitis is a rare and difficult condition to diagnose in children. The majority of patients make a full recovery with appropriate treatment. If treatment is delayed however consequences can be life-threatening.


Lateral and apical root resorption in teeth orthodontically moved into edentulous ridge areas.
The present study aimed at comparing the external lateral root resorption (ELRR) and external apical root resorption (EARR) between teeth moved through the atrophic edentulous ridge and those undergoing the usual orthodontic movement. Fifty-four premolars were evaluated, where 27 of them had been moved toward the edentulous ridge (Group 1) and 27 from the same patient, had not been translated, which comprised the control group (Group 2). ELRR was evaluated by 0-3 scores and EARR was evaluated by 0-4 scores, before and after movement. Measurements were compared by Kruskal-Wallis and Student-Newman-Keuls tests. ELRR increased statistically only in the Group 1 (p< 0.05). After orthodontic treatment, it was observed that almost 56% (n = 15) of teeth in Group 1 presented scores 2 and 3, while Group 2 presented scores 2 and 3 in about 11% (n= 3) of the teeth. EARR increased in both groups after orthodontic movement, however, statistically analyses showed no significant differences between groups (p> 0.05). Orthodontic movement into the atrophic edentulous ridge is subject to a greater lateral external root resorption.


[Principle military medical institution in Ukraine: history pages (to 250 anniversary of the foundation of Kiev Military Hospital)].
The article is dedicated to the history of the foundation, development of the principle military medical institution of Ukraine, which celebrates its 250 anniversary in June 2005. The authors proposed the periodization of the origin, main achievements and tendency of the development of the management of this military medical institution.


SNHG1/miR-556-5p/TCF12 feedback loop enhances the tumorigenesis of meningioma through Wnt signaling pathway.
Meningioma, as a sort of the malignantly intracranial tumors, has captured public attention for its second-highest morbidity all over the world. Long noncoding RNAs (lncRNAs), including lncRNA SNHG1, have been well known as essential players in the development of diverse cancers. However, the biological effect and regulatory mechanism of SNHG1 have not been mentioned in meningioma. In this work, it was discovered that SNHG1 was overexpressed in meningioma cell lines. SNHG1 deficiency restrained cell growth as well as accelerated apoptosis. Then mechanism experiments demonstrated that SNHG1 functioned as the role of sponging miR-556-5p and negatively regulated miR-556-5p expression. Moreover, it was verified that TCF12 is the direct downstream target of miR-556-5p. Furthermore, SNHG1/miR-556-5p/TCF12 axis promoted cell proliferation and suppressed cell apoptosis in meningioma via activating the Wnt signaling pathway. In the end, it was confirmed that TCF12 expression was positively regulated by SNHG1, and TCF12 could promote transcription of SNHG1 through binding with the promoter region of SNHG1. In conclusion, the SNHG1/miR-556-5p/TCF12 feedback loop promotes the tumorigenesis of meningioma through the Wnt signaling pathway.


Endocrine abnormalities of obesity.
Studies have shown that patients with central obesity have increased cortisol secretion, probably because they have increased activity of the hypothalamic-pituitary-adrenal (HPA) axis. A high waist-to-hip ratio (WHR) is associated with low production of sex steroids, such as testosterone in men, and a low rate of secretion of growth hormone. High levels of cortisol and insulin combined with low levels of growth hormone and sex steroid can cause lipid accumulation. These hormonal changes probably produce more deposition of visceral than subcutaneous fat. Patients who are deficient in either testosterone or growth hormone show a reduction in visceral adiposity when their hormone levels are normalized. Stress has been shown to activate the HPA axis and may cause the hormonal changes associated with obesity. Individuals with elevated WHR have indications of high levels of stress and anxiety. Monkeys that were stressed by social disruption were found to have increased cortisol levels and low sex steroid levels. Many of these animals had insulin resistance and visceral adiposity. Stimulants, such as alcohol and smoking, also increase the activity of the HPA axis.


Comparison of low ionic diluents for use with the Diamed antiglobulin gel test.
Microtube column systems, although widely used in transfusion serology for the detection of red cell antibodies, may not detect weak Fy(a), Jk(a), S and K antibodies. A number of low ionic diluents are used to shorten the incubation time required for red cell antibody detection in the antiglobulin test. However, there are no published reports to show whether these low ionic diluents vary in their ability to detect red cell antibodies using microcolumn detection systems. Three low ionic diluents, Diamed ID-CellStab, Diamed ID-Diluent2 and an in-house produced low ionic strength solution (LISS), were assessed using the Diamed-ID LISS/Coombs microtube column system (in accordance with the manufacturer's instructions), to ascertain whether the choice of diluent influences red cell antibody detection. Two hundred and seventy patient samples were screened for red cell antibodies. The reaction strength was increased in 50% of the samples with detectable red cell antibodies using LISS as the diluent compared with ID-CellStab. Of the 51 red cell antibodies directed against Rhesus, Duffy, Kidd or Kell antigens, 21% reacted more strongly in LISS compared with Diamed ID-CellStab with a difference in grading of > or =1. Minimal disparity was found between ID-Diluent2 and LISS. Biochemical analysis of pH, osmolality, sodium, potassium and phosphate were comparable for ID-CellStab, ID-Diluent2 and LISS. Measurement of conductivity in each low ionic diluent was performed as a measure of ionic strength in the final reactant mix, as the same amount of low ionic diluent was used for each test. The conductivity was 3 x 5 mS cm for LISS and ID-Diluent2, and 5 x 8 mS cm for ID-CellStab; the acceptable range being 3 x 7 +/- 0 x 3 mS cm as cited in the Guidelines for the Blood Transfusion Services in the United Kingdom. This evaluation suggests that ID-CellStab is a suboptimal low ionic diluent for red cell antibody detection using Diamed-ID LISS/Coombs gel cards. The poorer performance of ID-CellStab compared with LISS may be explained by its higher ionic strength.


HMGB1 is an independent predictor of death and heart transplantation in heart failure.
High-Mobility-Group Box 1 (HMGB1) has been established as an important mediator of myocardial inflammation and associated with progression of heart failure (HF). The aim of this study was to analyze the prognostic value of systemic HMGB1 levels in HF patients with ischemic and non-ischemic cardiomyopathy. We conducted an analysis (median follow-up time 2.5 years) of HMGB1 plasma concentration in 154 patients with systolic HF and correlated the results with disease severity and prognosis. HMGB1 in HF patients with severe symptoms (NYHA III/IV; 5.35 ng/ml; interquartile range (IQR) = 3.48-8.42 ng/ml) was significantly elevated compared with that in patients with mild symptoms (NYHA I/II; 3.37 ng/ml, IQR = 2.31-5.22 ng/ml, p < 0.0001) and with controls (3.25 ng/ml, IQR = 3.04-3.67 ng/ml, p < 0.0001). HMGB1 levels correlated with other markers of heart failure indicating an association of HMGB1 with disease severity in HF. In a univariate cox regression model for the combined endpoint of death and heart transplantation, HMGB1 proved to be a predictor at cut-off values based on HMGB1 terciles of either 3.4 or 6.1 ng/ml (p = 0.001 and p < 0.0001, respectively). In a multivariate cox regression model, which included NT-proBNP, creatinine, age, NYHA class, white blood cell count, anemia, and age, HMGB1 remained an independent predictor of the combined endpoint (hazard ratio (HR) = 2.48, 95% confidence interval (CI) = 1.06-5.83, p = 0.037 and HR = 2.48, 95% CI = 1.31-4.71, p = 0.005, respectively). Our findings demonstrate that HMGB1 plasma concentration is elevated in HF and correlates with disease severity and that is an independent predictor of the combined endpoint death and heart transplantation in HF patients.


Evidence for moiré intralayer excitons in twisted WSe2/WSe2 homobilayer superlattices.
Recent advances in twisted van der Waals heterostructure superlattices have emerged as a powerful and attractive platform for exploring novel condensed matter physics due to the interplay between the moiré potential and Coulomb interactions. The moiré superlattices act as a periodic confinement potential in space to capture interlayer excitons (IXs), resulting in moiré exciton arrays, which provide opportunities for quantum emitters and many-body physics. The observation of moiré IXs in twisted transition-metal dichalcogenide (TMD) heterostructures has recently been widely reported. However, the capture and study of the moiré intralayer excitons based on TMD twisted homobilayer (T-HB) remain elusive. Here, we report the observation of moiré intralayer excitons in a WSe2/WSe2 T-HB with a small twist angle by measuring PL spectrum. The multiple split peaks with an energy range of 1.55-1.73 eV are different from that of the monolayer WSe2 exciton peaks. The split peaks were caused by the trapping of intralayer excitons via the moiré potential. The confinement effect of the moiré potential on the moiré intralayer excitons was further demonstrated by the changing of temperature, laser power, and valley polarization. Our findings provide a new avenue for exploring new correlated quantum phenomena and their applications.


Evaluation of pelvic floor muscle function (PFMF) in cervical cancer patients with Querleu-Morrow type C hysterectomy: a multicenter study.
To evaluate the pelvic floor muscle function (PFMF) of cervical cancer patients after type QM-C hysterectomy and to explore the relationship between decreased PFMF and related factors. This was a multi-centered retrospective cohort study. 181 cervical cancer patients who underwent type QM-C hysterectomy were enrolled from 9 tertiary hospitals. Strength of PFMF were measured using neuromuscular apparatus (Phenix U8, French). Risk factors contributing to decreased PFMF were analyzed by univariate and multivariate ordinal polytomous logistic regression. Totally 181 patients were investigated in this study. 0-3 level of type I muscle fibre strength (MFSI) was 52.6% (95/181), 0-3 level of type IIA muscle fibre strength (MFSIIA) was 50% (91/181). Subjective stress urinary incontinence was 46% (84/181), urinary retention was 27.3% (50/181), dyschezia was 41.5% (75/181), fecal incontinence was 9% (18/181). ① MFSI: Multivariate ordinal polytomous logistic regression shows that the follow-up time (p < 0.05), chemotherapy and radiotherapy (p = 0.038) are independent risk factors of MFSI's reduction after type QM-C hysterectomy. ② MFSIIA: multivariate ordinal polytomous logistic regression shows that the follow-up time (p < 0.05) are independent risk factors of MFSIIA's reduction after type QM-C hysterectomy. The pelvic floor muscle strength (PFMS) increased after 9 months than in 9 months after operation, which showed that the PFMS could be recovered after operation. We advocate for more attention and emphasis on the PFMF of Chinese female patients with cervical cancer postoperation. PFMF after QM-C hysterectomy has not been analyzed by current study. The contribution is that patients with radical hysterectomy should do pelvic floor rehabilitation exercises in 3 months after operation. Clinical Trails NCT number of this study is 02492542.


The estimation of live weight based on linear traits in indigenous Tswana goats at various ages in Botswana.
The objective of this study was to determine the usefulness of some linear body measurements in predicting live weight in indigenous Tswana goats at various ages. Data for this study were obtained from 2,783 goats sampled from six agricultural regions of Botswana except for Tsabong and Ghanzi districts. Fifteen farmers keeping goats were randomly selected from each district and records taken on a random sample of 4-12 animals per farm depending on the 1999 Botswana Government average district flock size. Body measurements recorded were heart girth, height at withers, body length, shoulder width, and live weight. Information on age of each animal was estimated from dentition; flock size and sex of the animal were also recorded for each farm. Regression analysis using stepwise selection method in Statistical Analysis System was used to determine prediction equations for live weight with heart girth, height at withers, body length, and shoulder width as independent variables for male and female goats of different ages. In all models, heart girth contributed most in explaining variation in body weight as shown by high partial R(2) which ranged from 0.48 for female mature to 0.80 for mature male goats. Using mean square error, R(2), and Mallows' C(p), the best prediction equations were for female and male kids (R(2) = 0.83 and 0.82, respectively) and mature male goats (R(2) = 0.82). The poorest model was for mature female goats with R(2) = 0.57. Live body weight could be estimated with accuracy from linear body measurements in Tswana goats which are unique for each sex and age. More research is required to explore using these models to develop measuring tapes for use by resource poor farmers who keep indigenous Tswana goats.


Contributions of degradation and brain-to-blood elimination across the blood-brain barrier to cerebral clearance of human amyloid-β peptide(1-40) in mouse brain.
The purpose of the present study was to estimate the relative contributions of degradation and brain-to-blood elimination processes to the clearance of microinjected human amyloid-β peptide(1-40) (hAβ(1-40)) from mouse cerebral cortex, using a solid-phase extraction method together with a newly developed ultraperformance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS) quantitation method for intact hAβ(1-40). The clearance rate constant of hAβ(1-40) in mouse cerebral cortex was determined to be 3.21 × 10(-2)/min under conditions where the saturable brain-to-blood elimination process across the blood-brain barrier (BBB) was expected to be saturated. Thus, this clearance rate constant should mainly reflect degradation. The [(125)I]hAβ(1-40) elimination rate across the BBB under nonsaturating conditions was determined to be 1.48 × 10(-2)/min. Inhibition studies suggested that processes sensitive to insulin and phosphoramidon, which inhibit neprilysin, insulin-degrading enzyme, and endothelin-converting enzyme, are involved not only in degradation, but also in elimination of hAβ(1-40). In conclusion, our results suggest a dominant contribution of degradation to cerebral hAβ(1-40) clearance, and also indicate that a sequential process of degradation and elimination of degradation products is involved in cerebral hAβ(1-40) clearance.


Muscarinic type 1 receptors mediate part of nitric oxide's vagal facilitatory effect in the isolated innervated rat right atrium.
We investigated whether vagal cardiac cholinergic facilitation by nitric oxide (NO) is mediated by cardiac muscarinic receptor subtypes in the vagally innervated rat right atrium in vitro. Experiments were carried out in the presence of atenolol (4 microM). The right vagus was stimulated at 4, 8, 16, 32 Hz; pulse duration 1 ms at 20 V for 20s; vagal postganglionic activation was achieved using nicotine (0.1, 0.3, 0.5, 1mM) and the effect on cardiac interval (ms) assessed. Pirenzepine (1 microM), a M1 antagonist, attenuated vagally induced increase in cardiac interval. L-Arginine (0.34 mM) superfused with pirenzepine failed to reverse this attenuation, however, L-arginine applied alone reversed the reduction vagal cardiac slowing. Similarly, sodium nitroprusside (10 microM) applied alone, and not together with pirenzepine, was able to reverse the attenuation of vagal effects caused by pirenzepine. Synthetic MT7 (1 nM) toxin, a selective M1 antagonist confirmed these results. M3 antagonism using para-fluorohexahydrosiladifenidol (p-F-HHSiD) (300 nM) and M4 antagonism with PD 102807 (200 nM) did not affect the vagally induced increase in cardiac interval. Nicotine induced increase in cardiac interval was not altered by pirenzepine. These results show that antagonism of M1 receptors on cardiac vagal preganglionic fibres reduces vagal efficacy which can be recovered by either a nitric oxide synthase substrate or a NO donor.


Transitioning youths into care: linking identified HIV-infected youth at outreach sites in the community to hospital-based clinics and or community-based health centers.
To describe and analyze the process of transitioning HIV-infected youths from the point of HIV diagnosis into HIV treatment. Rather than simply offering HIV positive youths a list of referrals, the youth-focused SPNS grantees (AWAC) found it vital that youths were immediately assisted with linkage to a medical provider. From February 1997 to December 2000, 107 identified HIV-infected youths from the five adolescent SPNS projects were surveyed on needs and barriers. The time interval between HIV testing and youth linkage to care was also noted. Nine percent of youth reported perceived barriers to accessing health care. Perceived needs were identified as Mental Health (44.9%; n = 48); Alcohol and drug treatment (14%; n = 15); transportation to health care settings (40.2%; n = 43); and housing (46.7%; N = 50). At sites tracking linkage to care, the time of being transitioned into a medical setting ranged from 5 to 55 days (average 26 days). The period of transitioning identified HIV-infected youths into care can be reduced from 1-5 years to as short as 5-55 days. Success with linking these youth to care involves establishing a series of contacts at outreach sites wherein program staff seeks to build trusting relationships with youths, is able to track these youths and identify and address perceived needs.


Why Do We Talk To Ourselves?
Human beings talk to themselves; sometimes out-loud, other times in inner speech. In this paper, I present a resolution to the following dilemma that arises from self-talk. If self-talk exists then either, (i) we know what we are going to say and self-talk serves no communicative purpose, and must serve some other purpose, or (ii) we don't know what we are going to say, and self-talk does serve a communicative purpose, namely, it is an instance of us communicating with ourselves. Adopting (i) was the strategy taken by Bart Geurts, who claims that the primary purpose of self-talk is to entrain commitments, and is not (primarily) communicative. While accepting that self-talk can usefully play this role, I criticise the view that entraining commitments is self-talk's fundamental role. I argue that adopting the view that we are self-blind, at least to a significant degree, means that we can accept that self-talk does play a communicative role.


Human working memory capacity is 7+/-2 in a radial maze with distracting interruption: possible implication for neural mechanisms of declarative and implicit long-term memory.
Human participants were instructed to walk out along each of the arms of a 15-m in diameter, 8-arm radial maze once and only once. In order to approximate the circumstances under which laboratory rats remember visited sites, our human participants were asked to select arms in an unsystematic order. They scored an average of 7.6 to 7.8 correct choices, even if midway during a trial there was a 5-min interruption filled with a verbal-spatial interfering task (a scavenger hunt) or a 15-min interruption filled with a visuospatial task (a maze-running computer simulation). This finding extends our earlier research with humans in 13- or 17-arm radial mazes under nondelay conditions, in which we also found working memory (WM) capacity for about 7 to 9 places, the same as that of laboratory rats. We discuss earlier findings in other laboratories, showing that rats can successfully bridge long radial maze task interruptions of 5 or 8 h, and we compare our results also to those from studies in which human participants were not discouraged from reducing memory load by responding systematically in radial mazes. Because the radial maze task takes minutes to complete even under nondelay conditions its routine consideration as a working memory task in the animal literature alters the assumptions often made about the duration of WM in the human literature. Accumulating empirical findings about place-memory in humans, nonhuman mammals, and birds suggest it might be productive to reevaluate this theoretical issue with respect to present knowledge about the roles of the hippocampus and other brain structures in declarative memory and in procedural or implicit memory, while considering the hypothesis that some forms of information may exploit long-term memory in parallel with working memory.


The neuroprotective action of JNK3 inhibitors based on the 6,7-dihydro-5H-pyrrolo[1,2-a]imidazole scaffold.
Imidazole-based structures of p38 inhibitors served as a starting point for the design of JNK3 inhibitors. Construction of a 6,7-dihydro-5H-pyrrolo[1,2-a]imidazole scaffold led to the synthesis of the (S)-enantiomers, which exhibited p38/JNK3 IC50 ratio of up to 10 and were up to 20 times more potent inhibitors of JNK3 than the relevant (R)-enantiomers. The JNK3 inhibitory potency correlated well with inhibition of c-Jun phosphorylation and neuroprotective properties of the compounds in low K+-induced cell death of rat cerebellar granule neurones.


Adenovirus-mediated gene transfer of pathogen-associated molecular patterns for cancer immunotherapy.
The delivery of stimulatory signals to dendritic cells (DCs) in the tumor microenvironment could be an effective means to break tumor-induced tolerance. The work presented here evaluates the immunostimulatory properties of pathogen-associated molecular patterns (PAMPs), microbial molecules which bind Toll-like receptors and deliver activating signals to immune cells, when expressed in tumor cells using adenoviral (Ad) vectors. In vitro, transduction of A549 tumor cells with Ad vectors expressing either flagellin from Listeria monocytogenes or P40 protein from Klebsiella pneumoniae induced the maturation of human monocyte-derived DCs in co-cultures. In mixed lymphocyte reactions (MLRs), Ad-flagellin and Ad-P40 transduction of tumor cells stimulated lymphocyte proliferation and the secretion of IFN-gamma. In vivo, these vectors were used either as stand-alone immunoadjuvants injected intratumorally or as vaccine adjuvants combined with a tumor antigen-expressing vector. When Ad-PAMPs were administered intratumorally to mice bearing subcutaneous syngeneic B16F0-CAR (cocksackie-adenovirus receptor) melanomas, tumor progression was transiently inhibited by Ad-P40. In a therapeutic vaccine setting, the combination of Ad-MUC1 and Ad-PAMP vectors injected subcutaneously delayed the growth of implanted RenCa-MUC1 tumors and improved tumor rejection when compared with vaccination with Ad-MUC1 alone. These results suggest that Ad-PAMPs could be effective immunoadjuvants for cancer immunotherapy.


[Clinic and prevention of suicide in ederly. A literature review].
In almost all countries where reliable statistics researches are conducted, the rate of suicide among elderly people rises in people over 65 years old, reaching its highest point at 85 years old. Nevertheless, it seems that this phenomena is tended to be ignored. Although the evidence given by the epidemiology, many physicians and even of psychiatrits claim that the suicide among elderly is rare.


(E)-N'-(4-Hy-droxy-benzyl-idene)-3-nitro-benzohydrazide.
The mol-ecule of the title compound, C(14)H(11)N(3)O(4), assumes an E conformation about the C=N double bond. The benzene rings form a dihedral angle of 3.9 (2)°. The crystal structure is stabilized by N-H⋯O, O-H⋯N, O-H⋯O and C-H⋯O hydrogen bonds, forming layers parallel to (101). In addition, intra-layer π-π stacking inter-actions [centroid-centroid distance = 3.635 (2) Å] are observed.


Fatty acid nitration in human low-density lipoprotein.
Lipid nitration occurs during physiological and pathophysiological conditions, generating a variety of biomolecules capable to modulate inflammatory cell responses. Low-density lipoprotein (LDL) oxidation has been extensively related to atherosclerotic lesion development while oxidative modifications confer the particle pro-atherogenic features. Herein, we reviewed the oxidation versus nitration of human LDL protein and lipid fractions. We propose that unsaturated fatty acids present in LDL can be nitrated under mild nitration conditions, suggesting an anti-atherogenic role for LDL carrying nitro-fatty acids (NFA).


The changing epidemiological and complications profile of chronic suppurative otitis media in a developing country after two decades.
To evaluate the shift in epidemiological and complication profiles of chronic suppurative otitis media (CSOM) over two decades in a resource-limited economy. A retrospective analysis of the prevalence of CSOM and its complications in tertiary health institution a over a period of 2 years were compared between 203 cases seen in 2009 and December 2010 (present era); and 343 cases seen in 1990 and 1991 (previous era). Data were collected on the demographic characteristics of the patients, age of onset of otorrhoea, the disease type, laterality and complications of CSOM. The prevalence of CSOM significantly reduced from 9 % in the previous era to 5.1 % in the present era (P < 0.0001). The mean age of the recent group was 27.6 ± 19.3 years, and was 21.6 ± 18.2 years in the previous group. (P = 0.009). Although both group had peak age prevalence at 11-20 years, recent group saw more significant adult population >20 years of age (P ≤ 0.001). More significant cases of cholesteatoma were documented in the recent group (3.4 %) than previous era (0.9 %) (P = 0.02). Although no significant change was noted regarding the rates of complication of CSOM in the two eras (P = 0.18), there was a notable shift toward predominant extracranial pattern of complication in the recent group (P = 0.04). The prevalence of CSOM has shown a significant reduction after two decades and it appears that there has been a shift in the epidemiology of CSOM towards a predominantly adult population in the recent years. Despite an overall reduction in the incidence of intracranial complications of CSOM in our series, its complication rates have not changed remarkably after 20 years.


Familial Himalayan p wave and left ventricular hypertrabeculation/noncompaction.
"Himalayan P waves," are reported in congenital heart disease and cardiomyopathies. We report a family with hypertrophic cardiomyopathy, Himalayan P waves, extensive focal right atrial wall thickening and left ventricular hypertrabeculation/noncompaction (LVHT). The father received a pacemaker and underwent heart transplantation because of hypertrophic cardiomyopathy. His daughters showed Himalayan P waves and right atrial wall thickening. LVHT was diagnosed in sister A at age 23 years and developed in sister B between 42 and 46 years. In sister A the heart rate continuously declined. She refused implantation of a pacemaker and died with 49 years. Sister B, suffers from bradycardia. Himalayan P waves are due to focal right atrial wall thickening, may be familially and associated with LVHT.


Fully automated and super-rapid system for the detection of JAK2V617F mutation.
JAK2V617F is a common mutation in chronic myeloproliferative diseases (CMPDs). We have developed a system utilizing JAK2V617F-specific guanine quenching probe (QP-system) to detect JAK2V617F. With QP-system, results can be obtained from 100 microl of blood within 90 min. We compared QP-system with direct sequencing using 42 CMPD patients' specimens. JAK2V617F was detected in 25 specimens by QP-system, while direct sequencing failed to detect JAK2V617F in 7 of those 25. The presence of JAK2V617F mutation in these 7 specimens was confirmed by allele-specific PCR. These findings indicate that QP-system is more sensitive and useful than direct sequencing for diagnoses of CMPDs.


Arylsulphatase activity in the oviduct of the frog Rana esculenta. II. Progesterone-induced changes following ovariectomy and hypophysectomy.
The effects of progesterone treatment on arylsulphatase activity were studied histochemically and biochemically in the frog oviduct under different experimental conditions. In ovariectomized animals, the hormone induced a large increase in enzyme activity, while in hypophysectomized ones there was a large decrease in this activity. These results indicate that the facilitatory and inhibitory effects of progesterone on arylsulphatase activity are influenced by the presence in situ of the gonad. Hypotheses are advanced to explain different effects of the progesterone treatment.


The Influence of Baseline Clinical Status and Surgical Strategy on Early Good to Excellent Result in Spinal Lumbar Arthrodesis: A Machine Learning Approach.
The study aims to create a preoperative model from baseline demographic and health-related quality of life scores (HRQOL) to predict a good to excellent early clinical outcome using a machine learning (ML) approach. A single spine surgery center retrospective review of prospectively collected data from January 2016 to December 2020 from the institutional registry (SpineREG) was performed. The inclusion criteria were age ≥ 18 years, both sexes, lumbar arthrodesis procedure, a complete follow up assessment (Oswestry Disability Index-ODI, SF-36 and COMI back) and the capability to read and understand the Italian language. A delta of improvement of the ODI higher than 12.7/100 was considered a "good early outcome". A combined target model of ODI (Δ ≥ 12.7/100), SF-36 PCS (Δ ≥ 6/100) and COMI back (Δ ≥ 2.2/10) was considered an "excellent early outcome". The performance of the ML models was evaluated in terms of sensitivity, i.e., True Positive Rate (TPR), specificity, i.e., True Negative Rate (TNR), accuracy and area under the receiver operating characteristic curve (AUC ROC). A total of 1243 patients were included in this study. The model for predicting ODI at 6 months' follow up showed a good balance between sensitivity (74.3%) and specificity (79.4%), while providing a good accuracy (75.8%) with ROC AUC = 0.842. The combined target model showed a sensitivity of 74.2% and specificity of 71.8%, with an accuracy of 72.8%, and an ROC AUC = 0.808. The results of our study suggest that a machine learning approach showed high performance in predicting early good to excellent clinical results.


Triheptanoin - Novel therapeutic approach for the ultra-rare disease mitochondrial malate dehydrogenase deficiency.
Mitochondrial malate dehydrogenase (MDH2) deficiency (MDH2D) is an ultra-rare disease with only three patients described in literature to date. MDH2D leads to an interruption of the tricarboxylic acid (TCA) cycle and malate-aspartate shuttle (MAS) and results in severe early onset encephalopathy. Affected infants suffer from psychomotor delay, muscular hypotonia and frequent seizures. Laboratory findings are unspecific, including elevated lactate in blood and cerebrospinal fluid. Brain magnetic resonance imaging reveals delayed myelination and brain atrophy. Currently there is no curative therapy to treat this devastating disease. Here, we present a female patient diagnosed with MDH2D after a stroke-like episode at 18 months. Trio-whole exome sequencing revealed compound heterozygous missense variants in the MDH2 gene: c.398C>T, p.(Pro133Leu) and c.445delinsACA, p.(Pro149Hisfs*22). MDH2 activity assay and oxygraphic analysis in patient's fibroblasts confirmed the variants were pathogenic. At the age of 36 months, a drug trial with triheptanoin was initiated and well tolerated. The patient's neurologic and biochemical phenotype improved and she had no further metabolic decompensations during the treatment period suggesting a beneficial effect of triheptanoin on MDH2D. Further preclinical and clinical studies are required to evaluate triheptanoin treatment for MDH2D and other TCA cycle and MAS defects.


Gelatinous transformation of the bone marrow.
A study of so-called gelatinous transformation of the bone marrow in 14 patients is presented. The condition is characterized by the presence of extracellular "gelatinous" material, fat atrophy, and associated focal marrow hypoplasia and is often mistaken as representing edema, necrosis, or amyloid. Histochemical studies indicate that the extracellular substance is hyaluronic acid. Though the lesion is usually associated with severe weight loss and cachexia, it appears that factors other than malnutrition must play a significant role in the pathogenesis of gelatinous transformation. Anemia, frequently seen in these patients, does not appear to be a consequence of gelatinous transformation.


Disulfiram enhances subjective effects of dextroamphetamine in humans.
Disulfiram has shown promise in several clinical trials for cocaine addiction, but its potential utility in the treatment of amphetamine addiction has not been examined. The goal of this study was to determine the effects of disulfiram on acute physiological and subjective responses to dextroamphetamine in healthy volunteers. Five male and 5 female subjects participated in an outpatient double-blind, placebo-controlled, crossover study. Subjects were randomly assigned to a sequence of disulfiram (250 mg/day) or placebo treatments each lasting for 4 days. Day four of each treatment period was the experimental session, in which subjects orally ingested a single dose of dextroamphetamine (20 mg/70 kg). Outcome measures included heart rate, blood pressure, plasma cortisol and prolactin, subjective and performance on the Sustained Attention to Response Test (SART). Disulfiram did not affect dextroamphetamine-induced increases in heart rate, blood pressure, cortisol, or prolactin. Disulfiram did enhance some of the subjective effects of dextroamphetamine including ratings of "high," "anxious," "bad drug effects," "want more drug" and "drug liking" and was also associated with decreased performance in the SART test. How these enhanced subjective amphetamine responses affect cocaine use behavior remains to be determined in future clinical trials.


Are endogenous retroviruses involved in human autoimmune disease?
A role for viruses in the etiopathogenesis of human autoimmune diseases has long been suspected but has not yet been proven. In Sjögren's syndrome (SS), there is continuing experimental support for the possible involvement of Epstein-Barr virus. Since the advent of AIDS, there is also great interest in retroviruses and autoimmune disease. We previously reported that 30% of SS patients and 36% of systemic lupus erythematosus (SLE) patients have serum antibodies to the p24 gag protein of HIV-1. We now report that two mechanisms classic for retroviruses (molecular mimicry and immunosuppression) may be operative in SS and SLE. The p24 gag protein shares a proline-rich epitope with the Sm nucleoprotein to which many SLE patients have antibodies. The impaired lymphocyte activation seen in peripheral blood T cells in SS patients is also seen in a human T cell line infected with an A-type retroviral particle linked to SS. Many studies suggest that endogenous retroviral sequences are important in immunoregulation. We now suggest that endogenous retroviral sequences may also be important in the etiology and pathogenesis of SS and SLE.