Analysis of North Carolina Medicaid Claims Data to Simulate a Pediatric Accountable Care Organization.
Despite momentum for pediatric value-based payment models, little is known about tailoring design elements to account for the unique needs and utilization patterns of children and young adults. To simulate attribution to a hypothetical pediatric accountable care organization (ACO) and describe baseline demographic characteristics, expenditures, and utilization patterns over the subsequent year. This retrospective cohort study used Medicaid claims data for children and young adults aged 1 to 20 years enrolled in North Carolina Medicaid at any time during 2017. Children and young adults receiving at least 50% of their primary care at a large academic medical center (AMC) in 2017 were attributed to the ACO. Data were analyzed from April 2020 to March 2021. Primary outcomes were total cost of care and care utilization during the 2018 performance year. Among 930 266 children and young adults (377 233 children [40.6%] aged 6-12 years; 470 612 [50.6%] female) enrolled in Medicare in North Carolina in 2017, 27 290 children and young adults were attributed to the ACO. A total of 12 306 Black non-Hispanic children and young adults (45.1%), 6308 Hispanic or Latinx children and young adults (23.1%), and 6531 White non-Hispanic children and young adults (23.9%) were included. Most attributed individuals (23 133 individuals [84.7%]) had at least 1 claim in the performance year. The median (IQR) total cost of care in 2018 was $347 ($107-$1123); 272 individuals (1.0%) accounted for nearly half of total costs. Compared with children and young adults in the lowest-cost quartile, those in the highest-cost quartile were more likely to have complex medical conditions (399 individuals [6.9%] vs 3442 individuals [59.5%]) and to live farther from the AMC (median [IQR distance, 6.0 [4.6-20.3] miles vs 13.9 [4.6-30.9] miles). Total cost of care was accrued in home (43%), outpatient specialty (19%), inpatient (14%) and primary (8%) care. More than half of attributed children and young adults received care outside of the ACO; the median (IQR) cost for leaked care was $349 ($130-$1326). The costliest leaked encounters included inpatient, ancillary, and home health care, while the most frequently leaked encounters included behavioral health, emergency, and primary care. This cohort study found that while most children attributed to the hypothetical Medicaid pediatric ACO lived locally with few health care encounters, a small group of children with medical complexity traveled long distances for care and used frequent and costly home-based and outpatient specialty care. Leaked care was substantial for all attributed children, with the cost of leaked care being higher than the total cost of care. These pediatric-specific clinical and utilization profiles have implications for future pediatric ACO design choices related to attribution, accounting for children with high costs, and strategies to address leaked care.


Time trends in incidence, case fatality, and mortality of intracerebral hemorrhage.
To assess age- and sex-specific trends in incidence and 30-day and 1-year case fatality of intracerebral hemorrhage (ICH) in the Netherlands. Patients hospitalized for first ICH were identified through linkage of the national hospital discharge register and population register using ICD-9 code 431. We identified out-of-hospital deaths in the national cause of death register. Incidence, 30-day and 1-year case fatality, and total mortality rate were calculated by age and sex. We identified time trends by joinpoint regression analysis and Mann-Kendall tests. We identified 41,068 cases of ICH (51% men) between 1998 and 2010, of which 6% were out-of-hospital deaths. ICH incidence declined in men and women younger than 75 years (p ≤ 0.01, not significantly in men 35-54 years) but remained stable in patients 75 years and older. Thirty-day and 1-year case fatality declined in patients younger than 75 years (not significantly in women 35-54 years). In patients aged 35 to 54 years, ICH mortality remained stable until 2003 and then declined slightly (annual percentage change [APC] men: -7.09%; 95% confidence interval [CI] -11.39 to -2.59; women: -8.67%; 95% CI -15.18 to -1.66). In patients 55 to 74 years, mortality declined in men between 1995 and 2010 (APC -4.55%; 95% CI -5.49 to -3.59) and in women between 1992 and 2010 (APC -3.51%; 95% CI -4.16 to -2.85). Mortality did not decline in patients aged 75 years and older. In the Netherlands, ICH incidence, case fatality, and mortality rates have declined significantly in men and women younger than 75 years but remained stable in patients 75 years and older. The observed time trends may be explained by better prevention and treatment during the previous 2 decades of which the elderly do not seem to benefit.


Gene-centromere mapping of medaka sex chromosomes using triploid hybrids between Oryzias latipes and O. luzonensis.
Four sex-linked genetic markers (SL1, SL2, B2.38 and stsOPQ05-1) on the sex chromosomes of the medaka, O. latipes, were mapped in relation to the centromere by means of triploid hybrids between O. latipes and O. luzonensis. Female F1 hybrid O. latipes of two inbred strains, Hd-rR and HNI, were crossed with male O. luzonensis. Triploidization was induced by heat-shock treatment. Hatching rate of heat-shock treated eggs was 59%, and that of untreated hybrid eggs was 2%, indicating that most of the hatched fry were triploid. Using these triploid hatched fry, the map distances between the four loci and the centromere were examined. The order was SL2 - centromere - SL1 - B2.38 - stsOPQ05-1 and the map distances were: SL2 - centromere, 1%; centromere - SL1, 18%; SL1 - B2.38, 19%; B2.38 - stsOPQ05-1, 9%. Previous studies using FISH showed that SL2 is located on the short arm of large submetacentric chromosomes, and SL1 was closely linked to SDF (sex-determining factor). The results of gene-centromere mapping of this study show that SL1, B2.38 and stsOPQ05-1 are located on the long arm, and that. SDF is thus also on the long arm of the sex chromosomes.


Empagliflozin activates JAK2/STAT3 signaling and protects cardiomyocytes from hypoxia/reoxygenation injury under high glucose conditions.
The morbidity and mortality rates of cardiovascular disease are markedly higher in patients with diabetes than in non-diabetic patients, including patients with ischemia-reperfusion injury (IRI). However, the cardiovascular protective effects of Empagliflozin (EMPA) on IRI in diabetes mellitus have rarely been studied. In this study, we established a cardiomyocyte hypoxia/reoxygenation (H/R) injury model to mimic myocardial I/R injuries that occur in vivo. H9C2 cells were subjected to high glucose (HG) treatment plus H/R injury to mimic myocardial I/R injuries that occur in diabetes mellitus. Next, different concentrations of EMPA were added to the H9C2 cells and its protective effect was detected. STAT3 knockdown with recombinant plasmids was used to determine its roles. Our results showed that H/R injury-induced cell apoptosis, necroptosis, oxidative stress, and endoplasmic reticulum stress were further promoted by HG conditions, and HG treatment plus an H/R injury inhibited the activation of JAK2/STAT3 signaling. EMPA was found to protect against H/R-induced cardiomyocyte injury under HG conditions and activate JAK2/STAT3 signaling, while down-regulation of STAT3 reversed the protective effect of EMPA. When taken together, these findings indicate that EMPA protects against I/R-induced cardiomyocyte injury by activating JAK2/STAT3 signaling under HG conditions. Our results clarified the mechanisms that underlie the cardiovascular protective effects of EMPA in diabetes mellitus and provide new therapeutic targets for IRI in diabetes mellitus.


Reduced guanine nucleotide-stimulated polyphosphoinositide specific phospholipase C in platelet hyperaggregation in IDDM.
Platelet activity is increased in persons with insulin dependent diabetes mellitus (IDDM). Receptor-medicated phospholipase C (PLC) activation and hydrolysis of phosphatidylinositol bisphosphate (PIP2) accompanies platelet activation. Previous work from our laboratory has shown that PIP2 hydrolysis is decreased in platelets of persons with IDDM. PIP2 hydrolysis is mediated via a phosphoinositide(PI)-specific PLC. PI-PLC activity is regulated by guanine nucleotide(GTP)-binding proteins. We therefore examined the hypothesis that platelet aggregations and PI turnover in platelet from subjects with IDDM is linked to alterations in PI-specific PLC activity. We found thrombin induced platelet aggregation was increased in the IDDM group. Basal PI and PIP2-specific PLC activity was not statistically different for the two groups. Guanine-nucleotide stimulated PIP2-specific PLC activity was decreased in the IDDM platelets. The mechanism for the reduced PLC activity and its role in the platelet hyperaggregation requires further study.


Correlations between experimental chemotherapy in the murine glioma and effectiveness of clinical therapy regimens.
Intracerebral murine glioma 26 was used as a model system for evaluating two-drug combinations of antitumor agents, BCNU was combined with either procarbazine, dianhydrogalactitol, or ellipticine. CCNU was combined with procarbazine. All combinations were more active than the individual drugs alone. The most potent combinations achieved 85-100% tumor "cure" at 120 days, with combined toxicity indices of 0.25 (CCNU-procarbazine) to 1.30 (NCNU-dianhydrogalactitol). The experimental data were compared to clinical studies with CCNU, procarbazine, and vincristine, and BCNU-procarbazine.


Solid-cystic tumour of the pancreas. An endocrine neoplasm?
Immunohistochemical studies and DNA flow-cytometric investigations were performed in a case of solid-cystic tumour of the pancreas in a 35-year-old woman. All tumour cells were immunoreactive for the neuroendocrine cell markers chromogranin A and neuron-specific gamma-enolase. Moreover, about 10% of tumour cells were immunoreactive for insulin, while hypoglycaemia was absent. Few tumour cells (less than 1%) were immunoreactive for somatostatin, and no cells were found to be immunoreactive for pancreatic polypeptide or glucagon. No immunoreactivity was present for duct cell marker carcino-embryonic antigen and only individual cells were reactive for alpha 1-antitrypsin. Nuclear DNA content of the tumour cells was diploid and the proliferative activity was low. In confirmation of some reports on neuroendocrine markers in solid-cystic tumour of the pancreas, our findings support the theory that the lesion is a hormonally inactive neuroendocrine pancreatic tumour.


Magnetophotoselection of the triplet state of reaction centers from Rhodopseudomonas sphaeroides R-26.
Reaction centers of the photosynthetic bacterium Rhodopseudomonas sphaeroides R-26, give rise to large triplet state EPR signals upon illumination at low temperature (11 K). Utilizing monochromatic polarized light to generate the EPR spectra (magnetophotoselection) we have shown that the intensities of the observed triplet signals are strongly dependent upon the wavelength and polarization direction of the excitation. These data can be used to calculate the orientations of the excited transition moments with respect to each other and with respect to the triplet state principal magnetic axes system. Our quantitative approach is to follow the procedure outlined in a previous publication (Frank, H.A., Friesner, R., Nairn, J.A., Dismukes, G.C. and Sauer, K. (1979) Biochim. Biophys. Acta 547, 484-501) where computer simulations of the observed triplet state spectra were employed. The results presented in the present work indicate that the transition moment at 870 nm which is associated with the bacteriochlorophyll 'special pair' lies almost entirely along one of the principal magnetic axes of the triplet state. Aso, the 870 nm transition moment makes an angle of approx. 60 degrees with the 546 nm transition moment which is associated with a bacteriopheophytin. This latter result is in agreement with previous photoselection studies on the same bacterial species (Vermeglio, A., Breton, J., Paillotin, G. and Cogdell, R. (1978) Biochim. Biophys. Acta 501, 514-530).


Aqueous extraction of hemicelluloses from spruce--From hot to warm.
Aqueous extraction of hemicelluloses from spruce sapwood was performed at 90°C and 110°C. One of the main goals was to study if the same reaction mechanisms are valid at low temperatures as the ones observed previously at higher temperatures. An intensified cascade reactor system with a high liquid-solid ratio (∼ 180) was used in the experiments. Differences between the sugar specific extraction rates were observed especially in the beginning of the extraction processes. The experimental results fitted well to a kinetic model developed at higher temperatures, which confirms that the dissolution occurs with the same mechanisms at low temperature. Moreover, the correlation of the pH with the amount of sugars dissolved concurred with previous observations. The results contradict the assumption that low temperature dissolution would not occur and they help in studying the early stages of extraction as the kinetics are considerably slowed down.


A case report of Vibrio furnissii bacteremia and cellulitis in a malnourished patient without an apparent site of entry.
We herein report a case of Vibrio furnissii bacteremia with bilateral lower limb cellulitis. A 53-year-old Japanese man with a mood disorder presented to our hospital with fever and a complaint of an inability to walk. Two sets of blood cultures became positive for V. furnissii. The treatment regimen was modified to ceftazidime and doxycycline. The patient recovered without relapse. Despite thorough examinations, portal of entry of V. furnissii remained unclear. Although the bacteria was first misidentified as V. fluvialis by the phenotyping assay (API rapid ID 32E) and matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry, it was later confirmed as V. furnissii by dnaJ gene sequencing.


The effect of amrinone on liver regeneration in experimental hepatic resection model.
The phosphodiesterase inhibitors (PDEIs) have been proposed to improve hepatic reperfusion injury and hepatosplanchnic circulation, but the effects of these agents on liver regeneration have not been investigated thoroughly. The aim of this study was to investigate the effect of amrinone, a PDEI, on liver regeneration in rats. Sixty rats were divided into two groups, control and amrinone. Each group was then divided into three groups (n=10). An infusion of amrinone to the study group and of 0.9% NaCl to the control group was performed. Seventy percent liver resection was performed to the rats during the first hour of infusion. The infusion was maintained for 17 h after resection. A total of 18 h infusion was performed. Rats were allowed to survive for 24, 48, and 72 h, and then they were sacrificed. Biochemical, morphological, hematological, and histopathologic measurements and assessments were performed. There were statistically significant differences between the amrinone and control groups in alkaline phosphatase and relative liver weights at 24, 48, and 72 h (P<0.05). There also were statistically significant differences between the groups in AST, bilirubin, and albumine levels at 24 h, ALT and prothrombine time levels at 48 h, and aspartate aminotransferase and alanine aminotransferase levels at 72 h (P<0.05). Hepatic ATP levels, mitotic index, and proliferating cell nuclear antigen labeling index were significantly higher in amrinone group compared with control group at all three time intervals (P<0.05). Amrinone improves both morphological and functional liver regeneration after liver resection.


Potential Biomarkers in Diagnosis of Human Gastric Cancer.
The high incidence of gastric cancer (GC) and its consequent mortality rate severely threaten human's health. It is not frequently diagnosed until a relatively advanced stage. Surgery is the only potentially curative treatment. Thus, early screening and diagnosis are critical for patients with GC. The tumor marker assays used currently for detecting GC are simple and rapid, but the usage is limited by its low sensitivity and specificity. Here, we provide a brief description of some new potential markers and new biotechnological methods for the diagnosis of GC, hoping to find out more effective approaches for early detection of GC.


Understanding the effect of adverse childhood experiences on the risk of engaging in physical violence toward an intimate partner: The influence of relationship, social psychological, and sociodemographic contextual risk factors.
Adverse childhood experiences (ACEs) increase the risk of engaging in intimate partner violence (IPV) in later life. This study investigates the association between ACEs and engaging in physical violence toward a romantic partner in emerging adulthood while also accounting for proximal life experiences, including social psychological, intimate relationship, and sociodemographic characteristics. This study draws on two waves of data from the Toledo Adolescent Relationships Study, a 19-year population-based longitudinal cohort study of adolescents transitioning to adulthood from Lucas County, Ohio (United States). This investigation includes 878 (399 men and 479 women) emerging adults. To evaluate the association between ACEs and IPV perpetration, two waves of survey data were used, collected in 2001 and 2011-2012. ACEs had a cumulative effect on IPV, where each additional ACE increased the odds of engaging in IPV by 51.0 % (p < 0.001). However, current drug use (OR = 1.131, p < 0.05), arguments between partners (OR = 1.517, p < 0.01), partner mistrust (OR = 1.663, p < 0.001), and jealousy and control (OR = 1.412, p < 0.001) were also significant correlates of IPV reports. ACEs are a significant predictor of IPV perpetration among emerging adults, even when accounting for more proximal risk factors. These findings suggest that individuals working with clients who engage in IPV would do well to address the long-term trauma impacts of early life adversity in addition to more proximal risk factors to reduce the risk of continued violence.


Lumbar instability as an etiology of low back pain and its treatment by prolotherapy: A review.
Low back pain is a significant spinal disorder that affects much of the population at some point during their lives. While proper diagnosis is key, diagnosing the underlying cause of low back pain may often be unclear. In this review article, we discuss lumbar instability as an etiology of low back pain and its treatment by prolotherapy. Spinal ligaments may be an underlying culprit in the development of lumbar instability with resultant low back pain and associated disorders. In these cases, adequate treatment consisting of non-biologic prolotherapy or cellular prolotherapy, including platelet rich plasma (PRP), can be beneficial in restoring spinal stability and resolving chronic low back pain.


Chlamydia pneumoniae together with collagenase-2 (MMP-8) in periodontal lesions.
Dental infections may be associated with subsequent atherosclerosis. In this regard we wanted to study if traces of Chlamydia pneumoniae can be found from deep periodontal pockets characterized with elevated collagenase-2 [matrix metalloproteinase (MMP)-8] levels in gingival crevicular fluid (GCF). Chlamydia pneumoniae has not previously been found to infect the oral cavity. Subgingival samples of dental plaque were collected from 31 teeth in 12 adult periodontitis patients by means of sterile curettes and examined for C. pneumoniae using a quantitative PCR technique. GCF samples were also collected and assayed by an immunofluorometric assay (IFMA) for MMP-8. Chlamydia pneumoniae RNA was demonstrated in a sample from one of the patients studied. Periodontal treatment eliminated the C. pneumoniae from the patient's subgingival dental plaque as well as reduced GCF MMP-8 level. Our findings suggest that C. pneumoniae, which is not normally thought to be involved in periodontitis, can be found in dental plaque.


Comparison of collision-induced dissociation and electron-induced dissociation of phillyrin using FT-ICR MS.
Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry using collision-induced dissociation (CID) and electron capture dissociation (ECD) at high mass resolution was first applied to investigate the characteristic fragment ions of phillyrin. The CID experimental results demonstrated the elemental composition of fragment ions unambiguously, so a reasonable fragmentation pathway of phillyrin was proposed. The ECD fragmentation mechanism was believed to be fundamentally different from the CID method. ECD could be used not only in the biological field but also as a powerful complement to the structural identification of small molecular compounds. The characteristic fragmentation pathways were helpful in analyzing and interpreting the stability and property of the parent ion.


Autonomic activation and endogenous depression.
Psychophysiologic parameters were investigated in 10 patients with endogenous depression during the course of electroconvulsive therapy. Changes in the depressive mood were assessed by the patients at regular intervals using a general mood scale. EEG and heart rate response and habituation were recorded before, during, and after E.C.T. Prior to treatment a low variance and responsiveness and striking intra-individual stability were found in the physiologic parameters. At the start of E.C.T. there was a significant increase in the deviation values of heart rate and a significant increase in the occurrence of typical biphasic heart rate responses. The EEG showed increased amplitudes and a slowing of background activity. Treatment-dependent differences were also seen in the habituation of the EEG amplitude response. Regarding improvement in the depressive mood, differences were found between the retarded and the agitated patients. Agitated patients demonstrated a continous gradual decline of their depressive condition, while in retarded patients the general trend toward improvement passed through a period of increased mood lability. The results are interpreted with reference to the activation dependency of heart rate responses.


Application of case-based learning in psychology teaching: a meta-analysis.
Case-based learning (CBL) has been found to be effective for many subjects, but there is currently a lack of evidence regarding its utility in psychology education. The present study investigated whether CBL pedagogy can improve students' academic performance in psychology courses compared to the traditional teaching methods. A systematic review and meta-analysis were conducted to investigate the effectiveness of CBL in psychology teaching. Databases including PubMed, Embase, Web of Science, China National Knowledge Infrastructure (CNKI), the VIP database, and Wanfang data were searched to find eligible randomized controlled trials. Pooled effect estimates were calculated using Hedges' g under the random effects model, and a subgroup analysis was carried to investigate the heterogeneity among studies. Fifteen studies with 2172 participants, 1086 in the CBL group and 1086 in the traditional lecture-based teaching group, were included in the meta-analysis. Students in the CBL group scored significantly higher on exams than those in the lecture-based group [Hedges' g = 0.68, 95%CI (0.49, 0.88), p < 0.00]. Relatively high heterogeneity was noted among the included studies. Publication bias was examined by the funnel plot and Egger's test, but did not significantly influence the stability of the results. A subsequent evaluation using the trim-and-fill method confirmed that no single study was skewing the overall results. A qualitative review of the included studies suggested that most students in the CBL group were satisfied with the CBL teaching mode. This meta-analysis indicated that the CBL pedagogy could be effective in psychology education, and might help increase students' academic scores, while encouraging a more engaging and cooperative learning environment. At present, the application of CBL in psychology education is in its initial stage. Problems related to the curriculum itself, research methodology, and challenges faced by both teachers and learners have confined its practice. Fully tapping into the strengths of CBL in psychology teaching will require additional work and advancing research.


A 16-year trend of etiology in acute pancreatitis: The increasing proportion of hypertriglyceridemia-associated acute pancreatitis and its adverse effect on prognosis.
Acute pancreatitis (AP) is an acute inflammation of the pancreas, which caused increasing global health and economic burden. Longitudinal trends of disease etiology and its influence on prognosis remains unclear. The aim of this study was to analyze the trend of etiology in moderately severe acute pancreatitis (MSAP) and severe acute pancreatitis (SAP) and to evaluate the influence of the changing pattern of etiology. A 16-year cohort study was undertaken in a tertiary hospital. Patients who presented with MSAP or SAP from January 2001 to December 2016 were continuously enrolled. Demographic data, clinical manifestations, treatment strategy, and prognosis were recorded and verified. A total of 475 patients were included, among whom 173 (36.4%) had gallstone-associated pancreatitis and 108 (22.7%) had hypertriglyceridemia (HTG)-associated pancreatitis. The annual admission for MSAP and SAP rose steadily over the 16-year period (r = 0.907, P < .001), with an annually increasing proportion of HTG-associated pancreatitis (from 14.3% to 35.5%, r = 0.710, P = .015). Compared with gallstone-associated pancreatitis, hypertriglyceridemia-associated pancreatitis had significantly higher percentages of multiple organ dysfunction syndrome (MODS) (24.1% vs 12.1%, P = .009, FDR = 0.029) and cardiovascular failure (17.6% vs 4.6%, P < .001, FDR<0.001). Positive correlations were found between the annual proportion of HTG-associated pancreatitis and MODS (r = 0.574, P = .005) and between HTG-associated pancreatitis and cardiovascular failure (r = 0.512, P = .029). Regression analysis showed that HTG was an independent risk factor for both MODS (P = .004) and cardiovascular failure (P = .009). The number of annually admitted MSAP and SAP cases increased during the last 16 years along with an increasing proportion of HTG-associated AP. The increasing trend of HTG-associated AP might contribute to more frequently occurring MODS and cardiovascular failure.


Experience with a talker can transfer across modalities to facilitate lipreading.
Rosenblum, Miller, and Sanchez (Psychological Science, 18, 392-396, 2007) found that subjects first trained to lip-read a particular talker were then better able to perceive the auditory speech of that same talker, as compared with that of a novel talker. This suggests that the talker experience a perceiver gains in one sensory modality can be transferred to another modality to make that speech easier to perceive. An experiment was conducted to examine whether this cross-sensory transfer of talker experience could occur (1) from auditory to lip-read speech, (2) with subjects not screened for adequate lipreading skill, (3) when both a familiar and an unfamiliar talker are presented during lipreading, and (4) for both old (presentation set) and new words. Subjects were first asked to identify a set of words from a talker. They were then asked to perform a lipreading task from two faces, one of which was of the same talker they heard in the first phase of the experiment. Results revealed that subjects who lip-read from the same talker they had heard performed better than those who lip-read a different talker, regardless of whether the words were old or new. These results add further evidence that learning of amodal talker information can facilitate speech perception across modalities and also suggest that this information is not restricted to previously heard words.


Effects of multiple treatments of isoxicam on organ weights and serum SGPT, AP, and BUN in adjuvant-induced polyarthritic rats.
Isoxicam at 25 mg/kg/day x 28 days significantly inhibited the multiple alterations with the developing polyarthritis. In rats, this drug did not induce any changes in the relative liver weights or serum SGPT, AP, or BUN in polyarthritic rats. Isoxicam absorption was similar in normal and arthritic rats.


Identification of Conserved Proteomic Networks in Neurodegenerative Dementia.
Data-driven analyses are increasingly valued in modern medicine. We integrate quantitative proteomics and transcriptomics from over 1,000 post-mortem brains from six cohorts representing Alzheimer's disease (AD), asymptomatic AD, progressive supranuclear palsy (PSP), and control patients from the Accelerating Medicines Partnership - Alzheimer's Disease consortium. We define robust co-expression trajectories related to disease progression, including early neuronal, microglial, astrocyte, and immune response modules, and later mRNA splicing and mitochondrial modules. The majority of, but not all, modules are conserved at the transcriptomic level, including module C3, which is only observed in proteome networks and enriched in mitogen-activated protein kinase (MAPK) signaling. Genetic risk enriches in modules changing early in disease and indicates that AD and PSP have distinct causal biological drivers at the pathway level, despite aspects of similar pathology, including synaptic loss and glial inflammatory changes. The conserved, high-confidence proteomic changes enriched in genetic risk represent targets for drug discovery.


Simultaneous determination of amines and isocyanates in working atmospheres by gas-liquid chromatography.
A gas-liquid chromatographic method has been developed which permits trace analysis in a single run of amines and isocyanates occurring in relative concentrations of the order of 10,000:1. The amines were determined as free amines after alkalization of the sample and an extractive enrichment into toluene. The isocyanates were hydrolyzed to the corresponding amines during sampling in dilute sulphuric acid. This method gives the total isocyanate concentration. The use of a nitrogen sensitive detector and an alkali-treated packing, Pennwalt 223 with 4% KOH, are important prerequisites. Chromatographic parameters such as column liquid loading, injector impregnation and injector temperature were examined. Good separation between 2,4- and 2,6-toluenediamine isomers was obtained. The enrichment step was studied with respect to the degree of alkalization and to interferences from another amine present in large excess. The method was tested in the working atmospheres of two polyurethane foam factories. The substances determined were N-methylmorpholine, 1,4-diazabicyclo[2,2,2]octane and 2,4- and 2,6-toluenediisocyanates.


Preludes to retinal detachment in the aphakic eye.
The author reviews the characteristic differences in primary rhegmatogenous retina detachment occurring in phakic eyes and in aphakic eyes. The preoperative findings in aphakic detachment are explained on the basis of two factors: surgical trauma during cataract extraction and postoperative changes in the macromolecular compostion of the vitreous.


Compliance with wearing a thoracolumbar orthosis in nonoperative treatment of osteoporotic vertebral fractures: a prospective sensor-controlled study.
Hyperextension orthoses (HOs) for nonoperative treatment of osteoporotic vertebral fractures (OVFs) are widely prescribed. However, the compliance, how much an HO is worn after it has been prescribed, is widely unknown. This study was performed to assess the wearing time of HOs for OVFs in a prospective blinded, sensor-controlled manner. A prospective, single blinded observational study was performed. This study prospectively included 18 patients who were treated nonoperatively with an HO for OVFs. The true wearing time was measured using a hidden temperature-based sensor. The patients were invited to return for regular follow-up every 2 weeks for 6 weeks, at which time clinical evaluation (including a visual analog scale for pain and the Oswestry disability questionnaire) and radiographs of the spine were performed. Full compliance was defined as a wearing time of 15 hours per day. Correlation between compliance and demographic differences, patient reported outcomes and radiographic changes of the vertebral structures were calculated. The mean HO wearing time was 5.5±3.3 hours (37%±22% compliance). Female patients used the HO significantly longer per day than did male patients (6.5±3.2 vs 2.9±2.0 hours, p=.039). Age and body mass index had no influence on wearing behavior. Overall, compliance with wearing HOs is poor and shows great variability with significant gender-dependency but not associated with BMI, age, or pain-level. Further studies are required to confirm our results that the wearing time does not have an influence on kyphotic progression of the osteoporotic fractured segment, nor on clinical outcome at short term. I.


Implementation of a School Physical Activity Policy Improves Student Physical Activity Levels: Outcomes of a Cluster-Randomized Controlled Trial.
To assess the impact of a multistrategy intervention designed to improve teachers' implementation of a school physical activity (PA) policy on student PA levels. A cluster-randomized controlled trial was conducted in 12 elementary schools. Policy implementation required schools to deliver 150 minutes of organized PA for students each week via physical education, sport, or class-based activities such as energizers. Schools received implementation support designed using the theoretical domains framework to help them implement the current policy. A total of 1,502 children in kindergarten to grade 6 participated. At follow-up compared with control, students attending intervention schools had, measured via accelerometer, significantly greater increases in school day counts per minute (97.5; 95% confidence interval [CI], 64.5 to 130.4; P < .001) and moderate to vigorous physical activity (MVPA) (3.0; 95% CI, 2.2-3.8, P < .001) and a greater decrease in sedentary time (-2.1; 95% CI, -3.9 to -0.4, P = .02) per school day. Teachers in intervention schools delivered significantly more minutes (36.6 min) of PA to their students at follow-up (95% CI, 2.7-70.5, P = .04). Supporting teachers to implement a PA policy improves student PA. Additional strategies may be needed to support teachers to implement activities that result in larger gains in student MVPA.


Metachronous appearance of second malignancies in medullary thyroid carcinoma (MTC) patients: a diagnostic challenge and brief review of the literature.
Medullary thyroid carcinoma (MTC) is a rare tumour which frequently occurs in the context of the multiple endocrine neoplasia syndromes, where it coexists with other usually benign tumours. The clinical picture varies and distant metastases are frequently present at diagnosis. Calcitonin levels are elevated in the presence of metastatic disease. Two MTC cases are presented, which had elevated postoperative calcitonin levels. Imaging revealed lung lesions which were originally attributed to metastatic disease from the MTC. However, at follow-up, these cases presented unusual features. The rapid increase in the lung lesions and the development of hypercalcaemia in the first patient suggested a second unrelated tumour. Biopsy of the lung lesion was compatible with lung adenocarcinoma. In the second patient, the appearance of a liver mass, although calcitonin levels remained stable, led to biopsy of the lesion: this was negative for calcitonin and compatible with metastatic lung adenocarcinoma. These MTC cases show that further malignancies may coexist with MTC and may obscure the clinical picture and influence the therapeutic decisions, especially in the case of metastatic disease. Features such as unusual imaging characteristics and the development of hypercalcemia, never encountered in MTC outside the MEN2 syndromes, as well as 'disproportionately' low calcitonin levels, incompatible with extensive metastatic disease, were the factors that led to further work-up. Both the cases subsequently proved to carry an unsuspected second malignancy. It is crucial to discriminate the metastatic lesion attributed to MTC from another coexisting primary malignancy, because different therapeutic strategies are needed for each setting.


A STING-dependent innate-sensing pathway mediates resistance to corneal HSV-1 infection via upregulation of the antiviral effector tetherin.
Type 1 interferons (IFNs; IFNα/β) mediate immunological host resistance to numerous viral infections, including herpes simplex virus type 1 (HSV-1). The pathways responsible for IFNα/β signaling during the innate immune response to acute HSV-1 infection in the cornea are incompletely understood. Using a murine ocular infection model, we hypothesized that the stimulator of IFN genes (STING) mediates resistance to HSV-1 infection at the ocular surface and preserves the structural integrity of this mucosal site. Viral pathogenesis, tissue pathology, and host immune responses during ocular HSV-1 infection were characterized by plaque assay, esthesiometry, pachymetry, immunohistochemistry, flow cytometry, and small interfering RNA transfection in wild-type C57BL/6 (WT), STING-deficient (STING(-/-)), and IFNα/β receptor-deficient (CD118(-/-)) mice at days 3-5 postinfection. The presence of STING was critical for sustained control of HSV-1 replication in the corneal epithelium and resistance to viral neuroinvasion, but loss of STING had a negligible impact with respect to gross tissue pathology. Auxiliary STING-independent IFNα/β signaling pathways were responsible for maintenance of corneal integrity. Lymphatic vessels, mast cells, and sensory innervation were compromised in CD118(-/-) mice concurrent with increased tissue edema. STING-dependent signaling led to the upregulation of tetherin, a viral restriction factor we identify is important in containing the spread of HSV-1 in vivo.


Ultrastructure of oogenesis of two oviparous demosponges: Axinella damicornis and Raspaciona aculeata (Porifera).
We investigated the cytology of the oogenic cycle in two oviparous demosponges, Axinella damicornis and Raspaciona aculeata, during 2 consecutive years both by light and electron microscopy. Oocytes of both species were similar in their basic morphological features but differences were noticed in time required to complete oocyte maturation and mechanisms of acquisition of nutritional reserves. The oogenic cycle of A. damicornis extended for 7-8 months in autumn-spring, while that of R. aculeata did it for 3-5 months in summer-autumn. Yolk of A. damicornis was predominantly formed by autosynthesis. Oocytes endocytosed bacteria individually and stored them in groups in large vesicles. Bacteria were digested and lipidic material was added to the vesicles to produce a peculiar granular yolk hitherto unknown in sponges. Scarce cells carrying heterogeneous inclusions were observed in the perioocytic space, and were interpreted as putative nurse cells. Such cells were presumably releasing lipid granules to the perioocytic space. In contrast, large numbers of nurse cells were found surrounding the oocytes of R. aculeata. They transported both lipid granules and heterogeneous yolk bodies to the oocytes. R. aculeata also produced some of their yolk by autosynthesis. The involvement of nurse cells in the vitellogenesis of R. aculeata shortened the oocyte maturation, whereas a largely autosynthetic vitellogenesis in A. damicornis prolonged the duration of oogenesis.


Keap1 as Target of Genistein on Nrf2 Signaling Pathway Antagonizing Aβ induced Oxidative Damage of Cerebrovascular Endothelial Cells.
β-amyloid peptides (Aβ) induced oxidative damage contributes to the pathogenesis of neurodegenerative diseases, and the cerebrovascular system is more vulnerable to oxidative stress. Our earlier study showed a clue that Genistein (Gen) might activate the Nf-E2 related factor 2 (Nrf2) pathway to protect cerebrovascular cells from oxidative damage induced by Aβ, but the specific mechanisms and regulation targets are unclear. In this study, the anti-oxidative effects and the possible targets of Gen on regulating the Nrf2 pathway in bEnd.3 cells were investigated. Cells were divided into control, Aβ25-35, Gen, and Gen+Aβ25-35 groups. Cell viability, levels of malondialdehyde (MDA), Superoxide Dismutase (SOD) activity, and nitrotyrosine were evaluated. Moreover, mRNA and/or protein expressions of Nrf2 and kelchlike ECH-associated protein 1 (Keap1) were measured. Then we transfected Keap1 over-expressed plasmid into bEnd.3 cells and measured the protein expressions of Nrf2 pathway related factors. Data showed that Gen could inhibit the over-production of MDA and nitrotyrosine and activate SOD activity. Furthermore, we discovered that Gen could up-regulate Nrf2 mRNA and protein expression while down-regulating Keap1 protein expression. The Keap1 over-expressed plasmid study revealed that the up-regulation of Nrf2 protein expression induced by Gen pretreatment could be blocked by transfection of Keap1 over-expressed plasmid, and the same results were observed in Nrf2 downstream factors. Gen could alleviate the cerebrovascular cells' oxidative damage induced by Aβ25-35 by regulating the Nrf2 pathway, and Keap1 might be one of the targets of Gen in activating the Nrf2 pathway.


Contributions of genotoxic precursors from tributary rivers and sewage effluents to the Yodo River in Japan.
The cause of water pollution in the Yodo River was examined regarding the mutagenic potentials formed by chlorination of river waters and sewage effluents along the river. Mutagenicity of the extracts from chlorinated waters was measured by Ames test using S. typhimurium TA100 strain without metabolic activation. Mutagenic formation potentials of sewage effluents were 4-5 times higher than those of the three tributary river waters, the Kizu, Uji and Katsura Rivers. The contribution rates of the pollution loads from the three river waters and total of four sewage effluents to the Yodo River were revealed to be 10.2%, 42.2%, 22.1% and 25.5%, respectively. The rate of the sewage effluents occupied one-fourth and exceeded the rates of the Kizu and Katsura Rivers. The contribution rate of the two sewage effluents to the Katsura River was 46.3%. It could be confirmed that the relationship between COD and MFP in river waters and sewage effluents was very high with a correlation coefficient. These results indicated that the waters inflowing to the Yodo River were contaminated with numerous organic compounds, and the sewage effluents especially contribute to the formation of genotoxicity by the interaction between chlorine and organic components as genotoxic precursors in the purification plants.


Hemostasis vs. homeostasis: Platelets are essential for preserving vascular barrier function in the absence of injury or inflammation.
Platelets are best known for their vasoprotective responses to injury and inflammation. Here, we have asked whether they also support vascular integrity when neither injury nor inflammation is present. Changes in vascular barrier function in dermal and meningeal vessels were measured in real time in mouse models using the differential extravasation of fluorescent tracers as a biomarker. Severe thrombocytopenia produced by two distinct methods caused increased extravasation of 40-kDa dextran from capillaries and postcapillary venules but had no effect on extravasation of 70-kDa dextran or albumin. This reduction in barrier function required more than 4 h to emerge after thrombocytopenia was established, reverting to normal as the platelet count recovered. Barrier dysfunction was also observed in mice that lacked platelet-dense granules, dense granule secretion machinery, glycoprotein (GP) VI, or the GPVI signaling effector phospholipase C (PLC) γ2. It did not occur in mice lacking α-granules, C type lectin receptor-2 (CLEC-2), or protease activated receptor 4 (PAR4). Notably, although both meningeal and dermal vessels were affected, intracerebral vessels, which are known for their tighter junctions between endothelial cells, were not. Collectively, these observations 1) highlight a role for platelets in maintaining vascular homeostasis in the absence of injury or inflammation, 2) provide a sensitive biomarker for detecting changes in platelet-dependent barrier function, 3) identify which platelet processes are required, and 4) suggest that the absence of competent platelets causes changes in the vessel wall itself, accounting for the time required for dysfunction to emerge.


Klebsiella pneumoniae type 3 fimbria-mediated immunity to infection in the murine model of respiratory disease.
Type 3 fimbriae are expressed by most strains of Klebsiella pneumoniae and facilitate adherence to the basement membrane of human respiratory tissues. The ability of these appendages to stimulate a protective immune response in vivo has not been investigated. A murine model of acute pneumonia was used to determine whether the production of type 3 fimbria-specific antibodies correlated with protection against infection by K. pneumoniae. Purified fimbriae from several strains were used to immunize mice prior to challenge with a virulent strain. The immunized mice produced high titers of specific antibody and this was associated with protection against challenge with a low dose of bacteria that was lethal in unimmunized animals. However, challenge with a high number of bacteria resulted in no protection against infection.


Minute theropod eggs and embryo from the Lower Cretaceous of Thailand and the dinosaur-bird transition.
We report on very small fossil eggs from the Lower Cretaceous of Thailand, one of them containing a theropod embryo, which display a remarkable mosaic of characters. While the surficial ornamentation is typical of non-avian saurischian dinosaurs, the three-layered prismatic structure of the eggshell is currently known only in extant and fossil eggs associated with birds. These eggs, about the size of a goldfinch's, mirror at the reproductive level the retention of small body size that was paramount in the transition from non-avian theropods to birds. The egg-layer may have been a small feathered theropod similar to those recently found in China.


Inhibition of urea hydrolysis by free Cu concentration of soil solution in microbially induced calcium carbonate precipitation.
Urea hydrolysis is an initiating step of microbially induced calcium carbonate precipitation (MICP) which can be used as a stabilization technology in heavy metals contaminated soil. In this study, inhibition of urea hydrolysis was investigated in Cu-contaminated soil. At soil Cu concentration from 0 to 1000 mg/kg, the amount of urea hydrolyzed (i.e., initial urea 450 mM) ranged from 449.3 ± 1.4 to 10.5 ± 0.8 mM. Correspondingly, decrease in calcium carbonate precipitation was commensurate with the inhibition of urea hydrolysis. Interestingly, 2.75 times more urea were hydrolyzed in 350 days-aged soil than in freshly spiked soil even at the same soil Cu concentration of 250 mg/kg, suggesting the inhibitory effect of Cu in soil solution. Indeed, the concentrations of Cu in soil solution were 4.9 ± 0.1 and 21.0 ± 0.3 mg/L, respectively. Since MICP application involved an increase in Ca2+ concentration in soil, its effect was also determined. In the freshly spiked soil with 250 mg-Cu/kg, the Cu concentration in the soil solution increased from 7.6 ± 0.1 to 21.0 ± 0.3 mg/L as the calcium concentration increased from 0 to 450 mM. Accordingly, urea hydrolysis was significantly reduced from 217.5 ± 59.0 to 11.9 ± 0.2 mM. The effect of pH was also determined, showing that 32.8 ± 3.4 and 205.9 ± 32.5 mM of urea was hydrolyzed at soil pH of 4.5 and 7.8, respectively. The reason was attributed to the great difference in free Cu concentration in soil solution (i.e., 3.3 and 0.3 mg/L at pH 4.5 and 7.8, respectively). The relationship between amounts of urea hydrolyzed and free Cu concentrations was established and half-maximal inhibition concentration (IC50) of free Cu concentration in soil solution was predicted to be 0.39 mg/L.


CD69 prevents PLZFhi innate precursors from prematurely exiting the thymus and aborting NKT2 cell differentiation.
While CD69 may regulate thymocyte egress by inhibiting S1P1 expression, CD69 expression is not thought to be required for normal thymocyte development. Here we show that CD69 is in fact specifically required for the differentiation of mature NKT2 cells, which do not themselves express CD69. Mechanistically, CD69 expression is required on CD24+ PLZFhi innate precursors for their retention in the thymus and completion of their differentiation into mature NKT2 cells. By contrast, CD69-deficient CD24+ PLZFhi innate precursors express S1P1 and prematurely exit the thymus, while S1P1 inhibitor treatment of CD69-deficient mice retains CD24+ PLZFhi innate precursors in the thymus and restores NKT2 cell differentiation. Thus, CD69 prevents S1P1 expression on CD24+ PLZFhi innate precursor cells from aborting NKT2 differentiation in the thymus. This study reveals the importance of CD69 to prolong the thymic residency time of developing immature precursors for proper differentiation of a T cell subset.


An Exploration of the Risk Factors, Severity, and Sequelae of Vocal Fold Hemorrhage in a Population of Voice Patients Before and After Diagnosis.
Vocal fold hemorrhage (VFH) is the rupture (usually acute) of a blood vessel within the true vocal fold. The long-term sequelae of VFH on the mucosal wave (MW) and glottic gap on video stroboscopy remain understudied. The primary objective of this study was to investigate the short-term and long-term consequences of VFH through measured and rated analysis of the mucosal wave and glottic gap. The presence of VFH and its extent (limited/moderate vs. extensive VFH) were identified. The primary outcome of this study was mucosal wave, which was assessed on an ordinal scale by three blinded raters pre and posthemorrhage. Only patients who had undergone strobovideolaryngoscopy before sustaining VFH were included. Mucosal wave and glottic gap also were measured using image pixel analysis using the open-access tool, ImageJ (NIH, Bethesda, MD). Twenty-three subjects were included in this study (mean age 39.78 ± 15.54). Intra-rater reliability for MW ratings was 81.48% ± 6.150% (minimum 77.78%) for all evaluators (κ = 0.519 [0.267-0.772], P < 0.001). Inter-rater reliability analysis revealed 75.56% agreement between evaluators (κ = 0.524 [0.425-0.623], P < 0.001). MWMeasured extrapolated from ImageJ methodology correlated significantly with MWRated (n = 70, r = 0.448, P < 0.001). ΔMWMeasured from baseline to follow-up evaluation were compared for both the initial follow-up visit (FU1) and the second follow-up visit (FU2) [-4.135 ± 31.01 vs. 36.50 ± 39.97, P = 0.025]. Hence, ΔMWMeasured was significantly better by FU2 than FU1, with the larger positive change from baseline representing a greater improvement in the measured mucosal wave. Additionally, there were significant differences in ΔMWRated between those with limited/moderate VFH and those with extensive VFH at FU1. Duration of absolute voice rest correlated significantly with time to VFH resolution. Long-term change in mucosal wave after hemorrhage was assessed using both ΔMWMeasured and MWRated. Based on ΔMWRater, 35.0% of subjects demonstrated ongoing and worse mucosal wave restriction compared to baseline at their most recent follow-up visit. Based on ΔMWMeasured, 50.0% of the subjects showed ongoing and worse mucosal wave restriction compared to baseline at their most recent follow-up visit. Overall long-term restrictions in MW after hemorrhage were present in 35.0% of the subjects based on ratings and 50.0% of the patients based on the measured MW using ImageJ, demonstrating the importance of ongoing study into this pathology and how to prevent it, especially in PVU and professional singers. Patients presenting with extensive hemorrhage were at risk for more prominent, detectable changes in mucosal wave compared to those with limited/moderate hemorrhage in the short-term, defined by a mean FU time of 3 months, but not long-term, characterized by a mean FU time 6 months or greater. Whether the severity of VFH is a true indicator of mucosal wave alterations requires additional study, as does the reliability and validity of ImageJ mucosal wave and glottic gap measurement techniques.


[Construction for detecting of human papillomaviruses types by using a multiplexed Luminex assay].
To develop multiplex human papillomavirus genotyping, a quick and sensitive high-throughput procedure for the identification of multiple HPV genotypes by using a multiplexed Luminex array. Standard plasmids containing HPV genomic DNA were developed a standard multiplexed suspension array for detection 13 HPV types on Luminex analysis platform. the primer and probes were selected, HPV type-specific oligonucleotide probes were coupled to fluorescence-labeled polystyrene beads. The one or two of mixed HPV standard plasmids were identified HPV type by using 13 HPV genetypes multiplexed suspension array, different HPV types can be correctly detected simultaneously. The positives of HPV clinical samples were detected by using this method, and the genetypes of samples by detecting were the same results with using BLAST in NCBI. The proposed method allowed a high through-out, special, simple, rapid and economical identification of HPV DNA genotypes, It is expected to be an extremely useful tool in the identification of HPV genetypes for a lot of clinical samples.


Peripheral T-cell lymphoma of AILD (angioimmunoblastic lymphadenopathy with dysproteinemia) type involving gastrointestinal tract. A morphologic, phenotypic and genotypic study.
A case of angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) which showed widespread involvement of the gastrointestinal tract is reported. A lymph node biopsy specimen showed the characteristic histological features of AILD. During the progression of the illness, lymphomatous lesions developed in the gastrointestinal tract, complicated by cytomegalovirus infection. A double immunoenzymatic study using a combination of Ki-67 antibody and antibodies against surface antigens demonstrated that CD3+, CD4+, and/or T-cell receptor (TCR) beta+ cells were predominant (67-68%) among the population of proliferating Ki-67% cells, rather than CD8+ or CD22+ cells. Clonal rearrangement of the TCR beta chain gene was also detected. These findings provide further evidence for the neoplastic nature of lesions of this type, and the diagnosis of peripheral T-cell lymphoma.


[On the problem of psychological (psychometric) diagnosis of intelligence in children with developmental disorders].
A review includes the following sections corresponded with methods of investigation and groups of patients: Wechsler test for atypical autism and schizophrenia; Psycho Educational Profile (PEP) in endogenous mental pathology, the Kaufman Test of Educational Achievement in children schizophrenia, Edinburgh Picture Test in mental dysontogenesis. The results of the studies in these fields are presented.


[Treatment of alcohol dependence in primary health care can provide good results. The 15-method--a structured care model matched to patient needs].
The main obstacle for people with an alcohol problem to seek help is the stigma attached to alcohol problems and alcohol treatment. Seeking help in primary health care is less stigmatised. Alcohol dependence is mostly of mild or moderate severity and can consequently be managed by non-specialists to a large extent. The 15-method is a structured stepped care model with evidence-based components, where interventions are matched to patient need. The first step, to recognise the impact of alcohol on health, should be mastered by all health professionals.


A communication from the left ventricle to the right atrium: a defect in the central fibrous body.
An isolated defect in the membranous atrioventricular septum was found in a five-year-old girl who presented with a cystic lesion in the right atrium. This type of left ventricular-right atrial communication could result from a structural abnormality of the central fibrous body in combination with arrested maturation of the membranous ventricular septum.


Therapeutic effect of Artemia enriched with Escherichia coli expressing double-stranded RNA in the black tiger shrimp Penaeus monodon.
We exploited Artemia as a double-stranded (ds)RNA-delivery system to combat viral diseases in shrimp. First, the transformed Escherichia coli (E. coli) expressing red fluorescent protein (RFP) was tested in the Artemia enrichment process. RFP signals detectable in the gut of Artemia under confocal microscope were evident for the successful encapsulation. Second, the Artemia enrichment process was performed using E. coli producing Laem-Singh virus (LSNV)-specific dsRNA, which has been previously shown to inhibit the viral infection in the black tiger shrimp Penaeus monodon by intramuscular injection and oral administration. The enriched Artemia nauplii were confirmed to contain dsRNA-LSNV by RT-PCR, and were subjected to the feeding test with P. monodon postlarvae. Quantitative RT-PCR indicated that a number of LSNV copies in most of the treated shrimp were, at least, 1000-fold lower than the untreated controls. During 11-17weeks after feeding, average body weight of the treated group was markedly increased relative to the control group. A smaller differential growth rate of the treated group as compared to the control was also noticed. These results suggested that feeding shrimp with the dsRNA-enriched Artemia can eliminate LSNV infection, which is the cause of retarded growth in P. monodon. The present study reveals for the first time the therapeutic effect of dsRNA-enriched Artemia for shrimp disease control.


Building Specialized Nursing Practice Capacity in Bangladesh: An Educational Program to Prepare Nurses to Care for Oncology and Bone Marrow Transplant Patients in Dhaka, Bangladesh.
In 2012, the Minister of Health and other leaders in the Bangladesh government approached Massachusetts General Hospital to establish the country's first bone marrow transplant program at Dhaka Medical College Hospital to serve the needs of the people of Bangladesh. Stated goals of this collaboration included a broad focus on the care of oncology patients with a specific emphasis on care of patients with hematologic malignancies and of women with gynecologic cancers. The purpose of this article is to describe the international nursing collaboration between Massachusetts General Hospital, Simmons College, the AK Khan Healthcare Trust in Dhaka, and Dhaka Medical College Hospital that was established to share nursing knowledge and to build specialized professional nursing capacities to deliver high-quality cancer care in the public sector. Over the past 3 years, through the educational programs that have been developed within this collaboration-the Enhanced Specialized Nurse Training Program-the Bangladeshi nurses have received continuing professional development based on Western standards of nursing and have been offering nursing care to patients who have undergone chemotherapy and bone marrow transplantation. The challenges, opportunities, and outcomes of this international collaboration have been highly rewarding and mutually beneficial.


Development and validation of a risk score to predict unplanned hospital readmissions in ICU survivors: A data linkage study.
Intensive Care Unit (ICU) follow-up clinics are growing in popularity internationally; however, there is limited evidence as to which patients would benefit most from a referral to this service. The objective of this study was to develop and validate a model to predict which ICU survivors are most likely to experience an unplanned hospital readmission or death in the year after hospital discharge and derive a risk score capable of identifying high-risk patients who may benefit from referral to follow-up services. A multicentre, retrospective observational cohort study using linked administrative data from eight ICUs was conducted in the state of New South Wales, Australia. A logistic regression model was developed for the composite outcome of death or unplanned readmission in the 12 months after discharge from the index hospitalisation. 12,862 ICU survivors were included in the study, of which 5940 (46.2%) patients experienced unplanned readmission or death. Strong predictors of readmission or death included the presence of a pre-existing mental health disorder (odds ratio [OR]: 1.52, 95% confidence interval [CI]: 1.40-1.65), severity of critical illness (OR: 1.57, 95% CI: 1.39-1.76), and two or more physical comorbidities (OR: 2.39, 95% CI: 2.14-2.68). The prediction model demonstrated reasonable discrimination (area under the receiver operating characteristic curve: 0.68, 95% CI: 0.67-0.69) and overall performance (scaled Brier score: 0.10). The risk score was capable of stratifying patients into three distinct risk groups-high (64.05% readmitted or died), medium (45.77% readmitted or died), and low (29.30% readmitted or died). Unplanned readmission or death is common amongst survivors of critical illness. The risk score presented here allows patients to be stratified by risk level, enabling targeted referral to preventative follow-up services.


Coronary steal syndrome with coil embolization of a large LIMA side branch: radionuclide evidence for reversible ischemia.
This case presents the controversy over coronary artery steal syndrome following bypass surgery when a large branch of the left internal mammary artery (LIMA) is not ligated. A discussion of previous attempts to understand the physiology of this anatomy is compared with case reports of objective evidence for ischemia that resolves following occlusion of the LIMA side branch.


Two-Dimensional Device with Light-Controlled Capability for Treatment of Cancer-Relevant Infection Diseases.
Concurrent infection in cancer treatment is the leading cause of high cancer mortality that requires urgent action. Currently developed diagnostic methods are hindered by the difficulty of rapidly and reliably screening small amounts of pathogens in the blood and then release pathogens for downstream analysis, limiting the advance of cancer concurrent infection diseases diagnosis and targeted treatment. Herein, we present a near-infrared (NIR) light-responsive black phosphorus (BP)-based device that effectively captures and releases pathogen for downstream drug-resistance analysis. The aptamer-modified BP nanostructures exhibit enhanced topographical interactions and binding capabilities with pathogen, enabling highly efficient and selective capture of pathogen in serum. NIR light irradiation induces BP nanostructure to generate a local thermal effect, which regulates the three-dimensional structure of the aptamer and causes efficient release of pathogen from the substrate surface. The released pathogen is resistant to ampicillin as demonstrated by downstream genetic analysis. The design of the functionalized light-controlled device for monitoring pathogen behavior shows great potential for assisting in cancer therapy and promoting personalized healthcare.


Prototheca zopfii: natural, transient, occurrence in pigs and rats.
Domestic swine faeces and fresh faeces from trapped barnyard rats were heavily contaminated with Prototheca zopfii, a cause of dairy cow mastitis. When the pigs and rats were maintained on Prototheca-free diets, the transient intestinal population of P. zopfii decreased precipitously and disappeared. When combined with the information that other farm animals excrete P. zopfii, it was concluded that contaminated animal feed may be the source of large numbers of P. zopfii in the farm environment. We found P. zopfii in wet spoiled feed. Rats are logical vectors for contamination of feed.


An oligomeric switch controls the Mrr-induced SOS response in E. coli.
Mrr from Escherichia coli K12 is a type IV restriction endonuclease whose role is to recognize and cleave foreign methylated DNA. Beyond this protective role, Mrr can inflict chromosomal DNA damage that elicits the SOS response in the host cell upon heterologous expression of specific methyltransferases such as M.HhaII, or after exposure to high pressure (HP). Activation of Mrr in response to these perturbations involves an oligomeric switch that dissociates inactive homo-tetramers into active dimers. Here we used scanning number and brightness (sN&B) analysis to determine in vivo the stoichiometry of a constitutively active Mrr mutant predicted to be dimeric and examine other GFP-Mrr mutants compromised in their response to either M.HhaII activity or HP shock. We also observed in vitro the direct pressure-induced tetramer dissociation by HP fluorescence correlation spectroscopy of purified GFP-Mrr. To shed light on the linkages between subunit interactions and activity of Mrr and its variants, we built a structural model of the full-length tetramer bound to DNA. Similar to functionally related endonucleases, the conserved DNA cleavage domain would be sequestered by the DNA recognition domain in the Mrr inactive tetramer, dissociating into an enzymatically active dimer upon interaction with multiple DNA sites.


Light chain dependent effects of actin binding on the S-1/S-2 swivel in myosin.
The S-1/S-2 swivel in myosin provides a flexible link between the head and tail portions of the molecule. We have investigated the properties of the swivel by employing limited proteolysis methods. Our results indicate that the binding of actin to heavy meromyosin inhibits both the chymotryptic and papain cleavage of the S-1/S-2 swivel, and that this effect is dependent on the presence of intact LC-2 light chains. Actin did not slow digestions carried out using heavy meromyosin previously treated with proteases to nick the LC-2 chains to 17,000 or 14,000 Mr fragments. Although the integrity of the LC-2 light chain appears to be required to transmit the effects of actin binding from the myosin head to the S-1/S-2 swivel, the binding of Ca2+ to the 17,000 Mr LC-2 fragment can still affect the chemical reactivity of SH1 thiol groups. Both chymotryptic and papain digestions of heavy meromyosin containing intact or fragmented LC-2 light chain show substantial temperature sensitivity between 5 degrees C and 35 degrees C. Calculated apparent activation energies for this process indicate that the S-1/S-2 swivel in myosin can undergo temperature-dependent structural changes independently of the state of the LC-2 light chain. Thus, both actin binding and temperature variations can induce structural transitions in the S-1/S-2 swivel.


[Practical aspects of pacemaker and ICD-lead extractions].
Special tools for lead removal enables transvenous lead extractions without cardiac exposure. The risk of fatal complications during extraction of long-term implanted leads requires detailed knowledge and trained physicians. In addition to patients' age and gender, individual extraction risk factors are access and time since implantation, lead position, kind and number of leads. Locking stylets to anchor the lead within the lumen are necessary in all extraction procedures while the use of external sheaths is optional. Higher risk of cardiac or central vein perforation during lead mobilisation with external laser sheaths has to be respected, and these tools should be used with strong indication only. In cases of high risk, lead explantation with an open heart procedure should be considered as an alternative.


Phorbol ester induces intracellular translocation of phospholipid/Ca2+-dependent protein kinase and stimulates amylase secretion in isolated pancreatic acini.
Treatment of intact rat pancreatic acini with phorbol ester (12-O-tetradecanoyl-phorbol-13-acetate, TPA) resulted in a time- and concentration-dependent translocation of phospholipid/Ca2+-dependent protein kinase (PL/Ca-PK) from the soluble fraction. Redistribution of PL/Ca-PK was concurrent with stimulation of amylase secretion by TPA-treated acini. Polymyxin B, a potent and selective inhibitor of PL/Ca-PK completely inhibited TPA-induced amylase secretion. These findings are consistent with a role for PL/Ca-PK in the regulation of pancreatic exocrine secretion.


A potentiostatic study of oxygen transmissibility and permeability through hydrogel membranes.
The permeability characteristics of membranes prepared from hydrogels of poly(2-hydroxyethylmethacrylate) (PHEMA) and poly(2-hydroxyethylmethacrylate-co-N,N-dimethylacrylamide) (PHNDA) are described. True values of the permeability and transmissibility coefficients of oxygen in the membranes are determined by using electrochemical procedures involving the measurement of the steady state current either in membranes with different thickness or in a single membrane in which its thickness is varied with layers of moistened paper. Comparison of the results obtained for the transport properties in these hydrogels with others obtained in other hydrogels permit to conclude that the degree of swelling rather than the chemical nature of the hydrogels affects the permeation properties. The chemical structure presumably only affects in high degree the chemical stability and flexibility of the hydrogel membranes.


Slowing the progression of aortic stenosis.
Aortic stenosis (AS) is the most frequent valvular heart disease in the Western world. The only definitive treatment for this disorder is surgical replacement of the aortic valve. Recent histopathologic studies have provided important insights into the pathogenesis of calcific AS. These results suggest that this disease develops as the result of an active disease process at the cellular and molecular levels that shares many similarities with vascular atherosclerosis, ranging from endothelial dysfunction to, ultimately, calcification. In this review, we discuss the cellular and molecular mechanisms, natural history, diagnosis, and therapeutic options, including the potential for medical therapies, for the treatment of AS.


Unusual MRI findings in a patient with history of frontal fracture and skin infestation by fly larvae, as a possible sign of intracerebral myiasis.
We report a case of an 11-year-old columbian immigrant with mild non-specific cephalalgia. He had a previous history of frontal fracture and skin infestation caused by Dermatobia hominis larvae. MRI performed revealed multiple subependymal and intraventricular lesions with concentric blooming artifacts and moderate hydrocephalus. Based on his previous history, intracerebral myiasis diagnosis was suggested. His mother denied any kind of diagnostic surgery or treatment. To the best of our knowledge, this is the first MRI report of a possible intracerebral myiasis, an exceedingly rare entity.


Associations between psychiatric comorbidities and sleep disturbances in children with attention-deficit/hyperactivity disorder.
Children with attention-deficit/hyperactivity disorder (ADHD) often have sleep complaints and also higher rates of psychiatric comorbidities such as mood and anxiety disorders that may affect sleep. The authors hypothesized that children with ADHD and psychiatric comorbidities would have higher overall sleep disturbance scores as measured by a sleep questionnaire than children with ADHD without comorbidities. This cross-sectional analysis in an academic center studied 317 children with ADHD; 195 subjects had no comorbid conditions, 60 were anxious and 62 were depressed. Participants completed the Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present State, 4th Revised Edition and the Children's Sleep Habits Questionnaire. Median age (range) was 8.9 (6-18.7) years; 78% were male. Median (interquartile range) Total Sleep Disturbance Score (TSDS) on Children's Sleep Habits Questionnaire for subjects with no comorbidities was 44 (40-49); anxiety, 48 (43-54); and depression, 46 (41-52). Compared with subjects without comorbidities, TSDS in anxious subjects was greater (p = .008). TSDS in depressed subjects was not significantly different. Compared with subjects without comorbidities, anxious subjects had higher Bedtime Resistance, Sleep Onset Delay, and Night Wakings subscales (p = .03, .007, and .007, respectively); depressed subjects had higher Sleep Onset Delay and Sleep Duration subscales (p = .003 and .01, respectively). Anxiety in children with ADHD contributed to higher overall sleep disturbance scores, compared with children with ADHD alone. Both comorbidities were associated with higher Sleep Onset Latency subscale scores. Further study of the impact of psychiatric comorbidities on sleep in children with ADHD is warranted.


Temporal variability profiling of the default mode across epilepsy subtypes.
Epilepsies are a group of neurological disorders sharing certain core features, but also demonstrate remarkable pathogenic and symptomatic heterogeneities. Various subtypes of epilepsy have been identified with abnormal shift in the brain default mode network (DMN). This study aims to evaluate the fine details of shared and distinct alterations in the DMN among epileptic subtypes. We collected resting-state functional magnetic resonance imaging (MRI) data from a large epilepsy sample (n = 371) at a single center, including temporal lobe epilepsy (TLE), frontal lobe epilepsy (FLE), and genetic generalized epilepsy with generalized tonic-clonic seizures (GGE-GTCS), as well as healthy controls (HC, n = 150). We analyzed temporal dynamics profiling of the DMN, including edge-wise and node-wise temporal variabilities, and recurrent dynamic states of functional connectivity, to identify abnormalities common to epilepsies as well as those specific to each subtype. The analyses revealed that hypervariable edges within the specific DMN subsystem were shared by all subtypes (all PNBS < .005), and deficits in node-wise temporal variability were prominent in TLE (all t(243) ≤ 2.51, PFDR < .05) and FLE (all t(302) ≤ -2.65, PFDR < .05) but relatively weak in GGE-GTCS. Moreover, dynamic states were generally less stable in patients than controls (all P's < .001). Collectively, these findings demonstrated general DMN abnormalities common to different epilepsies as well as distinct dysfunctions to subtypes, and provided insights into understanding the relationship of pathophysiological mechanisms and brain connectivity.


Advances in tissue expansion.
Tissue expansion offers a versatile new technique in the reconstruction of many areas of the body. The authors discuss laboratory studies that provide insight into the mechanism of the technique and present examples from clinical experience acquired over the past seven years, including applications in breast reconstruction and reconstruction of the scalp, head and neck, and extremities.


Importance of lymphovascular invasion and invasive front on survival in floor of mouth cancer.
The floor of mouth (FOM) is a common site of oral squamous cell carcinoma (SCC). The purpose of this study was to investigate pathological predictors of survival in FOM SCC. We conducted a retrospective study of 54 consecutive patients undergoing surgery for FOM SCC. Pathological parameters were extracted from histological reports with original pathology slides re-reviewed by 2 pathologists for missing data. On univariate analysis, depth of invasion >10 mm (p = .009), lymphovascular invasion (LVI; p < .001), noncohesive invasive front (p = .006), perineural invasion (PNI; p = .003), and nodal metastases (p = .02) were significant predictors of overall survival. On multivariate analysis, LVI (p = .009) and invasive front (p < .001) remained significant. Postoperative radiotherapy improved survival in patients with LVI, PNI, and nodal metastases, and was just outside significance for noncohesive invasive front (p = .06). LVI is an adverse prognosticator in FOM SCC and indicates postoperative radiotherapy. Further study is required to investigate the importance of invasive front. © 2015 Wiley Periodicals, Inc. Head Neck 38: E1528-E1534, 2016.


Evolutionary hypervariability in the hinge region of the immunoglobulin alpha gene.
The hinge region of the immunoglobulin molecule is responsible for antigen-binding and cross-linking reactions, varying the distance between the two antigen-binding sites. As the amino acid sequence of the hinge region is identical among immunoglobulin molecules of the same (sub)class, it has been regarded as a constant region. By comparison of the nucleotide sequences among primate C alpha genes, it is clear that there is a wide variety of length among the hinge regions of hominoid C alpha genes, which basically consist of tandem repeats of a 15 base-pair sequence. This reiterated structure probably facilitates rapid evolutionary changes in the length of the hinge region. The hinge region of the Old World monkey C alpha gene has a non-reiterated structure whose nucleotide sequence is quite different from those of the hominoid C alpha genes, although its surrounding region is conserved during evolution. This unusual hypervariability reveals that the hinge region has evolved as a semi-variable region in contrast to its constant character from an ontogenic viewpoint.


[Amniotic membrane transplantation after surgical resection in conjunctival Kaposi's sarcoma].
To describe a case of isolated conjunctival Kaposi's sarcoma treated with surgical resection and amniotic membrane graft. After surgical resection of the lesion an amniotic membrane graft was used to reconstruct the conjunctival surface. Histological studies were performed. Rapid regeneration of the conjunctiva, with minimal scarring, occurred after the surgical procedure and no recurrence of the lesion was observed at 18 months of follow-up. Surgical resection with amniotic membrane grafting appears to be a useful therapeutic option in isolated conjunctival Kaposi's sarcoma.


Postoperative complications following tonsillectomy and adenoidectomy--who is at risk?
Postoperative complications following a large series of adenotonsillectomies (784 patients) at the Children's Hospital of Philadelphia fell into two major categories: hemorrhage and airway problems. Bleeding requiring operative intervention occurred in one child in the immediate postoperative period and in 16 children (2.0%) within 7-10 days. Eight patients (1.0%) had minor bleeding requiring only observation, and 9 others (1.2%) reported mild bleeding at home. Airway obstruction requiring the reinsertion of an airway or intensive care observation occurred in 10 children (1.3%). In general, these children were younger and most had significant underlying medical problems. We conclude that many complications encountered after adenotonsillar surgery are intrinsic to the patient's disease and overall medical condition. Improved surgical and anesthetic techniques have minimized the complications noted in earlier studies.


The L-cell and D-cell of mitosis.
The two daughter cells produced by the mitosis of an eukaryotic cell present some patterns of organization with mirror-symmetry. For example, during the metaphase the succession of chromosomes in the ring of equatorial plate develop in the clockwise direction for the daughter D-cell and in the counter clockwise direction for the daughter L-cell. A similar mirror-symmetry is manifested between the patterns formed by the molecular receivers of the two daughter cells. This is a topological information, which interferes in the cellular differentiation and in the structural development of the polycellular organisms.


Advances in the discovery and development of selective heme-displacing IDO1 inhibitors.
Indoleamine 2,3-dioxygenase 1 (IDO1) has been considered as an attractive intracellular target for the development of small-molecule cancer immunotherapy. Results in human clinical studies indicated that the first-generation IDO1 inhibitor epacadostat lacked anticancer activity when combined with the anti-PD-1 antibody pembrolizumab. Epacadostat inhibits IDO1 activity by forming a tertiary IDO1-heme-inhibitor complex. Recently, IDO1 inhibitors capable of displacing the heme group to form a binary high-affinity complex have been discovered and investigated in humans. Structures, mode of action, preclinical activities, and status of clinical development are discussed and compared between the two classes of IDO1 inhibitors that bind to IDO1 protein in the presence (holo-IDO1) or absence (apo-IDO1) of the heme group, respectively. By selectively displacing the heme from IDO1 enzymes, apo-IDO1 inhibitors demonstrate high target selectivity, durable target engagement, and exceptional potency in cellular assays. Data from publications, patent disclosures, and conference proceedings as recent as 2019 are analyzed and summarized. The outcomes in cancer patients for the first-generation IDO1 inhibitors were disappointing. However, the unique mode of action by the heme-displacing IDO1 inhibitors might help their successful clinical translation and provide a novel class of anticancer drugs for cancer immunotherapy.


Slow Long-Term Exhumation of the West Central Andean Plate Boundary, Chile.
We present a regional analysis of new low-temperature thermochronometer ages from the Central Andean fore arc to provide insights into the exhumation history of the western Andean margin. To derive exhumation rates over 10 million-year timescales, 38 new apatite and zircon (U-Th)/He ages were analyzed along six ~500-km long near-equal-elevation, coast parallel, transects in the Coastal Cordillera (CC) and higher-elevation Precordillera (PC) of the northern Chilean Andes between latitudes 18.5°S and 22.5°S. These transects were augmented with age-elevation profiles where possible. Results are synthesized with previously published thermochronometric data, corroborating a previously observed trenchward increase in cooling ages in Peru and northern Chile. One-dimensional thermal-kinematic modeling of all available multichronometer equal-elevation samples reveals mean exhumation rates of <0.2 km/Myr since ~50 Ma in the PC and ~100 Ma in the CC. Regression of pseudovertical age-elevation transects in the CC yields comparable rates of ~0.05 to ~0.12 km/Myr between ~40 and 80 Ma. Differences between the long-term mean 1-D rates and shorter-term age-elevation-derived rates indicate low variability in the exhumation history. Modeling results suggest similar background exhumation rates in the CC and PC; younger ages in the PC are largely a function of increased heat flow and consequently an elevated geothermal gradient near the arc. Slow exhumation rates are suggestive of semiarid conditions across the region since at least the Eocene and deformation and development of the Andean fore arc around this time.


The use of wheat germ agglutinin--horseradish peroxidase conjugates for studies of anterograde axonal transport.
Injection of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP), into the hemisected spinal cord of the rat, cat and monkey consistently resulted in the intense anterograde labeling of ascending spinal projections such as the spinothalamic tract and spinocerebellar tracts and their terminal fields. Injections of WGA-HRP in the dorsal column nuclei resulted in the anterograde labeling of the medial lemniscus and its terminal fields in the thalamus. Injection of similar amounts of horseradish peroxidase alone (HRP) in hemisected animals or the dorsal column nuclei resulted in little anterograde labeling. The rate of the anterograde transport of WGA-HRP in cut axons appears to be greater than 200 mm/day. Small amounts of transneuronal labeling appeared to occur after injection of WGA-HRP in both cut axons and undamaged cell bodies. These results suggest that the amount of anterograde labeling observed after injection of WGA-HRP into both cut axons and undamaged cell bodies is significantly greater than the anterograde labeling observed after injections of HRP alone. Therefore, in the central nervous system WGA-HRP appears to be a far more effective anterograde tracer than HRP alone.


Cyproterone acetate vs leuprolide acetate in combination with transdermal oestradiol in transwomen: a comparison of safety and effectiveness.
To retrospectively compare the effectiveness and safety of 1-year administration of transdermal oestradiol (TE) with cyproterone acetate (CPA) or leuprolide acetate (Leu) in transwomen. Forty transwomen received 50 mg of CPA daily orally (n = 20; CPA+E group) or Leu at a dose of 3·75 mg i.m. monthly (n = 20; Leu+E group) in combination with TE at a dose of 1 or 2 mg daily for 1 year. Reproductive hormones, biochemical parameters, body composition and bone mineral density were assessed. LH, FSH and total testosterone levels were significantly decreased by month three of hormone administration in both groups and continued to decrease until month 12; the decrease in LH levels in the first 12 months was significantly faster in the Leu+E group. Prolactin was significantly increased at month 12 in the CPA+E group only. Bone metabolism parameters and bone mineral density as detected at DEXA did not significantly change in either group, apart from a statistically significant increase in parathyroid hormone after 52 weeks of Leu administration. Total cholesterol and HDL-cholesterol were significantly increased in the Leu+E group and reduced in the CPA+E group. No major adverse effects were registered in either group. Psychological well-being parameters did not differ between the two groups. Preliminary results from this retrospective observational pilot study suggest that CPA and Leu in combination with TE are equally effective in the suppression of gonadotrophins and testosterone levels over 1 year. Whether the different effects on HDL-cholesterol may lead to long-term different cardiovascular safety profiles remains to be defined.


Biochemical changes in articular cartilage after joint immobilization by casting or external fixation.
Knees of mature dogs were immobilized for 6 weeks by long-leg casts allowing 8 degrees-15 degrees of motion, a model studied by others, or with external fixators, a new, more severe model that kept the joints rigid. Some animals were allowed to recover for 1 week after the immobilization period. Articular cartilage was examined histologically and biochemically. After 6 weeks of immobilization, water increased 7% in both casted and fixator-immobilized joints compared with normal knee cartilage, while hexuronic acid was 23 and 28% lower, respectively. The limited motion permitted by the casts resulted in a smaller depression of proteoglycan synthesis and less proteoglycan loss during immobilization than occurred in the rigid external fixator group. The protective effect of limited motion was shown clearly during the recovery period: as measured by hexuronic acid content, cartilage from the casted joints had almost recovered within 1 week, whereas the external fixator group experienced little or no recovery during the week after treatment. In contrast to previous studies by others with casted joints, both newly synthesized [35S]sulfate-labeled and accumulated unlabeled proteoglycans from both casted and fixator-immobilized cartilages were able to form complexes with exogenous hyaluronic acid to the same extent as those from control cartilage. Thus, in immobilized cartilage, failure of the newly synthesized proteoglycan to bind to hyaluronate is not a mechanism of accelerated proteoglycan loss. The accelerated proteoglycan turnover appears to be caused by a combination of decreased synthesis and increased proteolysis of the secreted proteoglycans.(ABSTRACT TRUNCATED AT 250 WORDS)


Differentiation of Plasmodium male gametocytes is initiated by the recruitment of a chromatin remodeler to a male-specific cis-element.
Plasmodium parasites, the causative agents of malaria, possess a complex lifecycle; however, the mechanisms of gene regulation involved in the cell-type changes remain unknown. Here, we report that gametocyte sucrose nonfermentable 2 (gSNF2), an SNF2-like chromatin remodeling ATPase, plays an essential role in the differentiation of male gametocytes. Upon disruption of gSNF2, male gametocytes lost the capacity to develop into gametes. ChIP-seq analyses revealed that gSNF2 is widely recruited upstream of male-specific genes through a five-base, male-specific cis-acting element. In gSNF2-disrupted parasites, expression of over a hundred target genes was significantly decreased. ATAC-seq analysis demonstrated that decreased expression of these genes correlated with a decrease of the nucleosome-free region upstream of these genes. These results suggest that global changes induced in the chromatin landscape by gSNF2 are the initial step in male differentiation from early gametocytes. This study provides the possibility that chromatin remodeling is responsible for cell-type changes in the Plasmodium lifecycle.


Oxytocin receptor ligands induce changes in cytoskeleton in neuroblastoma cells.
Aim of the present study was to evaluate effects of ligands of oxytocin receptors on gene expression of neurofilament proteins (nestin and microtubule-associated protein 2 (MAP2)) associated with neuronal differentiation and growth factors (brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF)) related to neuronal growth. Fluorescent staining of F-actin was used to observe morphology of cells. Co-treatment with oxytocin and oxytocin receptor antagonist--atosiban--resulted in significant increase of MAP2 gene expression in SK-N-SH cells. There was no effect of oxytocin on gene expression of growth factors BDNF and NGF. Surprisingly, oxytocin with atosiban significantly increased mRNA levels for both BDNF and NGF. Gene expression of vasopressin receptor (V1aR) significantly decreased in response to vasopressin. Atosiban decreased mRNA levels for oxytocin receptor (OXTR) and V1aR. Oxytocin significantly decreased OXTR and nestin mRNA levels and increased mRNA levels for BDNF and NGF in U-87 MG cells. The densest recruitment of F-actin filaments was observed in apical parts of filopodia in SK-N-SH cells incubated in oxytocin presence. Present data demonstrate complex role of ligands of oxytocin receptors in regulation of gene expression of intermediate filaments and thus, oxytocin might be considered as a growth factor in neuronal type of cells.


Fine structure and peroxidatic activity of rat blood monocytes.
Rat blood monocytes contain granules of two types, primary and secondary granules, which can be distinguished on morphological grounds. Secondary granules have a halo between the limiting membrane and the matrix; primary granules have no halo and are generally more elongate than the secondary granules. In addition, there are differences in the distribution of the diameters of these two types of granules. The cytochemical characteristics of the peroxidatic activity of these granules were investigated by using different prefixation conditions in combination with a number of DAB media differing in pH and H2O2 concentration. Prolongation of the prefixation and increasing the pH of the incubation medium increased the staining intensity of the secondary granules and decreased the staining intensity of the primary granules. In unfixed cells peroxidatic activity could be demonstrated only in the primary granules. It is concluded from the present findings that peroxidase occurs in primary granules and not in secondary granules, whereas catalase is present in secondary granules but probably not in primary granules. No evidence was found for the presence of microperoxisomes in rat blood monocytes.


Antiallergic effect of the root of Paeonia lactiflora and its constituents paeoniflorin and paeonol.
The root of Paeonia lactiflora PALL (PL, Family Paeoniaceae) has been used frequently as an antipyretic and anti-inflammatory agent in traditional medicines of Korea, China and Japan. To evaluate antiallergic effect of PL, we isolated its main constituents, paeoniflorin and paeonol, and evaluated in vivo their inhibitory effects against passive cutaenous anaphylaxis (PCA) reaction induced by IgE-antigen complex and scratching behaviors induced by compound 48/80. PL, paeoniflorin and paeonol potently inhibited PCA reaction and scratching behaviors in mice. Paeoniflorin exhibited the most potent inhibition against scratching behaviors and the acetic acid-induced writhing syndrome in mice. Paeonol most potently inhibited PCA reaction and mast cells degranulation. These findings suggest that PL can improve IgE-induced anaphylaxis and scratching behaviors, and may be due to the effect of its constituents, paeoniflorin and paeonol.


Breaking bad news: a guide for effective and empathetic communication.
Breaking negative news to patients is a common occurrence for nurse practitioners. This difficult task requires patience and refined communication skills, and must be approached with empathy for all parties involved. There are several ways to deliver bad news to patients successfully using patient-centered communication techniques and methods.


CRISPR-Cas9 Correction of Duchenne Muscular Dystrophy in Mice by a Self-Complementary AAV Delivery System.
Duchenne muscular dystrophy (DMD) is a fatal neuromuscular disorder, caused by mutations in the DMD gene coding dystrophin. Applying clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (CRISPR-Cas) for therapeutic gene editing represents a promising technology to correct this devastating disease through elimination of underlying genetic mutations. Adeno-associated virus (AAV) has been widely used for gene therapy due to its low immunogenicity and high tissue tropism. In particular, CRISPR-Cas9 gene editing components packaged by self-complementary AAV (scAAV) demonstrate robust viral transduction and efficient gene editing, enabling restoration of dystrophin expression throughout skeletal and cardiac muscle in animal models of DMD. Here, we describe protocols for cloning CRISPR single guide RNAs (sgRNAs) into a scAAV plasmid and procedures for systemic delivery of AAVs into a DMD mouse model. We also provide methodologies for quantification of dystrophin restoration after systemic CRISPR-Cas9-mediated correction of DMD.


Involvement of protein tyrosine phosphorylation and reduction of cellular sulfhydryl groups in cell death induced by 1' -acetoxychavicol acetate in Ehrlich ascites tumor cells.
Elucidation of the mechanisms underlying potential anticancer drugs continues and unraveling these mechanisms would not only provide a conceptual framework for drug design but also promote use of natural products for chemotherapy. To further evaluate the efficacy of the anticancer activity of 1'-acetoxychavicol acetate (ACA), this study investigates the underlying mechanisms by which ACA induces death of Ehrlich ascites tumor cells. ACA treatment induced loss of cell viability, and Western blotting analysis revealed that the compound stimulated tyrosine phosphorylation of several proteins with 27 and 70 kDa proteins being regulated in both dose- and time-dependent manner prior to loss of viability. Protein tyrosine kinase inhibitor herbimycin A moderately protected cells from ACA-induced toxicity. In addition, cellular glutathione and protein sulfydryl groups were also significantly reduced both dose- and time-dependently during evidence of cell death. Replenishing thiol levels by antioxidant, N-acetylcysteine (NAC), an excellent supplier of glutathione and precursor of glutathione, substantially recovered the viability loss, but the recovery being time-dependent, as late addition of NAC (at least 30 min after ACA addition to cultures) was, however, ineffective. Addition of NAC to ACA treated cultures also abolished tyrosine phosphorylation of the 27 kDa protein. These results, at least partly, identify cellular sulfhydryl groups and protein tyrosine phosphorylation as targets of ACA cytotoxicity in tumor cells.


Designing a multi-epitope vaccine to provoke the robust immune response against influenza A H7N9.
A new strain of Influenza A Virus (IAV), so-called "H7N9 Avian Influenza", is the first strain of this virus in which a human is infected by transmitting the N9 of influenza virus. Although continuous human-to-human transmission has not been reported, the occurrence of various H7N9-associated epidemics and the lack of production of strong antibodies against H7N9 in humans warn of the potential for H7N9 to become a new pandemic. Therefore, the need for effective vaccination against H7N9 as a life-threatening viral pathogen has become a major concern. The current study reports the design of a multi-epitope vaccine against Hemagglutinin (HA) and Neuraminidase (NA) proteins of H7N9 Influenza A virus by prediction of Cytotoxic T lymphocyte (CTL), Helper T lymphocyte (HTL), IFN-γ and B-cell epitopes. Human β-defensin-3 (HβD-3) and pan HLA DR-binding epitope (PADRE) sequence were considered as adjuvant. EAAAK, AAY, GPGPG, HEYGAEALERAG, KK and RVRR linkers were used as a connector for epitopes. The final construct contained 777 amino acids that are expected to be a recombinant protein of about ~ 86.38 kDa with antigenic and non-allergenic properties after expression. Modeled protein analysis based on the tertiary structure validation, docking studies, and molecular dynamics simulations results like Root-mean-square deviation (RMSD), Gyration, Root-mean-square fluctuation (RMSF) and Molecular Mechanics Poisson-Boltzmann Surface Area (MM/PBSA) showed that this protein has a stable construct and capable of being in interaction with Toll-like receptor 7 (TLR7), TLR8 and m826 antibody. Analysis of the obtained data the demonstrates that suggested vaccine has the potential to induce the immune response by stimulating T and Bcells, and may be utilizable for prevention purposes against Avian Influenza A (H7N9).