[Preventing dependency in the elderly.].
dependency, i.e. the need to depend on another person to perform activities of daily living, is the main concern and cause of suffering and poor quality of life in the elderly. The prevalence of dependency increases with age and is related to the presence of prior disease and fragility. Dependency is associated with increased morbidity, mortality and institutionalization, as well as with greater health and social resource utilization, all of which increases health costs. to create a consensus document on the main health recommendations for the prevention of dependency in the elderly, based on the scientific evidence available to date, with the collaboration of scientific societies and public health administrations (the Spanish Ministry of Health, Autonomous Communities and Cities). a) a preliminary consensus document was drafted by an expert group composed of representatives of various scientific societies and health administrations. This document was based on a review of the recommendations and guidelines published by the main organizations involved in health promotion and the prevention of disease, functional deterioration and dependency in the elderly; b) the consensus document was reviewed by the remaining experts assigned by the scientific societies and central and autonomous administrations; c) the final document was approved after a session in which the text was discussed and reviewed by all the experts participating in the working group (including the academic committee); d) the document was presented and discussed in the First National Conference on Prevention and Health Promotion in Clinical Practice in Spain. All participating experts signed a conflicts of interest statement. the document provides recommendations, with their grades of evidence, grouped in the following three categories: a) health promotion and disease prevention, with specific preventive activities for the elderly, including prevention of geriatric syndromes; b) prevention of functional deterioration, with clinical recommendations that can be applied in primary and specialized care; c) prevention of iatrogeny (drug prescription, inappropriate use of diagnostic and therapeutic modalities and healthcare). These recommendations were tailored to the characteristics of the older person (OP), categorized in five groups: healthy OP, OP with chronic disease, fragile or at risk OP, dependent OP, and OP at the end of life. these recommendations should be implemented by public health administrations to improve strategies for the prevention of dependency in the elderly in the XXI century.


Inhibitors of chymase as mast cell-stabilizing agents: contribution of chymase in the activation of human mast cells.
There has long been evidence that inhibitors of chymotryptic proteinases can inhibit the degranulation of rodent mast cells, but their actions on human mast cells and the contribution of mast cell chymase itself have received little attention. We investigated the ability of the selective chymase inhibitor Z-Ile-Glu-Pro-Phe-CO(2)Me and other proteinase inhibitors to inhibit chymase and cathepsin G activity, and we examined their potential to modulate the responsiveness of mast cells dispersed from human skin, lung, and tonsil tissues. IgE-dependent histamine release from skin mast cells was inhibited by up to about 80% after preincubation with Z-Ile-Glu-Pro-Phe- CO(2)Me (up to 0.1 microM), 70% with chymostatin (17 microM), and 60% with soybean trypsin inhibitor (0.5 microM). The mast cell-stabilizing properties of chymase inhibitors appeared to be greater for skin mast cells than for those from lung, whereas tonsil mast cells were relatively unresponsive. There were marked differences in the time course of responses to inhibitors, and the effect was dependent on the stimulus, with calcium ionophore-induced histamine release being unaffected. Incubation of dispersed skin, lung, or tonsil cells for up to 45 min with purified chymase failed to induce histamine release, although preincubation of cells with chymase was able to suppress IgE-dependent activation. Chymase could thus contribute to mast cell degranulation and after secretion could provide a feedback mechanism to limit this process. Nevertheless, inhibitors of chymase can be potent mast cell stabilizers, particularly in the skin.


Episode-based efficiency comparisons for physicians and nurse practitioners.
Most previous studies comparing the efficiency of new health practitioners with that of physicians have used the visit as the basic unit of output. Several researchers have noted, though, that the episode is a conceptually superior output unit in several respects, although it is more complex to deal with methodologically. This study demonstrates the application of episode-based methods for comparing the efficiency of physicians with that of nurse practitioners. Data are drawn from the information system of the Columbia Medical Plan and from observations of provider time inputs. The analysis is confined to care episodes for otitis media and sore throat in the Department of Pediatrics. Results indicate that per episode costs with nurse practitioners as the initial provider are approximately 20 per cent below the costs of episodes in which physicians are the initial provider. Examination of a limited amount of data on patient-reported measures of effectiveness indicates that while nurse practitioners' care is less costly, it is not less effective. These findings are particularly interesting in light of recent doubts expressed about cost-savings from using new health practitioners, and particularly nurse practitioners, in group practice settings.


Treatment of hemicrania continua with radiofrequency ablation and long-term follow-up.
The objective of this research is to describe novel procedural treatments for hemicrania continua that allow patients to remain off indomethacin. Case reports are presented. We describe four distinct patients with indomethacin-responsive hemicrania continua who were unable to discontinue the use of indomethacin without headache recurrence. No other medications were effective for their syndrome. Secondary causes of headache were ruled out in each case. Each patient underwent diagnostic blockade of either the atlanto-axial joint, C2 dorsal root ganglion or sphenopalantine ganglion depending on their clinical examination and presence of cranial autonomic symptoms. A positive response led to a radiofrequency ablation of the C2 ventral ramus, C2 dorsal root ganglion or sphenopalantine ganglion, which provided headache relief in all case patients as complete as indomethacin. Long-term follow-up of these patients has shown that all have remained essentially headache free without the need for indomethacin. One patient has needed repeat radiofrequency procedures with consistent response. Hemicrania continua is defined by its sensitivity to indomethacin but very few patients are able to discontinue the medication without headache recurrence. As the risks of chronic indomethacin use are substantial, alternative treatments are necessary to protect patient health. We are now able to suggest several radiofrequency ablation procedures as effective as indomethacin with long-term follow-up.


Expanding newborn screening: process, policy, and priorities.
In the 1960s, newborn screening programs tested for a single very rare but serious disorder. In recent years, thanks to the development of new screening technology, they have expanded into panels of tests; a federally sponsored expert group has recommended that states test for twenty-nine core disorders and twenty-five secondary disorders. By the standards used to decide whether to introduce new preventive health services into clinical use, the decision-making in newborn screening policy has been lax.


Multicenter comparison of the interobserver agreement of standard EUS scoring and Rosemont classification scoring for diagnosis of chronic pancreatitis.
EUS has less than optimal interobserver agreement for the diagnosis of chronic pancreatitis. The newly developed Rosemont consensus scoring system includes weighted criteria and stricter definitions for individual features. The primary aim was to compare the interobserver agreement of standard and Rosemont scoring. Multiple tertiary-care institutions. Fifty EUS videos were interpreted by 14 experts. Each expert interpreted the videos on two occasions: First, the videos were read by using standard scoring (9 criteria). Second, after viewing a presentation of the Rosemont classification, the same experts re-read the videos by using Rosemont scoring. Fleiss' kappa (K) statistics are reported with 95% confidence intervals (CI). The interobserver agreement was "substantial" (K = 0.65 [95% CI, 0.52-0.77]) for Rosemont scoring and "moderate" (K = 0.54 [95% CI, 0.44-0.66]) for standard scoring; however, the difference was not statistically significant (P = 0.12). The sample size does not allow detection of differences in K of <0.25. Use of the Rosemont classification did not significantly increase interobserver agreement for EUS diagnosis of chronic pancreatitis compared with standard scoring.


G-quadruplex structures within the 3' UTR of LINE-1 elements stimulate retrotransposition.
Long interspersed nuclear elements (LINEs) are ubiquitous transposable elements in higher eukaryotes that have a significant role in shaping genomes, owing to their abundance. Here we report that guanine-rich sequences in the 3' untranslated regions (UTRs) of hominoid-specific LINE-1 elements are coupled with retrotransposon speciation and contribute to retrotransposition through the formation of G-quadruplex (G4) structures. We demonstrate that stabilization of the G4 motif of a human-specific LINE-1 element by small-molecule ligands stimulates retrotransposition.


Expression of leukocyte alkaline phosphatase gene in normal and leukemic cells: regulation of the transcript by granulocyte colony-stimulating factor.
The levels of leukocyte alkaline phosphatase (LAP) messenger RNA (mRNA) are evaluated in B and T lymphocytes, monocytes, and polymorphonuclear cells (PMNs), and this transcript is found to be present only in PMNs. Precursors of the myelomonocytic pathway, represented by leukemic cells isolated from several cases of chronic myelogenous leukemia (CML) in its stable and blastic phase and acute myelogenous leukemia (AML), are devoid of LAP transcript. These data support the notion that LAP is a marker of the granulocyte terminal differentiation. Despite the absence of LAP mRNA in both the myeloid and the lymphoid precursors, nuclear run-on experiments show constitutive transcription of the LAP gene in leukemic cells obtained from AML, CML, as well as acute lymphoblastic leukemia (ALL) and B-cell chronic lymphocytic leukemia (B-CLL). In CML and in chronic myelo-monocytic leukemia (CMML) PMNs, granulocyte colony-stimulating factor (G-CSF) specifically accumulates LAP mRNA without showing a substantial increase in the rate of transcription of the LAP gene. Once increased by G-CSF, LAP mRNA is very stable, showing a half-life of more than 4 hours in the presence of actinomycin-D. G-CSF is suggested to play a pivotal role in the modulation of LAP transcript in PMNs.


SUVmax/THKmax as a biomarker for distinguishing advanced gastric carcinoma from primary gastric lymphoma.
Gastric carcinoma and primary gastric lymphoma (PGL) are the two most common malignancies in stomach. The purpose of this study was to screen and validate a biomarker of (18)F-fluorodeoxyglucose positron emission tomography/computed tomography ((18)F-FDG PET/CT) for distinguishing advanced gastric carcinoma (AGC) from PGL for clinical applications. We reviewed PET/CT scans collected from January 2008 to April 2012 of 69 AGC and 38 PGL (14 low-grade mucosa-associated lymphoid tissue [MALT], 24 non-MALT aggressive non-Hodgkin lymphoma [ANHL]) with a focus on FDG intensity (maximum standardized uptake value [SUVmax]) of primary lesions and its CT-detected abnormalities, including maximal gastrointestinal wall thickness (THKmax) and mucosal ulcerations. Gastric FDG uptake was found in 69 (100%) patients with AGC and 36 (95%, 12 MALT vs. 24 ANHL)with PGL. The presence of CT-detected abnormalities of AGC and PGL were 97% (67/69) and 89% (12 MALT vs. 22 ANHL), respectively. After controlling for THKmax, SUVmax was higher with ANHL than AGC (17.10 ± 8.08 vs. 9.65 ± 5.24, p<0.05) and MALT (6.20 ± 3.60, p<0.05). THKmax did not differ among MALT, ANHL and AGC. Mucosal ulceration was more common with AGC (n = 9) than PGL (n = 2),but the difference was not statistically significant (p>0.05). Cross-validation analysis showed that for distinguishing ANHL from AGC, the classifier with SUVmax as a feature achieved a correct classification rate of 81% with thresholds 13.40 ± 1.12 and the classifier with SUVmax/THKmax as a feature achieved a correct classification rate of 83% with thresholds 7.51 ± 0.63. SUVmax/THKmax may be as a promising biomarker of FDG-PET/CT for distinguishing ANHL from AGC. Structural CT abnormalities alone may not be reliable but can help with PET assessment of gastric malignancies. (18)F-FDG PET/CT have potential for distinguishing AGC from PGL at the individual level.


Clinical outcomes following treatment of human intrabony defects with GTR/bone replacement material or access flap alone. A multicenter randomized controlled clinical trial.
This prospective multicenter randomized controlled clinical trial was designed to compare the clinical outcomes of papilla preservation flap surgery with or without the application of a guided tissue regeneration (GTR)/bone replacement material. One hundred and twenty-four patients with advanced chronic periodontitis were recruited in 10 centers in seven countries. All patients had at least one intrabony defect of > or = 3 mm. The surgical procedures included access for root instrumentation using either the simplified or the modified papilla preservation flap in order to obtain optimal tissue adaptation and primary closure. After debridement, the regenerative material was applied in the test subjects, and omitted in the controls. At baseline and 1 year following the interventions, clinical attachment levels (CALs), probing pocket depths (PPDs), recession, full-mouth plaque scores and full-mouth bleeding scores (FMBS) were assessed. One year after treatment, the test defects gained 3.3 +/- 1.7 mm of CAL, while the control defects yielded a significantly lower CAL gain of 2.5 +/- 1.5 mm. Pocket reduction was also significantly higher in the test group (3.7 +/- 1.8 mm) when compared with the controls (3.2 +/- 1.5 mm). A multivariate analysis indicated that the treatment, the clinical centers, baseline PPD and baseline FMBS significantly influenced CAL gains. Odds ratios (ORs) of achieving above-median CAL gains were significantly improved by the test procedure (OR = 2.6, 95% CI 1.2-5.4) and by starting with deeper PPD (OR = 1.7, 1.3-2.2) but were decreased by receiving treatment at the worst-performing clinical center (OR = 0.9, 0.76-0.99). The results of this trial indicated that regenerative periodontal surgery with a GTR/bone replacement material offers an additional benefit in terms of CAL gains, PPD reductions and predictability of outcomes with respect to papilla preservation flaps alone.


Parasitic infection among residents of an institution for mentally retarded persons.
Data on occurrence of intestinal parasites among resident and entering populations in a South Carolina institution for mentally retarded persons were examined. Following an eradication effort in 1973 for Trichuris trichiura, the prevalence of this parasite remained below 5 percent for the resident population, with the exception of 1974 and 1975, when small numbers of stool examinations occurred. Significantly higher proportions of the entering population, compared to the residents, harbored T. trichiura, Ascaris lumbricoides, Giardia lamblia, and Entamoeba histolytica. We concluded that annual examination and treatment can control trichuriasis; however, complete eradication may require examination following home visits by the resident population.


Decreased STAMP2 expression in association with visceral adipose tissue dysfunction.
Six-transmembrane protein of prostate 2 (STAMP2) is a counter-regulator of inflammation and insulin resistance according to findings in mice. However, there have been contradictory reports in humans. We aimed to explore STAMP2 in association with inflammatory and metabolic status of human obesity. STAMP2 gene expression was analyzed in adipose tissue samples (171 visceral and 67 sc depots) and during human preadipocyte differentiation. Human adipocytes were treated with macrophage-conditioned medium, TNF-α, and rosiglitazone. In visceral adipose tissue, STAMP2 gene expression was significantly decreased in obese subjects, mainly in obese subjects with type 2 diabetes. STAMP2 gene expression and protein were significantly and inversely associated with obesity phenotype measures (body mass index, waist, hip, and fat mass) and obesity-associated metabolic disturbances (systolic blood pressure and fasting glucose). In addition, STAMP2 gene expression was positively associated with lipogenic (FASN, ACC1, SREBP1, THRSP14, TRα, and TRα1), CAV1, IRS1, GLUT4, and CD206 gene expression. In sc adipose tissue, STAMP2 gene expression was not associated with metabolic parameters. In both fat depots, STAMP2 gene expression in stromovascular cells was significantly higher than in mature adipocytes. STAMP2 gene expression was significantly increased during the differentiation process in parallel to adipogenic genes, being increased in preadipocytes derived from lean subjects. Macrophage-conditioned medium (25%) and TNF-α (100 ng/ml) administration increased whereas rosiglitazone (2 μM) decreased significantly STAMP2 gene expression in human differentiated adipocytes. Decreased STAMP2 expression (mRNA and protein) might reflect visceral adipose dysfunction in subjects with obesity and type 2 diabetes.


Ictal adipokines are associated with pain severity and treatment response in episodic migraine.
To evaluate ictal adipokine levels in episodic migraineurs and their association with pain severity and treatment response. This was a double-blind, placebo-controlled trial evaluating peripheral blood specimens from episodic migraineurs at acute pain onset and 30 to 120 minutes after treatment with sumatriptan/naproxen sodium vs placebo. Total adiponectin (T-ADP), ADP multimers (high molecular weight [HMW], middle molecular weight, and low molecular weight [LMW]), leptin, and resistin levels were evaluated by immunoassays. Thirty-four participants (17 responders, 17 nonresponders) were included. In all participants, pretreatment pain severity increased with every quartile increase in both the HMW:T-ADP ratio (coefficient of variation [CV] 0.51; 95% confidence interval [CI]: 0.08, 0.93; p = 0.019) and resistin levels (CV 0.58; 95% CI: 0.21, 0.96; p = 0.002), but was not associated with quartile changes in leptin levels. In responders, T-ADP (CV -0.98; 95% CI: -1.88, -0.08; p = 0.031) and resistin (CV -0.95; 95% CI: -1.83, -0.07; p = 0.034) levels decreased 120 minutes after treatment as compared with pretreatment. In addition, in responders, the HMW:T-ADP ratio (CV -0.04; 95% CI: -0.07, -0.01; p = 0.041) decreased and the LMW:T-ADP ratio (CV 0.04; 95% CI: 0.01, 0.07; p = 0.043) increased at 120 minutes after treatment. In nonresponders, the LMW:T-ADP ratio (CV -0.04; 95% CI: -0.07, -0.01; p = 0.018) decreased 120 minutes after treatment. Leptin was not associated with treatment response. Both pretreatment migraine pain severity and treatment response are associated with changes in adipokine levels. Adipokines represent potential novel migraine biomarkers and drug targets.


Expression of an ovarian cancer anti-idiotype antibody (6B11VLVHCH3) in Chinese hamster ovary (CHO) cells with improved immunoactivity and stability over proteins expressed in prokaryotic cells.
The 6B11VLVHCH3 is an anti-idiotype antibody to ovarian cancer. It can mimic the ovarian cancer-associated antigen OC166-9. We have previously expressed it in Escherichia coli. We now express the 6B11VLVHCH3 in a Chinese hamster ovary (CHO) cell system. The 6B11VLVHhCH3 sequence was amplified from the prokaryotic vector and cloned into the eukaryotic expression vector pSecTag2. The construct was transfected into CHO cells. Then the transfected monoclonal CHO cells were sequentially adapted to growth in serum-free medium. Functional differences between the antibody produced in E. coli or CHO cells were assessed. The antibody produced by CHO cells was able to both mimic ovarian cancer antigen and retain FC region functionality. Importantly, the antigen mimicry activity of 6B11VLVHhCH3 produced by CHO cells is approximately 20 times higher than antibody expressed by E. coli. This method of expression provides a safer and higher quality antibody for clinical drug development.


Familial correlates of social anxiety in children and adolescents.
Retrospective studies suggest a relationship between parental rearing practices and social phobia. The present study investigated whether socially anxious children perceive their current parental rearing as rejecting, overprotective, and lacking emotional warmth, and as emphasizing the importance of other's opinion, and de-emphasizing social initiatives and family sociability. Furthermore, we examined whether parents of socially anxious children report to rely on such rearing practices, and suffer themselves from social fears. A regression analysis as well as extreme group comparisons were applied. Little support was found for the presumed role of the assessed family rearing aspects in the development of social anxiety in children. Solely family sociability (children's and mothers' report) and children's perception of overprotection of the mother predicted social anxiety in the regression analysis. Given the influence of the mentioned rearing practices, social anxiety of the mother still significantly predicted social anxiety of the child. In the extreme group comparisons, differences in the expected direction were found between socially anxious and normal children on parental rejection, emotional warmth, and family sociability. However, the lack of differences between socially anxious and clinical control children suggests that these variables do not form a specific pathway to social fears.


Helicobacter trogontum sp. nov., isolated from the rat intestine.
A new Helicobacter species that colonizes the colonic mucosa of Wistar and Holtzman rats was isolated and characterized. This bacterium was gram negative, its cells were rod shaped with pointed ends, and its protoplasmic cylinder was entwined with periplasmic fibers. It was catalase and oxidase positive, rapidly hydrolyzed urea, and was susceptible to metronidazole and resistant to cephalothin and nalidixic acid. The new organism was microaerophilic and grew at 42 degrees C, a feature that differentiates it from two other murine intestine colonizers, Helicobacter hepaticus and Helicobacter muridarum. On the basis of 16S rRNA sequence analysis data, the new organism was identified as a Helicobacter species that is most closely related to H. hepaticus. This bacterium is named Helicobacter trogontum. The type strain is strain LRB 8581 (= ATCC 700114).


The clinical epidemiology and course of the spectrum of renal diseases associated with HIV infection.
While an understanding of the epidemiology and clinical course of HIV-associated nephropathy (HIVAN) is growing, little is known about the risk factors and clinical course of the other renal diseases that may also occur as a complication of HIV infection. This study was undertaken to compare HIVAN to the spectrum of other kidney diseases seen among HIV-infected patients. This retrospective cohort study included all HIV-infected patients who underwent renal biopsy during the course of their clinical care at six major medical centers. Demographic and clinical information were abstracted from each patient's clinical record. Time to initiation of renal replacement therapy was compared for patients with lesions other than HIVAN to patients with HIVAN using Cox proportional hazards regression. Eighty-nine patients (47 with lesions other than HIVAN and 42 with HIVAN) were available for inclusion. Patients with lesions other than HIVAN were less likely to be black (37/47 vs. 42/42, P= 0.02), more likely to have a positive hepatitis B surface antigen (10/37 vs. 4/42, P= 0.04), less likely to have the diagnosis of hypertension (24/46 vs. 31/42, P= 0.03), more likely to have a greater creatinine clearance at time of biopsy (60.6 vs. 39.0 cc/min, P= 0.008), and have a greater CD4 lymphocyte count at time of biopsy (287 vs. 187 cells/mL, P= 0.04) compared to patients with HIVAN. Lesions other than HIVAN were associated with a longer time to initiation of renal replacement therapy compared with HIVAN (HR 0.33, 95% CI 0.15-0.71, P= 0.005). Other factors associated with a longer time to renal replacement therapy included higher creatinine clearance at time of biopsy, greater CD4(+) lymphocyte count, the absence of hepatitis C antibody, and the use of an angiotensin-converting enzyme inhibitor or angiotensin receptor blocker. The type of renal disease (HIVAN vs. other) interacted significantly with HIV-1 RNA level and the use of antiretroviral therapy (P= 0.0001 and 0.006, respectively). Among patients with lesions other than HIVAN, the presence of nondetectable HIV-1 RNA was not associated with a greater risk of progression of renal disease (HR 0.27, P= 0.24). Among patients with HIVAN, because all patients had detectable virus at the time of institution of renal replacement therapy, this highly significant association could not be quantified. Among patients with lesions other than HIVAN, the use of antiretroviral therapy was not associated with the progression to renal replacement therapy (HR 3.29, P= 0.06). Among patients with HIVAN, the use of antiretroviral therapy was associated with a slower progression to renal replacement therapy (HR 0.24, P= 0.03). Among HIV-infected patients with renal disease other than HIVAN, viral suppression and the use of antiretroviral therapy are not associated with a beneficial effect on renal function; thus, additional therapeutic strategies may need to be utilized. Because renal histology is associated with prognostic differences, these data provide outcomes information that will improve the clinical utility of renal biopsy among HIV-infected patients with renal disease.


Binding to EGF receptor of a laminin-5 EGF-like fragment liberated during MMP-dependent mammary gland involution.
Extracellular matrix (ECM) fragments or cryptic sites unmasked by proteinases have been postulated to affect tissue remodeling and cancer progression. Therefore, the elucidation of their identities and functions is of great interest. Here, we show that matrix metalloproteinases (MMPs) generate a domain (DIII) from the ECM macromolecule laminin-5. Binding of a recombinant DIII fragment to epidermal growth factor receptor stimulates downstream signaling (mitogen-activated protein kinase), MMP-2 gene expression, and cell migration. Appearance of this cryptic ECM ligand in remodeling mammary gland coincides with MMP-mediated involution in wild-type mice, but not in tissue inhibitor of metalloproteinase 3 (TIMP-3)-deficient mice, supporting physiological regulation of DIII liberation. These findings indicate that ECM cues may operate via direct stimulation of receptor tyrosine kinases in tissue remodeling, and possibly cancer invasion.


Utilization of multigene panels in hereditary cancer predisposition testing: analysis of more than 2,000 patients.
The aim of this study was to determine the clinical and molecular characteristics of 2,079 patients who underwent hereditary cancer multigene panel testing. Panels included comprehensive analysis of 14-22 cancer susceptibility genes (BRCA1 and BRCA2 not included), depending on the panel ordered (BreastNext, OvaNext, ColoNext, or CancerNext). Next-generation sequencing and deletion/duplication analyses were performed for all genes except EPCAM (deletion/duplication analysis only). Clinical histories of ColoNext patients harboring mutations in genes with well-established diagnostic criteria were assessed to determine whether diagnostic/testing criteria were met. Positive rates were defined as the proportion of patients with a pathogenic mutation/likely pathogenic variant(s) and were as follows: 7.4% for BreastNext, 7.2% for OvaNext, 9.2% for ColoNext, and 9.6% for CancerNext. Inconclusive results were found in 19.8% of BreastNext, 25.6% of OvaNext, 15.1% of ColoNext, and 23.5% of CancerNext tests. Based on information submitted by clinicians, 30% of ColoNext patients with mutations in genes with well-established diagnostic criteria did not meet corresponding criteria. Our data point to an important role for targeted multigene panels in diagnosing hereditary cancer predisposition, particularly for patients with clinical histories spanning several possible diagnoses and for patients with suspicious clinical histories not meeting diagnostic criteria for a specific hereditary cancer syndrome.


Improving Quality of Chest Radiographs After Placement of Peripherally Inserted Central Catheters.
The goal of this study was to determine the best method for localizing peripherally inserted central catheters (PICCs) in postplacement portable chest radiographs. A retrospective analysis showed no significant difference in visualization of the PICC tip between different chest radiograph projections. Modifications were made to an institutional PICC protocol to obtain anteroposterior chest views with the guidewire present only. Repeat analysis demonstrated statistically significant increases in the frequency of anteroposterior radiographs performed, the number of chest radiographs with guidewire, and the localization of the catheter. By standardizing the acquisition of PICC placement chest radiographs, fewer variant projection radiographs were performed and the catheter tip was confidently localized in more examinations.


Knockdown of SP1/Syncytin1 axis inhibits the proliferation and metastasis through the AKT and ERK1/2 signaling pathways in non-small cell lung cancer.
Syncytin 1 is considered as an oncogene in various malignant tumors, but its effect on non-small cell lung cancer (NSCLC) has not been reported. We investigated the specific role of Syncytin 1 on NSCLC through the transfection of Syncytin 1 knockdown or overexpression plamids in A549 cells. Our results proved that knockdown of Syncytin 1 inhibited the proliferation, and blocked the cell cycle on G1 phase by inhibiting the expression of Nusap1, Cyclin D1, CDK6, and CDK4. Cell cycle arrest also leaded to increased apoptosis in Syncytin 1 knockdown cells. Suppression of Syncytin 1 inhibited the migration and invasion, as well as the expressions of epithelial-mesenchymal transition (EMT) makers, N-cadherin, β-catenin, and Vimentin, indicating that Syncytin 1 knockdown inhibited the metastasis via reversing the EMT process in A549 cells. The phosphorylation levels of Akt, mTOR, and Erk1/2 were all decreased in Syncytin 1 knockdown cells, suggesting the signaling pathways by which Syncytin 1 operated as an oncogene in NSCLC. Moreover, the underexpression of transcription factor SP1 downregulated the Syncytin 1 expression in A549 cells. The rescue experiment of Syncytin 1 in SP1 knockdown cells further proved that Syncytin 1 could block the inhibition of cell growth induced by SP1 knockdown. In conclusion, knockdown of SP1/Syncytin1 axis inhibited the progression of NSCLC by the reversion of tumor epithelial-mesenchymal transition process and suppression of Akt and Erk signaling pathways, suggesting that they are potential targets for targeted therapy of NSCLC.


[Monitoring of plasma disappearance rate of indocyanine green in a patient with necrotizing fasciitis and septic shock].
A 45-year old man with diabetes mellitus and arterial hypertension was transferred with fever of unknown origin, suspected diabetic angiopathy and sepsis. On admission the patient presented all signs of septic shock. Livid coloured injuries on his right hand and petechial bleeding in distal extremities were observed. A different origin of sepsis was not found in transesophageal ultrasound cardiography and computed tomography of cranium, chest and abdomen. Immediately after admission early goal-directed therapy was initiated. Apart from calculated antibiotic therapy intensive insulin therapy and hydrocortisone substitution was begun. The patient presented a multiple organ dysfunction syndrome. Plasma disappearance rate of indocyanine green (PDR (ICG)) on admission was 20,4 %/min (normal range > 18 %/min) and fell to 6,8 %/min within 12 hours, while central venous oxygen saturation remained normal. Despite therapy according to current guidelines for severe sepsis, the patient deteriorated. Surgical debridement was performed suspecting necrotizing fasciitis and application of recombinant human activated protein C (rhAPC) started. PDR (ICG) rapidly raised to normal values; the patient recovered and was discharged after 9 days. Monitoring of PDR (ICG) allows for improved bedside evaluation of liver perfusion of the critically ill and is not only able to predict prognosis but may help in decision making for supportive therapies.


High Dose Intravenous Vitamin C for Preventing The Disease Aggravation of Moderate COVID-19 Pneumonia. A Retrospective Propensity Matched Before-After Study.
Background: Coronavirus disease 2019 (COVID-19) pandemic is continuing to impact multiple countries worldwide and effective treatment options are still being developed. In this study, we investigate the potential of high-dose intravenous vitamin C (HDIVC) in the prevention of moderate COVID-19 disease aggravation. Methods: In this retrospective before-after case-matched clinical study, we compare the outcome and clinical courses of patients with moderate COVID-19 patients who were treated with an HDIVC protocol (intravenous injection of vitamin C, 100 mg/kg/day, 1 g/h, for 7 days from admission) during a one-month period (between March 18 and april 18, 2020, HDIVC group) with a control group treated without the HDIVC protocol during the preceding two months (January 18 to March 18, 2020). Patients in the two groups were matched in a 1:1 ratio according to age and gender. Results: The HDIVC and control groups each comprised 55 patients. For the primary outcomes, there was a significant difference in the number of patients that evolved from moderate to severe type between the two groups (HDIVC: 4/55 vs. control: 12/55, relative risk [RR] = 0.28 [0.08, 0.93], P = 0.03). Compared to the control group, there was a shorter duration of systemic inflammatory response syndrome (SIRS) (P = 0.0004) during the first week and lower SIRS occurrence (2/21 vs 10/22, P = 0.0086) on Day 7 (6-7 days after admission). In addition, HDIVC group had lower C-reactive protein levels (P = 0.005) and higher number of CD4+ T cells from Day 0 (on admission) to Day 7 (P = 0.04)." The levels of coagulation indicators, including activated partial thromboplastin time and D-dimer were also improved in the HDIVC compared to the control group on Day 7. Conclusion: HDIVC may be beneficial in limiting disease aggravation in the early stage of COVID-19 pneumonia, which may be related to its improvements on the inflammatory response, immune function and coagulation function. Further randomized controlled trials are required to augment these findings.


The standardized uptake value of 18-fluorodeoxyglucose positron emission tomography after chemoradiation and clinical outcome in patients with localized gastroesophageal carcinoma.
Empirical trimodality (TM) or bimodality (BM) approaches lead to unpredictable outcomes in patients with localized gastroesophageal carcinoma (LGC). We hypothesized that the standardized uptake value (SUV) of positron emission tomography (PET) after chemoradiation would define the clinical outcome in patients. We analyzed 272 (TM = 155 and BM = 117) LGC patients after chemoradiation PET and studied its association with overall survival (OS) and disease-free survival (DFS). The log-rank test, multivariate Cox proportional hazards model, and Kaplan-Meier survival plots were used to analyze the dichotomized data (using the median as a cutoff). The median survival of TM patients was 49 months (95% CI: 36 to not reached) and that of BM patients was 20 months (95% CI: 15-34; p < 0.0001); however, if the postchemoradiation SUV was <4.6 (4.6 = median SUV), the OS of TM and BM patients was not significantly different (p = 0.22). In the multivariate Cox model, postchemoradiation SUV (p = 0.02), baseline stage (p = 0.03), histology grade (p = 0.01), and having surgery (p = 0.004) were independent prognosticators for OS and DFS. These data document that the low postchemoradiation SUV in both TM and BM patients defines clinical aggression and has a similar clinical outcome. Systematic validation of PET changes, in this setting, can facilitate individualization of therapy of LGC patients.


[The perichromatin compartment of the cell nucleus].
In this review, the data on the structure and composition of the perichromatin compartment, a special border area between the condensed chromatin and the interchromatin space of the cell nucleus, are discussed in the light of the concept of nuclear functions in complex nuclear architectonics. Morphological features, molecular composition and functions of main extrachromosomal structures of the perichromatin compartment, perichromatin fibrils (PFs) and perichromatin granules (PGs) including nuclear stress-bodies (nSBs) that are derivates of the PGs under heat shock, are presented. A special attention was paid to the features of the molecular compositions of PFs and PGs in different cell types and at different physiological conditions.


Epstein-Barr virus-related persistent erythema multiforme in chronic fatigue syndrome.
Erythema multiforme (EM) has been rarely reported in Epstein-Barr virus (EBV)-associated diseases; this includes patients with chronic fatigue syndrome who have chronic or recurrent and disabling illness and an abnormal antibody reactivity to EBV. We describe a patient fulfilling the chronic fatigue syndrome diagnostic criteria who had developed an unusually persistent EM resistant to corticosteroids therapy. The EBV DNA was studied in skin EM lesions, throat washings, peripheral mononuclear cells, and plasma. The EBV antigens were studied in skin EM lesions and in mononuclear cells. The patient was followed up to 2 years. The patient had abnormal titers of antibodies against various EBV antigens and by immunofluorescence she disclosed the EBV nuclear antigen and the viral capsid antigen in the blood vessels of the affected skin. The dot blot hybridization assay detected viral DNA in throat washings and mononuclear cells, but not in plasma. The presence of the viral genomic content in lesional skin is suggested by the autoradiographic signal and by the difference from appropriate control specimens. Skin lesions and constitutional symptoms cleared after acyclovir sodium therapy and recurred after discontinuation of this therapy. This is the first EM case in which evidence of the EBV causal role has been provided. The association with chronic fatigue syndrome suggests the EBV role in selected cases of this syndrome.


[Anorexia nervosa and music therapy].
Supporting people suffering from anorexia nervosa remains a challenge. While traditional treatment focuses more on speech and its emergence for the patient and family, non-medicinal and non-verbal therapies are used more systematically, sometimes as a last resort. Yet, as a non-verbal mediated therapy, music therapy can find a legitimate place and complement all the psychiatric, psychotherapeutic, somatic, dietetic and nursing areas where speech already circulates.


Changes in heart rate, blood pressure and renal sympathetic nerve activity induced by microinjection of capsaicin into area postrema in rats.
The effects of capsaicin microinjection into area postrema (AP) on mean arterial pressure (MAP), heart rate (HR) and renal sympathetic nerve activity (RSNA) were investigated in 36 anesthetized Sprague-Dawley rats. The results obtained are as follows. (1) Following microinjection of capsaicin (10 micromol/L, 50 nl) into the AP, MAP, HR and RSNA were significantly increased from 12.34+/-0.53 kPa, 328.52+/-7.54 bpm and 100+/-0% to 15.17+/-0.25 kPa (P<0.001), 354.81+/-8.54 bpm (P<0.001) and 156.95+/-7.57% (P<0.001), respectively. (2) Ruthenium red (RR, 100 mmol/L, 0.2 ml, iv), a capsaicin receptor antagonist, significantly inhibited these effects of capsaicin. (3) Pretreatment with a NMDA receptor antagonist MK-801 (500 microgram/kg, 0.2 ml, iv) also reduced these effects of capsaicin. The above results indicate that microinjection of capsaicin into AP induces excitatory effects on MAP, HR and RSNA, which are mediated by capsaicin receptors with glutamate involvement.


Nationwide Effect of COVID-19 on Cases Performed During Pediatric Orthopaedic Surgery Fellowship Training in the United States.
The COVID-19 pandemic negatively affected surgical training in the United States. We hypothesized that reported case volume during pediatric orthopaedic surgery fellowship training would decrease markedly during the 2019 to 2020 academic year, which corresponded with the COVID-19 outbreak. The Accreditation Council for Graduate Medical Education provided nationwide case logs for accredited pediatric orthopaedic surgery fellows (2017 to 2021). Annual reported case volumes were extracted and summarized as means ± SD. Parametric tests were used to compare annual case volumes. A total of 149 pediatric orthopaedic fellows from 23 accredited fellowships were included. A 16% year-over-year (YoY) decrease was noted in the reported case volume during the 2019 to 2020 academic year (238 ± 80 vs. 255 ± 60, P < 0.001). Nonacute case categories had the most notable YoY percentage decreases: Soft Tissue: Transfer, Lengthen, Release (-42%); Clubfoot (-34%); and Foot and Ankle Deformity (-31%). Acute case categories had the most notable YoY percentage increases: Trauma Lower Limb (12%) and Trauma Upper Limb (10%). A subsequent 42% YoY increase was noted in the reported case volume during the 2020 to 2021 academic year. A 16% YoY decrease was noted in the reported case volume during the 2019 to 2020 academic year, which corresponded to widespread economic shutdowns during the initial COVID-19 outbreak. Nonacute cases experienced the greatest negative effect. The results from this study may inform the orthopaedic surgery community on the effect of future national emergencies, such as viral outbreaks.


[The structural functional analysis of beds stock of curative preventive organizations of the state public health system of the Russian Federation].
The article presents the results of comprehensive scientific analysis of size and structure of beds stock of medical curative preventive organizations of state and municipal health care systems of the Russian Federation. The issues of beds support of population on national, federal okrugs and federation subjects' levels including differentiation on different medical specialties are considered. The main indicators of functioning of hospitals, per capita consumption of hospital medical care and territorial characteristics and differences of these indicators are analyzed In conditions of on-going decrease of size of beds stock and amount of medical care in hospitals and against the background of stability of main indicators of beds use the expressed but not always objectively conditioned differences continue to be present concerning both population support with beds stock and indicators of consumption of medical care in hospitals. All these occurrences undoubtedly impact accessibility of this type of medical care to population and its resource capacity for the government. In 2012, beds support of population decreased from 85.7 to 84.1 beds per 10 000 of population. The value of indicator in federal subjects differs up to 2.9 times. In the structure of beds stock are prevailing specialized beds or groups of beds on such medical specialties as psychiatry, surgery, obstetrics and gynecology and therapy. The per capita use of medical care in hospitals decreased up to 2.609 beds-per-day that is 6.2% lower than standard value from the program of state guarantees of free-of-charge medical care support of citizen. The end values of indicator in federal subjects differ in 2.7 times. In federal subjects indicators of mean number of work of bed per year differ up to 1.2 times, of mean duration of treatment--up to 1.6 times, turn-over of bed--up to 1.6 times, hospital lethality--up to 5.9 times. The results of study confirm necessity of structural functional optimization of national beds stock and development of interaction between hospitals and out-patient services.


Hypothalamus-pituitary-adrenal axis of heroin addicts.
The hypothalamus-pituitary-adrenal axis of heroin addicts was investigated by evaluating plasma cortisol levels throughout the day in 37 heroin abusers (HA), 17 of whom showed detectable morphine levels, indicating heroin administration in the previous hours and in 12 controls. All HA showed lower cortisol levels in basal condition (100.7 +/- 61.7 ng/ml, M +/- S.D.) compared to the control group (159.7 +/- 40.6, P less than 0.05). Moreover all HA (65.1 +/- 28.9%), and in particular those taking heroin during the study (39.1 +/- 41.4%), show a reduced cortisol decrease in the evening, which was significantly lower than in controls (85.3 +/- 10.1%; P less than 0.01). As far as the acute effects of opiates are concerned morphine (0.1 mg/kg) significantly reduced plasma cortisol levels within 60 min in a group of 8 healthy subjects. The same pattern was displayed by only 3/8 HA whose morphine levels were higher than 1 ng/ml, i.e. in those reaching the highest plasma morphine concentrations. These data indicate that chronic opiate abuse leads to a hypoadrenalism which could be the result of morphine-induced changes at the hypothalamic level.


Sublobar resection for stage IA non-small cell lung cancer.
Advancements in the diagnosis, staging and management of lung cancer have all led to improvements in outcomes associated with sublobar resection. Lobectomy, for early stage lung cancers has been the treatment of choice for many years. However, there is mounting evidence that sublobar resection when applied to the appropriate patient population can provide not only excellent oncologic results but also equivalent survival to lobectomy. Therefore, it is time that we reevaluate the management of peripheral stage IA lung cancers.


Continuity of health anxiety from childhood to adolescence and associated healthcare costs: a prospective population-based cohort study.
Severe health anxiety (HA) is characterized by excessive and impairing worry and preoccupation with health issues and can cause increased and unnecessary medical examinations. HA in childhood and adolescence is scarcely explored, hindering the potential for prevention and early intervention. HA was assessed in 1,278 children/youths at two time points at ages 11 and 16 years in a general population-based birth cohort. Register-based data on costs related to nonhospital-based primary and secondary somatic health services were obtained over the follow-up period. The presence of functional somatic symptoms, emotional disorders and chronic somatic illness at baseline were included as covariates. High HA (top 10% score) at age 11 predicted high HA at age 16 (relative risk [RR] 2.03, 95% CI: 1.26-3.31). The group with persistent HA was small (n = 17, 1.3%), resulting in broad confidence intervals. The statistical effect of HA at age 11 on HA at age 16 was heavily reduced after adjustment for sex and all covariates (RR: 1.49, 95% CI: 0.85-2.60). In the adjusted model, somatic illness at age 11 (RR: 1.91, 95% CI: 1.22-2.98) and female sex (RR: 3.33, 95% CI: 2.01-5.50) were independently associated with HA at age 16. Persistent HA was associated with approximately doubled healthcare costs compared to the group with consistently low HA. Incident HA at age 16 was associated with increased costs over follow-up. The increased costs were not explained by chronic somatic illness. A small subgroup of children had persistent high levels of HA from late childhood to adolescence and displayed increased healthcare costs. Female sex and chronic somatic disorders at age 11 were independent risk factors of HA at age 16. These findings provide potential means of early identification and of therapeutic levers. Further intervention development and evaluation are needed.


Placement of a vena cava filter in term pregnancy: case report and review of the literature.
During pregnancy there are hemostatic changes that may result in a hypercoagulable state producing thrombotic consequences. This condition may be aggravated in women who are carriers of congenital thrombophilic factors. These factors may increase obstetric complications such as miscarriages, fetal growth restriction, placental abruption and preeclampsia. Trombophilic factors may also cause venous thromboembolism, which is the leading cause of maternal morbidity and mortality. We report a case of a 22-year-old woman with factor V Leiden mutation, whose pregnancy was complicated with deep venous thrombosis requiring placement of a vena cava filter.


Outcomes and factors associated with tolerance in infants with non-IgE-mediated cow's milk allergy with gastrointestinal manifestations.
To evaluate outcomes of oral food challenge (OFC) test to assess tolerance in infants with non-IgE-mediated cow's milk allergy (CMA) with gastrointestinal manifestations and explore clinical data predictive of these outcomes. Single-center retrospective study including infants (age < 12 months) who were referred for CMA between 2000 and 2018 and underwent OFC on follow-up. A univariate logistic regression test was performed to evaluate variables associated with the outcomes of the follow-up OFC test. Eighty-two patients were included, 50% were male. Eighteen patients had a positive OFC test (22%). Most patients had presented with hematochezia (77%). The median age of symptom onset was 30 days. Two-thirds of the patients were on appropriate infant formula (extensively hydrolyzed or amino acid-based formula), exclusively or in association with breastfeeding. The median time on an elimination diet before the OFC test was 8 months (Q1 6 - Q3 11 months). All cases with positive follow-up OFC tests (n = 18) had been exposed to cow's milk-based formula before the first clinical manifestation of CMA. Five out of eight cases with Food Protein-Induced Enterocolitis Syndrome (FPIES) had positive OFC tests. Exposure to cow's milk-based formula before diagnosis, a history of other food allergies, hematochezia and diarrhea were predictors of a positive OFC test. In infants with non-IgE-mediated CMPA with gastrointestinal manifestations, the use of cow's milk-based formula, a history of other food allergies, and hematochezia and diarrhea upon initial presentation were associated factors for the later achievement of tolerance.


Enterohemorrhagic Escherichia coli exploits EspA filaments for attachment to salad leaves.
Enterohemorrhagic Escherichia coli (EHEC) strains are important food-borne pathogens that use a filamentous type III secretion system (fT3SS) for colonization of the gut epithelium. In this study we have shown that EHEC O157 and O26 strains use the fT3SS apparatus for attachment to leaves. Leaf attachment was independent of effector protein translocation.


Experimental and theoretical investigation of overall energy deposition in surface-induced unfolding of protein ions.
Recent advances in native mass spectrometry have enabled its use to probe the structure of and interactions within biomolecular complexes. Surface-induced dissociation, in which inter- and intramolecular interactions are disrupted following an energetic ion-surface collision, is a method that can directly interrogate the topology of protein complexes. However, a quantitative relationship between the ion kinetic energy at the moment of surface collision and the internal energy deposited into the ion has not yet been established for proteins. The factors affecting energy deposition in surface-induced unfolding (SIU) of protein monomers were investigated and a calibration relating laboratory-frame kinetic energy to internal energy developed. Protein monomers were unfolded by SIU and by collision-induced unfolding (CIU). CIU and SIU cause proteins to undergo the same unfolding transitions at different values of laboratory-frame kinetic energy. There is a strong correlation between the SIU and CIU energies, demonstrating that SIU, like CIU, can largely be understood as a thermal process. The change in internal energy in CIU was modeled using a Monte Carlo approach and theory. Computed values of the overall efficiency were found to be approximately 25% and used to rescale the CIU energy axis and relate nominal SIU energies to internal energy. The energy deposition efficiency in SIU increases with mass and kinetic energy from a low of ∼20% to a high of ∼68%, indicating that the effective mass of the surface increases along with the mass of the ion. The effect of ion structure on energy deposition was probed using multiple stages of ion activation. Energy deposition in SIU strongly depends on structure, decreasing as the protein is elongated, due to decreased effective protein-surface collisional cross section and increased transfer to rotational modes.


Early diagnosis of solitary pulmonary nodules.
Early detection of solitary pulmonary nodules (SPNs) and early treatment are of great importance. However, patients with early SPNs always do not present with any symptoms or signs, only to demonstrate SPNs in radiology findings. So it is very critical to improve the ability to identify the SPNs, and with the development of sorts of diagnostic modalities, the accuracy in the evaluation of the SPNs has improved greatly. In this paper, the diagnostic methods and techniques of SPNs are reviewed.


Variation in Nordic Work-Related Cancer Risks after Adjustment for Alcohol and Tobacco.
Background: Alcohol and tobacco strongly increases the risk of cancers of the tongue, mouth, pharynx, larynx, and oesophagus, and are also established risk factors for cancer of the liver, colon, and rectum. It is well documented that these habits are unequally distributed among occupational groups. Most occupational cohort studies lack information on these potentially important confounders, and may therefore be prone to bias. Aim: The aim of the study is to present Nordic standardized incidence ratios (SIRs) for alcohol and tobacco related cancer by occupation, after adjustment for alcohol and tobacco, and to compare to the unadjusted SIRs. Material and Methods: The study is based on the Nordic Occupational Cancer (NOCCA) database. We used confirmatory factor analysis models for simultaneous analysis of the cancer sites related to alcohol and tobacco, to obtain factors that allow for computation of adjusted expected numbers from the reference rates. We then calculated adjusted SIRs for the relevant cancer sites for each occupation. Results: For some occupations and cancers, the changes of risk estimates were striking, from significantly high to significantly low and vice versa. Among Nordic farmers, unadjusted SIRs for cancer of the mouth and oesophagus were 0.56 (95% confidence interval (CI) 0.51⁻0.61) and 0.67 (CI 0.63⁻0.70), respectively. After adjustment, estimates changed to 1.10 (CI 1.01⁻1.21) and 1.16 (CI 1.10⁻1.22). Unadjusted SIR for pharynx cancer among wood workers was 0.83 (CI 0.75⁻0.91), adjusted SIR was 1.14 (CI 1.03⁻1.25). For larynx cancer, results in the opposite direction were seen: unadjusted SIR for economically inactive was 1.38 (CI 1.31⁻1.46) while the adjusted SIR was 0.91 (CI 0.86⁻0.96). Conclusions: Adjustment for the latent indicators of alcohol and tobacco consumption changed risk estimates for several occupations, gave a less confounded description of risk, and may guide in the identification of true risk factors.


Three multispectral configurations of a snapshot kaleidoscope-based camera in long wavelength infrared spectral band.
In the field of spectral imaging, numerous instruments use scanning-based technologies. However, the temporal dimension of these systems, whether to scan the spectrum or scan the scene, can be an issue for some applications. This is particularly the case when trying to observe and identify rapid temporal variations in a fixed scene or detecting objects of interest when moving. In this case, it is suitable to observe the desired spectral information of the scene simultaneously, and so-called snapshot systems have been thus investigated. In this paper, we study the ability of a kaleidoscope-based multiview camera to acquire multispectral information in the long wavelength infrared. Several strategies and technologies will be compared to add the spectral function inside the different blocks of a kaleidoscope-based camera: the front lens, the kaleidoscope, or the reimaging lens. The studied camera uses an uncooled infrared detector and thus must deal with the issue of having a large aperture.


SMAD4 haploinsufficiency in small intestinal neuroendocrine tumors.
Patients with small intestinal neuroendocrine tumors (SINETs) frequently present with lymph node and liver metastases at the time of diagnosis, but the molecular changes that lead to the progression of these tumors are largely unknown. Sequencing studies have only identified recurrent point mutations at low frequencies with CDKN1B being the most common harboring heterozygous mutations in less than 10% of all tumors. Although SINETs are genetically stable tumors with a low frequency of point mutations and indels, they often harbor recurrent hemizygous copy number alterations (CNAs) yet the functional implications of these CNA are unclear. Utilizing comparative genomic hybridization (CGH) arrays we analyzed the CNA profile of 131 SINETs from 117 patients. Two tumor suppressor genes and corresponding proteins i.e. SMAD4, and CDKN1B, were further characterized using a tissue microarray (TMA) with 846 SINETs. Immunohistochemistry (IHC) was used to quantify protein expression in TMA samples and this was correlated with chromosome number evaluated with fluorescent in-situ hybridization (FISH). Intestinal tissue from a Smad4+/- mouse model was used to detect entero-endocrine cell hyperplasia with IHC. Analyzing the CGH arrays we found loss of chromosome 18q and SMAD4 in 71% of SINETs and that focal loss of chromosome 12 affecting the CDKN1B was present in 9.4% of SINETs. No homozygous loss of chromosome 18 was detected. Hemizygous loss of SMAD4, but not CDKN1B, significantly correlated with reduced protein levels but hemizygous loss of SMAD4 did not induce entero-endocrine cell hyperplasia in the Smad4+/- mouse model. In addition, patients with low SMAD4 protein expression in primary tumors more often presented with metastatic disease. Hemizygous loss of chromosome 18q and the SMAD4 gene is the most common genetic event in SINETs and our results suggests that this could influence SMAD4 protein expression and spread of metastases. Although SMAD4 haploinsufficiency alone did not induce tumor initiation, loss of chromosome 18 could represent an evolutionary advantage in SINETs explaining the high prevalence of this aberration. Functional consequences of reduced SMAD4 protein levels could hypothetically be a potential mechanism as to why loss of chromosome 18 appears to be clonally selected in SINETs.


Multifunctional two-photon active silica-coated Au@MnO Janus particles for selective dual functionalization and imaging.
Monodisperse multifunctional and nontoxic Au@MnO Janus particles with different sizes and morphologies were prepared by a seed-mediated nucleation and growth technique with precise control over domain sizes, surface functionalization, and dye labeling. The metal oxide domain could be coated selectively with a thin silica layer, leaving the metal domain untouched. In particular, size and morphology of the individual (metal and metal oxide) domains could be controlled by adjustment of the synthetic parameters. The SiO2 coating of the oxide domain allows biomolecule conjugation (e.g., antibodies, proteins) in a single step for converting the photoluminescent and superparamagnetic Janus nanoparticles into multifunctional efficient vehicles for theranostics. The Au@MnO@SiO2 Janus particles were characterized using high-resolution transmission electron microscopy (HR-)TEM, powder X-ray diffraction (PXRD), optical (UV-vis) spectroscopy, confocal laser fluorescence scanning microscopy (CLSM), and dynamic light scattering (DLS). The functionalized nanoparticles were stable in buffer solution or serum, showing no indication of aggregation. Biocompatibility and potential biomedical applications of the Au@MnO@SiO2 Janus particles were assayed by a cell viability analysis by coincubating the Au@MnO@SiO2 Janus particles with Caki 1 and HeLa cells. Time-resolved fluorescence spectroscopy in combination with CLSM revealed the silica-coated Au@MnO@SiO2 Janus particles to be highly two-photon active; no indication for an electronic interaction between the dye molecules incorporated in the silica shell surrounding the MnO domains and the attached Au domains was found; fluorescence quenching was observed when dye molecules were bound directly to the Au domains.


Osteochondritis dissecans of the humeral head in two small-breed dogs.
Shoulder pathology consistent with osteochondritis dissecans of the humeral head was diagnosed in two small-breed dogs. In both cases, the diagnosis was made tentatively on the basis of radiography and confirmed by arthroscopy of the affected shoulder joint. Both dogs were successfully treated by surgical removal of a flap of cartilage from the affected area. Clinical and radiographic follow-up was available in both cases. The excised cartilage flaps were examined histopathologically and showed the same characteristics as cartilage flaps from dogs of large breeds prone to developing osteochondritis dissecans of the shoulder.


Cysts in malignant gliomas. Identification by computerized tomography.
The clinical course, computerized tomography (CT) scans, and postmortem reports for 265 patients treated for malignant brain tumors were reviewed. Forty-one patients underwent reoperation for tumor recurrence and one had needle aspiration as a diagnostic procedure; of these patients, seven (3% of 265 and 17% of 42) were diagnosed as harboring tumor cysts and 10 (25% of 41) as having necrotic tumor centers. The CT scans on the 17 patients harboring tumors with surgically confirmed cysts and necrotic centers were reviewed; criteria for distinguishing between cysts and central necrosis are suggested. The relative benefits of repeated aspiration and surgical therapy for these cystic lesions are discussed.


Behavioural effects of novel multitarget anticholinesterasic derivatives in Alzheimer's disease.
The current pharmacological approach to Alzheimer's disease (AD) treatment, mostly based on acetylcholinesterase inhibitors (AChEIs), is being revisited, especially in terms of the temporal frames and the potential benefits of their noncanonic actions, raising the question of whether inhibitors of AChE might also act in a disease-modifying manner. Besides, in the last decades, the pharmacophoric moieties of known AChEIs have been covalently linked to other pharmacophores in the pursuit of multitarget hybrid molecules that are expected to induce long-lasting amelioration of impaired neurotransmission and clinical symptoms but also to exert disease-modifying effects. Our research consortium has synthesized and defined the pharmacological profile of new AChEIs derivatives of potential interest for the treatment of AD. Among these, huprines and derivatives have been characterized successfully. Huprine X, a reversible AChE inhibitor, designed by molecular hybridization of tacrine and huperzine A, has been shown to affect the amyloidogenic process in vitro, and the AD-related neuropathology in vivo in mice models of the disease. More recently, we have shown that a group of donepezil-huprine heterodimers exerts a highly potent and selective inhibitory action on AChE both in vitro and ex vivo, simultaneously interacting with both peripheral and catalytic binding sites, and inhibiting the β-amyloid aggregation, whereas some levetiracetam-huprine hybrids have been shown to reduce epileptiform activity, neuroinflammation and amyloid burden in an animal model of AD. Here, we summarize the behavioural correlates of these noncanonic actions as assessed in three distinct biological scenarios: middle-age, cognitive deficits associated with ageing and AD-like phenotype in mice. Besides the improvement in the hallmark cognitive symptomatology without inducing side effects, these drugs have shown to be able to modulate emotional and anxiety-like behaviours or to reduce spontaneous seizures, all of them related to the so-called 'behavioural and psychological symptoms of dementia'. Overall, the studies show that these novel multitarget anticholinesterasics exert noncanonic actions providing symptomatic and disease-modifying benefits of potential interest for the management of AD.


Analysis of rat hemopoietic cells on the fluorescence-activated cell sorter. I. Isolation of pluripotent hemopoietic stem cells and granulocyte-macrophage progenitor cells.
A scheme is presented whereby pluripotent hemopoietic stem cells (PHSC) from rat bone marrow can be enriched 320-fold with the aid of the fluorescence- activated cell sorter. This scheme is based on the observations that PHSC are strongly positive for Thy-1 antigen (upper 10th percentile); have light- scattering properties (size distribution) between those of bone marrow lymphocytes and myeloid progenitor cells; and are relatively resistant to cortisone. It is estimated that PHSC may constitute 80 percent of the cells isolated according to these parameters. Candidate PHSC are described at the light and electron microscopic levels. At least two populations of accessory cells appear to influence the number and/or the nature of the hemopoietic colonies that form in the in vivo spleen colony-forming unit assay. Putative amplifier cells are strongly Thy-1(+) and cortisone sensitive; putative suppressor cells are weakly Thy-1(+) and cortisone resistant. Three subsets of granulocyte (G) -macrophage (M) progenitor cells (in vitro colony-forming cells [CFC]) are identified on the basis of relative fluorescence intensity for Thy-1 antigen: G-CFC are strongly Thy-l(+); M-CFC are weakly Thy-l(+); and cells that produce mixed G and M CFC have intermediate levels of Thy-1. GM-cluster-forming cells and mature G and M are Thy-1(-). The results suggest that G-CFC are bipotential cells that give rise to G and M-CFC; and that the latter produce mature M through a cluster- forming cell intermediate. Thy-1 antigen is also demonstrated on members of the eosinophil, megakaryocyte, erythrocyte, and lymphocyte cell series in rat bone marrow. In each instance, the relative concentration of Thy-1 antigen is inversely related to the state of cellular differentiation.


Effects of exercise training on heart rate variability in individuals with lower extremity arterial disease and claudication: A systematic review.
To perform a systematic review of studies assessing the effects of regular exercise on heart rate variability (HRV) in individuals with lower extremity arterial disease (LEAD) and symptoms of claudication. A systematic search in the electronic databases MEDLINE, Embase, and Scielo, was conducted and updated on January 21, 2023. Randomized clinical trials investigating patients with LEAD and IC, assessing ≥ 4 wk of exercise interventions, and reporting at least one HRV measure (e.g., time or frequency domains) at baseline and follow-up were included. Two reviewers independently screened studies for inclusion, performed data extraction, and quality assessment of included studies. Data from 7 trials were included (i.e., 5 walking, 1 resistance, and 1 isometric handgrip training), totaling 327 patients (66% males; range: 61 - 68 yr; ankle brachial index: 0.4 - 0.7). Following exercise training, three studies investigating walking training reported an increase in parasympathetic modulation indices and/or a decrease in sympathetic modulation indices (n = 2) as well as an increase in non-linear indices (n = 1). The current evidence is weak, and larger randomized controlled trials are needed to confirm the efficacy of exercise training in improving HRV. Additionally, the high divergence in the methodology of studies indicated the need for standard tools to improve the quality of HRV measurements in exercise trials. It is recommended to use standard procedures in future trials investigating HRV.


An integrated computer-controlled system for assisting researchers in cortical excitability studies by using transcranial magnetic stimulation.
Transcranial magnetic stimulation (TMS) is the most important technique currently available to study cortical excitability. Additionally, TMS can be used for therapeutic and rehabilitation purposes, replacing the more painful transcranial electric stimulation (TES). In this paper we present an innovative and easy-to-use tool that enables neuroscientists to design, carry out and analyze scientific studies based on TMS experiments for both diagnostic and research purposes, assisting them not only in the practicalities of administering the TMS but also in each step of the entire study's workflow. One important aspect of this tool is that it allows neuroscientists to specify research designs at will, enabling them to define any parameter of a TMS study starting from data acquisition and sample group definition to automated statistical data analysis and RDF data storage. It also supports the diagnosing process by using on-line support vector machines able to learn incrementally from the diseases instances that are continuously added into the system. The proposed system is a neuroscientist-centred tool where the protocols being followed in TMS studies are made explicit, leaving to the users flexibility in exploring and sharing the results, and providing assistance in managing the complexity of the final diagnosis. This type of tool can make the results of medical experiments more easily exploitable, thus accelerating scientific progress.


Neural activity and diurnal variation of cortisol: evidence from brain electrical tomography analysis and relevance to anhedonia.
The medial prefrontal cortex (mPFC), hippocampus, and amygdala are implicated in the regulation of affect and physiological processes, including hypothalamic-pituitary-adrenal (HPA) axis function. Anhedonia is likely associated with dysregulation of these processes. Dense-array resting electroencephalographic and cortisol were obtained from healthy and anhedonic groups. Low-resolution electromagnetic tomography was used to compute intracerebral current density. For the control group, voxelwise analyses found a relationship between current density in beta and gamma bands and steeper cortisol slope (indicative of more adaptive HPA axis functioning) in regions of the hippocampus, parahippocampal gyrus, and mPFC. For the anhedonic group, the mPFC finding was absent. Anhedonia may be characterized by disruptions of mPFC-mediated neuroendocrine regulation, which could constitute a vulnerability to the development of stress-related disorders.


Targeted Cancer Therapies.
Targeted cancer therapies involve chemotherapeutic agents that attack, directly or indirectly, a specific genetic biomarker found in a given cancer. Targeted oncology includes monoclonal antibodies, small molecule inhibitors, antibody-drug conjugates, and immunotherapy. For example, the monoclonal antibodies trastuzumab and pertuzumab target human epidermal growth factor receptor 2 (HER2) and are used when treating HER2-positive breast cancer. Although targeted oncology has improved survival by years for some incurable cancers such as metastatic breast and lung cancer, as few as 8% of patients with advanced cancer qualify for targeted oncology medications, and even fewer benefit. Other limitations include serious adverse events, illustrated by a 20% to 30% rate of heart attack, stroke, or peripheral vascular events among patients taking ponatinib, which is used in treating chronic myelogenous leukemia. Immune checkpoint inhibitor therapy-related adverse effects such as hypothyroidism are common, and more severe adverse events such as colitis and pneumonitis can be fatal and require immediate intervention. Drug interactions with widely prescribed medications such as antacids and warfarin are common. Additionally, financial toxicities are a problem for patients with cancer who are using costly targeted therapies. Future directions for targeted oncology include tumor-agnostic drugs, which target a given mutation and could be used in treating cancers from multiple organ types. An overview of indications, mechanism of action, and toxicities of targeted cancer therapies is offered here.


Physical Activity through a Classroom-Based Intervention: A Pragmatic Non-Randomized Trial among Swedish Adolescents in an Upper Secondary School.
Schools are an important arena to curb the decline in physical activity (PA) in youth. School-based interventions with accelerometer-measured PA are warranted. This study aimed to increase accelerometer-measured PA in adolescents following a 12-month school-based intervention. Two school-classes of 16-18-year-old Swedish students were allocated to intervention group and control group. Accelerometer-measured PA was gathered at baseline, 6- and 12-month follow-up. Mixed-effects linear regression was used to investigate between-group and within-group differences in mean minutes per day (min/day) of moderate to vigorous PA (MVPA), light PA (LPA) and sedentary time (ST). Fifty-seven students participated (intervention group = 31, control group = 26). At 12-month follow-up, the intervention group performed 5.9 (95% CI: -4.3, 16.2) min/day more in MVPA, 1.8 (95% CI: -17.9, 14.2) min/day less in LPA, and 4.1 (95% CI: -27.3, 19.2) min/day less in ST compared to the control group. Within the intervention group, there was no significant change in PA. Within the control group, LPA decreased (95% CI: -19.6, -0.2; p = 0.044) and ST increased (95% CI: 1.8, 30.8; p = 0.028). Although no between-group differences in PA were statistically significant, the within-group changes may suggest a preventive impact on the decline in PA during adolescence.


Innovative One-Shot Paradigm to Tune Filler-Polymer Matrix Interface Properties by Plasma Polymer Coating in Osteosynthesis Applications.
The present study aims to improve the interfacial bonding between hydroxyapatite particles (HAs) and polylactide (PLA) to enhance the mechanical performance and biocompatibility of bone implants based on HA/PLA. For this, one-shot surface functionalization of HA via plasma polymerization is developed. Taking advantage of acetylene plasma chemistry, the hydrophobicity of HA particles was finely tuned prior to their introduction into a PLA matrix via an extrusion process. The effect of the plasma power (20 or 100 W) on the composition of the plasma polymer film (PPF) formed on the HA surface was studied via Fourier transform infrared (FTIR) spectroscopy, time-of-flight secondary-ion mass spectrometry (ToF-SIMS), and X-ray photoelectron spectroscopy (XPS). The amount of PPF formed was evaluated via thermogravimetric analyses (TGA). Cytotoxicity of the modified HA particles was monitored by the WST-1 proliferation assay and lactate dehydrogenase (LDH) release and showed that independent on the studied conditions, cell viability remained above the 70% threshold and LDH accumulation changes were insignificant, suggesting good biocompatibility. Contact angle measurements and morphological and rheological analyses showed that the low working power promoted more hydrophobic surfaces and a better HA/PLA interface. Dynamic mechanical analyses revealed that the storage modulus at 37 °C increased for the composite containing functionalized particles by 1.5 times compared to the neat particle's composites. This work opens a route toward further one-shot development of improved scaffolds for bone tissue engineering.


Is thermosensing property of RNA thermometers unique?
A large number of studies have been dedicated to identify the structural and sequence based features of RNA thermometers, mRNAs that regulate their translation initiation rate with temperature. It has been shown that the melting of the ribosome-binding site (RBS) plays a prominent role in this thermosensing process. However, little is known as to how widespread this melting phenomenon is as earlier studies on the subject have worked with a small sample of known RNA thermometers. We have developed a novel method of studying the melting of RNAs with temperature by computationally sampling the distribution of the RNA structures at various temperatures using the RNA folding software Vienna. In this study, we compared the thermosensing property of 100 randomly selected mRNAs and three well known thermometers--rpoH, ibpA and agsA sequences from E. coli. We also compared the rpoH sequences from 81 mesophilic proteobacteria. Although both rpoH and ibpA show a higher rate of melting at their RBS compared with the mean of non-thermometers, contrary to our expectations these higher rates are not significant. Surprisingly, we also do not find any significant differences between rpoH thermometers from other gamma-proteobacteria and E. coli non-thermometers.


Activated Raf-1 causes growth arrest in human small cell lung cancer cells.
Small cell lung cancer (SCLC) accounts for 25% of all lung cancers, and is almost uniformly fatal. Unlike other lung cancers, ras mutations have not been reported in SCLC, suggesting that activation of ras-associated signal transduction pathways such as the raf-MEK mitogen-activated protein kinases (MAPK) are associated with biological consequences that are unique from other cancers. The biological effects of raf activation in small cell lung cancer cells was determined by transfecting NCI-H209 or NCI-H510 SCLC cells with a gene encoding a fusion protein consisting of an oncogenic form of human Raf-1 and the hormone binding domain of the estrogen receptor (DeltaRaf-1:ER), which can be activated with estradiol. DeltaRaf-1:ER activation resulted in phosphorylation of MAPK. Activation of this pathway caused a dramatic loss of soft agar cloning ability, suppression of growth capacity, associated with cell accumulation in G1 and G2, and S phase depletion. Raf activation in these SCLC cells was accompanied by a marked induction of the cyclin-dependent kinase (cdk) inhibitor p27(kip1), and a decrease in cdk2 protein kinase activities. Each of these events can be inhibited by pretreatment with the MEK inhibitor PD098059. These data demonstrate that MAPK activation by DeltaRaf-1:ER can activate growth inhibitory pathways leading to cell cycle arrest. These data suggest that raf/MEK/ MAPK pathway activation, rather than inhibition, may be a therapeutic target in SCLC and other neuroendocrine tumors.


The Effects of Natural Product-Derived Extracts for Longitudinal Bone Growth: An Overview of In Vivo Experiments.
Approximately 80% of children with short stature are classified as having Idiopathic Short Stature (ISS). While growth hormone (GH) treatment received FDA approval in the United States in 2003, its long-term impact on final height remains debated. Other treatments, like aromatase inhibitors, metformin, and insulin-like growth factor-1 (IGF-1), have been explored, but there is no established standard treatment for ISS. In South Korea and other Asian countries, East Asian Traditional Medicine (EATM) is sometimes employed by parents to potentially enhance their children's height growth, often involving herbal medicines. One such product, Astragalus membranaceus extract mixture HT042, claims to promote height growth in children and has gained approval from the Korean Food and Drug Administration (KFDA). Research suggests that HT042 supplementation can increase height growth in children without skeletal maturation, possibly by elevating serum IGF-1 and IGF-binding protein-3 levels. Preclinical studies also indicate the potential benefits of natural products, including of EATM therapies for ISS. The purpose of this review is to offer an overview of bone growth factors related to ISS and to investigate the potential of natural products, including herbal preparations, as alternative treatments for managing ISS symptoms, based on their known efficacy in in vivo studies.


Outcrossing between an agroforestry plantation and remnant native populations of Eucalyptus loxophleba.
Gene dispersal among populations of a species is an important force influencing their genetic structure. Dispersal may also occur between taxa that would normally be isolated when nonendemic, domesticated or transgenic species are planted within the natural range of interfertile taxa. Such a mosaic of populations is typical of many agricultural landscapes, and investigations are needed to assess the risks of genetic contamination of the endemic populations but a combination of approaches may be necessary because of the limitations of research in this landscape. This study used microsatellite markers and a range of analyses (mating system, paternity exclusion, Bayesian assignment) to examine gene dispersal between remnants of the endemic Eucalyptus loxophleba ssp. supralaevis and a plantation of a nonendemic subspecies. Our results indicate that remnant populations are connected by significant dispersal to pollen sources up to 1.94 km away including the plantation. The combined analyses showed that the pollen pool and outcrossing rates of individuals within remnants varied significantly probably because of asynchronous flowering and that the likelihood of paternity was not correlated with spatial proximity. More than half of all progeny had male parents from outside their stand with the largest proportions estimated to come from the plantation by exclusion (42.4%) or Bayesian analyses (18.8-76%). Fragmentation may not be associated with decreased gene dispersal between populations of tree species, natural or planted, so that the distances required to buffer endemic trees in fragmented rural landscapes are likely to be large.


Isolation of polysomes from Nostoc sp. MAC and translation of messenger RNA in a heterologous cell-free system.
A method has been established which isolated polysomes from the lysozyme/EDTA-shocked cyanobacterium, Nostoc sp. MAC. In a typical preparation the total recovery of RNA as polysomes was 83%, in which 77% of the polysome fraction was present at sizes greater than 5-mers and 23% as 2-4-mers. Messenger RNA isolated from such a preparation of polysomes produced a 10-fold stimulation in the incorporation of [35S]methionine into polypeptides by a cell-free system of Escherichia coli. The in vitro-synthesized polypeptides were analysed on an SDS-polyacrylamide gradient gel together with in vivo-labelled proteins of Nostoc sp. MAC: seven polypeptides co-migrated with the in vivo-synthesized products. This is the first report of the expression of cyanobacterial messenger RNA in a heterologous cell-free system from E. coli; the efficiency of the system is discussed.


Daily cognitive complaints and engagement in older adulthood: Personality traits are more predictive than cognitive performance.
Cognitive complaints and engagement in cognitive activities are two consistent predictors of cognitive aging outcomes, including risk for nonnormative decline. Though research has considered predictors of complaints and engagement in general, little work has attended to the fact that these fluctuate at the daily level. The current study examined individual difference predictors of means and variability for engagement and complaints across 10 days in a sample of older adults (n = 136; Mage = 70.45 years). When comparing personality traits to indicators of cognitive performance, personality differences appeared better unique predictors for these measures of daily cognitive life. Specifically, even when accounting for demographics, measures of cognitive performance, and the other personality traits investigated, older adults higher on openness to experience reported fewer daily cognitive complaints and more engagement on average, as well as greater daily variability in engagement. In addition, higher neuroticism predicted greater variability in reports of cognitive complaints across days. Implications are discussed with respect to how these findings advance our understanding of cognitive complaints and engagement in daily life. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Inferring disease-related pathways using a probabilistic epistasis model.
We present a probabilistic model called a Joint Intervention Network (JIN) for inferring interactions among a chosen set of regulator genes. The input to the method are expression changes of downstream indicator genes observed under the knock-out of the regulators. JIN can use any number of perturbation combinations for model inference (e.g. single, double, and triple knock-outs). RESUITS/CONCLUSIONS: We applied JIN to a Vibrio cholerae regulatory network to uncover mechanisms critical to its environmental persistence. V. cholerae is a facultative human pathogen that causes cholera in humans and responsible for seven pandemics. We analyzed the expression response of 17 V. cholerae biofilm indicator genes under various single and multiple knock-outs of three known biofilm regulators. Using the inferred network, we were able to identify new genes involved in biofilm formation more accurately than clustering expression profiles.


Global error minimization in image mosaicing using graph connectivity and its applications in microscopy.
Several applications such as multiprojector displays and microscopy require the mosaicing of images (tiles) acquired by a camera as it traverses an unknown trajectory in 3D space. A homography relates the image coordinates of a point in each tile to those of a reference tile provided the 3D scene is planar. Our approach in such applications is to first perform pairwise alignment of the tiles that have imaged common regions in order to recover a homography relating the tile pair. We then find the global set of homographies relating each individual tile to a reference tile such that the homographies relating all tile pairs are kept as consistent as possible. Using these global homographies, one can generate a mosaic of the entire scene. We derive a general analytical solution for the global homographies by representing the pair-wise homographies on a connectivity graph. Our solution can accommodate imprecise prior information regarding the global homographies whenever such information is available. We also derive equations for the special case of translation estimation of an X-Y microscopy stage used in histology imaging and present examples of stitched microscopy slices of specimens obtained after radical prostatectomy or prostate biopsy. In addition, we demonstrate the superiority of our approach over tree-structured approaches for global error minimization.


Human mediator enhances activator-facilitated recruitment of RNA polymerase II and promoter recognition by TATA-binding protein (TBP) independently of TBP-associated factors.
Mediator is a general cofactor implicated in the functions of many transcriptional activators. Although Mediator with different protein compositions has been isolated, it remains unclear how Mediator facilitates activator-dependent transcription, independent of its general stimulation of basal transcription. To define the mechanisms of Mediator function, we isolated two forms of human Mediator complexes (Mediator-P.5 and Mediator-P.85) and demonstrated that Mediator-P.5 clearly functions by enhancing activator-mediated recruitment of RNA polymerase II (pol II), whereas Mediator-P.85 works mainly by stimulating overall basal transcription. The coactivator function of Mediator-P.5 was not impaired when TATA-binding protein (TBP) was used in place of TFIID, but it was abolished when another general cofactor, PC4, was omitted from the reaction or when Mediator-P.5 was added after pol II entry into the preinitiation complex. Moreover, Mediator- P.5 is able to enhance TBP binding to the TATA box in an activator-dependent manner. Our data provides biochemical evidence that Mediator functions by facilitating activator-mediated recruitment of pol II and also promoter recognition by TBP, both of which can occur in the absence of TBP-associated factors in TFIID.


Molecular evolution of GPCRs: Melanocortin/melanocortin receptors.
The melanocortin receptors (MCRs) are a family of G protein-coupled receptors that are activated by melanocortin ligands derived from the proprotein, proopiomelanocortin (POMC). During the radiation of the gnathostomes, the five receptors have become functionally segregated (i.e. melanocortin 1 receptor (MC1R), pigmentation regulation; MC2R, glucocorticoid synthesis; MC3R and MC4R, energy homeostasis; and MC5R, exocrine gland physiology). A focus of this review is the role that ligand selectivity plays in the hypothalamus/pituitary/adrenal-interrenal (HPA-I) axis of teleosts and tetrapods as a result of the exclusive ligand selectivity of MC2R for the ligand ACTH. A second focal point of this review is the roles that the accessory proteins melanocortin 2 receptor accessory protein 1 (MRAP1) and MRAP2 are playing in, respectively, the HPA-I axis (MC2R) and the regulation of energy homeostasis by neurons in the hypothalamus (MC4R) of teleosts and tetrapods. In addition, observations are presented on trends in the ligand selectivity parameters of cartilaginous fish, teleost, and tetrapod MC1R, MC3R, MC4R, and MC5R paralogs, and the modeling of the HFRW motif of ACTH(1-24) when compared with α-MSH. The radiation of the MCRs during the evolution of the gnathostomes provides examples of how the physiology of endocrine and neuronal circuits can be shaped by ligand selectivity, the intersession of reverse agonists (agouti-related peptides (AGRPs)), and interactions with accessory proteins (MRAPs).


Robust Electrografting on Self-Organized 3D Graphene Electrodes.
Improving graphene-based electrode fabrication processes and developing robust methods for its functionalization are two key research routes to develop new high-performance electrodes for electrochemical applications. Here, a self-organized three-dimensional (3D) graphene electrode processed by pulsed laser deposition with thermal annealing is reported. This substrate shows great performance in electron transfer kinetics regarding ferrocene redox probes in solution. A robust electrografting strategy for covalently attaching a redox probe onto these graphene electrodes is also reported. The modification protocol consists of a combination of diazonium salt electrografting and click chemistry. An alkyne-terminated phenyl ring is first electrografted onto the self-organized 3D graphene electrode by in situ electrochemical reduction of 4-ethynylphenyl diazonium. Then the ethynylphenyl-modified surface efficiently reacts with the redox probe bearing a terminal azide moiety (2-azidoethyl ferrocene) by means of Cu(I)-catalyzed alkyne-azide cycloaddition. Our modification strategy applied to 3D graphene electrodes was analyzed by means of atomic force microscopy, scanning electron microscopy, Raman spectroscopy, cyclic voltammetry, and X-ray photoelectron spectroscopy (XPS). For XPS chemical surface analysis, special attention was paid to the distribution and chemical state of iron and nitrogen in order to highlight the functionalization of the graphene-based substrate by electrochemically grafting a ferrocene derivative. Dense grafting was observed, offering 4.9 × 10(-10) mol cm(-2) surface coverage and showing a stable signal over 22 days. The electrografting was performed in the form of multilayers, which offers higher ferrocene loading than a dense monolayer on a flat surface. This work opens highly promising perspectives for the development of self-organized 3D graphene electrodes with various sensing functionalities.


Blood Pressure Targets for Older Patients-Do Advanced Age and Frailty Really Not Matter?
In 2017, Hypertension Canada removed advanced age and frailty as considerations for caution when deciding on intensive therapy in their guidelines for the diagnosis, risk assessment, prevention, and treatment of hypertension in adults. Dementia is not mentioned. In this commentary, we review why advanced age and frailty were removed, and examine what is currently known about the relationship between hypertension and both incident and prevalent dementia. We make the case that the presence of frailty (especially when severe) and dementia should be considered when deciding on intensive therapy in future iterations of Hypertension Canada guidelines.


A "Prime and Deploy" Strategy for Universal Influenza Vaccine Targeting Nucleoprotein Induces Lung-Resident Memory CD8 T cells.
Lung-resident memory T cells (TRM) play an essential role in protecting against pulmonary virus infection. Parenteral administration of DNA vaccine is generally not sufficient to induce lung CD8 TRM cells. This study investigates whether intramuscularly administered DNA vaccine expressing the nucleoprotein (NP) induces lung TRM cells and protects against the influenza B virus. The results show that DNA vaccination poorly generates lung TRM cells and massive secondary effector CD8 T cells entering the lungs after challenge infection do not offer sufficient protection. Nonetheless, intranasal administration of non-replicating adenovirus vector expressing no Ag following priming DNA vaccination deploys NP-specific CD8 TRM cells in the lungs, which subsequently offers complete protection. This novel 'prime and deploy' strategy could be a promising regimen for a universal influenza vaccine targeting the conserved NP Ag.


Solid-state NMR studies of the structure, dynamics, and assembly of beta-sheet membrane peptides and alpha-helical membrane proteins with antibiotic activities.
beta-Sheet antimicrobial peptides and alpha-helical channel-forming colicins are bactericidal molecules that target the lipid membranes of sensitive cells. Understanding the mechanisms of action of these proteins requires knowledge of their three-dimensional structure in the lipid bilayer. Solid-state NMR has been used to determine the conformation, orientation, depth of insertion, oligomerization, mobility, and lipid interaction of these membrane peptides and proteins. We review the NMR methods developed and applied to study the structure and dynamics of these antibiotic membrane proteins. These studies shed light on how these peptides disrupt lipid membranes and provide fundamental insights into the folding of beta-sheet and alpha-helical membrane proteins.


Teratological risk evaluation and prevention of voluntary abortion.
Many women exposed to completely innocuous agents during pregnancy have a high perception of adverse effects to such an extent that they may interrupt their pregnancy. The objective of our study is to evaluate the importance of the perception of the risk level in making the decision to end the pregnancy and the relevance that a teratology consultation can have in preventing unmotivated terminations of pregnancy We carried out a survey on 350 women in Rome who voluntarily interrupted their pregnancy to evaluate the prevalence due to presumed teratogen. Contemporarily we studied the pregnancy outcomes, the clinical, the psychological and the socio-economic factors of 142 women who contacted our Teratology Information Service (TIS) in the 1(st)trimester of pregnancy because suspected of teratogen exposure: 72 decided to terminate their pregnancy, whereas 70 were used as a control group. On 350 women who voluntarily interrupted their pregnancy, 4 cases (1.4%) reported exposure to a suspected teratogen, but our evaluation determined only 1 case. On 72 women decided to terminate their pregnancy and who contacted our TIS, after counselling 73% continued their pregnancy with respect to 97% of the control group. Those women who interrupted their pregnancy did so because of personal reasons independently to or the type of exposure or the risk attributed by us. From our data it appears that a percentage of voluntary abortions is related to suspected teratogen exposure and that TIS are effective in the prevention of this kind of voluntary abortions caused by groundless fears.


Evaluation of the effects of gold nanoparticle shape and size on contrast enhancement in radiological imaging.
There has been increasing interest in the use of a nanoparticle-based media as a contrast-enhancement agent in medical imaging, particularly with gold Nanoparticles in radiography. Particularly attractive, is the prospect of modifying the surface of these materials with monoclonal antibodies to preferentially bind the nanoparticles to tumour sites. These materials differ from conventional molecular agents in their ability to be modified with cell specificity, or tailored for size and shape for maximum uptake. We investigated the consideration that quantum confinement electronic effects in nanometre-sized metals might have an effect on the integrated photon attenuation of gold atoms; in the same manner as these materials affect X-ray absorption and scattering as seen in X-ray absorption spectroscopy. This experiment has been designed to identify any effect on contrast enhancement that might result from employing gold nanoparticles with a variety of sizes. Spherical particles and nanorods were synthesised for this application. Image contrast enhancement was quantified by contrast-to-noise ratio in computed radiography. Results are consistent with existing measurements of gold nanoparticle contrast enhancement in radiography. No significant variation in attenuation depending on particle size was observed. Findings indicate that nanoparticle-based contrast agents in the size range 4-30 nm-can be synthesised for maximum stability or cell specificity (directed cellular uptake) without consideration of effect of size on contrast enhancement.