Fall in C-peptide during first 2 years from diagnosis: evidence of at least two distinct phases from composite Type 1 Diabetes TrialNet data.
Interpretation of clinical trials to alter the decline in β-cell function after diagnosis of type 1 diabetes depends on a robust understanding of the natural history of disease. Combining data from the Type 1 Diabetes TrialNet studies, we describe the natural history of β-cell function from shortly after diagnosis through 2 years post study randomization, assess the degree of variability between patients, and investigate factors that may be related to C-peptide preservation or loss. We found that 93% of individuals have detectable C-peptide 2 years from diagnosis. In 11% of subjects, there was no significant fall from baseline by 2 years. There was a biphasic decline in C-peptide; the C-peptide slope was -0.0245 pmol/mL/month (95% CI -0.0271 to -0.0215) through the first 12 months and -0.0079 (-0.0113 to -0.0050) from 12 to 24 months (P < 0.001). This pattern of fall in C-peptide over time has implications for understanding trial results in which effects of therapy are most pronounced early and raises the possibility that there are time-dependent differences in pathophysiology. The robust data on the C-peptide obtained under clinical trial conditions should be used in planning and interpretation of clinical trials.


Vein graft surveillance: is graft revision without angiography justified and what criteria should be used?
The objective of this study was to assess the accuracy of color-flow duplex surveillance parameters to detect infrainguinal vein graft stenoses and to investigate whether graft revision without angiography is justified. In a prospective study in which three centers participated, the data of graft surveillance in 300 patients were analyzed. For the evaluation of surveillance criteria all patients underwent a digital subtraction angiography if a graft stenosis was suspected. To create a control group, in patients with normal grafts a consented digital subtraction angiography was performed also. From these data the accuracy of seven duplex and three ankle blood pressure-derived variables was assessed. The relation between various surveillance criteria and continued graft patency was determined with life table analysis with the transient state method. The mean follow-up period was 20 months (range, 1 to 40 months). At univariate and multivariate analysis the peak systolic velocity (PSV) ratio provided the best correlation with angiographic stenoses > or = 70% (PSV ratio cutoff 3.0: sensitivity 80%, specificity 84%). This finding did not differ between the participating centers. With life table methods it was demonstrated that the best combination of efficacy (limitation of the number of unnecessary revisions), safety (minimal number of correctable lesions missed), and reduction of angiograms was obtained by a two-parameter surveillance algorithm. This algorithm included a PSV ratio < 2.5 to delineate patients in whom a conservative approach without angiography or revision was appropriate, a PSV ratio > or = 4.0 to indicate patients in whom vein graft revision without angiography could be scheduled, and a group with PSV ratios between 2.5 and 4.0 in whom angiography was to be performed to determine clinical management on the basis of the stenosis severity. This algorithm had a positive predictive value of 93% and a negative predictive value of 89%. In addition, it resulted in a reduction of the number of angiograms of 49% compared with a policy of angiographies in all patients with a PSV ratio > or = 2.5. The best criterion to identify a failing graft is the PSV ratio. With a two-parameter algorithm for vein graft surveillance, the incidence of unnecessary revisions and of missed high-grade lesions was acceptably low, whereas the number of angiograms was reduced by one half.


Unexpected formation of 1-diethylaminobutadiene in photosensitized oxidation of triethylamine induced by 2,3-dihydro-oxoisoaporphine dyes. A 1H NMR and isotopic exchange study.
[reaction: see text] Photoreduction of oxoisoaporphine dyes occurs via a stepwise mechanism of electron-proton-electron transfer that leads to the N-hydrogen oxoisoaporphine anion. When triethylamine, TEA, was used as the electron donor in anaerobic conditions, 1-diethylaminobutadiene, DEAB, was one of the oxidation products of TEA, among diethylamine and acetaldehyde. DEAB was identified by (1)H NMR and GC-MS experiments by comparison with the authentic 1-diethylaminobutadiene. This is the first report of a butadienyl derivative formed in the dye-sensitized photooxidation of TEA. In addition, isotopic exchange experiments with TEA-d(15) and D(2)O show that the hydrogens at carbon-2 and carbon-4 of the butadienyl moiety are exchangeable. The observed isotopic exchange pattern could be explained by the head-to-tail coupling of an N,N-diethylvinylamine intermediate that exchanges hydrogens at the C-beta via the enammonium ion.


Ubiquitin Ser65 phosphorylation affects ubiquitin structure, chain assembly and hydrolysis.
The protein kinase PINK1 was recently shown to phosphorylate ubiquitin (Ub) on Ser65, and phosphoUb activates the E3 ligase Parkin allosterically. Here, we show that PINK1 can phosphorylate every Ub in Ub chains. Moreover, Ser65 phosphorylation alters Ub structure, generating two conformations in solution. A crystal structure of the major conformation resembles Ub but has altered surface properties. NMR reveals a second phosphoUb conformation in which β5-strand slippage retracts the C-terminal tail by two residues into the Ub core. We further show that phosphoUb has no effect on E1-mediated E2 charging but can affect discharging of E2 enzymes to form polyUb chains. Notably, UBE2R1- (CDC34), UBE2N/UBE2V1- (UBC13/UEV1A), TRAF6- and HOIP-mediated chain assembly is inhibited by phosphoUb. While Lys63-linked poly-phosphoUb is recognized by the TAB2 NZF Ub binding domain (UBD), 10 out of 12 deubiquitinases (DUBs), including USP8, USP15 and USP30, are impaired in hydrolyzing phosphoUb chains. Hence, Ub phosphorylation has repercussions for ubiquitination and deubiquitination cascades beyond Parkin activation and may provide an independent layer of regulation in the Ub system.


Elimination of a human T-cell region in staphylokinase by T-cell screening and computer modeling.
Staphylokinase is a potent highly fibrin-selective thrombolytic agent, but it induces a humoral immune response in most treated patients. Staphylokinase-specific T-lymphocytes can be found in normal healthy individuals, from whom a large panel of staphylokinase-specific T-cells were cloned. The staphylokinase amino acid sequence 71-87 was widely recognized, as it induced proliferation of T-cell clones isolated from 90% of the donors. Computer modeling of this area, threaded as 11-mer peptides within the peptide-binding groove of the major HLA-DR alleles, indicated two putative partially overlapping binding sequences. The region-(71-87)-specific T-cell clones recognized either one or the other minimal peptide, confirming that both sequences could be functional T-cell epitopes. Furthermore, to guide the mutagenesis to eliminate T-cell reactivity, the contribution of each residue to the HLA-DR-anchoring and T-cell receptor exposure was evaluated for both binding motifs. Computer calculations combined with functional assays resulted in the design of staphylokinase-variants, including 2 to 4 amino acid substitutions in the region 71-87. These variants were no longer recognized by the region-(71-87)-specific T-cell clones, and importantly no new staphylokinase-variant-specific cellular immune response could be measured.


Correlation between resistin gene polymorphism and clinical aspects of lung cancer.
Lung cancer is one of the most common cancers and is associated with a poor survival rate in the Chinese Han population. Analysis of genetic variants could lead to improvements in prognosis following lung cancer treatment. Resistin (RETN) is an important mediator of metabolic diseases and tumor progression. In this study, we explored the effects of RETN single nucleotide polymorphisms (SNPs) on the susceptibility and clinicopathological characteristics of patients with lung cancer. Four RETN SNPs (rs7408174, rs1862513, rs3745367, and rs3219175) were analyzed using TaqMan SNP genotyping in 371 patients with lung cancer and 451 cancer-free controls. The results showed that the RETN SNP rs3219175 with AG or at least 1 A allele was associated with a higher risk of lung cancer than wild-type (GG) carriers. Moreover, the RETN SNP rs3219175 with AG or AG + AA alleles was associated with a higher risk of distant metastasis than that in patients carrying GG alleles. We also used genotype-tissue expression datasets to compare the correlation of the RETN SNP rs3219175 in lung tissue and whole blood. In conclusion, our study demonstrated, for the first time, that RETN polymorphisms were correlated with lung cancer progression in the Chinese Han population.


A Novel Bedside-Focused Ward Surveillance and Response System.
Rapid response systems (RRSs) have been universally adopted in much of the developed world; yet, despite broad implementation, their success has often been limited. Even with successful systems, there is a small body of evidence regarding effective organizational elements that are responsible for improved outcomes. New organizational processes were implemented that restructured the existing RRS, and the impact on the number of rapid response team (RRT) alerts, cardiac arrest, and mortality rates was evaluated. A prospective five-year before-and-after comparison of adult ward patient outcomes was conducted at a community regional medical center. The key intervention was expanded administrative oversight of the system, which led to (1) restructuring the content and depth of ward nurse education regarding early recognition of at-risk patients; (2) system changes empowering prompt mobilization of the RRT; (3) development of RRT treatment protocols; and (4) a more frequent and comprehensive data collection and analysis for system compliance and performance improvement. Some 28,914 patients were observed in the 24-month control period, and 39,802 patients were observed in the 33-month intervention period. RRT activations increased from 10.2 to 48.8/1,000 discharges (p <0.001), ward cardiac arrest decreased from 3.1 to. 2.4/1000 discharges (p = 0.04), hospital mortality decreased from 3.8% to 3.2% (p <0.001), and the observed-to-expected ratio decreased from 1.5 to 1.0 (p <0.001). Expanded administrative involvement of an existing RRS that focused on early recognition of patient deterioration by the bedside nurse led to improved performance of the system, with a significant increase in number of RRTs and decreases in cardiac arrests and hospital mortality.


Distinctive characteristics of granulation tissue in laparotomy wounds with underlying oncological processes.
This study aimed to investigate the effects of malignant neoplasms on the morphological characteristics of laparotomy wound granulation tissue in the muscular-aponeurotic layer. The study involved a sample of 34 deceased individuals who had undergone abdominal organ surgery. Biopsy samples were taken from the muscular-aponeurotic layer of the anterior abdominal wall and subjected to histological examination, including staining with hematoxylin and eosin and methylene blue/Chromotrope 2B using N.Z. Slinchenko's method. Descriptive methods and morphometry were used to evaluate pathomorphological changes. The results suggest that malignant neoplasms significantly impede and decelerate the maturation of laparotomy wound granulation tissue. Surgeries performed at the late stages of abdominal organ malignant neoplasms result in an uneven and slow maturation of the tissue, characterized by a higher prevalence of lymphoid cells, increased blood vessel volume, reduced optical density of stained collagen fibers, and pronounced chromotropophilia of collagen fibers. These distinct features should be considered to prevent postoperative eventration, a complication that is more likely to occur in this patient group. Clinicians should be aware of the possible consequences of malignant neoplasms on laparotomy wound granulation tissue, which may require additional measures to prevent postoperative complications in these patients.


Pd(II)-Catalyzed, Picolinamide-Assisted, Z-Selective γ-Arylation of Allylamines To Construct Z-Cinnamylamines.
Investigations of Pd(II)-catalyzed, picolinamide-assisted, γ-C(sp2)-H activation and Z-selective arylation of allylamines are reported. The reactions of N-allylpicolinamides with various aryl iodides in the presence of the catalyst Pd(OAc)2 and additive AgOAc have led to the selective γ-arylation of allylamines to construct various cinnamylamines with moderate to good yields and good to high E/Z ratios. To obtain good E/Z ratios, the Pd(II)-catalyzed arylation reaction of N-allylpicolinamides was probed using different additives, directing groups, and reaction conditions. The Pd(II)-catalyzed arylation of an allylamine containing both γ-C(sp2)-H and γ-C(sp3)-H bonds afforded moderate yields of the γ-C(sp2)-H and γ-C(sp3)-H bisarylated cinnamylamines. Although Heck-type γ-arylations of allylamines have generally afforded the E-cinnamylamines, the bidentate directing group picolinamide-directed arylations of allylamines were found to be Z-selective. A plausible mechanism was proposed for the observed regioselectivity and Z-selective arylation of N-allylpicolinamides. Additionally, the Pd(II)-catalyzed arylation of an N-allyl-5-methylisoxazole-3-carboxamide afforded the E-cinnamylamines plausibly via a ligand-free Heck-type reaction mechanism.


Crystal structure of enolase from Drosophila melanogaster.
Enolase is an important enzyme in glycolysis and various biological processes. Its dysfunction is closely associated with diseases. Here, the enolase from Drosophila melanogaster (DmENO) was purified and crystallized. A crystal of DmENO diffracted to 2.0 Å resolution and belonged to space group R32. The structure was solved by molecular replacement. Like most enolases, DmENO forms a homodimer with conserved residues in the dimer interface. DmENO possesses an open conformation in this structure and contains conserved elements for catalytic activity. This work provides a structural basis for further functional and evolutionary studies of enolase.


Involvement of STAT3 in bladder smooth muscle hypertrophy following bladder outlet obstruction.
We examined the involvement of the signal transducer and activator of transcription 3 (STAT3) in bladder outlet obstruction (BOO)-induced bladder smooth muscle hypertrophy using a rat in vivo and in vitro study. BOO induced increases in bladder weight and bladder smooth muscle thickness 1 week after the operation. By using antibody microarrays, 64 of 389 proteins blotted on the array met our selection criteria of an INR value between > or = 2.0 and < or = 0.5. This result revealed up-regulation of transcription factors, cell cycle regulatory proteins, apoptosis-associated proteins and so on. On the other hand, down-regulation (INR value < or = 0.5) of proteins was not found. In a profiling study, we found an increase in the expression of STAT3. A significant increase in nuclear phosphorylated STAT3 expression was confirmed in bladder smooth muscle tissue by immunohistochemistry and Western blot analysis. Cyclical stretch-relaxation (1 Hz) at 120% elongation significantly increased the expression of STAT3 and of alpha-smooth muscle actin in primary cultured bladder smooth muscle cells. Furthermore, the blockade of STAT3 expression by the transfection of STAT3 small interfering RNA (siRNA) significantly prevented the stretch-induced increase in alpha-smooth muscle actin expression. These results suggest that STAT3 has an important role in the induction of bladder smooth muscle hypertrophy.


Cardiac computed tomography in patients with symptomatic new-onset atrial fibrillation, rule-out acute coronary syndrome, but with intermediate pretest probability for coronary artery disease admitted to a chest pain unit.
Atrial fibrillation (AF) and coronary artery disease (CAD) may be encountered coincidently in a large portion of patients. However, data on coronary artery calcium burden in such patients are lacking. Thus, we sought to determine the value of cardiac computed tomography (CCT) in patients presenting with new-onset AF associated with an intermediate pretest probability for CAD admitted to a chest pain unit (CPU). Calcium scores (CS) of 73 new-onset, symptomatic AF subjects without typical clinical, electrocardiographic, or laboratory signs of acute coronary syndrome (ACS) admitted to our CPU were analyzed. In addition, results from computed tomography angiography (CTA) were related to coronary angiography findings whenever available. Calcium scores of zero were found in 25%. Median Agatston score was 77 (interquartile range: 1-270) with gender- and territory-specific dispersal. CS scores above average were present in about 50%, high (> 400)-to-very high (> 1000) CS scores were found in 22%. Overall percentile ranking showed a relative accordance to the reference percentile distribution. Additional CTA was performed in 47%, revealing stenoses in 12%. Coronary angiography was performed in 22% and resulted in coronary intervention or surgical revascularization in 7%. On univariate analysis, CS > 50th percentile failed to serve as an independent determinant of significant stenosis during catheterization. Within a CPU setting, relevant CAD was excluded or confirmed in almost 50%, the latter with a high proportion of coronary angiographies and subsequent coronary interventions, underlining the diagnostic value of CCT in symptomatic, non-ACS, new-onset AF patients when admitted to a CPU.


Insights into the Role of Commensal-Specific T Cells in Intestinal Inflammation.
Trillions of microorganisms exist in the human intestine as commensals and contribute to homeostasis through their interactions with the immune system. In this review, we use previous evidence from published papers to elucidate the involvement of commensal-specific T cells (CSTCs) in regulating intestinal inflammatory responses. CSTCs are generated centrally in the thymus or peripherally at mucosal interfaces and present as CD4+ or CD8+ T cells. Bacteria, fungi, and even viruses act commensally with humans, warranting consideration of CSTCs in this critical relationship. Dysregulation of this immunological balance can result in both intestinal inflammation or damaging autoimmune responses elsewhere in the body. Given the relative novelty of CSTCs in the literature, we aim to introduce the importance of their role in maintaining immune homeostasis at barrier sites such as the intestine.


Prognostic role of overexpressed Bromodomain and extra-terminal family in ovarian cancer.
Background: BET family proteins have a role as epigenetic readers to accelerate the transcription of target genes. Several studies have shown that the BET protein family played important roles in several biological processes. Although, the prognostic influence of individual BET genes family in ovarian cancer patients remains unclear. Methods: We investigated BET mRNA prognostic roles subtypes in ovarian cancer patients by means of the KM plotter database. The BET mRNA expression and protein in cancer and normal ovarian cells was determined using qRTPCR and western blot. We used the HPA database to look at the protein expression profiles in normal and cancer tissues for this study. Results: Among BET members, mRNA expression BRD2 showed improve OS in all the ovarian malignancy patients, serous patients, stage III and IV, grade II and grade III, TP53 mutated ovarian cancer patients, as well as all patients treated with Platin based chemotherapy. As for BRD3, we found that BRD3 expression was related to better OS in endometrioid ovarian carcinoma and stage III+IV ovarian carcinoma patients, as well as all patients managed with Taxol and concurrent Taxol+Platin based chemotherapy. In addition, BRDT was associated with better OS in all ovarian carcinoma patients, grade I and grade III, all clinical stage (I+II, III+IV) patients, as well as all patients cured with Taxol and concurrent Taxol+Platin chemotherapy. Conclusion: We conclude that high expression of BRD2, BRD3, and BRDT predicted a better prognosis. mRNA expression of BET family is considerably associated with the prognosis of ovarian carcinoma and individual BET family gene could act as a predictive prognostic indicator in ovarian carcinoma.


Decreased expression of autophagy markers in culture-positive patients with chronic otitis media.
Abnormal autophagy plays a role in the pathogenesis of various diseases. This study aimed to evaluate associations between the clinical manifestations of chronic otitis media (COM) and expression of autophagy markers. Associations between presence of bacteria, otorrhea, and conductive and sensorineural hearing loss and levels of autophagy-related mRNAs were investigated in 47 patients with COM. Autophagy-related mRNAs were detected in all inflammatory tissues of COM patients. LC3-II showed the highest level of expression, followed by Beclin-1, P13KC3, Rubicon, and mTOR. Beclin-1 mRNA levels were significantly lower in culture-positive than in culture-negative patients. Autophagy is involved in the pathogenesis of COM. The finding that expression of autophagy markers, especially Beclin-1, was lower in culture-positive than in culture-negative patients suggested that these markers are closely associated with the clinical features of COM.


Sequential morphological changes of the constrictive basilar artery in a canine model of experimental cerebral vasospasm by talc injection.
To demonstrate the possible role of foreign-body reaction to extravasated blood in provoking chronic cerebral vasospasm, talc (crystallized hydrous magnesium silicate) was injected as a non-biologic foreign material into the canine-cisterna cerebellomedullaris, and pathologic changes were followed. Angiographically, this cisternal talc injection induced delayed and prolonged constriction of the basilar artery, without any evidence of so-called early phase cerebral vasospasm that should occur shortly after an insult. Pathologically, around the spastic artery in the subarachnoid space with talc injection, the appearance of a moderate cellular migration coincided with cerebral vasoconstriction, which took place 2 days after the talc injection. In the spastic basilar arterial wall, marked constrictive and degenerative changes including myonecrosis and subintimal proliferation were induced by cisternal talc injection as early as on day 2, and the changes were progressive with time. These pathologic changes were extremely analogous to those of the human autopsy cases with chronic cerebral vasospasm (chronic VS) but more prominent than those observed in the experimental autologous blood-induced model. The present study demonstrated that a foreign-body reaction to talc alone could induce chronic VS in the absence of extravasated blood. Thus, it is possible to consider that inflammatory reactions to extravasated autologous blood in subarachnoid hemorrhage may give rise to chronic VS.


Functional alterations in resting-state networks for Theory of Mind in Parkinson's disease.
In Parkinson's disease (PD), impairment of Theory of Mind (ToM) has recently attracted an increasing number of neuroscientific investigations. If and how functional connectivity of the ToM network is altered in PD is still an open question. First, we explored whether ToM network connectivity shows potential PD-specific functional alterations when compared to healthy controls (HC). Second, we tested the role of the duration of PD in the evolution of functional alterations in the ToM network. Between-group connectivity alterations were computed adopting resting-state functional magnetic resonance imaging (rs-fMRI) data of four groups: PD patients with short disease duration (PD-1, n = 72); PD patients with long disease duration (PD-2, n = 22); healthy controls for PD-1 (HC-1, n = 69); healthy controls for PD-2 (HC-2, n = 22). We explored connectivity differences in the ToM network within and between its three subnetworks: Affective, Cognitive and Core. PD-1 presented a global pattern of decreased functional connectivity within the ToM network, compared to HC-1. The alterations mainly involved the Cognitive and Affective ToM subnetworks and their reciprocal connections. PD-2-those with longer disease duration-showed an increased connectivity spanning the entire ToM network, albeit less consistently in the Core ToM network, compared to both the PD-1 and the HC-2 groups. Functional connectivity within the ToM network is altered in PD. The alterations follow a graded pattern, with decreased connectivity at short disease duration, which broadens to a generalized increase with longer disease duration. The alterations involve both the Cognitive and Affective subnetworks of ToM.


Intravenous parecoxib sodium as an analgesic alternative to morphine in acute trauma pain in the emergency department.
Parecoxib sodium is the first parenteral COX-2 inhibitor used for pain management licensed for postoperative pain. However, no study has assessed the usage of parecoxib for acute traumatic pain in the emergency department (ED). The objective of this study was to investigate a potential alternative analgesic agent in the ED by determining the mean reduction of pain score between acute traumatic pain patients who were administered with intravenous (IV) parecoxib sodium versus IV morphine sulfate. The onset of perceptible analgesic effect and side effects were also evaluated. A randomized, double-blinded study comparing IV parecoxib 40 mg versus IV morphine at 0.10 mg/kg was conducted in adult patients presented with acute traumatic pain with numeric rating scale (NRS) of 6 or more within 6 hours of injury. Patients were randomized using a computer-generated randomization plan. Drug preparation and dispensing were performed by a pharmacist. Periodic assessment of blood pressure, pulse rate, oxygen saturation, and NRS were taken at 0, 5, 15, and 30 minute intervals after the administration of the study drug. The primary outcome was the reduction of NRS. Side effect and drug evaluation was conducted within 30 minutes of drug administration. There was no statistically significant difference in the reduction of mean NRS between patients in the IV parecoxib group or IV morphine group (P = 0.095). The mean NRS for patients treated with IV morphine were 7.1 at 0 minutes, 4.5 at 5 minutes, 3.1 at 15 minutes, and 2.0 at 30 minutes. Whereas mean NRS for patients who received IV parecoxib were 7.8 at 0 minutes, 5.7 at 5 minutes, 4.7 at 15 minutes, and 3.9 at 30 minutes. The onset of perceptible analgesic effects could be seen as early as 5 minutes. Dizziness was experienced in 42.9% of patients who received IV morphine compared to none in the parecoxib group. There was non-significant trend toward superiority of IV morphine over IV parecoxib. Looking at its effectiveness and the lack of opioid-related side-effects, the usage of IV parecoxib sodium may be extended further to a variety of cases in the ED.


Substitution of modified starch with hydrogen peroxide-modified rice bran in salad dressing formulation: physicochemical, texture, rheological and sensory properties.
Rice bran samples were treated under different conditions including hydrogen peroxide content (1, 4, and 7 wt%) and media pH (10.5, 11.5, and 12.5). Water holding capacity and color measurement results showed acceptable improvements compared with the untreated native bran confirmed by Fourier transform infrared analysis. Optimization of modification conditions upon characterization results suggested the introduction of 7% hydrogen peroxide at pH = 12.5. Accordingly, 1, 2 and 3 wt% of the rice bran treated under the optimized conditions, was used in salad dressing formulation; as for .3 wt% of modified starch in the formulation of blank sample, 1 wt% of treated rice bran dietary fiber was substituted. Biopolymer swelling and formation of a stable viscous gel network promoted by the chemical treatment of lignocellulosic rice bran restrict the mobility of oil droplets dispersed in the continuous phase which would consequently retard the emulsion instability phenomena. This effect was also confirmed by flow behavior and viscoelastic characterization results. Salad dressing samples containing 1 and 2 wt% treated rice bran showed acceptable physicochemical, rheological and organoleptic properties besides superior nutritional characteristics compared with the commercial modified starch traditionally used in salad dressing formulations. Despite recommended consumption of dietary fibers, addition of unprocessed lignocellulosic materials to food products usually raise negative effects in sensory, color, and texture quality. This study investigates the modification of rice bran, the byproduct of brown rice milling, to substitute modified starch traditionally used in salad dressing formulations to achieve optimum properties desirable for the final product. Optimization of modification conditions upon characterization of the formulated samples in this study would suggest new improved formulation for the commercial product.


cDNAs for a chymotrypsinogen-like protein from two strains of Plodia interpunctella.
Gut proteinases are involved in the solubilization and activation of insecticidal toxins produced by Bacillus thuringiensis and may also be involved in resistance development. Approximately threefold lower chymotrypsin-like enzyme activity was observed in a Bt(entomocidus)-resistant strain of the Indianmeal moth, Plodia interpunctella, than that in the Bt-susceptible strain. Because chymotrypsin-like proteinases are involved in Bt protoxin activation in P. interpunctella, we compared cDNA sequences, mRNA expression levels, and genomic DNA for chymotrypsin-like enzymes in Bt-susceptible and Bt-resistant strains of P. interpunctella. To isolate cDNA coding for chymotrypsinogen-like proteinases, a probe was developed using polymerase chain reaction (PCR) amplification of a cDNA library from the Bt-susceptible strain using a vector primer and a degenerate primer corresponding to a conserved sequence in the active site of serine proteinases. This probe was used to screen cDNA libraries from resistant and susceptible strains. Predicted amino acid sequences from cDNA clones of each strain share similarity with sequences of chymotrypsin-like proteinases and are most similar to a chymotrypsin-like proteinase from the tobacco hornworm, Manduca sexta. cDNAs for putative chymotrypsinogen-like proteins, from both Bt-susceptible and Bt-resistant strains of P. interpunctella share an identical open reading frame of 846 nucleotides. The encoded proteins contain amino acid sequence motifs of serine proteinase active sites, disulfide-bridge cysteine residues, and both zymogen activation and signal peptides. A difference between these cDNAs was observed only in the untranslated region where a substitution of guanine for adenine occurred in the Bt-resistant strain. Southern and Northern blotting analyses indicated that there are no major differences in chymotrypsinogen-like genomic organization and mRNA expression in the two strains. These data suggest that chymotrypsinogen-like proteinase genes and their transcription are similar in the Bt-susceptible and Bt-resistant strains of P. interpunctella.


PET/CT Variants and Pitfalls in Bone and Soft Tissue Sarcoma.
Sarcomas are rare tumors of mesenchymal origin and comprise only around 1% of adult cancers. The abundance of sarcoma histiotypes, with distinct imaging characteristics, biology, clinical behavior and treatment strategy, result in a complex disease presentation, requiring management by multidisciplinary specialized sarcoma centers. Oncologic and musculoskeletal radiology guidelines provide minimal guidance and only fragmentary information on the indications of 18F-FDG PET/CT in sarcoma. Therefore, knowledge of various phenotypes with preference for bone and lymph node metastases or higher incidence of local and distant recurrence is essential to select the appropriate diagnostic imaging tests and its interpretation. Benign and malignant soft tissue and bone tumors often share common radiographic and metabolic imaging characteristics. In addition, metastases of various histiotypes might exhibit a spectrum of atypical imaging appearances. Therefore, imaging specialists need to be aware of these variants and associated pitfalls of sarcoma imaging.


Cost-effectiveness of sorafenib compared to best supportive care in second line renal cell cancer from a payer perspective in Cyprus.
The objective of this study is to assess the cost effectiveness of sorafenib as a second line treatment of advanced renal cell carcinoma compared to standard best supportive care (BSC) in Cyprus. A probabilistic Decision analytic Markov Model was created to simulate disease progression and data from landmark trials were used. Actual local costs were set according to current guidelines in Cyprus. The incremental cost per quality adjusted life year of sorafenib versus BSC was €102,059. The probability of sorafenib to be cost effective at the threshold of €60,000 was 0%. Total costs were sensitive to the price of product, its effectiveness and to a lesser degree to the utility values. Sorafenib demonstrated superior clinical effectiveness compared to BSC, but it's not cost effective under current willingness to pay threshold. Its orphan status along with solidarity principle may justify reimbursement on an individual patient basis.


Determination of glyphosate in biological fluids by 1H and 31P NMR spectroscopy.
Identification of glyphosate in four cases of poisoning, using nuclear magnetic resonance spectroscopy of biological fluids is reported. It has been performed by using a combination of 1H and 31P NMR analyses. Characterization of the N-(phosphonomethyl) glycine herbicide was achieved by chemical shift considerations and coupling constant patterns: CH2-(P) presents specific resonance at 3.12 ppm and appears as a doublet with a H-P characteristic coupling constant of 12.3 Hz. Moreover, resonances due to isopropylamine were present, confirming the ingestion of the considered trade formulation. After a calibration step, quantitation was performed by 1H and 31P NMR spectroscopy. The benefit and reliability of NMR investigations of biological fluids are discussed, particularly when the clinical picture is quite confusing.


Diabetic foot screening: why is it neglected?
Understanding reasons for the neglect of foot screening during the annual review of people with diabetes enables the development of solutions for this omission. This study explores the reasons within the context of health care delivery systems in terms of the professional, social, political and economic aspects of this screening. Information was obtained through reviewing publications on diabetic foot and health care reform. The omission of annual foot examination for people with diabetes is attributed to the nature of diabetes-related foot problems, people with diabetes, health care professionals and the current structure of health care delivery systems. Increasing the adherence to foot screening for those with diabetes requires short- and long-term strategies. Short- and long-term strategies for reminding patients and staff about foot screening are suggested.


Efficacy of opioid-free anesthesia in modified radical mastectomy: a cross-sectional observational study.
Perioperative management of female patients undergoing breast surgeries differs from other patients due to chronic pain and postoperative nausea and vomiting. The anesthesia could consist of opioid-free general anesthesia (OFA) or non-opioid-free general anesthesia (NOFA). OFA relies on multimodal analgesia preoperatively and postoperatively. However, it is not yet established whether OFA could replace NOFA as a standard regimen for the management of breast surgeries. The aim of this study is to evaluate the efficacy of OFA for breast surgeries in female patients. Patients undergoing modified radical mastectomy were retrospectively recruited. Two groups were defined: group 1, consisting of treated patients using OFA and group 2, consisting of treated patients using NOFA. Mean time to extubate and mean dose of morphine after recovery were computed. Postoperative morphine and antiemetic use were assessed for up to 24 h. A comparison of the computed data was conducted between both groups. A total of 116 patients were included with a mean age of 53±13 years. Group 1 consisted of 56 (mean age was 54±14 years). Group 2 consisted of 60 patients (mean age was 51±12 years). Demographic parameters and time to extubate did not yield significant differences. We noticed morphine sparing at T0 and T12 with statistically significant differences P=0.043 and P=0.006, respectively. OFA could be considered in modified radical mastectomy management in female patients; nerve block seemed to act as morphine sparing in operative and postoperative settings by providing significantly less time to extubate with less postoperative morphine requirement.


The effect of flowering time and distance between pollen source and recipient on maize.
Field experiments were conducted in Central Taiwan for 2 crop seasons to examine the effect of non-coincidence flowering on the cross-pollination (CP) rate of maize at various distances. Four local maize hybrid varieties with different flowering dates and one local maize variety were sown as the pollen sources and recipient, respectively. All varieties were sown on the same day to simulate the real situation of coexistence in which adjacent fields are sown with different genetically modified (GM) and non-GM varieties of maize. The CP rate was <0.2% at a distance of 3 m for the first crop season when the flowering time for the recipient was 5 d later than that of the pollen source variety. The CP rate was <0.02% at all distances for the second season when the flowering time for the recipient was 7 d later than that of the pollen source variety. The CP rate was <1% at a distance of 0.75 m when the flowering time was 3 d later. However, varieties with closer synchrony may result in a CP rate of >1% at a distance of 1.5 m and <1% at 2.25 m. Temporal separation and isolation distances can work together in Taiwan with fragmented landscapes to minimize the adventitious presence of one crop with another.


Family caregiver expectations and management of the stroke trajectory.
Less than 20% of stroke survivors enter rehabilitation or long-term care facilities after their stroke. Stroke recovery is extremely variable and the resulting uncertainty places a heavy burden on the survivors' family caregivers. According to the trajectory framework, chronic conditions have a defined course that can be shaped and managed. This grounded theory study, part of a larger research project, explored the expectations of family caregivers of the stroke trajectory and their management strategies. Thirteen family caregivers of stroke patients in a sparsely populated area participated in semi-structured interviews. The caregivers were without ideas about what the recovery of their loved ones would be like and had difficulty making projections about the trajectory. They used several strategies, however, in attempts to manage the stroke trajectory. They constructed a positive recovery, reconstituted family life, maintained family routines, created a safety net, and redoubled self-reliance. The findings have implications for how nurses support family caregivers of stroke survivors.


Spin current generation and relaxation in a quenched spin-orbit-coupled Bose-Einstein condensate.
Understanding the effects of spin-orbit coupling (SOC) and many-body interactions on spin transport is important in condensed matter physics and spintronics. This topic has been intensively studied for spin carriers such as electrons but barely explored for charge-neutral bosonic quasiparticles (including their condensates), which hold promises for coherent spin transport over macroscopic distances. Here, we explore the effects of synthetic SOC (induced by optical Raman coupling) and atomic interactions on the spin transport in an atomic Bose-Einstein condensate (BEC), where the spin-dipole mode (SDM, actuated by quenching the Raman coupling) of two interacting spin components constitutes an alternating spin current. We experimentally observe that SOC significantly enhances the SDM damping while reducing the thermalization (the reduction of the condensate fraction). We also observe generation of BEC collective excitations such as shape oscillations. Our theory reveals that the SOC-modified interference, immiscibility, and interaction between the spin components can play crucial roles in spin transport.


Functional characterization of human CTC1 mutations reveals novel mechanisms responsible for the pathogenesis of the telomere disease Coats plus.
Coats plus is a rare recessive disorder characterized by intracranial calcifications, hematological abnormalities, and retinal vascular defects. This disease results from mutations in CTC1, a member of the CTC1-STN1-TEN1 (CST) complex critical for telomere replication. Telomeres are specialized DNA/protein structures essential for the maintenance of genome stability. Several patients with Coats plus display critically shortened telomeres, suggesting that telomere dysfunction plays an important role in disease pathogenesis. These patients inherit CTC1 mutations in a compound heterozygous manner, with one allele encoding a frameshift mutant and the other a missense mutant. How these mutations impact upon telomere function is unknown. We report here the first biochemical characterization of human CTC1 mutations. We found that all CTC1 frameshift mutations generated truncated or unstable protein products, none of which were able to form a complex with STN1-TEN1 on telomeres, resulting in progressive telomere shortening and formation of fused chromosomes. Missense mutations are able to form the CST complex at telomeres, but their expression levels are often repressed by the frameshift mutants. Our results also demonstrate for the first time that CTC1 mutations promote telomere dysfunction by decreasing the stability of STN1 to reduce its ability to interact with DNA Polα, thus highlighting a previously unknown mechanism to induce telomere dysfunction.


Mesometrial smooth muscle in the mouse: its control of uterine blood flows.
The form and function of the mesometrial smooth muscle and the interposed mesometrial branches and tributaries of the uterine vessels were studied in the nonpregnant and pregnant mouse to see whether contractions of mesometrial muscle alter uterine blood flows. Histological sections of mouse uterine horn demonstrated that the outer longitudinal layer of myometrium extends onto the mesometrium and sandwiches the mesometrium and its vessels as a bilaminar myometrial extension (BME). The BME ends midway across the mesometrium as a free edge. Cleared specimens, perfused with silicone rubber, revealed that the mesometrial branches and tributaries of the uterine vessels formed longitudinally communicating looping arcades between adjacent mesometrial vessels. Comparisons with human dissections and uterine histological sections revealed similar patterns of mesometrial smooth muscle and mesometrial blood vessels. BME activity and its control of mesometrial blood flow were studied by transillumination of the surgically exposed mesometria of anesthetized day 12 pregnant mice. Observed contractions of the BME coincided precisely with uterine contractions measured at the cervix and the BME contractions diminished or stopped venous outflow particularly in the midhorn regions. Arterial flows seemed to be unaffected and were diminished or halted only during infrequent forceful and sustained contractions. Trapped venous outflow passed up or down the venous arcades to escape through less restricted mesometrial veins. Uterine and BME contractions normally take place throughout gestation. It is possible that abnormally long and forceful contractions may compromise the embryo or fetus.


The novel cardiac z-disc protein CEFIP regulates cardiomyocyte hypertrophy by modulating calcineurin signaling.
The z-disc is a structural component at the lateral borders of the sarcomere and is important for mechanical stability and contractility of both cardiac and skeletal muscles. Of note, the sarcomeric z-disc also represents a nodal point in cardiomyocyte function and signaling. Mutations of numerous z-disc proteins are associated with cardiomyopathies and muscle diseases. To identify additional z-disc proteins that might contribute to cardiac disease, we employed an in silico screen for cardiac-enriched cDNAs. This screen yielded a previously uncharacterized protein named cardiac-enriched FHL2-interacting protein (CEFIP), which exhibited a heart- and skeletal muscle-specific expression profile. Importantly, CEFIP was located at the z-disc and was up-regulated in several models of cardiomyopathy. We also found that CEFIP overexpression induced the fetal gene program and cardiomyocyte hypertrophy. Yeast two-hybrid screens revealed that CEFIP interacts with the calcineurin-binding protein four and a half LIM domains 2 (FHL2). Because FHL2 binds calcineurin, a phosphatase controlling hypertrophic signaling, we examined the effects of CEFIP on the calcineurin/nuclear factor of activated T-cell (NFAT) pathway. These experiments revealed that CEFIP overexpression further enhances calcineurin-dependent hypertrophic signal transduction, and its knockdown repressed hypertrophy and calcineurin/NFAT activity. In summary, we report on a previously uncharacterized protein CEFIP that modulates calcineurin/NFAT signaling in cardiomyocytes, a finding with possible implications for the pathogenesis of cardiomyopathy.


Clinical Application of Targeted Deep Sequencing in Solid-Cancer Patients and Utility for Biomarker-Selected Clinical Trials.
Molecular profiling of actionable mutations in refractory cancer patients has the potential to enable "precision medicine," wherein individualized therapies are guided based on genomic profiling. The molecular-screening program was intended to route participants to different candidate drugs in trials based on clinical-sequencing reports. In this screening program, we used a custom target-enrichment panel consisting of cancer-related genes to interrogate single-nucleotide variants, insertions and deletions, copy number variants, and a subset of gene fusions. From August 2014 through April 2015, 654 patients consented to participate in the program at Samsung Medical Center. Of these patients, 588 passed the quality control process for the 381-gene cancer-panel test, and 418 patients were included in the final analysis as being eligible for any anticancer treatment (127 gastric cancer, 122 colorectal cancer, 62 pancreatic/biliary tract cancer, 67 sarcoma/other cancer, and 40 genitourinary cancer patients). Of the 418 patients, 55 (12%) harbored a biomarker that guided them to a biomarker-selected clinical trial, and 184 (44%) patients harbored at least one genomic alteration that was potentially targetable. This study demonstrated that the panel-based sequencing program resulted in an increased rate of trial enrollment of metastatic cancer patients into biomarker-selected clinical trials. Given the expanding list of biomarker-selected trials, the guidance percentage to matched trials is anticipated to increase. This study demonstrated that the panel-based sequencing program resulted in an increased rate of trial enrollment of metastatic cancer patients into biomarker-selected clinical trials. Given the expanding list of biomarker-selected trials, the guidance percentage to matched trials is anticipated to increase.


Organocatalytic Regiodivergent Ring Expansion of Cyclobutanones for the Enantioselective Synthesis of Azepino[1,2-a]indoles and Cyclohepta[b]indoles.
A regiodivergent organocatalytic enantioselective Michael addition/three-atom ring expansion sequence of electron-withdrawing group activated cyclobutanones with 2-nitrovinylindoles was developed. A series of azepino[1,2-a]indoles were obtained with exclusive regioselectivities and high diastereo- and enantioselectivities (up to >20:1 dr, 96% ee) with the application of the N1 nucleophilic site of the indole nucleus. Meanwhile, various cyclohepta[b]indoles could be accessed with high enantiopurity (up to 96% ee) through the Michael addition/boron-trifluoride-etherate-promoted indole C3-attack ring expansion process.


Swedish healthcare under pressure.
Swedish health care, run by local governments at both the regional (county) and the municipal levels, has been under pressure during the last 15 years, following increased scrutiny of performance and demand for cost-containment. Health-care expenditures per capita and levels of resource inputs have grown, but more slowly than in other EU countries. At the same time, the number of elderly people has increased, as have options for medical treatment. In the late 1980s, several local governments referred to long waiting-lists for elective treatment and anecdotal evidence of inefficiency and poor responsiveness when arguing for market-oriented reforms. A purchaser-provider split followed, and so did changes in the payment systems for health-care providers. According to the available evidence, these reforms yielded an increased volume of services in the short run; but traditional hierarchical management soon replaced the new incentives. Moreover, evidence suggests that changes introduced by the national government, and the deteriorating funding conditions together with a continued use of new medical technology, have had more far-reaching effects on health-care output and outcome than local-government reforms.


Targeting MDSC for Immune-Checkpoint Blockade in Cancer Immunotherapy: Current Progress and New Prospects.
Immune-checkpoint blockade (ICB) demonstrated inspiring effect and great promise in anti-cancer therapy. However, many obstacles, such as drug resistance and difficulty in patient selection, limited the efficacy of ICB therapy and awaited to be overcome. By timely identification and intervention of the key immune-suppressive promotors in the tumor microenvironment (TME), we may better understand the mechanisms of cancer immune-escape and use novel strategies to enhance the therapeutic effect of ICB. Myeloid-derived suppressor cell (MDSC) is recognized as a major immune suppressor in the TME. In this review, we summarized the roles MDSC played in the cancer context, focusing on its negative biologic functions in ICB therapy, discussed the strategies targeted on MDSC to optimize the diagnosis and therapy process of ICB and improve the efficacy of ICB therapy against malignancies.


Biological estimates of dose to inhabitants of Belarus and Ukraine following the Chernobyl accident.
The purpose of this paper is to investigate how well various assays on blood can detect radiation dose to people exposed many years previously and, if possible, to estimate that dose. The assays were applied to persons resident close to Chernobyl in 1986. Blood samples were taken 13-15 years after the reactor accident. The assays used were the frequencies of lymphocyte chromosomal translocations, micronuclei, HPRT mutations and apoptotic cells. Translocation yields in the exposed groups were marginally higher than in their respective controls, leading to dose estimates of about 0.2 Gy but with large uncertainties. All other assays showed inconsistency from person to person or other variations apparently not related to dose. The measurement of translocations, it is concluded, is the biological method of choice for retrospective dosimetry.


Characterization of Pseudomonas aeruginosa fatty acid profiles in biofilms and batch planktonic cultures.
The fatty acid composition of Pseudomonas aeruginosa PAO1 was compared between biofilm and batch planktonic cultures. Strain PAO1 biofilms were able to maintain a consistent fatty acid profile for up to 6 days, whereas strain PAO1 batch planktonic cultures showed a gradual loss of cis-monounsaturated fatty acids over 4 days. Biofilms exhibited a greater proportion of hydroxy fatty acids but a lower proportion of both cyclopropane fatty acids and saturated fatty acids (SAFAs). SAFAs with >=16 carbons, in particular, decreased in biofilms when compared with that in batch planktonic cultures. A reduced proportion of SAFAs and a decline in overall fatty acid chain length indicate more fluidic biophysical properties for cell membranes of P. aeruginosa in biofilms. Separating the biofilms into 2 partitions and comparing their fatty acid compositions revealed additional trends that were not observed in the whole biofilm: the shear-nonremovable layer consistently showed greater proportions of hydroxy fatty acid than the bulk liquid + shear-removable portion of the biofilm. The shear-nonremovable portion demonstrated a relatively immediate decline in the proportion of monounsaturated fatty acids between days 2 and 4; which was offset by an increase in the proportion of cyclopropane fatty acids, specifically 19:0cyc(11,12). Simultaneously, the shear-removable portion of the biofilm showed an increase in the proportion of trans-monounsaturated fatty acids and cyclopropane fatty acids.


Projections Onto Order Simplexes and Isotonic Regression.
Isotonic regression is the problem of fitting data to order constraints. This problem can be solved numerically in an efficient way by successive projections onto order simplex constraints. An algorithm for solving the isotonic regression using successive projections onto order simplex constraints was originally suggested and analyzed by Grotzinger and Witzgall. This algorithm has been employed repeatedly in a wide variety of applications. In this paper we briefly discuss the isotonic regression problem and its solution by the Grotzinger-Witzgall method. We demonstrate that this algorithm can be appropriately modified to run on a parallel computer with substantial speed-up. Finally we illustrate how it can be used to pre-process mass spectral data for automatic high throughput analysis.


Expression of the COVID-19 receptor ACE2 in the human conjunctiva.
SARS-CoV-2 is assumed to use angiotensin-converting enzyme 2 (ACE2) and other auxiliary proteins for cell entry. Recent studies have described conjunctival congestion in 0.8% of patients with laboratory-confirmed severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), and there has been speculation that SARS-CoV-2 can be transmitted through the conjunctiva. However, it is currently unclear whether conjunctival epithelial cells express ACE2 and its cofactors. In this study, a total of 38 conjunctival samples from 38 patients, including 12 healthy conjunctivas, 12 melanomas, seven squamous cell carcinomas, and seven papilloma samples, were analyzed using high-throughput RNA sequencing to assess messenger RNA (mRNA) expression of the SARS-CoV-2 receptor ACE2 and its cofactors including TMPRSS2, ANPEP, DPP4, and ENPEP. ACE2 protein expression was assessed in eight healthy conjunctival samples using immunohistochemistry. Our results show that the SARS-CoV-2 receptor ACE2 is not substantially expressed in conjunctival samples on the mRNA (median: 0.0 transcripts per million [TPM], min: 0.0 TPM, max: 1.7 TPM) and protein levels. Similar results were obtained for the transcription of other auxiliary molecules. In conclusion, this study finds no evidence for a significant expression of ACE2 and its auxiliary mediators for cell entry in conjunctival samples, making conjunctival infection with SARS-CoV-2 via these mediators unlikely.


[A method for studying the bone mass: dual-photon densitometry].
The assessment of bone mineral mass during growth, periods diseases or therapy which modify bone mineralization in the child is of great interest. Single-photon absorptiometry carried out on forearm bones informs on the mineral density of the compact bone of the appendicular skeleton. It is easy to perform in children and infants. Dual-photon absorptiometry appears to be the most accurate for measuring bone mineral content in endocrine and metabolic bone disorders. The low dose of radiation and the non-invasive nature of this high-precision method render it well-adapted and suitable for measuring bone mineralization in childhood.


The incidence and prevalence of inflammatory bowel disease among U.S. veterans: a national cohort study.
Temporal trends in incidence and prevalence of Crohn's disease (CD) and ulcerative colitis (UC) in the United States have been reported only in regional populations. The Veterans Affairs (VA) health care system encompasses a national network of clinical care facilities. The aim of this study was to identify temporal trends in the incidence and prevalence of CD and UC among VA users using national VA data sets. Veterans with CD and UC were identified during fiscal years 1998 to 2009 in the national VA outpatient and inpatient files. Incident and prevalent cases were identified by diagnosis code, and age-standardized and gender-standardized annual prevalence and incidence rates were estimated using the VA 1998 population as the standard population. The total of unique incident cases were 16,842 and 26,272 for CD and UC, respectively; 94% were men. The average annual age-standardized and gender-standardized incidence rate of CD was 33 per 100,000 VA users (range, 27-40), whereas the average for UC was 50 per 100,000 VA users (range, 36-65). In 2009, the age-standardized and gender-standardized point prevalence rate of CD was 287 per 100,000 VA users, whereas the point prevalence of UC was 413 per 100,000 VA users. Prevalence of CD and UC increased 2-fold to 3-fold among VA users between 1998 and 2009. The incidence of UC decreased among VA users from 1998 to 2004 but has remained stable from 2005 to 2009. The incidence of CD has remained stable during the observed time period.


Human adenovirus type 35: nucleotide sequence and vector development.
In this report, we describe the complete 34,794 base pair genomic sequence of the human adenovirus serotype 35 (Ad35) Holden strain. The viral genome exhibits a compact organization similar to other adenoviral serotypes, with overlapping genes on both strands. In all, 47 open reading frames (ORFs) were identified, including early (E1, 2, 3, 4) and late (L1, 2, 3, 4, 5) regions conserved among the adenoviridae family. In addition, 14 ORFs were identified that do not encode known adenoviral genes. Comparison of the predicted translational products of the conserved genes with those of other adenoviruses revealed that Ad35 has high homology to Ad7, Ad3, Ad21, Ad17, and simian Ads25. Based on the complete Ad35 DNA sequence, E3-, E1-, and E1/E3-deleted Ad35-based vector systems were developed. An HEK293-derived cell line was established for the propagation of the E1-deleted Ad35 vector, avoiding the emergence of replication-competent adenovirus. Moreover, production of the E1-deleted recombinant Ad35 vector was achieved by transient transduction of a plasmid encoding the Ad35 E1B gene in HEK293 cells. Testing showed that the Ad35-based vector efficiently infects both human and rhesus macaque dendritic cells. Our novel Ad35-based vectors and their corresponding packaging cell lines will provide a versatile and powerful system for DNA-based vaccine development and gene therapy applications.


Genotoxic, cytotoxic, and neurotoxic responses in Anodonta cygnea after complex metal mixture treatment.
Environmental effects associated with the release of various metals even at maximum permissible concentrations (MPC) to the aquatic ecosystems are evident. In the present work, time-dependent increase in accumulated metals amount in gills of Anodonta cygnea after exposure to complex metal (Zn 0.1, Cu 0.01, Ni 0.01, Cr 0.01, Pb 0.005, and Cd 0.005 mg/L, MPC accepted for the inland waters in EU) mixture at various time points (1, 2, 4, 7, 14, and 28 days) was investigated. Statistically significant increase of Cu and Cd was determined in mussel's gills after 7-day exposure, in comparison to control group; moreover, significantly elevated concentration of Cu was measured and after 14-day treatment (in comparison to control and pre-exposure group). Concentrations of five (Cu, Ni, Cr, Pb, and Cd) out of 6 investigated metals were statistically increased in gills tissue after 28-day treatment. Moreover, complex metal mixture has demonstrated tissue- and time-dependent genotoxicity (∑Gentox) and cytotoxicity (∑Cytox) responses in mussels. After 4-day exposure, there were found the highest ∑Gentox levels in gills cells and haemocytes. Two-day treatment of mussels resulted in the highest and statistically significant induction of ∑Cytox level (in gills). Furthermore, after short-term (4 days) exposure, statistically significant inhibition of AChE activity in hemolymph of metal mixture-exposed mussels, in comparison to control and pre-exposure group, was found. Comparison of investigated responses in different tissue of A. cygnea discloses new information about metal mixture (at MPC) impacts at different treatment time. According to the obtained geno- and cytotoxicity data, it is suggested that gills are more sensitive tissue. Environmentally relevant trace metal concentrations when existing in mixture are able to cause adverse effects in A. cygnea; therefore, biological effects at different levels of organism are expected as a realistic scenario.


Transmission Efficiency, Preference and Behavior of Bemisia tabaci MEAM1 and MED under the Influence of Tomato Chlorosis Virus.
Tomato chlorosis virus (ToCV, genus Crinivirus, family Closteroviridae) is an economically important virus in more than 20 countries. In China, ToCV was first detected in 2013 and has already spread throughout the country. ToCV is transmitted in a semi-persistent manner by the whitefly, Bemisia tabaci, but not seed. In the past two decades, the most invasive MEAM1 and MED have replaced the indigenous B. tabaci in China, and currently MED is the most dominant cryptic species. To better understand the prevalence of ToCV with their vectors, we tested the hypothesis that the rapid spread of ToCV in China is closely related to the dominance of MED. ToCV acquisition and accumulation rate following transmission was significantly higher by MED than MEAM1. In addition, ToCV persisted for more than 4 days in MED but only 2 days in MEAM1. Viruliferous MED preferred non-infected over virus-infected plants, although MED performed better on infected than on non-infected plants. Our combined results support the initial hypothesis that the rapid spread of ToCV is associated with the spread of B. tabaci MED in China.


Measuring Cohesion and Self-Disclosure in Psychotherapy Groups for Patients with Advanced Cancer: An Analysis of the Psychometric Properties of the Group Therapy Experience Scale.
Understanding the group process informs group interventions. However, there is little systematic research on group process variables in psychotherapy groups for patients with cancer. We analyzed the psychometric properties of the Group Therapy Experience Scale and evaluated its potential importance in advanced cancer therapy groups. The GTES demonstrated good internal consistency (coefficient alpha=.84). An exploratory factor analysis with varimax rotation yielded four factors. Although all four models were explored, the 1- and 2-factor models appeared to provide the best fit for the data. The GTES total score was negatively correlated with group size and positively correlated with number of groups attended by participants. Furthermore, the GTES total score was correlated with post-intervention spiritual well-being, benefit finding, post-traumatic growth and quality of life.


[Rapid identification of human testis spermatocyte apoptosis-related gene, TSARG2, by nested PCR and draft human genome searching].
Cloning apoptosis-related novel genes is a key to further understanding of apoptosis mechanism and the biology process of germ cells, and is of momentous significance on clarifying physiological and pathological process of spermatogenesis. To rapidly attain human novel gene full-length cDNA sequence, the gene-specific primers and the vector-specific primers were designed for nested PCR, and draft human genome searching was performed to rapidly identify the TSARG2 (GenBank accession number AY040204) 5' end from a human testis cDNA library, by using a cDNA fragment (GenBank accession number BE644542) as an electronic probe, which was significantly changed in cryptorchidism and represented a novel gene. Furthermore, a mouse homologue of this gene was identified (GenBank accession number AF395083) by lab on-line. TSARG2 with a 1 233 bp length was composed of 6 exons and spanned about 115 kb of genomic DNA, The putative protein encoded by this gene was 305 amino acid with a theoretical molecular weight of 34 751 dalton and did not share significant homology with any known protein in databases. TSARG2 was expressed in many tissues and mapped to chromosome 4q33-34.1 by database analyses. Therefore, we propose that nested-PCR and draft human genome searching are rapid, sensitive, accurate and efficient method for isolating gene 5' end, even full-length gene from cDNA library.


Intramolecular O···H Hydrogen Bonding of Salicylic Acid: Further Insights from O 1s XPS and 1H NMR Spectra Using DFT Calculations.
Intramolecular hydrogen bonding (HB) is a complex phenomenon that extends beyond a simple valence event, affecting the core electrons of a molecule. Salicylic acid (SA) and its conformers provide an excellent model compound for studying intramolecular HB as the proton donor (H) and acceptor (O) can be toggled by rotating the C-O and C-C bonds to form up to seven potential conformers through various HB. In this study, we computationally investigated intramolecular interactions in SA conformers with and without such HB, by examining their calculated O 1s core electron-binding energy (CEBE) and 1H NMR chemical shifts validated using recent measurements. The quantum mechanically stable SA conformers are fully defined by three rotatable bonds in the compound, which are abstracted as SA(η1η2η3) digital structures, where ηi = 0 if the ηi angles match the most stable SA conformer (000) and ηi = 1 otherwise. Our findings suggest that the stability is dominated by the appearance of the intergroup intramolecular HB of Hp···O (where O is in the carboxylic acid functional group and Hp is the phenolic proton in -OHp), and η3 serves as a switch of such HB. As a result, the (η1η20) SA conformers containing such Hp···O HB are more stable than other SA conformers (η1η21) without such the Hp···O HB. The present density functional theory calculations reveal that this Hp···O HB results in splitting of the O 1s CEBEs of two hydroxyl groups (-OH) by up to 1 eV and deshielding the Hp proton 1H NMR (δHp) up to 11.68 ppm for the (η1η20) conformers. Without such Hp···O HB, the O 1s XPS binding energies of two -OH groups will be closely located in the same band, and the 1H NMR chemical shift of the Hp atom will be as small as an 4.09 ppm SA conformer [SA-G(101)]. The present study indicates that the O 1s CEBE splitting between two -OH groups serves as an indicator of the presence of the Hp···O HB in SA conformers, which is also supported by the 1H NMR results.


[Influence of the prosthodontic construction on the marginal gingiva nearby dental crowns and bridges].
Along with the functional and aesthetic results, prosthodontic constructions must correspond to biological needs as well. They don't have to irritate the periodontal tissue, alveolar bone and gingiva. The aim of the study was to investigate gingival status under modified artificial teeth using cytomorphology (PAP) methods. Cytological investigations were carried out in 30 patients. 19 of them were using modified crowns and 11--modified bridges. Gingival status was marked using CD I and II indexes. Cytomorphometric investigations showed that modified prosthodontic constructions do not harm gingivas which are under the modification. Dynamic cytological changes found at the time of treatment represent individual biological reaction to the artificial teeth.


Airborne dispersal as a novel transmission route of coagulase-negative staphylococci: interaction between coagulase-negative staphylococci and rhinovirus infection.
To investigate whether rhinovirus infection leads to increased airborne dispersal of coagulase-negative staphylococci (CoNS). Prospective nonrandomized intervention trial. Wake Forest University School of Medicine, Winston-Salem, North Carolina. Twelve nasal Staphylococcus aureus-CoNS carriers among 685 students screened for S. aureus nasal carriage. Participants were studied for airborne dispersal of CoNS in a chamber under three conditions (street clothes, sterile gown with a mask, and sterile gown without a mask). After 2 days of pre-exposure measurements, volunteers were inoculated with a rhinovirus and observed for 14 days. Daily quantitative nasal and skin cultures for CoNS and nasal cultures for rhinovirus were performed. In addition, assessment of cold symptoms was performed daily, mucous samples were collected, and serum titers before and after rhinovirus inoculation were obtained. Sneezing, coughing, and talking events were recorded during chamber sessions. All participants had at least one nasal wash positive for rhinovirus and 10 developed a symptomatic cold. Postexposure, there was a twofold increase in airborne CoNS (P = .0004), peaking at day 12. CoNS dispersal was reduced by wearing a gown (57% reduction, P < .0001), but not a mask (P = .7). Nasal and skin CoNS colonization increased after rhinovirus infection (P < .05). We believe this is the first demonstration that a viral pathogen in the upper airways can increase airborne dispersal of CoNS in nasal S. aureus carriers. Gowns, gloves, and caps had a protective effect, whereas wearing a mask did not further reduce airborne spread.


Master's Degrees Among Academic Plastic Surgeons and Plastic Surgery Residents: What Are the Trends?
As more plastic surgery clinicians pursue advanced degrees and strive to become stronger physician-scientists, an objective understanding of how such degrees influence careers becomes important. We hypothesized that having a master's degree is associated with higher scholarly activity, research funding, academic progression, and leadership appointments. Accreditation Council for Graduate Medical Education-accredited integrated plastic surgery residency program Web sites were queried to create a data set of current academic plastic surgeons (APSs) and plastic surgery residents (PSRs). Scholarly metrics such as publications, citations, and H-indices were extracted from the Scopus database. National Institutes of Health and Plastic Surgery Foundation funding information was collected through their respective Web sites. Our cohort comprised 799 APSs and 922 PSRs, of whom 8% and 7.4%, respectively, had at least one master's degree. Academic plastic surgeons with master's of public health degrees had a significantly higher median number of publications and citations than APSs without a master's of public health. There was no association between any master's degree and academic rank or being a department chairman or program director. Academic plastic surgeons with master of science degrees were more likely to receive National Institutes of Health grants. Among PSRs, master's of science graduates had a higher median number of publications. Other master's degrees did not significantly influence scholarly productivity or funding. Certain master's degrees had an impact on scholarly productivity, with no significant effect on academic rank or leadership positions. The value of master's degrees in programs focusing on healthcare management, leadership skills, and business acumen likely extends beyond the scope of this study.


Non-HPV-Related Head and Neck Squamous Cell Carcinoma in a Young Patient Cohort.
Head and neck squamous cell carcinoma (HNSCC) is rare in patients younger than 40 years. Many practitioners suspect HNSCC is a more aggressive disease in this age group, and perhaps increasing in incidence; however, there are scant and conflicting data to support this assertion. We sought to compare outcomes for young patients with non-human papillomavirus (HPV)-related HNSCC to those of older patients. A retrospective chart review of patients with HNSCC treated from 2004 to 2016 at 2 tertiary referral centers. Patients aged 18 to 40 with p16-negative HNSCC were included in the young patient cohort (n = 59). A randomly selected stage- and subsite-matched cohort aged 55 to 65 was analyzed for comparison (n = 114). When considering all patients with HNSCC, patients younger than 40 were more likely to have oral tongue cancer (62.7%) compared to patients age 55 to 65 (16.9%). When an older patient cohort was stage- and subsite-matched to the young patient cohort, there were more never smokers (49.2% vs 17.5% of older patients, P < .01) and females (40.7% vs 24.6% of older patients, P = .028) in the young patient group. The young patient cohort had better average overall survival than the older group (14.4 vs 8.1 years, respectively, P = .02), but similar average disease-free survival (6.2 years vs 6.6 years, respectively, P = .67); 50.9% of young patients had tumors with adverse histologic features versus 42.0% of older patients (P = .28). The young patients demonstrated a superior average conditional survival after recurrence (9.8 years vs 3.2 years for older patients, P < .01). Despite the limitations of study design, these data suggest that young patients who develop non-HPV-related HNSCC tend to have similarly aggressive disease, but longer overall survival and better survival after recurrence. These findings may be attributable to better overall health as evidenced by fewer comorbidities.


Relationship between HER2 status and proliferation rate in breast cancer assessed by immunohistochemistry, fluorescence in situ hybridisation and standardised AgNOR analysis.
Lack of standardisation and inaccuracy in HER2 test results may adversely influence patient evaluation and therapy selection. In the present study we applied immunohistochemistry (IHC) using the A0485 and CB11 antibodies and fluorescence in situ hybridisation (FISH) for detection of HER2 in 74 routinely processed breast carcinoma specimens. The rapidity of cellular proliferation was assessed by standardised AgNOR analysis and compared with HER2 status. Protein over-expression was found in 30/74 cases by A0485 and in 20/74 by CB11 antibodies, while amplification was detected in 22/74 carcinomas by FISH. Twenty-seven of 74 tumours were high-level AgNOR expressors (mean AgNOR area >3.369 microm2), 19 of which revealed amplification. The highest concordance between results was achieved by FISH and CB11-IHC (97%), the concordance between FISH and A0485-IHC was 89 and 84% between FISH and AgNOR quantity, respectively. The overall concordance between A0485 and CB11-IHC was 85% with 10 incongruent cases, all scored 2+ by A0485 and 0/1+ by CB11. Eight of the discordant tumours were non-amplified by FISH and 7 were low AgNOR-expressors. Our results indicate that using CB11 antibody, a nearly complete agreement can be achieved between HER2 IHC and FISH in diagnostic paraffin material. Moreover, in 2+ positive IHC cases, the AgNOR analysis may represent an additional tool to select patients as candidates for Herceptin therapy due to the strong negative predictor value.


Assessment of the elastic stiffness of human cardiac fibres after an apical infarction using finite element simulation.
Several research works in the literature have focused on understanding the post-infarction ventricular remodelling phenomenon, but few works have considered the evaluation of the elastic behaviour of the cardiac tissue after a myocardial infarction. This paper presents an investigation focused on predicting the elastic performance of the human heart after a left ventricular apical infarction. The aim is to understand the elastic alterations of the cardiac fibres at different periods after an apical infarct. For this purpose, a hybrid method based on pressure and volume measurements of the left ventricle (LV) at different periods of ventricular remodelling, and the Finite Element Method (FEM), is developed. In addition, several performance indexes are defined to evaluate the heart performance during the ventricular remodelling process. The results show that during the first 2 weeks after a heart infarction, the cardiac fibres must support a much higher structural overload than during normal conditions. This structural overload is proportional to the aneurysm size but diminishes with the time, together with a significant reduction of the ventricular pumping capacity.


Azide⋅⋅⋅Oxygen Interaction: A Crystal Engineering Tool for Conformational Locking.
We have designed, synthesized, and crystallized 36 compounds, each containing an azide group and an oxygen atom separated by three bonds. Crystal structure analysis revealed that each of these molecules adopts a conformation in which the azide and oxygen groups orient syn to each other with a short O⋅⋅⋅Nβ contact. Geometry-optimized structures [using M06-2X/6-311G(d,p) level of theory] also showed the syn conformation in all 36 of these cases, suggesting that this is not merely a crystal packing effect. Quantum topological analysis using Bader's Atoms in Molecules (AIM) theory revealed bond paths and bond critical points (BCP) in these structures suggesting its nature and energetics to be similar to weak hydrogen bonding. The NCI-RDG plot clearly revealed the attractive interaction consisting of electrostatic or dispersive components in all the 36 systems. NBO analysis suggested a weak orbital-relaxation (charge-transfer) contribution of energy for a few (sp2) O-donor systems. Natural population analysis (NPA) and molecular electrostatic potential mapping (MESP) of these crystal structures further revealed the existence of favorable azide-oxygen interaction. A CSD search indicated the frequent and consistent occurrence of this interaction and its role dictating the syn conformation of azide and oxygen in molecules where these groups are separated by 2-4 bonds.


Management Patterns of Male Urethral Stricture Disease among Urologists: What Do the Guidelines Say? What Do Urologists Do?
The aim of the present study is to evaluate and analyze the daily clinical practice for male urethral stricture disease (MUSD) among urologists. Considering the latest guidelines on urethral stricture disease, a survey was developed regarding the various treatment options and preferences in different sites of male urethral stricture disease. The survey was sent to urologists via e-mail and phone application. A total of 266 urologists completed the survey and were included in the final analysis. In regard to workplace, 62 (23.3%), 58 (21.8%), 71 (26.7%), and 75 (28.2%) respondents worked in university hospitals, training and research hospitals, state hospitals, and private practice hospitals, respectively. In regard to the diagnostic method used in male urethral strictures, 88.7% of the participants would choose uroflowmetry + postvoiding residual (UF + PVR), and 64.6% would choose retrograde urethrography (RUG). Direct vision internal urethrotomy (DVIU) was the most frequently chosen method in penile urethral strictures (PUS), being chosen by 72.9%. Direct vision anterior internal urethrotomy was the most common method for both ≤2 cm and >2 cm strictures, 63.1%, and 30.8%, respectively. The most preferred graft for augmentation urethroplasty was buccal mucosa (75.8%). Endoscopic incision/resection (transurethral resection (TUR)) is the most frequently applied treatment method for posterior urethral/vesicourethral anastomotic strictures (86.4%). The present study clearly shows that most urologists still prefer DVIU and urethral dilatation to urethroplasty in MUSD, which contradicts current guidelines. Urologists should be encouraged to perform urethroplasty and/or refer patients to experienced centres for recurrent MUSD.


Corneal Haze and Densitometry in Keratoconus after Collagen Cross-Linking by Three Different Protocols.
To evaluate postoperative corneal haze and corneal densitometry following three different corneal cross-linking (CXL) protocols; standard, accelerated, and trans-epithelial (TE). The study recruited 104 eyes (53 patients) with progressive keratoconus divided into three groups: Group I were subjected to standard CXL, Group II to TE-CXL, and Group III to accelerated CXL (A-CXL) (10 mW/cm2 for 9 min). Subjective and objective corneal haze measures were evaluated before and 3, 6, and 12 months post-CXL using slit-lamp biomicroscopy and Pentacam Sheimpflug camera. There was a significant difference in corneal densitometry between the three groups at 3 and 6 months post-CXL (P < 0.0001). By the 12th month, a significant statistical difference was observed only in zones (0-2 mm) and (2-6 mm) in both the anterior and the central layers. In Group I, the densitometry value of the preoperative anterior stromal layer (anterior 120 μm) was 19.42 ± 1.81. Then, it peaked at 23.12 ± 1.21 at 3 months (P < 0.0001), reached 19.82 ± 1.19 at 6 months (P = 0.007), and decreased to 19.33 ± 3.23 (P > 0.05) at 12 months. In Group II, the preoperative densitometry value of the anterior layer was 19.41 ± 1.21, peaked at 19.72 ± 1.12 at 3 months (P = 0.02), reached 19.04 ± 1.18 at 6 months (P = 0.052), and increased to 19.13 ± 1.37 at 12 months (P = 0.84). In Group III, the preoperative densitometry value of the anterior stromal layer was 19.53 ± 2.23. Then, it peaked at 24.80 ± 1.08 at 3 months (P < 0.0001), decreased to 21.75 ± 1.11 at 6 months (P < 0.0001), and reached 19.77 ± 2.26 at 12 months (P = 0.047). There was no significant correlation between the visual acuity changes and the total corneal densitometry. The TE-CXL group showed a better and earlier recovery from the haze, while the A-CXL group showed a delay in recovering and persistent increased corneal densitometry, mainly in the anterior 120 μ.


The struggle of translating science into action: Foundational concepts of implementation science.
RATIONALE, AIMS, AND OBJECTIVES: "Implementation science," the scientific study of methods translating research findings into practical, useful outcomes, is contested and complex, with unpredictable use of results from routine clinical practice and different levels of continuing assessment of implementable interventions. The authors aim to reveal how implementation science is presented and understood in health services research contexts and clarify the foundational concepts: diffusion, dissemination, implementation, adoption, and sustainability, to progress knowledge in the field. Implementation science models, theories, and frameworks are critiqued, and their value for laying the groundwork from which to implement a study's findings is emphasised. The paper highlights the challenges of turning research findings into practical outcomes that can be successfully implemented and the need for support from change agents, to ensure improvements to health care provision, health systems, and policy. The paper examines how researchers create implementation plans and what needs to be considered for study outputs to lead to sustainable interventions. This aspect needs clear planning, underpinned by appropriate theoretical paradigms that rigorously respond to a study's aims and objectives. Researchers might benefit from a return to first principles in implementation science, whereby applications that result from research endeavours are both effective and readily disseminated and where interventions can be supported by appropriate health care personnel. These should be people specifically identified to promote change in service organisation, delivery, and policy that can be systematically evaluated over time, to ensure high-quality, long-term improvements to patients' health.


[Brucellosis: clinical presentation, diagnosis, complications and therapeutic options].
Brucellosis is a zoonotic disease that causes systemic symptoms and can involve many organs and tissues. The major sources of infection are consumption of unpasteurized diary products and occupational contact. Brucella is a small, gram-negative coccobacillus that grows slowly in vitro. There are four species of brucella that are pathogenic for humans; in Israel Brucella melitensis is still the most frequent organism. There are several methods to identified the organism and make the diagnosis (1) isolation of brucella from blood, tissue specimens, body fluids and bone marrow; (2) agglutination test and (3) polymerase chain reaction (PCR) that has recently been shown to be a promising tool for the diagnosis of acute disease. Involvement of the musculoskeletal system is the most common complication of brucellosis, while meningitis and endocarditis are life-threatening complications. The standard treatment for acute and chronic brucellosis is a combination of doxycycline with a second drug such as rifampicin or gentamicin, in order to cure, prevent complications and relapse. Although the rate of occurrence is ever-decreasing due to vaccination of animals, nonetheless, the disease has not been eradicated in Israel. This review focuses on the clinical presentation, diagnosis, and mainly on complications of brucellosis and the available therapeutic options.


Complications of cerebrospinal fluid drainage after thoracic aortic surgery: a review of 504 patients over 5 years.
Cerebrospinal fluid drainage, a well-established means of preventing paraplegia after thoracic aortic aneurysm surgery, can result in serious, sometimes lethal complications. In a large group of patients who underwent surgical thoracic aortic aneurysm repair with cerebrospinal fluid drainage, we examined the incidences of and potential risk factors for these outcomes. The records were reviewed of 504 patients who underwent surgical thoracic aortic aneurysm repair with cerebrospinal fluid drainage at the Texas Heart Institute at St. Luke's Episcopal Hospital between February 2005 and December 2009. All historical data, inpatient records, and billing data were searched for evidence of complications. Of the 504 patients, 14 (2.8%) had intracranial hemorrhage, of whom 10 (72%) had subdural hematoma. Postdural puncture headache developed in 49 patients (9.7%), of whom 17 (34.6%) required epidural blood patch placement for resolution. Multivariable analysis identified having a connective tissue disorder (odds ratio, 3.08; 95% confidence interval, 1.33-7.13) as an independent predictor of postdural puncture headache, but not age less than 40 years (odds ratio, 0.97; 95% confidence interval, 0.94-0.99). Cerebrospinal fluid drainage, as performed by our method, seems to be associated with a modest rate of intracranial bleeding in patients who undergo surgical thoracic aortic aneurysm repair. In contrast, postdural puncture headache is not uncommon, particularly in patients with connective tissue disease. Clinicians caring for these patients should consider the likelihood of postdural puncture headache, and any such patient with postoperative headache should be assessed for epidural blood patch placement.


4D intravital imaging studies identify platelets as the predominant cellular procoagulant surface in a mouse model of hemostasis.
Interplay between platelets, coagulation/fibrinolytic factors, and endothelial cells (ECs) is necessary for effective hemostatic plug formation. This study describes a novel four-dimensional (4D) imaging platform to visualize and quantify hemostatic plug components with high spatiotemporal resolution. Fibrin accumulation following laser-induced endothelial ablation was observed at the EC-platelet plug interface, controlled by the antagonistic balance between fibrin generation and breakdown. Phosphatidylserine (PS) was first detected in close physical proximity to the fibrin ring, followed by exposure across the endothelium. Impaired PS exposure in cyclophilinD -/- mice resulted in a significant reduction in fibrin accumulation. Adoptive transfer and inhibitor studies demonstrated a key role for platelets, but not ECs, in fibrin generation during hemostatic plug formation. Inhibition of fibrinolysis with tranexamic acid (TXA) led to increased fibrin accumulation in WT mice, but not in cyclophilinD -/- mice or WT mice treated with antiplatelet drugs. These studies implicate platelets as the functionally dominant procoagulant surface during hemostatic plug formation. In addition, they suggest that impaired fibrin formation due to reduced platelet procoagulant activity is not reversed by TXA treatment.


Prostaglandin D2 signaling mediated by the CRTH2 receptor is involved in MK-801-induced cognitive dysfunction.
Chemoattractant receptor-homologous molecule expressed on T helper type 2 cells (CRTH2), which is a second receptor for prostaglandin (PG) D2, is involved in inflammatory responses in peripheral tissue; however, its role in cognitive function remains unclear. Here, we demonstrate that CRTH2 is involved in cognitive function using a well-established animal model of cognitive dysfunction induced by MK-801, an N-methyl-d-aspartate receptor antagonist. Genetic deletion and pharmacological inhibition of CRTH2 suppressed MK-801-induced cognitive dysfunction. Pharmacological inhibition of cyclooxygenase-1, a rate-limiting enzyme in PG synthesis, also suppressed MK-801-induced cognitive dysfunction. Moreover, an MK-801-induced increase in c-Fos expression in the paraventricular nucleus (PVN) was abolished in the CRTH2-deficient mice. Together, these results suggest that PGD2-CRTH2 signaling is involved in both MK-801-induced cognitive dysfunction and neuronal activity regulation in the PVN. Furthermore, genetic association studies suggest that CRTH2 is weakly associated with cognitive function in humans. Our study provides evidence that PGD2-CRTH2 signaling is involved in cognitive function and may represent a potential therapeutic target for cognitive dysfunction in patients with psychiatric disorders.


Regulation of food intake by inflammatory cytokines in the brain.
A number of inflammatory cytokines are synthesized and released after activation of the immune system. In addition to other biological effects, these cytokines can potently inhibit food intake. Cytokine-mediated inhibition of food intake is of particular importance because excessive production of peripheral inflammatory cytokines is often associated with the cachexia-anorexia syndrome seen in some chronic diseases. The weight loss in cachexia is associated with an increase in morbidity and mortality. Understanding how cytokines regulate food intake may be crucial in enhancing quality of life and facilitating recovery in patients exhibiting cachexia. This review describes the main inflammatory cytokines that influence food intake and explores how peripheral cytokines communicate with hypothalamic nuclei to influence feeding.


Biofortified cassava increases β-carotene and vitamin A concentrations in the TAG-rich plasma layer of American women.
Biofortification of cassava with the provitamin A carotenoid β-carotene is a potential mechanism for alleviating vitamin A deficiency. Cassava is a staple food in the African diet, but data regarding the human bioavailability of β-carotene from this food are scarce. The objective of the present study was to evaluate provitamin A-enhanced cassava as a source of β-carotene and vitamin A for healthy adult women. The study was a randomised, cross-over trial of ten American women. The subjects consumed three different porridges separated by 2 week washout periods. Treatment meals (containing 100 g cassava) included: biofortified cassava (2 mg β-carotene) porridge with added oil (15 ml peanut or rapeseed oil, 20 g total fat); biofortified cassava porridge without added oil (6 g total fat); unfortified white cassava porridge with a 0·3 mg retinyl palmitate reference dose and added oil (20 g total fat). Blood was collected six times from - 0·5 to 9·5 h post-feeding. TAG-rich lipoprotein (TRL) plasma was separated by ultracentrifugation and analysed using HPLC with coulometric array electrochemical detection. The AUC for retinyl palmitate increased after the biofortified cassava meals were fed (P< 0·05). Vitamin A conversion was 4·2 (sd 3·1) and 4·5 (sd 3·1) μg β-carotene:1 μg retinol, with and without added oil, respectively. These results show that biofortified cassava increases β-carotene and retinyl palmitate TRL plasma concentrations in healthy well-nourished adult women, suggesting that it is a viable intervention food for preventing vitamin A deficiency.


Drugs that influence tubulin polymerization modulate thyrotropin-releasing hormone receptor number in AtT-20 cells.
In order to examine the role of cytoskeleton in modulating the cell surface receptors, AtT-20 cells (stably expressing thyrotropin-releasing hormone receptors) were incubated with drugs that are known to modify the tubulin-microtubule system. The binding of [3H]methyl thyrotropin-releasing hormone ([3H]mTRH) to intact cells increased as a function of time, and was linear from 1.25 x 10(6) to 6.25 x 10(6) cells/ml. Cells incubated with colchicine, vinblastine, and taxol for 16 hr were harvested and the cell concentration was determined using a haemocytometer. Because the drugs inhibited the cell proliferation at 100 nM, it was decided to examine the effect of 100 nM of each of the three drugs on the ability of [3H]mTRH to bind cell surface receptors. Cells were incubated with the drugs for 16 hr at 37 degrees. After the incubation, cells (5 x 10(6) cells/ml) from each group were assayed for [3H]mTRH binding. Colchicine, vinblastine, and taxol stimulated [3H]mTRH binding by up to 27, 27, and 21%, respectively, without altering the Ka of the ligand to the receptor. These results suggest that perturbation of cytosolic microtubules leads to a reorganization of the spatial location of hormone receptors.


Trauma and Delayed Memory: A Review of the "Repressed Memories" Literature.
This review aims to draw balanced conclusions about trauma and memory from the intensely polarized debate currently raging over "repressed" memories, or better, delayed memories (hereafter DM). Research suggests that: emotion impacts memory; psychogenic amnesia can be a reaction to unusual levels of trauma; memory is malleable and delayed memories are prone to errors; however, inaccuracies in traumatic memories are more likely to be in peripheral details than central details. Also reviewed are infantile amnesia, clinical surveys on DM, and two psychoanalytic perspectives on DM. Treatment recommendations are culled from the literature. In order that the debate over adult DM not divert attention from the reality of child abuse and its damage, child abuse issues begin and end the review.


Shape and stability of two-dimensional lipid domains with dipole-dipole interactions.
We study the general energy and shape of the two-dimensional solid monolayer domains with the dipole-dipole interactions. Compared with the domain energy without tilted dipole moments [M. Iwamoto and Z. C. Ou-Yang, Phys. Rev. Lett. 93, 206101 (2004)], the general dipolar energy is not only shape and size but also boundary orientation dependent. The general shape equation derived by this energy using variational approach predicts a circular solution and an equilibrium shape grown from this circle. In particular, the latter is composed of two branches: a translation-induced growth of all odd harmonic modes and a pressure-induced cooperative deformation by all even harmonic modes. The good qualitative agreement between our prediction and the experimental observations shows the validity of the present theory.


The postprandial effects of dietary antioxidants in humans.
Traditional risk factors as measured in the fasted individual are reported to be responsible for the prediction of only half of the incident cases of cardiovascular disease. However, many complex and deleterious reactions occur in the postprandial state. The consequences of oxidative reactions occurring during this time represent major risk for fatal and nonfatal heart disease, ischemia, and stroke, and include oxidative modifications to low-density lipoproteins (LDL), decreased production and bioactivity of nitric oxide (NO) in endothelial cells, and reduced endothelial function. Supplementation with antioxidants may prevent or reduce many of these risks. Antioxidants have been shown to reduce oxidative modification to LDL cholesterol, prevent glucose auto-oxidation, improve the bioactivity of NO, and attenuate or prevent the decrease in endothelial function associated with the postprandial state. Because many nonfasting reactions represent major risk for disease, postprandial risk analysis must form a larger part of the diagnostic strategy for disease prevention in the future.


Multiple Myeloma Relapse as Intracranial Plasmacytoma: A Rare Presentation.
Multiple myeloma (MM) infrequently involves the central nervous system (CNS). The usual sites of involvement are skull and meninges; however, intracranial tumors are exceedingly rare. We report the case of a 60-year-old female who presented to our center for the complaint of recurrent syncope. The patient was diagnosed with MM approximately one and a half years ago and had received chemotherapy followed by an allogeneic bone marrow transplant and was in remission afterward. We initiated workup for syncope and a brain MRI revealed an intracranial mass. Histopathological studies of the intracranial mass confirm the diagnosis of plasmacytoma and further testing shows relapse of MM. This is a unique case of MM relapse with isolated intracranial plasmacytoma. It usually carries a poor prognosis. Early diagnosis and management are imperative to improve survival.