Implication of high grade intraepithelial neoplasia in adverse pathology after radical prostatectomy.
The implication of high grade prostatic intraepithelial neoplasia (HGPIN) to prostate cancer aggressiveness and prognosis is conflicted. The aim of the present study was to evaluate the role of HGPIN in prediction of adverse pathology in patients undergoing a radical prostatectomy. We retrospectively analysed patients who underwent a radical prostatectomy between January 2005 and December 2010. The relationship between HGPIN and the presence of upgrade, positive surgical margins (PSM), extracapsular disease (ECD), seminal vesicle invasion (SVI) and lymph node invasion (LNI) was analysed. HGPIN predictive ability was estimated by using receiver operating characteristic curves. HGPIN was found in 160 (53.3%) specimens. A statistically significant correlation was found between HGPIN and preoperative prostate specific antigen (p=0.020) and patients' age (p=0.025). No significant differences were found, regarding the presence of adverse pathological findings, between the patients with or without HGPIN, irrespective of the preoperative risk stratification. HGPIN did not reach significance for the prediction of upgrade, PSM, ECD, SVI and LNI. The presence of concomitant HGPIN and prostate cancer found not to be related with tumor aggressiveness in patients undergoing a radical prostatectomy and should not be considered as a parameter for the operative outcome prediction.


IgG1, IgG4 and IgE antibody reactivity to mutant forms of the major dust mite allergen Lep d 2 among atopic and nonatopic subjects naturally exposed to Lepidoglyphus destructor.
Lepidoglyphus destructor is a common dust mite causing IgE-mediated sensitization. The major allergen, Lep d 2, has previously been cloned and expressed as several double Cys to Ser mutants with the purpose of producing hypoallergenic variants for immunotherapy. Our aim was to investigate the reactivity pattern of IgG1, IgG4 and IgE antibodies to wild-type (wt) rLep d 2 and four mutants among atopic and nonatopic subjects in relation to sensitization and exposure to L. destructor. Inhibition and sandwich ELISA were used to compare IgG1, IgG4 and IgE antibody reactivities to rLep d 2 variants in serum of 20 atopic and 18 nonatopic farmers naturally exposed to L. destructor. A group of 22 urban subjects served as controls. Atopic farmers demonstrated correlating IgE and IgG4 levels to rLep d 2(wt) (rs = 0.70; p < 0.0001) which were significantly (p < 0.0001) higher than those of nonatopic farmers and urban controls. No IgG4 antibodies were detected in nonatopic farmers despite chronic allergen exposure. A parallel reactivity pattern of IgE and IgG4 to all rLep d 2 mutants was observed. The mutant lacking all 3 disulfide bonds, rLep d 2.6Cys, demonstrated neither any IgE nor IgG4 reactivity. In contrast, IgG1 antibodies had a different reactivity pattern and were detected among most subjects irrespective of atopy, exposure to L. destructor or disulfide impairments in rLep d 2. Moreover, IgG1 levels to rLep d 2(wt) and rLep d 2.6Cys correlated (n = 60; rs = 0.65; p < 0.0001). IgE/IgG4 Ab to rLep d 2 were restricted to atopic farmers and demonstrated parallel recognition patterns of conformational epitopes. In contrast, IgG1 antibodies are ubiquitously found and mainly recognize sequential structures. The observed isotypic difference and interindividual variation in antibody specificities among atopic and nonatopic subjects imply careful investigation of hypoallergenic variants destined for immunotherapy.


Flagellar Synchronization Is a Simple Alternative to Cell Cycle Synchronization for Ciliary and Flagellar Studies.
The unicellular green alga Chlamydomonas reinhardtii is an ideal model organism for studies of ciliary function and assembly. In assays for biological and biochemical effects of various factors on flagellar structure and function, synchronous culture is advantageous for minimizing variability. Here, we have characterized a method in which 100% synchronization is achieved with respect to flagellar length but not with respect to the cell cycle. The method requires inducing flagellar regeneration by amputation of the entire cell population and limiting regeneration time. This results in a maximally homogeneous distribution of flagellar lengths at 3 h postamputation. We found that time-limiting new protein synthesis during flagellar synchronization limits variability in the unassembled pool of limiting flagellar protein and variability in flagellar length without affecting the range of cell volumes. We also found that long- and short-flagella mutants that regenerate normally require longer and shorter synchronization times, respectively. By minimizing flagellar length variability using a simple method requiring only hours and no changes in media, flagellar synchronization facilitates the detection of small changes in flagellar length resulting from both chemical and genetic perturbations in Chlamydomonas. This method increases our ability to probe the basic biology of ciliary size regulation and related disease etiologies. IMPORTANCE Cilia and flagella are highly conserved antenna-like organelles that found in nearly all mammalian cell types. They perform sensory and motile functions contributing to numerous physiological and developmental processes. Defects in their assembly and function are implicated in a wide range of human diseases ranging from retinal degeneration to cancer. Chlamydomonas reinhardtii is an algal model system for studying mammalian cilium formation and function. Here, we report a simple synchronization method that allows detection of small changes in ciliary length by minimizing variability in the population. We find that this method alters the key relationship between cell size and the amount of protein accumulated for flagellar growth. This provides a rapid alternative to traditional methods of cell synchronization for uncovering novel regulators of cilia.


Cross-linking membrane IgM induces production of inositol trisphosphate and inositol tetrakisphosphate in WEHI-231 B lymphoma cells.
The addition of anti-IgM to the immature B lymphoma cell line WEHI-231 resulted in breakdown of phosphatidylinositol 4,5-bisphosphate, generating diacylglycerol and inositol 1,4,5-trisphosphate (Ins(1,4,5)P3). These reactions have recently been demonstrated in mature resting B cells stimulated with anti-IgM, as well. In addition to Ins(1,4,5)P3, inositol tetrakisphosphate (InsP4) and inositol 1,3,4-trisphosphate (Ins(1,3,4)P3) were rapidly generated in WEHI-231 cells upon stimulation of the antigen receptor with anti-IgM. These two inositol polyphosphates are probably generated from Ins(1,4,5)P3 by phosphorylation to yield InsP4 and removal of the 5-phosphate from InsP4 to yield Ins(1,3,4)P3. It is possible that these inositol polyphosphates play a second messenger role in mediating the biologic effects of antigen-receptor signaling. It had previously been shown that anti-IgM also causes an increase in cytoplasmic free calcium. Therefore, the relationship between Ca2+ elevation and phosphoinositide breakdown was investigated. Although elevation of cytoplasmic Ca2+ with ionophores can trigger phosphoinositide breakdown, this required levels of Ca2+ well beyond those normally seen in response to anti-IgM. Thus, the Ca2+ elevation seen in response to anti-IgM cannot be the event controlling phosphoinositide breakdown. WEHI-231 cells have been shown to have a calcium storage compartment that releases Ca2+ in the presence of Ins(1,4,5)P3; therefore, it is likely that anti-IgM stimulates phosphoinositide breakdown as a primary event and this leads to the elevation of cytoplasmic Ca2+.


Haemophilus influenzae b polysaccharide revaccination: a continued role for the unconjugated vaccine.
In a study of the issue of revaccination with Haemophilus influenzae b polysaccharide vaccine, 36 children who had been vaccinated with polysaccharide vaccine at age 18 to 20 months were stratified according to their 3-week postvaccination anticapsular antibody concentration determined by radioantigen binding assay, then sequentially assigned to one of three groups so as to have comparable distributions of peak antibody concentrations. These children were revaccinated with polysaccharide vaccine at 24 to 27, 30 to 33, or 36 to 39 months of age. The post-reimmunization geometric mean antibody concentrations in these three groups were 3.1, 3.0 and 7.8 micrograms/ml, respectively, and the percentages rising to greater than 1 microgram/ml were 75, 80 and 93%. The geometric means and antibody response rates were nearly identical to age-matched control groups not previously vaccinated. Thus there was no evidence of priming (memory induction) or of tolerance induction by the early (18 months old) primary vaccination.


An evaluation of multidimensional fingerprinting in the context of clinical proteomics.
Multidimensional fingerprinting (MDF) utilizes measurable peptide characteristics to identify proteins. In this study, 3-D fingerprinting, namely, parent protein molecular weight, peptide mass, and peptide retention time on RPLC, is used to identify 331 differentially expressed proteins between normal and human colon cancer plasma membrane samples. A false discovery rate (FDR) procedure is introduced to evaluate the performance of MDF on the colon cancer dataset. This evaluation establishes a false protein identification rate below 15% for this dataset. Western blot analysis is performed to validate the differential expression of the MDF-identified protein VDAC1 on the original tissue samples. The limits of MDF are further assessed by a simulation study where key parameters such as database size, query size, and mass accuracy are varied. The results of this simulation study demonstrate that fingerprinting with three dimensions yields low FDR values even for large queries on the complete human proteome without the need for prior peptide sequencing by tandem mass spectrometry. Specifically, when mass accuracy is 10 ppm or lower, full human proteome searches can achieve FDR values of 10% or less.


RANKL-Induced Btn2a2 - A T Cell Immunomodulatory Molecule - During Osteoclast Differentiation Fine-Tunes Bone Resorption.
Butyrophilins, which are members of the extended B7 family of immunoregulators structurally related to the B7 family, have diverse functions on immune cells as co-stimulatory and co-inhibitory molecules. Despite recent advances in the understanding on butyrophilins' role on adaptive immune cells during infectious or autoimmune diseases, nothing is known about their role in bone homeostasis. Here, we analyzed the role of one specific butyrophilin, namely Btn2a2, as we have recently shown that Btn2a2 is expressed on the monocyte/macrophage lineage that also gives rise to bone degrading osteoclasts. We found that expression of Btn2a2 on monocytes and pre-osteoclasts is upregulated by the receptor activator of nuclear factor κ-B ligand (RANKL), an essential protein required for osteoclast formation. Interestingly, in Btn2a2-deficient osteoclasts, typical osteoclast marker genes (Nfatc1, cathepsin K, TRAP, and RANK) were downregulated following RANKL stimulation. In vitro osteoclast assays resulted in decreased TRAP positive osteoclast numbers in Btn2a2-deficient cells. However, Btn2a2-deficient osteoclasts revealed abnormal fusion processes shown by their increased size. In vivo steady state µCT and histological analysis of bone architecture in complete Btn2a2-deficient mice showed differences in bone parameters further highlighting the fine-tuning effect of BTN2a2. Moreover, in rheumatoid arthritis patients and experimental arthritis, we detected significantly decreased serum levels of the secreted soluble Btn2a2 protein. Taken together, we identified the involvement of the immunomodulatory molecule Btn2a2 in osteoclast differentiation with potential future implications in basic and translational osteoimmunology.


Problems in using high performance liquid chromatography for drug analysis.
Some problems encountered in using high performance liquid chromatography (HPLC) for drug analysis are discussed. These include high procurement and operational costs, lengthy training required, excessive downtimes , lower precision in HPLC versus gas chromatography (GC) and lack of a universal sensitive detector. Some solutions to these difficulties are presented.


Impact of season on liver mitochondrial oxidative stress and the expression of HSP70 in grey mullets from contaminated estuary.
Heat shock proteins (HSPs) are the ubiquitous feature of cells in which these proteins cope with stress induced denaturation of other proteins. HSP70 is found to play a primary role in cellular defense under stress condition. In the present investigation, the seasonal impact on environmental stress induced mitochondrial HSP70 (mtHSP70) expression in the liver mitochondria was examined in grey mullets, Mugil cephalus living in the Ennore estuary (polluted site) was compared with the Kovalam estuary (unpolluted site) over the course of two seasons viz monsoon and summer from April 2006 to March 2008. Oxidative stress was determined along with mtHSP70 expression studies in fish liver mitochondria collected from these two estuaries for both the seasons. The liver mitochondria of grey mullet fish collected from polluted Ennore estuary showed increased levels of lipid peroxide and mtHSP70 expression along with decrease in total antioxidant capacity and glutathione redox ratio levels ((c) P < 0.05) when compared to unpolluted Kovalam estuary fish. In the fish liver mitochondria of Ennore estuary, there was significant seasonal variation ((b) P < 0.05) in both oxidative stress marker levels (34% increase) and mitochondrial HSP70 expression (33% increase) with increased level during summer season but the Kovalam estuary fish did not show any significant seasonal variation. In the Ennore estuary fish that are exposed to chronic environmental stress, the overexpression of liver mtHSP70 particularly during summer season may confer differential effects on the cell survival by protection against oxidative stress induced changes.


Genetic variability and relationships among seventeen Trichoderma isolates to control dry root rot disease using RAPD markers.
Trichoderma spp. has been identified as potential antagonist of Fusarium solani, which is causing dry root rot of Citrus. A random amplified polymorphic DNA (RAPD) marker was used to estimate the genetic variation among 17 isolates of Trichoderma. These isolates were characterized using 20 random primers of the OPM series, out of which 16 primers gave a total of 145 DNA fragments, showing 91.8% polymorphism. The genetic distance between each isolate was calculated, and cluster analysis was used to generate a dendrogram showing the relationship among them. The isolates grouped into two major clusters, the first major cluster consisted of TCT14, TCT17, TCT13, TCT12 and TCT16. The remaining isolates in the second major cluster separated in two sub-clusters; the first cluster consisted of TCT4, TCT10, TCT2, TCT3, TCT8, TCT6, TCT9, and the second sub-cluster consisted of TCT1 TCT15 TCT5, TCT11, and TCT7. The similarity matrix indicated that TCT6 and TCT13 were genetically distinct as they showed only 22.6% similarity followed by TCT5 and TCT16; TCT6 and TCT16 (25%), while the isolates TCT4 and TCT10 were found to be genetically similar, as 66.7% similarity was observed between the isolates followed by 61.3% similarity between the TCT2 and TCT4 isolates.


Limitations and pitfalls of 99mTc-EDDA/HYNIC-TOC (Tektrotyd) scintigraphy.
Tektrotyd kit was developed by Polatom company for 99mTc labeling to make an alternative tracer of somatostatin receptor scintigraphy available. Since 2005, 99mTc-EDDA/HYNIC-Tyr3-Octreotide has been used in clinical imaging and achieved high impact in management of patients with neuroendocrine tumors. Knowing the limitations and pitfalls is essential to provide ac-curate diagnosis. Therefore, the potential pitfalls associated with the use of 99mTc-EDDA/HYNIC-TOC are reviewed on the basis of own experience. Data were analyzed of 310 patients who underwent somatostatin receptor scintigraphy with 99mTc-Tektrotyd. Pitfalls during radiolabeling process or acquisition can worsen the sensitivity of SRS (somatostatin receptor scintigraphy). Recognizing physi-ological and clinical pitfalls, the diagnostic accuracy will improve.


Migration within Spain, level of education, and cervical cancer screening.
Migration has been related to the utilization of preventive care services. We analysed the relation between cervical cancer screening and migration within the same country and socioeconomic status in a context in which there is no organized screening. The health survey of Barcelona (Spain) was the data source. Barcelona, a city in north eastern Spain, has experienced important migration from the south and other regions of Spain. Variables studied were the migrant women's year of arrival, age at arrival, educational attainment and Pap smear test uptake. Multivariate logistic regression analysis was performed to examine the correlations among the variables of interest. Just over 30% of the women had never had a Pap smear test. Uptake was higher among younger, educated women born either in Barcelona or abroad. After adjusting for age, migrant women were at higher risk for not participating in screening tests (odds ratio: 1.23; 1.09-1.39); but after adjusting for educational attainment and age, the odds ratio was no longer significant. This study shows that migrants within Spain have less access to preventive services, such as cervical cancer screening in an opportunistic setting. However, this association is almost completely explained by socioeconomic status. Migration could be seen as a social factor that puts people at risk of falling into lower socioeconomic status associated with poor access to screening.


A case-control evaluation of 143 single nucleotide polymorphisms for breast cancer risk stratification with classical factors and mammographic density.
Panels of single nucleotide polymorphisms (SNPs) stratify risk for breast cancer in women from the general population, but studies are needed assess their use in a fully comprehensive model including classical risk factors, mammographic density and more than 100 SNPs associated with breast cancer. A case-control study was designed (1,668 controls, 405 cases) in women aged 47-73 years attending routine screening in Manchester UK, and enrolled in a wider study to assess methods for risk assessment. Risk from classical questionnaire risk factors was assessed using the Tyrer-Cuzick model; mean percentage visual mammographic density was scored by two independent readers. DNA extracted from saliva was genotyped at selected SNPs using the OncoArray. A predefined polygenic risk score based on 143 SNPs was calculated (SNP143). The odds ratio (OR, and 95% confidence interval, CI) per interquartile range (IQ-OR) of SNP143 was estimated unadjusted and adjusted for Tyrer-Cuzick and breast density. Secondary analysis assessed risk by oestrogen receptor (ER) status. The primary polygenic risk score was well calibrated (O/E OR 1.10, 95% CI 0.86-1.34) and accuracy was retained after adjustment for Tyrer-Cuzick risk and mammographic density (IQ-OR unadjusted 2.12, 95% CI% 1.75-2.42; adjusted 2.06, 95% CI 1.75-2.42). SNP143 was a risk factor for ER+ and ER- breast cancer (adjusted IQ-OR, ER+ 2.11, 95% CI 1.78-2.51; ER- 1.81, 95% CI 1.16-2.84). In conclusion, polygenic risk scores based on a large number of SNPs improve risk stratification in combination with classical risk factors and mammographic density, and SNP143 was similarly predictive for ER-positive and ER-negative disease.


Intracellular localization and epitope mapping of feline herpesvirus type 1 glycoproteins.
Monoclonal antibodies (MoAbs) were used to characterize feline herpesvirus type 1 (FHV-1) glycoproteins (gp). Intracellular localization and transport of these proteins as revealed by a sequential indirect immunofluorescence assay (IFA) on fixed infected cells showed slight differences between FHV-1 gp143/108 and gp113. Antibodies against gp143/108 first showed membrane fluorescence at 4 hrs post-infection (PI) followed by a pronounced perinuclear and cytoplasmic staining from 8 hrs PI onwards. Those reacting with gp113 showed the same pattern but fluorescence did not appear until 8 hrs PI. In contrast, MoAbs against gp60 first showed para- and perinuclear staining at 12 hrs PI which became intranuclear at 16 hrs PI, followed by intracytoplasmic staining at 20 hrs PI. Sequential IFA of unfixed infected cells revealed that the three glycoproteins were expressed on the cell surface membrane as well. Topographical mapping of the functional epitopes of gp113 by ELISA additivity test indicated the presence of 2 antigenic domains--a neutralizing domain consisting of 3 overlapping epitopes and a non-neutralizing domain. On the other hand, gp143/108 contained only one antigenic site consisting of 5 similar or overlapping epitopes, one of which seemed to be a conserved region recognized by all MoAbs reacting to this protein.


Inhibitory Effects of Trapping Agents of Sulfur Drug Reactive Intermediates against Major Human Cytochrome P450 Isoforms.
In some cases, the formation of reactive species from the metabolism of xenobiotics has been linked to toxicity and therefore it is imperative to detect potential bioactivation for candidate drugs during drug discovery. Reactive species can covalently bind to trapping agents in in vitro incubations of compound with human liver microsomes (HLM) fortified with β-nicotinamide adenine dinucleotide phosphate (NADPH), resulting in a stable conjugate of trapping agent and reactive species, thereby facilitating analytical detection and providing evidence of short-lived reactive metabolites. Since reactive metabolites are typically generated by cytochrome P450 (CYP) oxidation, it is important to ensure high concentrations of trapping agents are not inhibiting the activities of CYP isoforms. Here we assessed the inhibitory properties of fourteen trapping agents against the major human CYP isoforms (CYP1A2, 2C9, 2C19, 2D6 and 3A). Based on our findings, eleven trapping agents displayed inhibition, three of which had IC50 values less than 1 mM (2-mercaptoethanol, N-methylmaleimide and N-ethylmaleimide (NEM)). Three trapping agents (dimedone, N-acetyl-lysine and arsenite) did not inhibit CYP isoforms at concentrations tested. To illustrate effects of CYP inhibition by trapping agents on reactive intermediate trapping, an example drug (ticlopidine) and trapping agent (NEM) were chosen for further studies. For the same amount of ticlopidine (1 μM), increasing concentrations of the trapping agent NEM (0.007-40 mM) resulted in a bell-shaped response curve of NEM-trapped ticlopidine S-oxide (TSO-NEM), due to CYP inhibition by NEM. Thus, trapping studies should be designed to include several concentrations of trapping agent to ensure optimal trapping of reactive metabolites.


Synthesis and evaluation of trimethoxyphenyl isoxazolidines as inhibitors of secretory phospholipase A2 with anti-inflammatory activity.
A series of trimethoxyphenyl isoxazolidine derivatives, 5a(i-v) and 5b(i-v), bearing different constituents at the 5th position of the isoxazolidine ring were synthesized and evaluated in vitro and in vivo for their inhibitory activity against purified group I and II phospholipase A2 (PLA2) enzymes from snake venom and human inflammatory synovial fluid. Irrespective of modification to the pharmacophore (isoxazolidine ring), they exhibited greater specificity for group II PLA2. The length of alkyl or aryl group at the 5th position, which alters the hydrophobic and aromatic property, was responsible for enhancing the inhibition towards PLA2 enzymes. All of the compounds quench the fluorescent property of the purified PLA2 enzyme, and quenching increases with the increase in length of alkyl or aryl group. The inhibitory effect of compounds appeared to be due to the direct interaction of compounds with the enzyme. Inhibition is substrate-dependent, and the inhibitor likely competes with the substrate for the same binding site of the enzyme. The IC50 value for the most potent interacting inhibitor 5b(v) was 54.8 microM. The most active interacting compounds 5a(v) and 5b(v) from in vitro inhibition of PLA2 activity showed similar potency in in vivo neutralization of PLA2-induced mouse paw edema and hemolytic activity.


Corneal dermoid in two laboratory beagle dogs.
Two male laboratory beagle dogs used in toxicity studies, one 7 months old and the other 9 months old, showed the evidence of corneal dermoid. Grossly, the dermoid was observed in both cases as hair growth from the cornea. In one case, the hairs had been removed from the cornea, but regrowth was observed about 70 days later. Histopathologically, melanocytes, melanin granules, hairs, hair bulbs, adipose tissue and sebaceous and sweat glands were observed in the corneal epithelium and propria. According to the information obtained from 4 breeders, the incidence of corneal dermoid was extremely rare in laboratory beagle dogs.


18F-FDG PET/CT and histology for diagnosing recurrent/remnant tumors in head and neck cancer patients treated with radiotherapy.
The aim of this study was to assess the diagnostic performance of fluorine-18-fluoro-2-deoxyglucose (18F-FDG) positron emission tomography (PET)/computed tomography (CT) for locoregional recurrent/residual tumor in patients with head and neck cancer (HNC) who underwent previous radiotherapy (RT). 18F-FDG PET/CT images from patients with HNC who previously underwent RT were retrospectively reviewed. Only cases with histological confirmation within 4 weeks of PET/CT imaging were included. Standardized uptake values were obtained for lesions and PET/CT findings were compared with histological results. Of 181 cases, 114 (63%) were histologically confirmed as malignant and 67 (37%) as benign. The sensitivity, specificity, and accuracy of PET/CT were 93%, 64%, and 82%, respectively. Inflammation was the most common cause of false positives and small tumor volume and low 18F-FDG avidity were the causes of false negatives. PET/CT had 100% sensitivity and 56% specificity for detecting recurrent or residual disease within 12 weeks after RT and 93% sensitivity and 64% specificity, more than 12 weeks after RT. The frequency of false positives in PET/CT images within 12 weeks of RT was similar to the results obtained 12 weeks after RT (15% vs. 14%). False positives were more frequent in PET/CT cases after two-dimensional or three-dimensional conformal RT than in those after intensity-modulated RT, although not statistically significant (15% vs. 9%, p>0.05). 18F-FDG PET/CT might aid the diagnosis of locoregional residual/recurrent tumors in patients with HNC previously treated with RT. Inflammation was the main cause of false positives regardless of the interval between RT and PET/CT, even several years after RT. Therefore, histological verification of positive PET/CT findings should be conducted during follow-up of HNC patients treated with RT.


Analysis of CTG repeat length variation in the DMPK gene in the general population and the molecular diagnosis of myotonic dystrophy type 1 in Malaysia.
The lack of epidemiological data and molecular diagnostic services in Malaysia has hampered the setting-up of a comprehensive management plan for patients with myotonic dystrophy type 1 (DM1), leading to delayed diagnosis, treatment and support for patients and families. The aim of this study was to estimate the prevalence of DM1 in the 3 major ethnic groups in Malaysia and evaluate the feasibility of a single tube triplet-primed PCR (TP-PCR) method for diagnosis of DM1 in Malaysia. We used PCR to determine the size of CTG repeats in 377 individuals not known to be affected by DM and 11 DM1 suspected patients, recruited from a tertiary hospital in Kuala Lumpur. TP-PCR was performed on selected samples, followed by Southern blot hybridisation of PCR amplified fragments to confirm and estimate the size of CTG expansion. The number of individuals not known to be affected by DM with (CTG)>18 was determined according to ethnic group and as a whole population. The χ2 test was performed to compare the distribution of (CTG)>18 with 12 other populations. Additionally, the accuracy of TP-PCR in detecting CTG expansion in 11 patients with DM1 was determined by comparing the results with that from Southern blot hybridisation. Of the 754 chromosomes studied, (CTG)>18 frequency of 3.60%, 1.57% and 4.00% in the Malay, Chinese and Indian subpopulations, respectively, was detected, showing similarities to data from Thai, Taiwanese and Kuwaiti populations. We also successfully detected CTG expansions in 9 patients using the TP-PCR method followed by the estimation of CTG expansion size via Southern blot hybridisation. The results show a low DM1 prevalence in Malaysia with the possibility of underdiagnosis and demonstrates the feasibility of using a clinical and TP-PCR-based approach for rapid and cost-effective DM1 diagnosis in developing countries.


Serological biomarkers in triage of FIT-positive subjects?
FIT-based colorectal cancer screening has been implemented in many countries including Denmark, where 916 colorectal cancer and 4468 high- or medium-risk adenoma patients were identified within April-December 2014, among 16,806 subjects with a positive FIT test. Screening increases the overall requirements for colonoscopy, which may challenge the current capacity. Some countries have increased their initial FIT cut-off level in order to comply with lack of colonoscopy capacity. Many patients with neoplasia will not be detected, however, by using increased FIT cut-off levels. The number of patients with neoplastic lesions missed by increased cut-off levels appears to be much higher than expected. Therefore, tests that identify those patients missed by increased FIT cut-off levels must be developed. Preliminary results of determination of one of several biomarker entities currently under investigation show that nucleosome blood tests may be one option for identifying some of these patients. Implementation of a triage test consisting of FIT, blood-based biomarkers and plus/minus colonoscopy is suggested to identify subjects with FIT levels between the initial and the increased cut-off level that must be offered colonoscopy. In addition, triage may reduce the frequency of unnecessary colonoscopies by 25%.


Does the sexually transmitted infections foundation course deliver and change practice? Feedback from delegates 2002-2006.
Attendees at Newcastle sexually transmitted infections foundation (STIF) courses since 2002 were sent a postal questionnaire to ascertain views about the course, its effect on practice and the desire for further education. Totally 156 forms were returned (48% response). The majority 97% were satisfied with the course, 97% wanted updates, 57% annually and 33% biennially. Following a STIF course, 69% provided HIV testing (only 14% of those, prior to attendance). However, only 39% routinely offered HIV testing and only 34% routinely offered syphilis testing to patients whom they considered to be at risk of a sexually transmitted infection. Common reasons for not offering testing were lack of time for counselling, lack of confidence, no perceived need or anonymity concerns resulting in referral to genitourinary medicine. This was despite training, which encourages routine HIV testing with a pretest discussion rather than 'counselling' and education about recent outbreaks of syphilis.


Telephone requests for donation: concerns expressed by families and the impact of the donor registry.
The request process for eye and tissue donation is different from the process that families of organ donors experience, but the research into eye/tissue-only decision making has been sparse. To determine the concerns of families approached over the phone for eye/tissue donations and to study the impact of the donor registry on those decisions. Written instrument filled out by family services coordinators while speaking with families about donation via phone in the San Diego, California, area during 2011.Main Outcome Measures-Responses marked on a 21-item instrument, contextual notes, donor registrations, and decisions made. With a 60% refusal rate, the most common reasons reported for declining donation are that the potential donor said during life he/she did not want to donate (26%) or that the family, not knowing the potential donor's wishes, opted not to donate (13%). Other specific reasons for not donating were as follows: wanting the body buried whole (8%), concerns about age and prior health (7%), and incompatibility with religion or culture (7%). Consenting families had different concerns: worries about delays (36%), the potential donors' age and health (25%), how the body might look for viewing (14%), the amount of paperwork (10%), for-profit status of beneficiaries (9%), international distribution (8%), and family disunity (8%). Registry-related refusals weighted against donations of tissues from registered donors amounted to a deficit of 92 potential donations. When donations lost to faulty assumptions based on public education are added, the deficit increases to 122 lost donations.


A conversational system for the computer analysis of nucleic acid sequences.
We present a conversational system for the computer analysis of nucleic acid and protein sequences based on the well-known Queen and Korn program (1). The system can be used by persons with only minimal knowledge of computers.


Phenotypic modulation of human articular chondrocytes by bistratene A.
Chondrocytes undergo phenotypic alterations following extended periods in monolayer culture, i.e., they become bipolar and flattened, proliferate, and synthesise type I as opposed to type II collagen. This process has been termed chondrocyte dedifferentiation. Bistratene A is a macrolide polyether that specifically activates the delta isoform of protein kinase C (PKCdelta) in some cell types. Here, we show that dedifferentiated human articular chondrocytes became rounded and underwent cell growth arrest after treatment with bistratene A. In addition, bistratene A-treated chondrocytes became more immunopositive for type II collagen, but less immunopositive for type I collagen. These phenotypic changes were associated with a prior and extensive disruption of actin microfilaments and translocation of PKCdelta to the nuclear membrane. Concurrent treatments of chondrocytes with a specific inhibitor of PKCdelta, rottlerin, partially blocked the morphological effects of bistratene A.


A biophysical, morphological and morphometrical survey of the eye of the small shark (Hemiscyllium plagiosum).
The biophysical and morphological characteristics of the small shark's ( Hemiscyllium plagiosum ) eye were studied. The focal lengths of the lens were found to be very short. A small proportion of cones were present in the retina and this proportion varied with the weights of the sharks. The topographic distribution of cones in different segments of the retina was also presented.


Orthotopic liver transplantation: nonoperative management of early, acute portal vein thrombosis.
We report a case of acute portal vein thrombosis that occurred 1 week after orthotopic liver transplantation in a patient with sclerosing cholangitis. Unlike other patients reported in the literature who were first seen with variceal bleeding or acute hepatic failure, this patient initially had mild clinical signs, consisting of an abnormal prothrombin time, an increase in liver function test values, and enlarging but nonbleeding gastroesophageal varices. Whereas patients with more extreme symptoms often die or require retransplantation, this patient was managed nonoperatively. Spontaneous lysis of the portal vein thrombus occurred over the ensuing 2 weeks. The diagnosis and management of this milder form of early, acute portal vein thrombosis are discussed.


Non-fatal suicidal behaviours in French Polynesia: Results of the WHO/START study and its implications for prevention.
This is the first research article examining non-fatal suicidal behaviours (NFSB) in French Polynesia. The study was conducted in the frames of the WHO/START Study in 2008-2010. The main objective of the investigation was to obtain reliable data in order to develop evidence-based suicide prevention strategies. Interviews with people presenting with NFSB at the Emergency Department (ED) of the French Polynesia Hospital (CHPF) were conducted by emergency and psychiatry departments' staff examining socio-demographic and clinical information and motives triggering suicidal behaviour. Odds ratios and rate ratios with 95% confidence intervals were calculated. There were 556 presentations of NFSB by 515 persons at the ED of CHPF (ratio 1.08) with the average rate of 75 per 100,000 for those treated in the hospital. An estimate of cases occurring in remote islands might bring the rate for the whole French Polynesia around 94 per 100,000. NFSB was more prevalent in females; rates for both genders were highest in the age group 25-35 years and 15-24 years. The main suicide method was drug poisoning by psychotropic drugs. The most frequent psychiatric disorder was mood disorder (45.3%); however, 26% of subjects had no 'major' psychiatric disorders, with 14.5% without a psychiatric diagnosis or only reactive disorders (F43-11.7%). There was a high prevalence of previous NFSB (52.1%). Study includes only NFSB seeking medical help from the biggest hospital in the country. Suicide prevention activities specific to the findings and the socio-cultural context of French Polynesia should be considered.


Polysynaptic olfactory pathway to the ipsi- and contralateral entorhinal cortex mediated via the hippocampus.
Interactions between olfactory cortices and the hippocampus support sensory discrimination and spatial learning functions. The olfactory input accesses the hippocampal formation via a polysynaptic pathway mediated by the lateral and rostral entorhinal cortex (EC). We recently demonstrated that following repetitive stimulation of the lateral olfactory tract (LOT) at 2-8 Hz, a delayed response (onset at circa 60 ms) was evoked in the caudal portion of the EC, identified as medial EC, that does not receive a direct olfactory input. By performing simultaneous laminar profile analysis in the EC and in different hippocampal subfields, we conclusively demonstrate that the delayed EC response evoked by repetitive ipsilateral LOT stimulation is headed by the sequential activation of the dentate gyrus and the CA3/CA1 subfields in the septal and temporal hippocampus. Repetitive stimulation of the contralateral LOT also induced an EC response that peaked at 76.28+/-2.42 ms (n=15). Current source density analysis and time-delay analysis of simultaneous field potential laminar profiles performed from the EC and from DG, CA3 and CA1 hippocampal subfields suggested that the contralateral EC response is mainly carried by an intrahippocampal CA3-CA3 commissural pathway. Contralateral LOT stimulation also induced a later EC component (delay >100 ms) generated in the superficial layers, mediated either by local associative interactions or by extrahippocampal circuits. The opportunity to activate the ipsi- and contralateral olfactory pathways in the same experiment and to record field potentials profiles simultaneously in different structures of both hemispheres in the isolated guinea-pig brain confirms that this preparation is unique and is particularly suitable for investigating the system physiology of the limbic region. The present study demonstrates that patterned stimulation of the olfactory input that mimics sniffing patterns during odor discrimination induces a diffuse activation of both ipsi- and contralateral hippocampi and ECs. The findings contribute to the understanding the physiological mechanisms that underlie associative interactions between olfactory and non-olfactory cortical inputs converging into the mesial temporal region.


N-Linked Glycosylation of the p24 Family Protein p24δ5 Modulates Retrograde Golgi-to-ER Transport of K/HDEL Ligands in Arabidopsis.
The K/HDEL receptor ERD2 mediates the transport of soluble endoplasmic reticulum (ER)-resident proteins containing a C-terminal K/HDEL signal from the Golgi apparatus back to the ER via COPI (COat Protein I)-coated vesicles. Sorting of ERD2 within COPI vesicles is facilitated by p24 proteins. In Arabidopsis, p24δ5 has been shown to interact directly with ERD2 via its luminal GOLD (GOLgi Dynamics) domain and with COPI proteins via its cytoplasmic C-terminal tail at the acidic pH of the Golgi apparatus. Several members of the p24 family in mammals and yeast have been shown to be glycosylated, but whether Arabidopsis p24 proteins are glycosylated and the role of the sugar moiety in p24 function remain unclear. Here, we show that Arabidopsis p24δ5 protein is N-glycosylated in its GOLD domain. Furthermore, we demonstrate that this post-translational modification is important for its coupled transport with p24β2 at the ER-Golgi interface, for its interaction with the K/HDEL receptor ERD2, and for retrograde transport of ERD2 and K/HDEL ligands from the Golgi apparatus back to the ER.


The Hydrodynamics of a Swirling Blood Flow in the Left Heart and Aorta.
This paper proposes a new approach to the quantitative analysis of the hydrodynamic structure of a blood flow in the flow channel running from the left atrium to the end of the aorta. This approach is based on the concept of the structural organization of tornado-like swirling jets in channels with a given geometric configuration. Considering the large amount of experimental data in our possession, it was shown that along the entire length of the flow channel, conditions exist for the generation and maintenance of a swirling structure of the jet throughout the entire cardiac cycle. This study has given rise to a new direction in research in fundamental physiology and medicine, which is of great practical importance for diagnosing and treating circulatory disorders accompanied by changes in the geometric configuration and biomechanical characteristics of the heart and great vessels.


Role of Rho in rabbit parietal cell.
Rho is known as an important regulator of actin microfilament formation. We were led to study it because a dynamic rearrangement of actin filaments occurs during activation of gastric acid secretion. In order to use specific probes, the rabbit gastric gland culture system was employed and the various genes were expressed using adenovirus vector. When the constitutive active mutant of Rho (RhoAV14) was expressed, histamine- or carbachol-stimulated acid secretion monitored by (14)C-aminopyrine accumulation was inhibited. Conversely, expression of C3 toxin, the specific inhibitor of Rho, and expression of G(12/13)-specific regulator of G-protein signaling domain, the specific inhibitor of G(12/13) which is considered to be an upstream mediator of Rho, both potentiated acid secretion stimulated by the agonists. F-actin staining of parietal cell expressing RhoAV14 revealed that the microfilament supporting the intracellular canaliculi (not on the basolateral membrane) almost disappeared. No clear changes in the intracellular localization of Rho were observed during stimulation of parietal cell. In resting glands, the endogenous active form of Rho was relatively high, and it decreased during histamine stimulation. The finding that any treatment which inhibit Rho augment acid secretion whereas those that activate Rho inhibit secretion strongly suggests that the Rho-pathway conducts a negatively regulating signal in parietal cell activation, possibly via site-specific regulation of actin microfilaments.


5-HT1A receptor activation before acute stress counteracted the induced long-term behavioral effects.
The long-term behavioral consequences of acute immobilization (IMMO) in rats and the effects of 5-HT(1A) receptor activation (8-OH-DPAT: 0.3 mg/kg, sc) were studied. Corticosterone levels after IMMO with previous 8-OH-DPAT treatment were also studied. Twenty-four hours after IMMO (3 h), rats performed conditioned (passive avoidance) and unconditioned (escape behavior) anxiety tests in the elevated T maze. Pre-exposure to IMMO induces long-term behavioral changes in contrast with control rats. These behavioral alterations include an increase of anxiogenic responses, such as exploratory behavior and passive avoidance response. This effect was counteracted by 8-OH-DPAT pretreatment and reversed by WAY-100635 when administered before 8-OH-DPAT. Serum corticosterone levels increased during the first hour of stress and after 8-OH-DPAT administration. Our results support the hypothesis that involvement of acute stress is crucial in the anxiety-like behaviors and in the potentiation of fear. The activation of 5-HT(1A) receptors counteracted the long-term effects induced by IMMO.


Efficacy and Safety of Once-Daily Inhaled Umeclidinium in Asian Patients with COPD: Results from a Randomized, Placebo-Controlled Study.
Previous studies demonstrating efficacy and safety of once-daily umeclidinium (UMEC) in patients with chronic obstructive pulmonary disease (COPD) have included few Asian patients. This study evaluated efficacy and safety of UMEC 62.5 mcg versus placebo in Asian patients with COPD. A Phase III, randomized, double-blind, parallel-group study. Patients (aged ≥40 years with COPD, pre-, and post-albuterol forced expiratory volume in 1 s [FEV1]/forced vital capacity ratio <0.70 and low risk of exacerbations) were randomized 2:1 to once-daily UMEC 62.5 mcg or placebo via the ELLIPTA inhaler for 24 weeks. Primary endpoint was change from baseline (CFB) in trough FEV1 on Day 169. Secondary endpoints were weighted mean FEV1 over 0-6 hrs post-dose on Day 1 and CFB in Transition Dyspnea Index (TDI) focal score on Day 168. A total of 306 patients were included in the modified intent-to-treat population (UMEC: 205; placebo: 101). UMEC versus placebo provided a statistically significant improvement in least squares (LS) mean trough FEV1 between baseline and Day 169 (154 mL [95% confidence interval (CI): 113, 194]; p<0.001). A clinically meaningful difference of 125 mL in favor of UMEC (95% CI: 103, 147; p<0.001) was also seen in LS weighted mean FEV1 0-6 hrs post-dose on Day 1. A LS mean treatment difference in TDI focal score of 0.9 units in favor of UMEC was seen on Day 168 (95% CI: 0.3, 1.5; p=0.004). Incidence of on-treatment adverse events (AEs) was lower in the placebo (55%) versus UMEC arm (60%); non-fatal serious AEs, drug-related AEs, and AEs leading to withdrawal were similar with UMEC and placebo. Once-daily UMEC 62.5 mcg resulted in statistically significant and clinically meaningful improvements in lung function and dyspnea, compared with placebo, in Asian patients with COPD, with no new safety concerns observed.


Technical Note: Flattening filter free beam from Halcyon linac: Evaluation of the profile parameters for quality assurance.
The use of flattening filter free (FFF) beams generated by standard linear accelerators is increasing in the clinical practice. The radiation intensity peaked toward the beam central axis is properly managed in the optimization process of treatment planning through intensity modulation. Specific FFF parameters for profile analysis, as unflatness and slope for FFF beams, based on the renormalization factor concept has been introduced for quality assurance purposes. Recently, Halcyon, an O-ring based linear accelerator equipped with a 6 MV FFF beam only has been introduced by Varian. Renormalization factors and related fit parameters according to Fogliata et al. ["Definition of parameters for quality assurance of FFF photon beams in radiation therapy," Med. Phys. 39, 6455-6464 (2012)] have been evaluated for the 6 MV FFF beam generated by Halcyon units. The Halcyon representative beam data provided by Varian were used. Dose fall-off at the field edges was matched with an unflattened beam generated by a 6 MV from a TrueBeam linac. Consistency of the results was evaluated against measurements on a clinical Halcyon unit, as well as a TrueBeam 6 MV FFF for comparison. The five parameters in the analytical equation for estimating the renormalization factor were determined with an R2 of 0.997. The comparison of the unflatness parameters between the Halcyon representative and hospital beam data was consistent within a range of 0.6%. Consistently with the computed parameters, the Halcyon profiles resulted in a less pronounced peak than TrueBeam. Renormalization factors and related fit parameters from the 6 MV FFF beam generated by the Varian Halcyon unit are provided.


Resonance modulation, annihilation and generation of anti-resonance and anti-phasonance in 3D neuronal systems: interplay of resonant and amplifying currents with slow dynamics.
Subthreshold (membrane potential) resonance and phasonance (preferred amplitude and zero-phase responses to oscillatory inputs) in single neurons arise from the interaction between positive and negative feedback effects provided by relatively fast amplifying currents and slower resonant currents. In 2D neuronal systems, amplifying currents are required to be slave to voltage (instantaneously fast) for these phenomena to occur. In higher dimensional systems, additional currents operating at various effective time scales may modulate and annihilate existing resonances and generate antiresonance (minimum amplitude response) and antiphasonance (zero-phase response with phase monotonic properties opposite to phasonance). We use mathematical modeling, numerical simulations and dynamical systems tools to investigate the mechanisms underlying these phenomena in 3D linear models, which are obtained as the linearization of biophysical (conductance-based) models. We characterize the parameter regimes for which the system exhibits the various types of behavior mentioned above in the rather general case in which the underlying 2D system exhibits resonance. We consider two cases: (i) the interplay of two resonant gating variables, and (ii) the interplay of one resonant and one amplifying gating variables. Increasing levels of an amplifying current cause (i) a response amplification if the amplifying current is faster than the resonant current, (ii) resonance and phasonance attenuation and annihilation if the amplifying and resonant currents have identical dynamics, and (iii) antiresonance and antiphasonance if the amplifying current is slower than the resonant current. We investigate the underlying mechanisms by extending the envelope-plane diagram approach developed in previous work (for 2D systems) to three dimensions to include the additional gating variable, and constructing the corresponding envelope curves in these envelope-space diagrams. We find that antiresonance and antiphasonance emerge as the result of an asymptotic boundary layer problem in the frequency domain created by the different balances between the intrinsic time constants of the cell and the input frequency f as it changes. For large enough values of f the envelope curves are quasi-2D and the impedance profile decreases with the input frequency. In contrast, for f ≪ 1 the dynamics are quasi-1D and the impedance profile increases above the limiting value in the other regime. Antiresonance is created because the continuity of the solution requires the impedance profile to connect the portions belonging to the two regimes. If in doing so the phase profile crosses the zero value, then antiphasonance is also generated.


Tilt testing results are influenced by tilt protocol.
It is unknown how the return to supine position influences duration of loss of consciousness (LOC) and cardioinhibition during tilt test. Retrospective analysis of two datasets containing records of patients who underwent tilt testing for unexplained syncope in two centres was performed. Patients, totalling 1232, were included in the study: 262 in a Swedish centre and 970 patients in a Polish centre. In Sweden, tilt table with tilt-down time (TDT) of 18 s was used (Group II). In Poland, two different tilt tables were used, one of them with TDT of 10 s (Group I, n = 325), and the other with TDT of 47 s (Group III, n = 645). Cardioinhibitory reflex occurred most frequently in Group III, whereas number of pauses >3 s, frequency of very long asystole ≥30 s, and the total duration of pauses >3 s demonstrated a trend to increase from Group I to III. Duration of LOC in Groups II and III was significantly longer compared with Group I (32.0 and 33.7 s vs. 16.4 s). In the multivariate-adjusted regression model, cardioinhibitory reflex was predicted by tilt-table model (odds ratio per model with increasing TDT: 1.40; 95% confidence interval, 1.19-1.64; P < 0.0001), whereas LOC duration was longer with increasing TDT (P < 0.0001) and age (P < 0.0001). Longer TDT during induced vasovagal syncope increases the prevalence of cardioinhibitory reflex and prolongs the duration of LOC. Tilt-down time does not affect asystolic pause duration but delay may lead to occurrence of multiple pauses, higher frequency of very long asystole, and longer total asystole duration.


Feasibility of basic emergency medical technicians to perform selected advanced life support interventions.
Emergency medical technician-basic (EMT-B) providers often provide the initial care to victims of out-of-hospital cardiac arrest. While automated external defibrillators enable EMT-B providers to deliver rescue shocks, patients in cardiac arrest may require additional interventions that EMT-B providers may not presently deliver. We sought to evaluate the feasibility of training EMT-B providers to provide additional cardiac resuscitation procedures using the laryngeal mask airway (LMA) and intraosseous (IO) access. In this prospective observational study, we trained 18 EMT-B providers to use the LMA and IO drill (EZ-IO) in a three-hour educational session. Working in two-person teams, the rescuers performed a simulated ventricular fibrillation resuscitation. We evaluated placement success as well as elapsed time to placement of the LMA and EZ-IO. EMT-B providers successfully placed the LMA in 14 of 18 scenarios (78%; 95% confidence interval, 52% to 94%), with a mean of two attempts for placement. Subjects successfully placed the EZ-IO in 17 of 18 scenarios (94%; 95% confidence interval, 73% to 100%), all on the first attempt. The median time to LMA placement following the third shock was 109 seconds (interquartile range, 58-158) and the median time to EZ-IO placement was 72 seconds (interquartile range, 50-93) after LMA placement. EMT-B providers demonstrated moderate success in performing advanced-level cardiac resuscitation interventions. These observations suggest potential for expanding the role of basic-level rescuers in cardiopulmonary resuscitation.


Overexpression of Coptis japonica norcoclaurine 6-O-methyltransferase overcomes the rate-limiting step in Benzylisoquinoline alkaloid biosynthesis in cultured Eschscholzia californica.
Benzylisoquinoline alkaloids are one of the most important secondary metabolite groups, and include the economically important analgesic morphine and the antimicrobial agent berberine. To improve the production of these alkaloids, we investigated the effect of the overexpression of putative rate-limiting step enzymes in benzylisoquinoline alkaloid biosynthesis. We introduced two O-methyltransferase [Coptis japonica norcoclaurine 6-O-methyltransferase (6OMT) and 3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase (4'OMT)] expression vectors into cultured California poppy cells to avoid the gene silencing effect of endogenous genes. We established 20 independent lines for 6OMT transformants and 15 independent lines for 4'OMT transformants. HPLC/liquid chromatography-mass spectrometry (LC-MS) analysis revealed that the overexpression of C. japonica 6OMT was associated with an average alkaloid content 7.5 times greater than that in the wild type, whereas the overexpression of C. japonica 4'OMT had only a marginal effect. Further characterization of 6OMT in California poppy cells indicated that a 6OMT-specific gene is missing and 4OMT catalyzes the 6OMT reaction with low activity in California poppy, which supports the notion that the 6OMT reaction is important for alkaloid biosynthesis in this plant species. We discuss the importance of 6OMT in benzylisoquinoline alkaloid biosynthesis and the potential for using a rate-limiting step gene to improve alkaloid production.


The genetic base of Brazilian soybean cultivars: evolution over time and breeding implications.
Genetic diversity is essential for crop breeding and one way to estimate it is through the concept of genetic base, which can be defined as the number of ancestors and their relative genetic contributions (RGC) to each cultivar. The RGC can be estimated through the coefficient of parentage between the ancestors and cultivars. Previous studies determined that the genetic base of Brazilian soybean was very narrow. The objective of this work was to evaluate the pedigree of 444 Brazilian soybean cultivars to estimate their genetic base. The cultivars were divided according to their release dates and according to their origin (public or private), and the genetic base for each group was also estimated. We found 60 ancestors, of which the top four (CNS, S-100, Roanoke and Tokyo, respectively) contribute 55.3% of the genetic base. Only 14 ancestors have an RGC over 1.0%, and they represent 92.4% of the genetic base. Analysis of the release dates indicated that there has been an increase in the number of ancestors over time, but the four main ancestors were the same over all periods, and their cumulative RGC increased from 46.6% to 57.6%, indicating a narrowing of the genetic base.


Capacity planning seeing the forest for the trees.
Strategies for responding to today's rise in demand for inpatient services are many. The following are a few "tried-and-true" strategies: Build new and replacement hospitals Spin off specialty services Convert space occupied by less-profitable specialty services Improve patient management Reclaim mothballed space Implement urgent care tracks for appropriate ED patients.


Induction Chemotherapy is Not Superior to a Surgery-First Strategy for Clinical N1 Non-Small Cell Lung Cancer.
Guidelines recommend primary surgical resection for non-small cell lung cancer (NSCLC) patients with clinical N1 disease and adjuvant chemotherapy if nodal disease is confirmed after resection. We tested the hypothesis that induction chemotherapy for clinical N1 (cN1) disease improves survival. Patients treated with lobectomy or pneumonectomy for cT1-3 N1 M0 NSCLC from 2006 to 2011 in the National Cancer Data Base were stratified by treatment strategy: surgery first vs induction chemotherapy. Propensity scores were developed and matched with a 2:1 nearest neighbor algorithm. Survival analyses using Kaplan-Meier methods were performed on the unadjusted and propensity-matched cohorts. A total of 5,364 cN1 patients were identified for inclusion, of which 565 (10.5%) were treated with induction chemotherapy. Clinical nodal staging was accurate in 68.6% (n = 3,292) of patients treated with surgical resection first, whereas 16.3% (n = 780) were pN0 and 10.7% (n = 514) were pN2-3. Adjuvant chemotherapy was given to 60.9% of the surgery-first patients who were pN1-3 after resection. Before adjustment, patients treated with induction chemotherapy were younger, with lower comorbidity burden, were more likely to be treated at an academic center and to have private insurance (all p < 0.001), but were significantly more likely to have T3 tumors (28.7% vs 9.9%, p < 0.001) and to require pneumonectomy (23.5% vs 18.5%, p = 0.005). The unadjusted and propensity-matched analyses found no differences in short-term outcomes or survival between groups. Induction chemotherapy for cN1 NSCLC is not associated with improved survival. This finding supports the currently recommended treatment paradigm of surgery first for cN1 NSCLC.


Changes in tumor vascularity depicted by contrast-enhanced EUS as a predictor of prognosis and treatment efficacy in patients with unresectable pancreatic cancer (PEACE): A study protocol.
Patients with unresectable pancreatic cancer have a poor prognosis. The analysis of prognostic factors before treatment may be helpful in determining the best therapeutic strategies. The aim of the PEACE study is to assess the vascularity of pancreatic malignant tumors using contrast-enhanced harmonic EUS (CEH-EUS) and to clarify the prognostic value of tumor vascularity in patients with locally advanced and metastatic pancreatic cancer. Hereby, we present the protocol of a prospective, nonrandomized, single-arm, multicenter study aiming to assess changes in tumor vascularity using CEH-EUS before and 2 months after treatment initiation in patients with unresectable, locally advanced/metastatic pancreatic cancer and to examine the correlation between vascular changes and treatment response, progression-free survival, and overall survival.


Triglyceride-Glucose Index and New-Onset Atrial Fibrillation in ST-Segment Elevation Myocardial Infarction Patients After Percutaneous Coronary Intervention.
New-onset atrial fibrillation (NOAF) is associated with worse prognostic outcomes in cases diagnosed with ST-segment elevation myocardial infarction (STEMI) patients after percutaneous coronary intervention (PCI). The triglyceride-glucose (TyG) index, as a credible and convenient marker of insulin resistance, has been shown to be predictive of outcomes for STEMI patients following revascularization. The association between TyG index and NOAF among STEMI patients following PCI, however, has not been established to date. To assess the utility of the TyG index as a predictor of NOAF incidence in STEMI patients following PCI, and to assess the relationship between NOAF and long-term all-cause mortality. This retrospective cohort research enrolled 549 STEMI patients that had undergone PCI, with these patients being clustered into the NOAF group and sinus rhythm (SR) group. The predictive relevance of TyG index was evaluated through logistic regression analyses and the receiver operating characteristic (ROC) curve. Kaplan-Meier curve was employed to explore differences in the long-term all-cause mortality between the NOAF and SR group. NOAF occurred in 7.7% of the enrolled STEMI patients after PCI. After multivariate logistic regression analysis, the TyG index was found to be an independent predictor of NOAF [odds ratio (OR): 8.884, 95% confidence interval (CI): 1.570-50.265, P = 0.014], with ROC curve analyses further supporting the predictive value of this parameter, which exhibited an area under ROC curve of 0.758 (95% CI: 0.720-0.793, P < 0.001). All-cause mortality rates were greater for patients in the NOAF group in comparison with the SR group over a median 35-month follow-up period (log-rank P = 0.002). The TyG index exhibits values as an independent predictor of NOAF during hospitalization, which indicated a poorer prognosis after a relatively long-term follow-up.


Suspecting the Figures: What Church Leaders Think About Government's Commitment to Combating COVID-19 in Nigeria.
Public trust in government can significantly determine the outcome of health policies in any society. Hence, studies have been gauging peoples' level of trust in their governments' commitment and capacity to win the fight against COVID-19. However, these studies have omitted religious leaders. This is despite the fact that religious leaders play key roles in the area of health in many societies. The present study, therefore, explored the opinions church leaders have about the credibility of the COVID-19 statistics and other government responses in Nigeria. Data were collected through semi-structured interviews with 18 church leaders drawn from Anglican, Catholic, and Pentecostal churches in Nigeria. A descriptive narrative approach was employed in the thematic organization and analysis of data. Findings show that only one participant expressed confidence in the credibility of the COVID-19 statistics and other government's responses. The rest, with the exception of one participant who was uncertain, was distributed between those who believe the statistics and other government efforts are exaggerated and those who believe they are false. The study also found that denominational affiliation mattered with respect to the perceptions about the credibility of the COVID-19 statistics and other government responses. Implications of findings for policy and research are discussed.


Osteopontin involvement in granuloma formation and in the severity of Paracoccidioides brasiliensis infection.
The participation of osteopontin (OPN) in Paracoccidioides brasiliensis infected mice, its association to granulomatogenesis, severity of infection, pattern of lesions, nitric oxide (NO) levels and fungal load were evaluated in this investigation. Immunohistochemistry analysis showed marked OPN staining in extracellular matrix and in macrophages and multinucleated giant cells at the center of lesions, suggesting a possible role of OPN in the distribution of these cells within the granulomas. At 15 days post-infection with a virulent P. brasiliensis isolate, OPN+ cells were more numerous and intensely immunostained in the loose granulomas of susceptible mice than in those of resistant mice. In addition, high fungal loads and low NO levels were observed in susceptible mice. At 120 days after infection, resistant mice had increased total OPN levels (ELISA) and OPN positivity in compact granulomas, higher NO levels and lower fungal loads than susceptible mice. Residual lesions associated with low OPN levels, high NO and control of fungal dissemination were observed in both mouse strains at 120 days post-infection with the slightly virulent fungal isolate. Therefore, OPN could be associated with higher severity of the disease in an early phase of infection and with a degree of control of the progressive infection.


[Perioperative Blood Glucose Control in Patients with Diabetic Mellitus].
Current criteria used for the diagnosis of diabetic mellitus are (1) HbA(1c) level > 6.5% (National Glycohemoglobin Standardization Program certified methods), (2) fasting glucose concentration > 126 mg x dl(-1) (7.0 mmol x l(-1)), (3) 2-h plasma glucose concentration > 200 mg x dl(-1) (11.1 mmol x l(-1)) during an oral glucose tolerance test, or (4) classic symptoms of hyperglycemia or hyperglycemic crisis with a random plasma glucose concentration > 200 mg x dl(-1) (11.1 mmol x l(-1)). It is reported that 15-20% of perioperative patients were with the presence of diabetic mellitus. However, there is little information to guide perioperative glycemic control in these patients so far. In this review, we summarize the current available evidence to assist physician to provide the better glycemic control in perioperative period.


Inhaled Pulmonary Vasodilator Therapy for Management of Right Ventricular Dysfunction after Left Ventricular Assist Device Placement and Cardiac Transplantation.
Right ventricular failure (RVF) after cardiac transplant (CTX) or implantation of a continuous-flow left ventricular assist device (CF-LVAD) is associated with significant postoperative morbidity and mortality. A variety of modalities have been used to treat postoperative RVF, including management of volume status, intravenous inotropes and vasodilators, and right-sided mechanical support. Inhaled vasodilator agents are a unique treatment option aimed at minimizing systemic absorption by delivering therapy directly to the pulmonary vasculature. Current LVAD and CTX guidelines endorse inhaled vasodilators for managing postoperative RVF; however, no guidance is offered regarding agent selection, dosing, or administration. A review of the current literature confirms that inhaled pulmonary vasodilator agents have been shown to decrease pulmonary artery pressure when used in the perioperative period of CF-LVAD implant or CTX. However, the literature regarding the potential impact on clinical outcomes (e.g., survival or risk of developing RVF) is lacking with these medications. Based on our assessment of the literature, we suggest that when RVF occurs in the setting of a normal pulmonary vascular resistance (PVR), traditional inotropic therapy (e.g., dobutamine) should be used. Conversely, if the PVR is elevated (> 250 dynes/sec/cm5 or 3 Wood units), or the patient has other evidence of a high right ventricular afterload (i.e., a transpulmonary gradient > 12 mm Hg), then an inhaled pulmonary vasodilator would be the preferred initial pharmacologic agent. Drug selection depends largely on the institution's capacity to safely prepare and administer the medication, along with formulary considerations, such as the high costs associated with inhaled iloprost and inhaled nitric oxide.


A genome wide analysis of alternative splicing events during the osteogenic differentiation of human cartilage endplate-derived stem cells.
Low back pain is a prevalent disease, which leads to suffering and disabilities in a vast number of individuals. Degenerative disc diseases are usually the underlying causes of low back pain. However, the pathogenesis of degenerative disc diseases is highly complex and difficult to determine. Current therapies for degenerative disc diseases are various. In particular, cell-based therapies have proven to be effective and promising. Our research group has previously isolated and identified the cartilage endplate‑derived stem cells. In addition, alternative splicing is a sophisticated regulatory mechanism, which greatly increases cellular complexity and phenotypic diversity of eukaryotic organisms. The present study continued to investigate alternative splicing events in osteogenic differentiation of cartilage endplate‑derived stem cells. An Affymetrix Human Transcriptome Array 2.0 was used to detect splicing changes between the control and differentiated samples. Additionally, molecular function and pathway analysis were also performed. Following rigorous bioinformatics analysis of the data, 3,802 alternatively spliced genes were identified, and 10 of these were selected for validation by reverse transcription‑polymerase chain reaction. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway analysis also revealed numerous enriched GO terms and signaling pathways. To the best of our knowledge, the present study is the first to investigate alternative splicing mechanisms in osteogenic differentiation of stem cells on a genome‑wide scale. The illumination of molecular mechanisms of stem cell osteogenic differentiation may assist the development novel bioengineered methods to treat degenerative disc diseases.


Oral injury and mouthguard usage by athletes in Japan.
The use of mouthguards in contact sports effectively prevents oral injury and preserves oral structures. We investigated oral injuries and awareness concerning mouthguards among Japanese high school soccer and rugby players. Athletes were asked a series of questions concerning their history of oral injury while participating in sports, and their pattern of mouthguard use was determined. The data were evaluated statistically using chi-square analysis. The incidence of oral injuries was 32.3% for soccer athletes and 56.5% for rugby athletes, with 0.8% and 24.1% of the respective groups having mouthguards. There were significant differences between the soccer and rugby groups (P < 0.001). Although 81.8% of soccer athletes thought mouthguards were unnecessary, only 26.2% of rugby athletes shared this opinion and there was a significant difference between the soccer and rugby groups. Many soccer athletes had insufficient knowledge about mouthguards and were not concerned about preventing oral injury, although it was in fact a common problem in their sport. Athletes as well as coaches must be made aware of the high risk of oral injury when playing soccer, rugby, and other contact sports.


[Determination of the activity of delta-aminolevulinic acid dehydratase--a method for the diagnosis of lead poisoning in birds].
In avian patients enteral, parenteral and aerogenous lead intoxications are frequently observed. Ingestion of non-metallic lead may not be diagnosed by radiography. Alternatively, estimation of the delta-aminolevulinic acid dehydratase (delta-ALA Dehydratase) activity is a suitable diagnostic test for lead intoxications. The enzyme delta-ALA Dehydratase, which plays a central role in erythropoiesis, is inhibited by lead and is therefore a specific indicator for lead intoxications. The activity of delta-ALA Dehydratase is measured directly in the blood; not the amount of excreted delta-ALA in the urine as in mammals. Blood volumes of 0.45 ml are necessary and thus allow to perform the test in birds as small as 45 g body weight (e.g. Budgerigars). Possible ranges of application as well as a suitable therapy for lead intoxication in birds are presented.


Structure and function of μ-conotoxins, peptide-based sodium channel blockers with analgesic activity.
μ-Conotoxins block voltage-gated sodium channels (VGSCs) and compete with tetrodotoxin for binding to the sodium conductance pore. Early efforts identified µ-conotoxins that preferentially blocked the skeletal muscle subtype (NaV1.4). However, the last decade witnessed a significant increase in the number of µ-conotoxins and the range of VGSC subtypes inhibited (NaV1.2, NaV1.3 or NaV1.7). Twenty µ-conotoxin sequences have been identified to date and structure-activity relationship studies of several of these identified key residues responsible for interactions with VGSC subtypes. Efforts to engineer-in subtype specificity are driven by in vivo analgesic and neuromuscular blocking activities. This review summarizes structural and pharmacological studies of µ-conotoxins, which show promise for development of selective blockers of NaV1.2, and perhaps also NaV1.1,1.3 or 1.7.


Glycosylated antitumor ether lipids are more effective against oncogene-transformed fibroblasts than alkyllysophospholipids.
We have investigated the antiproliferative effects of different types of antitumor ether lipids (AELs) against non-transformed and transformed fibroblasts. The compounds examined were choline phosphate-containing alkyllysophospholipids (1-O-octadecyl-2-O-methyl glycerophosphocholine (ET18-OCH3), 2'-(trimethylammonio)ethyl 3-(hexadecyloxy)-2-(methoxymethyl)propylphosphate (oxo-BM 41.440), 2'-(triethylammonio)ethyl 4-(hexadecyloxy)-3-methoxybutane phosphonate (ET16-OCH3-phosphonocholine)), and glycosylated ether-linked diglycerides (1-O-hexadecyl-2-O-methyl-3-S-(beta-D-1'-thioglucopyranosyl-sn-gly cerol) [ET16-OCH3-beta-thio-Glc] and 1-O-hexadecyl-2-O-methyl-3-O-(2'amino-2'-deoxy-beta-D-glucopyranosyl)-sn -glycerol (ET16-OCH3-Gln)). The choline phosphate-containing alkyllysophospholipids (ALPs) had little or moderate effect on the proliferation and none on the viability of NIH 3T3 clone 7, and sublines transformed by raf (NIH/9IV #5), fes (Fes 1), src (Src 1) and mos (Mos 1) oncogenes. The glycosylated ether-linked diglycerides were more effective than the choline phosphate-containing ALPs. Of the two ether-linked diglycerides, ET16-OCH3-beta-thio-Glc did not affect the viability of the cells at any of the concentrations examined while ET16-OCH3-Gln was cytotoxic to all the transformed cell lines at concentrations equal to or greater than 9 microM. The IC50 for ET16-OCH3-Gln was 6.4 microM for Mos 1, 6.5 microM for NIH/9IV #5, 7.5 microM for Src 1, 8.2 microM for Fes 1 and 8.4 microM for NIH 3T3 clone 7. These results suggest that the ether-linked diglycerides may be more effective against fibrosarcomas than the cholinephosphate containing ALPs. Also, with the exception of ET16-OCH3-Gln there was no significant difference in the effect of the compounds on the transformed and untransformed cell lines, suggesting that the selectivity displayed by AELs may depend on both the type of compound and transformation in the cell.


Antemortem Health System Utilization in People Experiencing Homelessness Who Died in the Emergency Department.
People experiencing homelessness (PEH) have high rates of mortality, medical and psychiatric comorbidities, and emergency department utilization. In this study, a health system's emergency department encounters were evaluated to identify PEH who died in the emergency department. Patient demographics, medical history, prehospital and emergency department characteristics, and health care utilization patterns were collected. Descriptive statistics were calculated. We identified 48 PEH pronounced dead in the emergency department; mean age at death was 46.5. Forty-four (92%) decedents presented in cardiac arrest, 12 (25%) of which were substance use-related; 4 (8%) presented with trauma. Out of 44 patients presenting in cardiac arrest, (20.5%) had bystander cardiopulmonary resuscitation (CPR) performed before arrival of emergency medical services. In the year prior to death, 15 (32%) decedents had no documented health care utilization, while 16 (33%) had 10 or more emergency department/outpatient visits. Our study is the first to characterize PEH who died in the emergency department, analyzing the pre-hospital and in-hospital characteristics and antemortem health system utilization in this population. A sizeable proportion of deceased PEH had no health system contact in the 12 months prior to death, suggesting that those with high mortality risk may underutilize health services. Conversely, a similar proportion of decedents had extensive (more than 10) health system utilization in the year prior to death, representing possible opportunities to reduce mortality.


Mouse mast cell activation and desensitization for immune aggregate-induced histamine release.
Immune aggregate-induced histamine release and desensitization were studied in mouse mast cells. Maximal histamine release was rapid, occurred at 37 degrees C, and required the addition of alpha-L-phosphatidyl-L-serine and Ca2+. The amount of histamine released varied with the composition of the immune aggregates and was dependent on the antibody concentration. Saturation of mast cell Fc epsilon receptors with rat or mouse IgE had no effect on subsequent immune aggregate-induced release. The incubation of mouse mast cells with immune aggregates in the absence of cations of alpha-L-phosphatidyl-L-serine did not stimulate the release of histamine but resulted in desensitization of the cells for release with the addition of the same or unrelated immune aggregates. Such cells are capable, however, of IgE-mediated histamine release. Mast cells desensitized for IgE-mediated histamine release by incubation with anti-IgE were capable of immune aggregate-induced release. These data suggest that IgE-mediated and immune aggregate-induced triggering of mouse mast cells occurs through separate receptors.


Controlling bead and cell mobility in a recirculating hanging-drop network.
Integrating flowing cells, such as immune cells or circulating tumour cells, within a microphysiological system is crucial for body-on-a-chip applications. However, ensuring unimpeded recirculation of cells is a significant challenge. Closed microfluidic devices have a no-slip boundary condition along channel walls and a defined chip geometry (laminar flow) that hinders the ability to freely control cell flow. Open microfluidic devices, where the bottom device boundary is an air-liquid interface (ALI), e.g., hanging drop networks (HDNs), offer the advantage of an easily-actuatable fluid-phase geometry, where cells can either flow or stagnate. In this paper, we optimized a hanging-drop-integrated pneumatic-pump system for closed-loop recirculation of particles (i.e., beads or cells). Experiments with both beads and cells in cell culture medium initially resulted in particle stagnation, which was suggestive of a pseudo-no-slip boundary condition at the ALI. Transmission electron microscopy and dynamic light scattering measurements of the ALI suggested that aggregation of submicron-scale cell-culture-medium components is the cause of the pseudo-no-slip boundary condition. We used the finite element method to study the forces on particles at the ALI and to optimize HDN design (drop aperture) and operation (drop height) parameters. Based on this analysis, we report a phase diagram delineating the conditions for free flow or stagnation of particles at the ALI of hanging drops. Using our experimental setup with 3.5 mm drop apertures, we conducted particle flow experiments while actuating drop heights. We confirmed the ability to control the flow or stagnation of particles by actuating the height of hanging drops: a drop height over 300 μm led to particle stagnation and a drop height under 300 μm allowed for particle flow. This particle-flow control, combined with the ease of integrating scaffold-free organ models (microtissues or organoids) in HDNs, constitutes the basis for an experimental setup enabling the control of the residence time of single cells around 3D organ models.


Hyoscine for polyp detection during colonoscopy: A meta-analysis and systematic review.
To assess the role of hyoscine for polyp detection during colonoscopy. Studies (randomized controlled trials or RCTs) that compared the use of hyoscine vs no hyoscine or placebo for polyp detection during colonoscopy were included in our analysis. A search on multiple databases was performed in September 2013 with search terms being "hyoscine and colonoscopy", "hyoscine and polyp", "hyoscine and adenoma", "antispasmotic and colonoscopy", "antispasmotic and adenoma", and "antispasmotic and polyp". Jadad scoring was used to assess the quality of studies. The efficacy of hyoscine was analyzed using Mantel-Haenszel model for polyp and adenoma detection with odds ratio (OR). The I (2) measure of inconsistency was used to assess heterogeneity (P < 0.05 or I (2) > 50%). Statistical analysis was performed by RevMan 5.1. Funnel plots was used to assess publication bias. The search of the electronic databases identified 283 articles. Of these articles, eight published RCTs performed at various locations in Europe, Asia, and Australia were included in our meta-analysis, seven published as manuscripts and one published as an abstract (n = 2307). All the studies included patients with a hyoscine and a no hyoscine/placebo group and were of adequate quality (Jadad score ≥ 2). Eight RCTs assessed the polyp detection rate (PDR) (n = 2307). The use of hyoscine demonstrated no statistically significant difference as compared to no hyoscine or placebo for PDR (OR = 1.06; 95%CI: 0.89-1.25; P = 0.51). Five RCTs assessed the adenoma detection rate (ADR) (n = 2015). The use of hyoscine demonstrated no statistically significant difference as compared to no hyoscine or placebo for ADR (OR = 1.12; 95%CI: 0.92-1.37; P = 0.25). Furthermore, the timing of hyoscine administration (given at cecal intubation or pre-procedure) demonstrated no differences in PDR compared to no hyoscine or placebo. Publication bias or heterogeneity was not observed for any of the outcomes. Hyoscine use in patients undergoing colonoscopy does not appear to significantly increase the detection of polyps or adenomas.


Time for clients: temporal aspects of community psychiatric nursing.
Time is an integral element in all actions, organizations and relationships. Temporality has an effect on people's lives, and in turn people manipulate temporality as a means of imposing social order. From a semiotic perspective the amount of time spent on an action and the timing of its conveys messages about such things as importance and status. This holds equally true of organizations as well as individuals. This paper concentrates on a community psychiatric nursing department, its staff and clients. Sections of speech taken from interviews with nurses and clients are presented and discussed from the perspective of temporality to show how relationships are managed on an individual level and how work is organized around different client groups at an organizational level.


Microfilament Depolymerization Is a Pre-requisite for Stem Cell Formation During In vitro Shoot Regeneration in Arabidopsis.
De novo shoot regeneration is widely used in fundamental studies and agricultural applications. Actin microfilaments are involved in many aspects of plant cell division, cell morphogenesis and cell signal transduction. However, the function of actin microfilaments during de novo shoot regeneration is poorly understood. Here, we investigated the organization of actin microfilaments during this process and found that stem cell formation was associated with microfilament depolymerization. Furthermore, inhibition of microfilament depolymerization by phalloidin treatment or downregulation of actin depolymerizing factors (ADFs) restrained stem cell initiation and shoot regeneration. Inhibition of ADF expression affected the architecture of microfilaments during stem cell formation, and the polar transport and distribution of auxin were also disrupted. Together, our results demonstrate that organization of the microfilament cytoskeleton play important roles in stem cell formation and shoot meristem induction during shoot regeneration.


Proteome profiling reveals changes in energy metabolism, transport and antioxidation during drought stress in Nostoc flagelliforme.
Drought is an important abiotic stress that constrains the growth of many species. Despite extensive study in model organisms, the underlying mechanisms of drought tolerance in Nostoc flagelliforme remain elusive. We characterized the drought adaptation of N. flagelliforme by a combination of proteomics and qRT-PCR. A total of 351 differentially expressed proteins involved in drought stress adaptation were identified. It was found that the expression of several nutrient influx transporters was increased, including molybdate ABC transporter substrate binding protein (modA), sulfate ABC transporter substrate-binding protein (sbp) and nitrate ABC transporter (ntrB), while that of efflux transporters for toxic substances was also increased, including arsenic transporting ATPase (ArsA), potassium transporter (TrkA) and iron ABC transporter substrate-binding protein (VacB). Additionally, photosynthetic components were reduced while sugars built up during drought stress. Non-enzymatic antioxidants, orange carotenoid protein (OCP) homologs, cytochrome P450 (CYP450), proline (Pro) and ascorbic acid (AsA) were all altered during drought stress and may play important roles in scavenging reactive oxygen species (ROS). In this study, N. flagelliforme may regulates its adaptation to drought stress through the changes of protein expression in photosynthesis, energy metabolism, transport, protein synthesis and degradation and antioxidation. • A total of 351 DEPs involved in adaptation to drought stress were identified. • Changes in the expression of six OCP homologs were found in response to drought stress. • Differential expression of transporters played an important role in drought stress adaptation. • Most PSII proteins were downregulated, while PSI proteins were unchanged in response to drought stress. • Sugar metabolism was upregulated in response to drought stress.


Effects of QA 208-199 and its metabolite 209-668 on embryonic development in vitro after microinjection into the exocoelomic space or into the amniotic cavity of cultured rat conceptuses.
The objective of this study was to determine the direct embryotoxic effects in vitro of N-hydroxy-N-methyl-7-propoxy-2-naphthalene-ethanamine (QA 208-199, QAB) and of one of its metabolites, 7-propoxy-naphthalene-2-ylacetic acid (209-668, QAA), after circumventing the bioconverting conceptual membranes. The compounds were, therefore, microinjected either into the exocoelomic space or into the amniotic cavity of rat conceptuses of 10 d at prenatal age at doses of up to 84.9 ng (QAA) and 180 ng (QAB) per conceptus respectively. The conceptuses were subsequently cultured for 28 h after which their development was assessed. QAB produced marginal effects on embryonic differentiation only after microinjection of the compound into the amniotic cavity. Dysmorphogenic effects, however, occurred in a dose-dependent fashion after either exocoelomic or intraamniotic microinjections of the compound. The frequencies and types of anomalies were similar after either exposure route and consisted predominantly of anomalies associated with axial rotation. QAA also impaired embryonic differentiation at only the high dose level of 84.9 ng per embryo and after intraamniotic injections only. Dysmorphogenic effects were observed in all experimental groups, although the differences were not statistically significant when compared with the concomitant controls. An increased proportion of anomalies observed were in the cephalic region as compared to the defects produced by QAB. These data suggest that QAA most probably is not the QAB metabolite responsible for the embryotoxic action of QAB in vitro. Furthermore, the results tend to confirm the suggested involvement of the visceral yolk sac membrane in mediating QAB embryotoxicity.


Idiopathic interstitial pneumonia: a clinicopathological perspective.
Idiopathic interstitial pneumonia (IIP) represents a diverse group of lung disorders with variable prognoses and responses to therapy. As the name implies, the etiology is unknown. In the more severe forms of IIP, such as idiopathic pulmonary fibrosis and acute interstitial pneumonia, no effective therapies have been identified. In this perspective, the spatiotemporal variability in the histopathology of these disorders is discussed. It is proposed that common etiologies or injurious agents may produce variable histopathological "reactions" in the lung due to complex interactions between the host (genetic/epigenetic factors, age) and environmental factors. Accurate clinical and biological phenotyping may be necessary to stratify or group patients who are most likely to respond to specific modes of therapy.


Recent Advances in Polymeric Nanomedicines for Cancer Immunotherapy.
Immunotherapy has emerged as a promising approach to treat cancer, since it facilitates eradication of cancer by enhancing innate and/or adaptive immunity without using cytotoxic drugs. Of the immunotherapeutic approaches, significant clinical potentials are shown in cancer vaccination, immune checkpoint therapy, and adoptive cell transfer. Nevertheless, conventional immunotherapies often involve immune-related adverse effects, such as liver dysfunction, hypophysitis, type I diabetes, and neuropathy. In an attempt to address these issues, polymeric nanomedicines are extensively investigated in recent years. In this review, recent advances in polymeric nanomedicines for cancer immunotherapy are highlighted and thoroughly discussed in terms of 1) antigen presentation, 2) activation of antigen-presenting cells and T cells, and 3) promotion of effector cells. Also, the future perspectives to develop ideal nanomedicines for cancer immunotherapy are provided.


Posttranscriptional and Posttranslational Regulation of Actin.
Actin is one of the most abundant intracellular proteins, essential in every eukaryotic cell type. Actin plays key roles in tissue morphogenesis, cell adhesion, muscle contraction, and developmental reprogramming. Most actin studies have focused on its regulation at the protein level, either directly or through differential interactions with over a hundred intracellular binding partners. However, numerous studies emerging in recent years demonstrate specific types of nucleotide-level regulation that strongly affect non-muscle actins during cell migration and adhesion and are potentially applicable to other members of the actin family. This regulation involves zipcode-mediated actin mRNA targeting to the cell periphery, proposed to mediate local synthesis of actin at the cell leading edge, as well as the recently discovered N-terminal arginylation that specifically targets non-muscle β-actin via a nucleotide-dependent mechanism. Moreover, a study published this year suggests that actin's essential roles at the organismal level may be entirely nucleotide-dependent. This review summarizes the emerging data on actin's nucleotide-level regulation. Anat Rec, 301:1991-1998, 2018. © 2018 Wiley Periodicals, Inc.


Use of MRI-Guided Biopsy for Selection and Follow-up of Men Undergoing Hemi-gland Cryoablation of Prostate Cancer.
To investigate safety, efficacy, and quality of life impact of hemi-gland cryotherapy for clinically-significant prostate cancer (CaP), when patient selection and follow-up includes MRI-guided biopsy. Twenty-nine men with unilateral CaP (all clinically significant with prostate volume <60 cc) were enrolled in a prospective observational trial of hemi-gland cryotherapy. Mean patient age was 68.7 years. Median prostate-specific antigen (PSA) was 6.6 ng/mL. MRI-guided biopsy (3T-MRI, Artemis US fusion) was used for diagnosis and repeated at 6-month follow-up in all men. Treatment was under general anesthesia using the BTG/Galil system. Validated questionnaires were used to determine effects of treatment on urinary and sexual function and quality of life. Cryotherapy was completed satisfactorily in all 29 cases in <60 minutes with no intraoperative complications. Significant decreases in PSA (median decrease 5.6 ng/mL) and PSA density (median decrease 0.14 ng/mL/cc) were observed (P < .01). At 6 months, 23 patients (79%) demonstrated no residual cancer on follow-up MRI-guided biopsy of the treated side. Three patients (10%) revealed micro-residual disease. Three patients (10%) had residual cancer and underwent further treatment. Ipsilateral MRI lesions were present before treatment in 26 patients and after treatment in only 2, reflecting the gross ablative effect; however, MRI showed disappearance of lesions in 4 patients with residual tumor on biopsy. The single complication was 1 case of transient urinary retention; 85% of men who were sexually active continued without change after treatment. Voiding function was unchanged. Hemi-gland cryoablation for clinically-significant CaP is well-tolerated, and when patients are selected and followed by MRI/US fusion biopsy, cancer control appears promising at 6 months.


Diagnostic Accuracy of Posterior Circulation Stroke by Paramedics: A Systematic Review.
This systematic review aims to identify the diagnostic accuracy of posterior circulation stroke (PCS) by paramedics and the causes and duration of delay in its recognition. A systematic search using CINAHL Plus, MEDLINE, Scopus, and PubMed was performed. All databases were searched up to May 25, 2022. Studies were included where patients were adults, assessed by paramedics, and PCS was the primary diagnosis. Bias was assessed using the Newcastle-Ottawa Scale and the Effective Practice and Organization of Care tool. Results have been described by proportions, and both sensitivity calculations and subgroup analysis were performed utilizing MedCalc. A total of 797 titles/abstracts and a subsequent 87 full texts were screened, of which 15 were included. There were 5395 patients who were assessed by paramedics and had a confirmed diagnosis of PCS. Among five studies containing both true positive and false negative data, there were 98 (45.8%) true positives. PCS patients lost an average of 27 min (p < 0.001) compared to anterior stroke patients in the prehospital setting. One study revealed that educational intervention, including implementing the finger-to-nose test, increased the sensitivity for diagnosis from 45.8 to 74.1% (p = 0.039) and decreased the time from door to computed tomography from 62 to 41 min (p = 0.037). There is a substantial lack of evidence regarding the diagnosis of PCS by paramedics. Despite the low quality of evidence available, overall, the sensitivity for paramedic PCS diagnosis appears to be poor. Further investigation is required into paramedics' diagnosis of PCS and the use of educational interventions.Prospective Register of Systematic Reviews Registration Number: CRD42022324675.


Calcium entry blocking, beta 1-adrenoceptor blocking and hypotensive effects of YM-16151-4.
The calcium entry blocking, beta 1-adrenoceptor blocking and hypotensive effects of YM-16151-4 were studied in pithed rats and in conscious normotensive rats. In pithed rats, intravenous injection of YM-16151-4 inhibited a YC-170-induced pressor response and an isoproterenol-induced positive chronotropic response with DR10 values of 4.20 and 0.31 mg/kg, respectively. Similarly, a single oral dose of YM-16151-4 inhibited YC-170 and isoproterenol-induced responses with DR10 values of 109.94 and 9.14 mg/kg, respectively. On the other hand, both intravenous and single oral doses of YM-16151-4 produced a dose-dependent hypotensive effect with no increase in heart rate in conscious normotensive rats. The onset of this effect was slow but long-lasting. During repeated oral administration of YM-16151-4 (30 mg/kg) for 5 days, the hypotensive effect of the compound did not change, indicating that this effect did not increase and tolerance did not develop. The time course of the calcium entry blocking activity of YM-16151-4 was very similar to that of its beta 1-adrenoceptor blocking activity, suggesting that the activities of calcium antagonism and beta 1-adrenoceptor blockade of the compound were almost constant at any time after oral dosing. We conclude that YM-16151-4 combines calcium entry and beta 1-adrenoceptor blocking activities in vivo, and produces a hypotensive effect without an increase in heart rate.


Anisotropy of an 192iridium high dose rate source measured with a miniature ionization chamber.
The anisotropy of a high dose rate (HDR) 192Ir source was measured in air and in water using a miniature (0.147 cm3) ionization chamber. Measurements were made at a distance of 5 cm from the source center at polar angles from 10 degrees-170 degrees. The anisotropy was found to be less pronounced in water, and the anisotropy is asymmetric about the transverse axis. The results agree with previous ionization chamber and TLD measurements to within +/- 4%. Mean anisotropy factors were determined at each angle from all existing data at 5 cm distance, and compared to published Monte Carlo calculations, and to the values used in the microSelectron HDR brachytherapy planning system (BPS). The Monte Carlo photon transport code appears to systematically underestimate the anisotropy factor by up to 4% in the forward direction and overestimate it by up to 3% in the backward direction. The mean anisotropy factors also indicate that the BPS systematically underestimates the anisotropy factor by up to 3% in the forward direction, and overestimates it by up to 15% in the backward direction. However, the 15% difference occurs at 180 degrees where it is not likely to be clinically significant.