Transient receptor potential melastatin 7 (TRPM7) contributes to H2O2-induced cardiac fibrosis via mediating Ca(2+) influx and extracellular signal-regulated kinase 1/2 (ERK1/2) activation in cardiac fibroblasts.
Transient receptor potential melastatin 7 (TRPM7), a Ca(2+)-nonselective cation channel, plays a key role in the pathophysiological response of multiple cell types. However, the role of TRPM7 channels in hydrogen peroxide (H2O2)-induced cardiac fibrosis remains unclear. This study aimed to explore whether TRPM7 channels are involved in H2O2-induced cardiac fibrosis and the underlying mechanisms. Our results showed that 2-aminoethoxydiphenylborate (2-APB), which is commonly used to block TRPM7 channels, inhibited H2O2-induced cardiac fibrosis via attenuating the overexpression of important fibrogenic biomarkers and growth factors in cardiac fibroblasts, including collagen type I (Col I), fibronectin (FN), smooth muscle α-actin (α-SMA), connective tissue growth factor (CTGF), and transforming growth factor-β1 (TGF-β1). In addition, 2-APB also decreased H2O2-mediated elevation of the concentration of intracellular Ca(2+) ([Ca(2+)]i). Meanwhile, silencing TRPM7 channels by shRNA interference also impaired the increased [Ca(2+)]i and upregulation of Col I, FN, α-SMA, CTGF, and TGF-β1 induced by H2O2. Furthermore, we found that H2O2-mediated activation of extracellular signal-regulated kinase 1/2 (ERK1/2) decreased in TRPM7-shRNA cells and Ca(2+)-free culture media. These results demonstrated that TRPM7 channels contributed to H2O2-induced cardiac fibrosis and suggested that this contribution may be through mediating Ca(2+) influx and phosphorylation of ERK1/2.


[Sources of error in clinical practice].
The clinical setting is everything that contact us with the patient's reality, including physical, instrumental and laboratory exploration, and we use it for diagnostic and treatment purposes. Diagnosis is the synthesis by which physicians conclude an elaborated analysis of subjective and objective data, obtained by physical exam and technology. This procedure involves data collection about the patient and his disease, the results of various tests and laboratory findings, the interpretation of the data and a diagnostic summary. Medical errors may be derived from insufficiency in any of these phases. It is evident that technology will never replace the intellectual data recollection, analysis and reasoning that is the physician's art. This contribution underlines some of the defects frequently observed in clinical practice that may cause medical errors.


The stability of baseline-defined categories of alcohol consumption during the adult life-course: a 28-year prospective cohort study.
Studies that report the relationship between alcohol consumption and disease risk have predominantly operationalized drinking according to a single baseline measure. The resulting assumption of longitudinal stability may be simplistic and complicate interpretation of risk estimates. This study aims to describe changes to the volume of consumption during the adult life-course according to baseline categories of drinking. A prospective observational study. United Kingdom. A cohort of British civil servants totalling 6838 men and 3372 women aged 34-55 years at baseline, followed for a mean 19.1 (standard deviation = 9.5) years. The volume of weekly alcohol consumption was estimated from data concerning the frequency and number of drinks consumed. Baseline categories were defined: non-current drinkers, infrequent drinkers, 0.1-50.0 g/week, 50.1-100.0 g/week, 100.1-150.0 g/week, 150.1-250.0 g/week and >250.0 g/week. For women, the highest category was defined as > 100.0 g/week. Baseline frequency was derived as 'daily or almost daily' and 'not daily or almost daily'. Trajectories were estimated within baseline categories using growth curve models. Trajectories differed between men and women, but were relatively stable within light-to-moderate categories of baseline consumption. Drinking was least stable within the highest categories of baseline consumption (men: > 250.0 g/week; women: > 100.0 g/week), declining by 47.0 [95% confidence interval (CI) = 40.7, 53.2] and 16.8 g/week (95% CI = 12.6, 21.0), respectively, per 10-year increase in age. These declines were not a consequence of sudden transitions to complete abstention. Rates of decline appear greatest in older age, with trajectories converging toward moderate volumes. Among UK civil servants, consumption within baseline drinking categories is generally stable during the life-course, except among heavier baseline drinkers, for whom intakes decline with increasing age. This shift does not appear to be driven by transitions to non-drinking. Cohorts of older people may be at particular risk of misclassifying former heavy drinkers as moderate consumers of alcohol.


Should a moratorium be placed on sublingual nifedipine capsules given for hypertensive emergencies and pseudoemergencies?
Over the past 2 decades, nifedipine in the form of capsules has become widely popular in the treatment of hypertensive emergencies. Unlike other agents, such as sodium nitroprusside, nicardipine hydrochloride, diazoxide, and nitroglycerin--which require intravenous administration and monitoring of blood pressure--nifedipine can be given orally, and close monitoring is said not to be necessary. Although administration of nifedipine capsules has been reported to be expedient and safe, it has not been approved by the Food and Drug Administration for labeling for treatment of hypertensive emergencies or of any other form of hypertension because of lack of outcome data. A review of the literature revealed reports of serious adverse effects such as cerebrovascular ischemia, stroke, numerous instances of severe hypotension, acute myocardial infarction, conduction disturbances, fetal distress, and death. Sublingual absorption of nifedipine has been found to be poor; most of the drug is absorbed by the intestinal mucosa. Given the seriousness of the reported adverse events and the lack of any clinical documentation attesting to a benefit, the use of nifedipine capsules for hypertensive emergencies and pseudoemergencies should be abandoned.


Functional and Clinical Outcomes of Nonsurgically Managed Tibial Plateau Fractures.
This study sought to assess and compare long-term functional and clinical outcomes in patients with tibial plateau fractures that are treated nonsurgically. Over a period of 8 years, 305 consecutive tibial plateau fractures were treated by three surgeons at a single institution and followed prospectively in an Institutional Review Board-approved study. Overall, 41 patients (13%) were treated nonsurgically and 37 were available for follow-up. Indications for nonsurgical management were minimal fracture displacement or preclusion of surgery because of comorbidities. A series of univariate retrospective analyses were used to identify individual risk factors potentially predictive of Short Musculoskeletal Functional Assessment scores. Thirty-seven patients were included with a mean follow-up of 21 ± 14.9 months. Overall, 59% of patients (n = 22) attained good to excellent functional outcomes. In patients for whom surgery was precluded because of comorbidities, outcome scores were significantly poorer (38.8 ± 23.0 versus 12.7 ± 14.2; P = 0.001). Surgery precluded by a factor other than minimal fracture displacement predicted poor outcome (P = 0.002). Carefully selected patients with minimally displaced tibial plateau fractures can expect good to excellent outcomes when treated nonsurgically. Level III, retrospective comparative study.


Hepatitis B and C in Europe: an update from the Global Burden of Disease Study 2019.
In 2016, the World Health Assembly adopted the resolution to eliminate viral hepatitis by 2030. This study aims to provide an overview of the burdens of hepatitis B virus (HBV) and hepatitis C virus (HCV) in Europe and their changes from 2010 to 2019 using estimates from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2019. We used GBD 2019 estimates of the burden associated with HBV-related and HCV-related diseases: acute hepatitis, cirrhosis and other chronic liver diseases, and liver cancer. We report total numbers and age-standardised rates per 100 000 for mortality, prevalence, incidence, and disability-adjusted life-years (DALYs) from 2010 to 2019. For each HBV-related and HCV-related disease and each measure, we analysed temporal changes and percentage changes for the 2010-19 period. In 2019, across all age groups, there were an estimated 2·08 million (95% uncertainty interval [UI] 1·66 to 2·54) incident cases of acute hepatitis B and 0·49 million (0·42 to 0·57) of hepatitis C in Europe. There were an estimated 8·24 million (7·56 to 8·88) prevalent cases of HBV-related cirrhosis and 11·87 million (9·77 to 14·41) of HCV-related cirrhosis, with 24·92 thousand (19·86 to 31·03) deaths due to HBV-related cirrhosis and 36·89 thousand (29·94 to 45·56) deaths due to HCV-related cirrhosis. Deaths were estimated at 9·00 thousand (6·88 to 11·62) due to HBV-related liver cancer and 23·07 thousand (18·95 to 27·31) due to HCV-related liver cancer. Between 2010 and 2019, the age-standardised incidence rate of acute hepatitis B decreased (-22·14% [95% UI -35·44 to -5·98]) as did its age-standardised mortality rate (-33·27% [-43·03 to -25·49]); the age-standardised prevalence rate (-20·60% [-22·09 to -19·10]) and mortality rate (-33·19% [-37·82 to -28·13]) of HBV-related cirrhosis also decreased in this time period. The age-standardised incidence rate of acute hepatitis C decreased by 3·24% (1·17 to 5·02) and its age-standardised mortality rate decreased by 35·73% (23·48 to 47·75) between 2010 and 2019; the age-standardised prevalence rate (-6·37% [-8·11 to -4·32]), incidence rate (-5·87% [-11·24 to -1·01]), and mortality rate (-11·11% [-16·54 to -5·53]) of HCV-related cirrhosis also decreased. No significant changes were observed in age-standardised rates of HBV-related and HCV-related liver cancer, although we observed a significant increase in numbers of cases of HCV-related liver cancer across all ages between 2010 and 2019 (16·41% [2·81 to 30·91] increase in prevalent cases). Substantial reductions in DALYs since 2010 were estimated for acute hepatitis B (-27·82% [-36·92 to -20·24]), acute hepatitis C (-27·07% [-15·97 to -39·34]), and HBV-related cirrhosis (-30·70% [-35·75 to -25·03]). A moderate reduction in DALYs was estimated for HCV-related cirrhosis (-6·19% [-0·19 to -12·57]). Only HCV-related liver cancer showed a significant increase in DALYs (10·37% [4·81-16·63]). Changes in age-standardised DALY rates closely resembled those observed for overall DALY counts, except for HCV-liver related cancer (-2·84% [-7·75 to 2·63]). Although decreases in some HBV-related and HCV-related diseases were estimated between 2010 and 2019, HBV-related and HCV-related diseases are still associated with a high burden, highlighting the need for more intensive and coordinated interventions within European countries to reach the goal of elimination by 2030. Bill & Melinda Gates Foundation.


Estradiol transfer from male big brown bats (Eptesicus fuscus) to the reproductive and brain tissues of cohabiting females, and its action as a pheromone.
The powerful estrogen, 17β-estradiol, has been found to pass from male excretions to the reproductive organs, brain, and other tissues of cohabiting females in laboratory mice. The current studies were designed to examine whether this phenomenon also occurs in big brown bats (Eptesicus fuscus), a mammal appropriate for testing cross-species generality because of its phylogenetic distance from mice. When tritiated estradiol ((3)H-E2) was administered directly on the nasal area of adult female bats, radioactivity was reliably observed in the uterus and ovaries, and also in the brain and other tissues. When (3)H-E2 was applied to the skin, radioactivity was observed in reproductive and other peripheral tissues. We injected male bats with minute quantities of (3)H-E2 and housed each of them directly with groups of adult females for 48h. We then measured radioactivity in male and female bat tissues. In each of several replications of one male housed with three females, radioactivity was reliably observed in the uterus of all females, and in many other tissues in almost every female. Measurement in the organs of males directly exposed to (3)H-E2 showed high levels of radioactivity in the testes and especially the epididymides. These data indicate that estradiol is transferred from males to females, likely via absorptions from males' excretions and potentially also via intravaginal exposure during mating. Given the potency of estradiol in regulating female reproductive physiology and behavior, our data strongly suggest the potential for pheromonal action whereby male mammals induce sexual receptivity and ovulation in females.


Idiopathic thrombocytopenic purpura complicating chronic lymphocytic leukemia.
Three patients had severe, symptomatic idiopathic thrombocytopenic purpura (ITP) complicating chronic lymphocytic leukemia (CLL) that was refractory to prednisone treatment but that responded to splenectomy alone or in addition to immunosuppressive-cytotoxic treatment. Evidence of high titer of an antiplatelet factor, most likely an antibody, was corroborated in one case by the 14C-platelet serotonin release assay. This experience suggests that the treatment of ITP complicating CLL is similar to that of ITP not associated with lymphoproliferative disorders.


Training in pathology informatics: implementation at the University of Pittsburgh.
Pathology informatics is generally recognized as an important component of pathology training, but the scope, form, and goals of informatics training vary substantially between pathology residency programs. The Training and Education Committee of the Association for Pathology Informatics (API TEC) has developed a standard set of knowledge and skills objectives that are recommended for inclusion in pathology informatics training and may serve to standardize and formalize training programs in this area. The University of Pittsburgh (Pittsburgh, Pa) core rotation in pathology informatics includes most of these goals and is offered as an implementation model for pathology informatics training. The core rotation in pathology informatics is a 3-week, full-time rotation including didactic sessions and hands-on laboratories. Topics include general desktop computing and the Internet, but the primary focus of the rotation is vocabulary and concepts related to enterprise and pathology information systems, pathology practice, and research. The total contact time is 63 hours, and a total of 19 faculty and staff contribute. Pretests and posttests are given at the start and end of the rotation. Performance and course evaluation data were collected for 3 years (a total of 21 residents). The rotation implements 84% of the knowledge objectives and 94% of the skills objectives recommended by the API TEC. Residents scored an average of about 20% on the pretest and about 70% on the posttest for an average increase during the course of 50%. Posttest scores did not correlate with pretest scores or self-assessed computer skill level. The size of the pretest/posttest difference correlated negatively with the pretest scores and self-assessed computing skill level. Pretest scores were generally low regardless of whether residents were familiar with desktop computing and productivity applications, indicating that even residents who are computer "savvy" have limited knowledge of pathology informatics topics. Posttest scores showed that all residents' knowledge increased substantially during the course and that residents who were computing novices were not disadvantaged. In fact, novices tended to have higher pretest/posttest differences, indicating that the rotation effectively supported initially less knowledgeable residents in "catching up" to their peers and achieving an appropriate competency level. This rotation provides a formal training model that implements the API TEC recommendations with demonstrated success.


[The results of medicolegal proceedings involving dental care services].
The paper deals with the results of medicolegal proceedings involving inappropriate dental care service in both criminal and civil cases. The authors also propose measures for prevention of pitfalls described in the study.


HIV-1 expression protects macrophages and microglia from apoptotic death.
Macrophages and microglia are the predominant cells infected with HIV-1 in the brain, yet the effects of productive HIV infection on the fate of these cells are poorly understood. In this study, we tested the hypothesis that HIV-1 expression influences cell death in infected macrophages and microglial cells. We detected apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) in the cerebral white matter of control and HIV encephalitis (HIVE) brains, and quantitatively analysed apoptotic cells with respect to their location (vessel-associated vs. parenchymal), CD68 expression, and HIV-1 p24 expression. There were more vessel-associated, but not more parenchymal, TUNEL+ cells in HIVE cases as compared to controls. Vessel-associated TUNEL+ cells were primarily endothelial cells (von Willebrand factor+) or macrophages (CD68+). TUNEL+/CD68+ cells were present in both control and HIVE cases in similar frequencies (2.1 +/- 0.7% vs. 1.9 +/- 0.7% of total CD68+ populations, respectively). In HIVE, TUNEL+/p24+ cells were 0.4 +/- 0.2% of the total p24+ cell population, which was lower than the frequency of TUNEL+/CD68+ cells (1.9 +/- 0.7%) in the total CD68+ macrophage population. These results suggest that HIV-1-infected macrophages and microglia are resistant to apoptosis, and may contribute to the formation of a central nervous system viral reservoir.


Molecular weight and architectural dependence of well-defined star-shaped poly(lysine) as a gene delivery vector.
A series of well-defined star-shaped polypeptides were successfully synthesised by the ring opening polymerisation (ROP) of the N-carboxyanhydride (NCA) of ε-carbobenzyloxy-l-lysine (ZLL) using a range of generations of polypropylene imine (PPI) dendrimers as multifunctional initiators. The monomer feed ratio and dendrimer generation were varied to afford a series of polypeptide dendrimer hybrids with superior structural versatility and functionality. Subsequent protecting group removal yielded star-shaped poly(lysine) of controlled variation in polypeptide chain length and arm multiplicity. Star-shaped PLL polymers were used to prepare pDNA and siRNA to form "polyplexes" to determine their ability to complex different nucleic acid cargoes and were compared with linear PLL polyplex controls. Significant differences in size and surface charge were seen between star-shaped PLL polyplexes and linear PLL polyplexes for both cargoes. The star-shaped polypeptides were capable of more effective complexation of both nucleic acids at low N/P ratios compared to linear PLL as evidenced by zeta potential and electrophoretic data. This was particularly evident in siRNA polyplexes as linear PLL failed to completely complex siRNA into nanocomplexes of appropriate size for cell transfection i.e. <200 nm in size, while star poly(lysine) formed siRNA polyplexes <100 nm at certain N/P ratios, albeit strongly dependent on the particular molecular weight and architecture, as analysed by dynamic light scattering (DLS). Atomic force microscopy (AFM) identified discrete spherically shaped polyplexes for all star-shaped polypeptide-based polyplexes while linear PLL formed elongated irregular shaped complexes. This difference in morphology may go some way towards explaining the 300-fold increase in luciferase expression seen for star-shaped PLL polyplexes G5(64)-PLL40 compared to linear PLL pGLuc polyplexes in epithelial cells. Each of the PPI-PLL polymers appeared to be capable of protecting the nucleic acid cargoes from degradation by the relevant nuclease enzyme as effectively as the positive control polyethyleneimine (PEI) polyplexes. Overall the promising nucleic acid complexation, sizing, morphology and protection capacity of two different genetic "cargoes" highlight the potential of polypeptide dendrimer hybrids as gene delivery vectors.


Prognostic Classification System for Implant Recession Defects.
The etiology and progression of periimplant mucogingival defects are multifactorial. As such, the aim of this study was to discern and discuss the key long-term prognostic factors that change the balance of homeostasis/regeneration in periimplant mucogingival and recession defects. This report provides cases and a discussion of anatomical factors that affect the long-term maintenance of periimplant soft tissue. The factors guiding long-term maintenance of the periimplant soft tissue apparatus are increasingly complex due to the additive combination of patient-related factors, anatomical variations, and unique material-host interactions. Severity and number of these contributing factors should be considered before implant placement and/or in the treatment of periimplant defects to achieve the best esthetic and functional outcome. In addition, assessment of prognostic factors should be used to provide the patient a realistic, long-term assessment of the esthetic and functional stability of both implant and the surrounding periimplant tissues.


Water soluble, multifunctional antibody-porphyrin gold nanoparticles for targeted photodynamic therapy.
Photodynamic therapy (PDT) is a treatment of cancer by which tumour cells are destroyed using reactive oxygen species produced by photosensitizers following activation with visible or near infrared light. Successful PDT depends on the solubility and the targeting ability of the photosensitizers. In this work, the synthesis of a porphyrin-based water soluble nanoparticle conjugate containing a targeting agent that recognizes the erbB2 receptor overexpressed on the surface of particular cancer cells is reported. The nanoparticle conjugates were synthesized following two different protocols, viz. a biphasic and a monophasic method, with the aim to determine which method yielded the optimal nanosystem for potential PDT applications. The nanoparticles were characterized using UV-Vis absorption and fluorescence spectroscopies together with transmission electron microscopy and zeta potential measurements; and their ability to produce singlet oxygen following irradiation was investigated following the decay in absorption of a singlet oxygen probe. The nanoparticles synthesized using the monophasic method were shown to produce the highest amount of singlet oxygen and were further functionalized with anti-erbB2 antibody to target the erbB2 receptors expressed on the surface of SK-BR-3 human breast cancer cells. The water soluble, antibody-porphyrin nanoparticle conjugates were shown to elicit targeted PDT of the breast cancer cells.


Homogeneous and sensitive detection of microRNA with ligase chain reaction and lambda exonuclease-assisted cationic conjugated polymer biosensing.
A simple and homogeneous microRNA assay is developed by integration of ligase chain reaction (LCR) and lambda exonuclease-assisted cationic conjugated polymer (CCP) biosensing. LCR is utilized for exponential amplification of microRNA, and lambda exonuclease is introduced to degrade excess fluorescein-labeled probes in LCR for eliminating background signal. After addition of CCP, efficient fluorescence resonance energy transfer from CCP to fluorescein in LCR products occurs. The method is sensitive enough to detect 0.1 fM target microRNA and specific to discriminate one-base difference of microRNAs, which paves a new way for homogeneous microRNA detection and molecular diagnosis.


Proliferation of leukemic blood cells in short-term liquid cultures. Relevance to the differential diagnosis of acute myeloblastic and lymphoblastic leukemia.
In short-term liquid cultures of leukemic blood cells from 60 untreated patients with acute lymphoblastic leukemia (ALL) or acute myeloblastic leukemia (AML) the 3H-thymidine incorporation was measured. After 24, 48 and 72 h of culture, leukemic cells from AML patients consistently showed much higher incorporation values than those from ALL patients. In addition, the cultured cells were studied with the pulse cytophotometer to measure the cell phase distribution of the leukemic cell population. The percentages of cells in S, G2, and M phases in cultures from AML patients were also clearly higher than those from ALL patients. These test methods provide an additional diagnostic aid, especially for those forms of acute leukemia, which remain unclassified by conventional morphological and histochemical criteria.


Protective actions of a leukotriene B4 antagonist in splanchnic ischemia and reperfusion in rats.
Pentobarbital-anesthetized rats were subjected to occlusion of both the celiac and superior mesenteric arteries for 90 min followed by reperfusion for 2 h. All seven rats given only the vehicle died within 2 h of reperfusion, whereas rats treated with LY-255283 (3 or 10 mg/kg iv), a leukotriene B4 (LTB4) receptor antagonist given 10 min before reperfusion, exhibited significantly higher survival rates of 57% (4 out of 7) and 75% (6 out of 8), respectively, 2 h after reperfusion. Rats given 1 mg/kg of LY-255283 showed no significant improvement in survival. Splanchnic artery occlusion (SAO)-shock rats treated with LY-255283 (3 or 10 mg/kg) exhibited significantly attenuated accumulation of plasma free amino-nitrogenous compounds and of a myocardial depressant factor. Treatment with LY-255283 (10 mg/kg) markedly (P less than 0.01) ameliorated the deficits of endothelium-dependent relaxation of isolated superior mesenteric artery (SMA) rings in untreated SAO-shock rats. LY-255283 at 10 mg/kg significantly attenuated the increased myeloperoxidase activity in the intestinal tissue of SAO-shock rats. Moreover, LY-189444, a closely related compound having no LTB4 antagonist activity, did not protect rats in SAO shock, whereas a lipoxygenase inhibitor confirmed protection in SAO shock. These results suggest that LTB4 plays a pivotal role in endothelial dysfunction occurring in SAO-shock rats by chemoattraction and activation of neutrophils on the surface of vascular endothelial cells. Moreover, LY-255283 but not LY-189444 inhibited the adherence of rat neutrophils to isolated SMA endothelium.(ABSTRACT TRUNCATED AT 250 WORDS)


Localization of the complement-component-C3b-binding site and the cofactor activity for factor I in the 38kDa tryptic fragment of factor H.
Trypsin treatment of human factor H (H160) [enzyme/substrate ratio 1:100 (w/w), 30 min, 37 degrees C] generated a 38 kDa (H38) and a 142 kDa (H142) fragment linked by disulphide bonds (H38/142). The fragments were purified by reduction with 2-mercapto-ethanol, gel filtration on a Sephadex G-200 column and affinity chromatography with monoclonal anti-(factor H) antibody coupled to Sepharose 4B. This monoclonal antibody bound to a site in the 38 kDa fragment. To localize the C3b binding site in factor H we used two enzyme-linked immunosorbent assays (e.l.i.s.a.). For the first test, e.l.i.s.a. plates were coated with C3b; H160, H38/142, H38 and H142 were added, and their binding was monitored by goat anti-(factor H) and peroxidase-labelled rabbit anti-goat antibodies. Only intact factor H bound to the C3b-coated plates. For the second test, e.l.i.s.a. plates were coated with comparable amounts of factor H or its fragments, and C3b was offered at several dilutions. In contrast with the results from the first assay, C3b bound to intact factor H, H38/142 and H38 but not to H142, thus characterizing H38 as the fragment carrying the C3b-binding site. To identify the fragment responsible for the cofactor activity of factor H (cleavage of fluid-phase C3b by factor I), 125I-C3b was incubated with either H38 or H142 and factor I. H142 had no cofactor activity, whereas H38 had the same cofactor function as intact H. To further investigate the relationship between the C3b-binding site and the site of factor H essential for its cofactor activity, we made use of monoclonal antibodies directed against the H38. Those antibodies inhibiting the binding of C3b to H160 also inhibited the cofactor function, whereas those without effect on the C3b binding also did not interfere with the cofactor activity. This suggests that the C3b-binding site and the site essential for the cofactor activity of factor H are both localized in the 38 kDa tryptic fragment of factor H in close proximity or are identical.


[Antiviral activity of mutant interferons. A new approach to the study of structural-functional organization of interferons].
The developed approach to investing the structure-functional organization of interferon has been developed consisting in: 1) fusing genes of interferon and alpha-peptide of beta-galactosidase, the resultant protein having the interferon properties and being determined by the beta-galactosidase alpha-complementation test; 2) constructing mutant genes of interferon by the localized mutagenesis; 3) determining the mutant interferon activity; 4) deducing the amino acid sequence of mutant interferon by sequencing mutant genes; 5) analyzing structure-functional organization of interferon. In accordance with this approach, ten mutant interferons with up to 15 changes of amino acid substitutions are obtained and their antiviral activity is determined. The role of some amino acid residues in antiviral activity of interferon alpha 2 is revealed.


A Latent Class Analysis of Mental Health Beliefs Related to Military Sexual Trauma.
Military veterans with histories of military sexual trauma (MST) are at risk for several negative mental health outcomes and report perceived barriers to treatment engagement. To inform interventions to promote gender-sensitive access to MST-related care, we conducted an exploratory, multiple-group latent class analysis of negative beliefs about MST-related care. Participants were U.S. veterans (N = 1,185) who screened positive for MST within the last 2 months and reported a perceived need for MST-related treatment. Associations between class membership, mental health screenings, logistical barriers, difficulty accessing care, and unmet need for MST-related care were also examined. Results indicated a four-class solution, with classes categorized as (a) low barrier, with few negative beliefs; (b) high barrier, with pervasive negative beliefs; (c) stigma-related beliefs; and (d) negative perceptions of care (NPC). Men were significantly less likely than women to fall into the low barrier class (27.9% vs. 34.5%). Relative to participants in the low barrier class, individuals in all other classes reported more scheduling, ps < .001; transportation, p < .001 to p = .014; and work-related barriers, p < .001 to p = .031. Participants in the NPC class reported the most difficulty with access, p < .001, and those in the NPC and high barrier classes were more likely to report unmet needs compared to other classes, ps < .001. Brief cognitive and behavioral interventions, delivered in primary care settings and via telehealth, tailored to address veterans' negative mental health beliefs may increase the utilization of mental health treatment related to MST.


MS-Net: Multi-Site Network for Improving Prostate Segmentation With Heterogeneous MRI Data.
Automated prostate segmentation in MRI is highly demanded for computer-assisted diagnosis. Recently, a variety of deep learning methods have achieved remarkable progress in this task, usually relying on large amounts of training data. Due to the nature of scarcity for medical images, it is important to effectively aggregate data from multiple sites for robust model training, to alleviate the insufficiency of single-site samples. However, the prostate MRIs from different sites present heterogeneity due to the differences in scanners and imaging protocols, raising challenges for effective ways of aggregating multi-site data for network training. In this paper, we propose a novel multi-site network (MS-Net) for improving prostate segmentation by learning robust representations, leveraging multiple sources of data. To compensate for the inter-site heterogeneity of different MRI datasets, we develop Domain-Specific Batch Normalization layers in the network backbone, enabling the network to estimate statistics and perform feature normalization for each site separately. Considering the difficulty of capturing the shared knowledge from multiple datasets, a novel learning paradigm, i.e., Multi-site-guided Knowledge Transfer, is proposed to enhance the kernels to extract more generic representations from multi-site data. Extensive experiments on three heterogeneous prostate MRI datasets demonstrate that our MS-Net improves the performance across all datasets consistently, and outperforms state-of-the-art methods for multi-site learning.


Quantitative Epstein-Barr virus (EBV) serology in lung transplant recipients with primary EBV infection and/or post-transplant lymphoproliferative disease.
The Epstein-Barr virus (EBV)-specific antibody response was studied in lung transplant patients to assess their value in the diagnosis and prognosis of post-transplant lymphoproliferative disease. Recently developed synthetic peptides representing Epstein-Barr nuclear antigen-1 (EBNA-1), diffuse early antigen (EA(D)), and virus capsid antigen (VCA) were studied in a semiquantitative enzyme-linked immunosorbent assay (ELISA) to study antibody patterns in 12 seronegative lung transplant patients, of whom four developed a post-transplant lymphoproliferative disease, and seven seropositive lung transplant patients, all of whom developed a post-transplant lymphoproliferative disease. Immunoblot technique was used as a control. All 12 EBV-seronegative patients had a very limited antibody response that was restricted mainly to VCA antibodies. EA(D) antibodies became detectable in only two patients. Antibody response never preceded clinical diagnosis of post-transplant lymphoproliferative disease in the four EBV-seronegative patients who developed post-transplant lymphoproliferative disease. In the seven seropositive lung transplant patients with post-transplant lymphoproliferative disease, we found a rise in antibody titer in only two patients. Immunoblot analysis confirmed the serological results. In conclusion, EBV-specific antibody patterns after lung transplantation are highly restricted and variable and of limited value for the diagnosis or prognosis of post-transplant lymphoproliferative disease.


p53 Binding to the p21 promoter is dependent on the nature of DNA damage.
p53 Is a tumor suppressor that integrates signals from different stress induced signalling pathways, regulates cell cycle arrest, senescence, apoptosis and DNA repair. How p53 dictates cell fate is unclear. As a major transcriptional target of p53 in response to cellular stress, p21 is a key component in cell cycle control and apoptosis, directing an anti-apoptotic response following DNA damage. It is therefore likely that p53-dependent regulation of p21 contributes, at least in part, how p53 influences cellular outcome upon DNA damage. Here we compare the p53-dependent transcriptional regulation of p21 in response to DNA damage by ultraviolet (UV) radiation and ionizing radiation (IR). We demonstrate that despite comparable levels of p53 accumulation by both types of DNA damage, IR causes significant, early accumulation of p21 not seen in UV-damaged cells, with a substantially different cell cycle profile. Whereas UV and IR both target p21 protein for degradation immediately after DNA damage, differential post-damage p21 transcription is accountable for the disparity in p21 protein levels. Chromatin immunoprecipitation studies reveal that p53 displays a clear bias against the p21 promoter in UV-damaged cells compared to IR-damaged cells. We note differential post-translational modifications of nuclear p53 between UV and IR treatment. Furthermore we demonstrate that this disparity correlated with reduced histone acetylation on the TATA box within the p21 promoter following UV treatment. This suggests that the nature of DNA damage enables p53 to selectively discriminate between promoters in the induction of target genes, thereby regulating their expression and subsequent cellular outcome.


A cross-reactive human IgA monoclonal antibody blocks SARS-CoV-2 spike-ACE2 interaction.
COVID-19 caused by SARS-CoV-2 has become a global pandemic requiring the development of interventions for the prevention or treatment to curtail mortality and morbidity. No vaccine to boost mucosal immunity, or as a therapeutic, has yet been developed to SARS-CoV-2. In this study, we discover and characterize a cross-reactive human IgA monoclonal antibody, MAb362. MAb362 binds to both SARS-CoV and SARS-CoV-2 spike proteins and competitively blocks ACE2 receptor binding, by overlapping the ACE2 structural binding epitope. Furthermore, MAb362 IgA neutralizes both pseudotyped SARS-CoV and SARS-CoV-2 in 293 cells expressing ACE2. When converted to secretory IgA, MAb326 also neutralizes authentic SARS-CoV-2 virus while the IgG isotype shows no neutralization. Our results suggest that SARS-CoV-2 specific IgA antibodies, such as MAb362, may provide effective immunity against SARS-CoV-2 by inducing mucosal immunity within the respiratory system, a potentially critical feature of an effective vaccine.


Increased left atrial volume index: potent biomarker for first-ever ischemic stroke.
To establish the incidence and correlation of increased left atrial volume index (LAVI) in patients with first-ever ischemic stroke. Using our institution's epidemiological database, we defined a cohort of 432 patients (cases) who underwent transthoracic echocardiography within 60 days of first ischemic stroke between January 1, 1985, and December 31, 1994. Left atrial volume was measured with the biplane area-length method, indexed to body surface area (LAVI, expressed as mL/m(2)). The control group consisted of 416 community residents who underwent transthoracic echocardiography as participants in a stroke risk factor study. Increased LAVI was defined as 28 mL/m(2) or higher. Survival in patients was compared with expected survival among white Minnesotans and was further modeled as a function of age, sex, LAVI, and clinical risk factors. Among the included 306 patients, 230 (75%) had increased LAVI (mean+/-SD, 49+/-21 mL/m(2)). Patients with increased LAVI were older than those with normal LAVI (mean+/-SD age, 76+/-11 vs 71+/-13 years; P=.003) and had more cardiovascular risk factors (mean+/-SD number, 1.8+/-0.07 vs 1.3+/-0.89; P<.001). Mean LAVI was higher in cases than in age- and sex-matched controls (P<.001). At 5-year follow-up, cases showed excess mortality compared with age-matched controls (P=.001). After variables were adjusted for age, sex, and clinical risk factors, LAVI was independently associated with mortality. A useful index for prediction of adverse cardiovascular events, LAVI might also predict first ischemic stroke and subsequent mortality.


Increased biosynthesis of Alzheimer amyloid precursor protein in the cerebral cortex of rats with lesions of the nucleus basalis of Meynert.
The nucleus basalis of Meynert was lesioned by infusion of N-methyl-D-aspartate (NMDA) unilaterally in adult rat brain. Seven days post lesion we observed that polysomes isolated from the cerebral cortex affected by the lesion synthesized 2.6-fold greater amounts of the Alzheimer amyloid precursor protein (AAPP) compared to the nonlesioned side of the same rat brain. This increase exhibited specificity to AAPP in that overall protein synthesis was not altered by the lesion. The increase of AAPP did not alter the ratio of AAPP isotypes in rat brain (in which AAPP 695, which is lacking the protease inhibitor insert remains the predominant form). The increased synthesis did not result in the apparent accumulation of mature AAPP. These results indicate that a cholinergic lesion which models many of the neurochemical changes observed in Alzheimer's disease induces the expression of AAPP in a major projection region, the cerebral cortex.


Effects of a Coaching-based Childbirth Program on Anxiety and Childbirth Self-efficacy among Primigravida Women.
Childbirth self-efficacy plays an important role in women's ability to cope with labor and delivery. Coaching has been gaining popularity as a way to promote cognitive, emotional and behavioral change. This study aimed to test the effects of a Coaching-based childbirth program on anxiety and childbirth self-efficacy among primigravida women. The study design was a quasi-experimental pre-post design. A coaching-based childbirth program was developed synthesizing concepts and techniques from the literature on coaching and was verified by an expert panel. It consisted of four weekly 2-hour small group sessions. Pregnant women were recruited from H hospital in Seoul. Childbirth self-efficacy and state anxiety were measured before and after the program. Although there was a no significant reduction in anxiety, there were statistically significant increase for childbirth self-efficacy in the experimental group when compared to the control group. The program appears to increase childbirth self-efficacy for pregnant women. Future studies may benefit from using mixed coaching modalities and consider measuring health behaviors and obstetric outcomes to gain insights on its long-term impact.


Musculocontractural Ehlers-Danlos syndrome and neurocristopathies: dermatan sulfate is required for Xenopus neural crest cells to migrate and adhere to fibronectin.
Of all live births with congenital anomalies, approximately one-third exhibit deformities of the head and face. Most craniofacial disorders are associated with defects in a migratory stem and progenitor cell population, which is designated the neural crest (NC). Musculocontractural Ehlers-Danlos syndrome (MCEDS) is a heritable connective tissue disorder with distinct craniofacial features; this syndrome comprises multiple congenital malformations that are caused by dysfunction of dermatan sulfate (DS) biosynthetic enzymes, including DS epimerase-1 (DS-epi1; also known as DSE). Studies in mice have extended our understanding of DS-epi1 in connective tissue maintenance; however, its role in fetal development is not understood. We demonstrate that DS-epi1 is important for the generation of isolated iduronic acid residues in chondroitin sulfate (CS)/DS proteoglycans in early Xenopus embryos. The knockdown of DS-epi1 does not affect the formation of early NC progenitors; however, it impairs the correct activation of transcription factors involved in the epithelial-mesenchymal transition (EMT) and reduces the extent of NC cell migration, which leads to a decrease in NC-derived craniofacial skeleton, melanocytes and dorsal fin structures. Transplantation experiments demonstrate a tissue-autonomous role for DS-epi1 in cranial NC cell migration in vivo Cranial NC explant and single-cell cultures indicate a requirement of DS-epi1 in cell adhesion, spreading and extension of polarized cell processes on fibronectin. Thus, our work indicates a functional link between DS and NC cell migration. We conclude that NC defects in the EMT and cell migration might account for the craniofacial anomalies and other congenital malformations in MCEDS, which might facilitate the diagnosis and development of therapies for this distressing condition. Moreover, the presented correlations between human DS-epi1 expression and gene sets of mesenchymal character, invasion and metastasis in neuroblastoma and malignant melanoma suggest an association between DS and NC-derived cancers.


New Molecular Insights into the Inhibition of Dipeptidyl Peptidase-4 by Natural Cyclic Peptide Oxytocin.
Protease inhibition has led to treating many diseases and has been successful in producing many commercial drugs by pharmaceutical companies. Among many proteases, serine protease has been attractive in treating metabolic disorder diabetes mellitus (DM). Gliptins have been proven to inhibit dipeptidyl peptidase-4 (DPP4), a serine protease, and are an emerging therapeutic drug target to reduce blood glucose levels, but until now there is no natural cyclic peptide proven to inhibit serine protease DPP4. This study demonstrates the potential mechanism of natural cyclic peptide oxytocin (OXT) as a DPP4 inhibitor. To achieve this, initially, activity atlas and field-based models of DPP4 inhibitors were utilized to predict the possible features of positive and negative electrostatic, hydrophobic, and activity shapes of DPP4 inhibition. Oxytocin binding mode, flexibility, and interacting residues were studied using molecular docking simulations studies. 3D-RISM calculations studies revealed that the stability of water molecules at the binding site are favorable. Finally, an experimental study using fluorescence assay revealed OXT inhibits DPP4 in a concentration-dependent manner in a significant way (p < 0.05) and possess IC50 of 110.7 nM. These new findings significantly expand the pharmaceutical application of cyclic peptides, and in specific OXT, and implicate further optimization of OXT inhibition capacity to understand the effect of DPP4 inhibition. This work highlights the development of natural cyclic peptides as future therapeutic peptides to reduce glucose levels and treat diabetes mellitus.


Neuroimaging of mirtazapine enantiomers in humans.
Mirtazapine is a racemic antidepressant with a multireceptor profile. Previous studies have shown that the enantiomers of mirtazapine have different pharmacologic effects in the brain of laboratory animals. In the present study, we used positron emission tomography (PET) and autoradiography to study effects of (R)- and (S)-[(11)C]mirtazapine in the human brain. Detailed brain imaging by PET using three methods of kinetic data analysis showed no reliable differences between regional binding potentials of (R)- and (S)-[(11)C]mirtazapine in healthy subjects. Autoradiographic studies carried out in whole hemispheres of human brain tissue showed, however, that (R)- and (S)-mirtazapine differ markedly as inhibitors of [(3)H]clonidine binding at alpha(2)-adrenoceptors. The multireceptor binding profiles of mirtazapine enantiomers, along with individual differences between subjects, may preclude PET neuroimaging from demonstrating reliable differences between the regional distribution and binding of (R)- and (S)-[(11)C]mirtazapine in the living human brain.


A brief review of skull base chordomas.
Skull base chrodomas are slow growing neoplasms usually located along the midline. They display a locally invasive nature with possibilities of extracranial metastasis. Presentation is usually late and depends upon the location and extent of the tumour. Management aims at gross total resection via open microsurgical or endoscopic approach followed by adjuvant radiotherapy. Prognosis may be good for the classical and chondroid subtypes but remains poor for de-differentiated type.


Sugar transporters enable a leaf beetle to accumulate plant defense compounds.
Many herbivorous insects selectively accumulate plant toxins for defense against predators; however, little is known about the transport processes that enable insects to absorb and store defense compounds in the body. Here, we investigate how a specialist herbivore, the horseradish flea beetle, accumulates glucosinolate defense compounds from Brassicaceae in the hemolymph. Using phylogenetic analyses of coleopteran major facilitator superfamily transporters, we identify a clade of glucosinolate-specific transporters (PaGTRs) belonging to the sugar porter family. PaGTRs are predominantly expressed in the excretory system, the Malpighian tubules. Silencing of PaGTRs leads to elevated glucosinolate excretion, significantly reducing the levels of sequestered glucosinolates in beetles. This suggests that PaGTRs reabsorb glucosinolates from the Malpighian tubule lumen to prevent their loss by excretion. Ramsay assays corroborated the selective retention of glucosinolates by Malpighian tubules of P. armoraciae in situ. Thus, the selective accumulation of plant defense compounds in herbivorous insects can depend on the ability to prevent excretion.


Visibility of the urethral meatus and risk of urinary tract infections in uncircumcised boys.
Uncircumcised boys are at higher risk for urinary tract infections than circumcised boys. Whether this risk varies with the visibility of the urethral meatus is not known. Our aim was to determine whether there is a hierarchy of risk among uncircumcised boys whose urethral meatuses are visible to differing degrees. We conducted a prospective cross-sectional study in one pediatric emergency department. We screened 440 circumcised and uncircumcised boys. Of these, 393 boys who were not toilet trained and for whom the treating physician had requested a catheter urine culture were included in our analysis. At the time of catheter insertion, a nurse characterized the visibility of the urethral meatus (phimosis) using a 3-point scale (completely visible, partially visible or nonvisible). Our primary outcome was urinary tract infection, and our primary exposure variable was the degree of phimosis: completely visible versus partially or nonvisible urethral meatus. Cultures grew from urine samples from 30.0% of uncircumcised boys with a completely visible meatus, and from 23.8% of those with a partially or nonvisible meatus (p = 0.4). The unadjusted odds ratio (OR) for culture growth was 0.73 (95% confidence interval [CI] 0.35-1.52), and the adjusted OR was 0.41 (95% CI 0.17-0.95). Of the boys who were circumcised, 4.8% had urinary tract infections, which was significantly lower than the rate among uncircumcised boys with a completely visible urethral meatus (unadjusted OR 0.12 [95% CI 0.04-0.39], adjusted OR 0.07 [95% CI 0.02-0.26]). We did not see variation in the risk of urinary tract infection with the visibility of the urethral meatus among uncircumcised boys. Compared with circumcised boys, we saw a higher risk of urinary tract infection in uncircumcised boys, irrespective of urethral visibility.


Bacteriological survey of tuberculous lymphadenitis in South-east England: 1973-80.
During the eight years 1973-80 the Public Health Laboratory Service Regional Centre for Tuberculosis Bacteriology received cultures of mycobacteria from 2339 patients with tuberculous lymphadenitis. Of these, 2272 were M tuberculosis (2207 human and 65 bovine strains) and 67 were other mycobacterial species, usually M avium and its intracellulare variant. Disease due to the human strains of M tuberculosis occurred most often in young women of Asian ethnic origin. Many bovine strains isolated from Asian patients differ from the classical bovine type in being sensitive to pyrazinamide: the origin of these strains, whether from other people or from cattle, is unknown. Lymphadenitis due to bovine strains tended to occur in an older age group than the human strains and probably include relatively more cases of reactivation diseases. Infection caused by the other mycobacterial species occurred mainly in young children of European origin. Tuberculosis, therefore, remains an important cause of lymphadenopathy in Britain.


[Cerebrospinal fluid sorption in the system of complex treatment of chronic cerebral ischemia].
The cerebrospinal fluid was investigated in 16 patients with chronic cerebral ischemia. Reactions of the local immune system of the liquor was shown to change by the autoimmune type. Medical efficiency of cerebrospinal fluid sorption was proved and it can be considered a method of detoxication aimed at breaking the pathogenetic chain: formation of abundance of the autoantibodies--increased amount of the circulating immunocomplexes--damage of the cell membranes--discharge of deep antigens--appearance of a new generation of autoantibodies. Using cerebrospinal fluid sorption as a test for the detection of latent functional reserves of the neurons not changed irreversibly in the zone of reduced perfusion of the cerebral tissue is thought to be a perspective method.


endoscope-i: an innovation in mobile endoscopic technology transforming the delivery of patient care in otolaryngology.
Digital and mobile device technology in healthcare is a growing market. The introduction of the endoscope-i, the world's first endoscopic mobile imaging system, allows the acquisition of high definition images of the ear, nose and throat (ENT). The system combines the e-i Pro camera app with a bespoke engineered endoscope-i adaptor which fits securely onto the iPhone or iPod touch. Endoscopic examination forms a salient aspect of the ENT work-up. The endoscope-i therefore provides a mobile and compact alternative to the existing bulky endoscopic systems currently in use which often restrict the clinician to the clinic setting. Areas covered: This article gives a detailed overview of the endoscope-i system together with its applications. A review and comparison of alternative devices on the market offering smartphone adapted endoscopic viewing systems is also presented. Expert commentary: The endoscope-i fulfils unmet needs by providing a compact, highly portable, simple to use endoscopic viewing system which is cost-effective and which makes use of smartphone technology most clinicians have in their pocket. The system allows real-time feedback to the patient and has the potential to transform the way that healthcare is delivered in ENT as well as having applications further afield.


Distance Matters: Geographic barriers to long acting reversible and permanent contraception for homeless women Veterans.
Women Veterans who experience homelessness are at high risk of unintended pregnancy and adverse outcomes. Contraception could mitigate risks, yet access barriers exist across the Veterans Health Administration (VHA). We identified all US women Veterans, age 18-44y with evidence of homelessness in VHA administrative data between fiscal years 2002-2015, in order to document the geographic distribution of ever-homeless women Veterans in relation to VA Medical Centers (VAMCs) and assess geographic associations between long acting reversible contraceptives (LARC) or permanent contraception (PC) use. We calculated VAMC travel distance from last known ZIP Code. We used multivariate logistic regression models to explore contraceptive method associations. We included 41,722 ever-homeless women Veterans; 9.2% had LARC exposure and 7.5% PC. We found 29% of ever-homeless women Veterans resided >40miles from the nearest VAMC and increasing drive distance was negatively correlated with contraceptive exposure, especially for Veterans residing >100miles from a VAMC. Increasing distance to the nearest VAMC results in a geographic barrier to the most effective contraceptive options for women Veterans. The VHA is uniquely positioned to leverage its rural and homeless healthcare expertise to address geographic barriers and integrate comprehensive contraceptive services into established programs for high-risk Veterans.


[Additional administration of dutasteride in patients with benign prostatic hyperplasia who did not respond sufficiently to α1-adrenoceptor antagonist : investigation of clinical factors affecting the therapeutic effect of dutasteride].
We performed additional administration of dutasteride in patients who did not respond sufficiently to α1-adrenoceptor antagonist treatment for lower urinary tract symptoms (LUTS) associated with benign prostatic hyperplasia (BPH) (LUTS/BPH). Among 76 registered patients, efficacy was analyzed in 58 patients. International Prostate Symptom Score (IPSS), subscores for voiding and storage symptoms and quality of life (QOL) on the IPSS, and Overactive Bladder Symptom Score (OABSS) were all significantly improved from the third month of administration compared to the time of initiating additional administration of dutasteride. Additional administration of dutasteride also significantly reduced prostate volume, and residual urine with the exception of the sixth month after administration. Age at initiation of administration and voiding symptom subscore on the IPSS were clinical factors affecting the therapeutic effects of dutasteride. The rate of improvement with treatment decreased with increasing age at initiation of dutasteride administration, and increased as voiding symptom subscore on the IPSS increased. Therefore, additional administration of dutasteride appears useful for cases of LUTS/BPH in which a sufficient response is not achieved with α1-adrenoceptor antagonist treatment. Because patients who have severe voiding symptoms or begin dutasteride at an early age may be expected to respond particularly well to dutasteride in terms of clinical efficacy, they were considered to be suitable targets for additional administration.


Sedimenting pairs of elastic microfilaments.
The dynamics of two identical elastic filaments settling under gravity in a viscous fluid in the low Reynolds number regime is investigated numerically. A large family of initial configurations symmetric with respect to a vertical plane is considered, as well as their non-symmetric perturbations. The behaviour of the filaments is primarily governed by the elasto-gravitational number, which depends on the filament's length and flexibility, and the strength of the external force. Flexible filaments usually converge toward horizontal and parallel orientation. We explain this phenomenon and show that it occurs also for curved rigid particles of similar shapes. Once aligned, the two fibres either converge toward a stationary, flexibility-dependent distance, or tend to collide or continuously repel each other. Rigid and straight rods perform periodic motions while settling down. Apart from very stiff particles, the dynamics is robust to non-symmetric perturbations.


Neutron generator based on intense lithium beam driver.
We are proposing a compact neutron generator based on a Li beam driver. The proposed neutron generator comprises a laser ion source, a radio-frequency quadrupole linear accelerator (RFQ linac), a drift tube linac, and a target containing protons. In the generator, the lithium ion is used as a projectile instead of protons to utilize the kinematic focusing technique. The technique enables us to enhance the neutron flux without increasing the beam energy, which is important to develop a clean compact neutron generator. Moreover, the combination of a laser ion source and a RFQ linac with the direct plasma injection scheme will provide several tens of mA of a fully ionized lithium beam, which is much higher than that of conventional heavy ion sources comparable with proton drivers. Neutrons are generated by the nuclear reaction of the lithium ions and protons in the beam target. In this paper, we reported the current status of the development. For RFQ, we designed the RFQ rods to accelerate 40 mA of 7Li3+. We fabricated and installed the rods into a cavity, and, as a first test, accelerated 10 mA of C6+ successfully.


Product quality for nanomaterials: current U.S. experience and perspective.
In recent years, there has been an increased focus on developing novel drug delivery systems and targeted therapies through the use of nanotechnology and nanomaterials. Such focus is translating to an increasing number of investigational new drug (IND) applications, new drug applications (NDAs), and abbreviated new drug applications (ANDAs) for drug products containing nanomaterials to the United States Food and Drug Administration (FDA). Although subject to the same rigorous regulatory standards and regulatory pathways as any drug product, unique properties that arise from the small size, large surface area, and polydispersity of nanomaterials may lead to additional scientific considerations when following current FDA guidelines and practices for drug evaluation. This review article will discuss these scientific considerations based on the experience with FDA-approved drug products containing nanomaterials.


First study of Salmonella in meat in Romania.
The increasing antimicrobial resistance of Salmonella isolates is of major public health concern, but information regarding these aspects is still lacking in Romania. This study focused on a detailed and accurate investigation concerning prevalence, serotypes, and antimicrobial resistance patterns of Salmonella strains, isolated from pork and chicken meat, collected from all regions of Romania in 2011. The research was conducted on 650 samples of chicken and pork meat collected from production units and retail markets located in various regions of Romania. A total of 149 Salmonella isolates were recovered (22.92%), serotyped, confirmed by PCR, and further tested for antimicrobial susceptibility. Thirteen Salmonella serovars were identified; predominant serovars included Infantis, Typhimurium, Derby and Colindale. Multiple resistance was found in 83.22% (n = 124) of the isolates. The isolates were frequently resistant to tetracycline (80.53%), streptomycin (81.21%), sulfamethoxazole (87.25%), nalidixic acid (65.10%), and ciprofloxacin (42.95%). Additionally, a markedly lower resistance rate was observed for ampicillin (20.81%), chloramphenicol (16.78%), and ceftazidime (11.41%). Among 137 resistant Salmonella isolates, 35 different resistance patterns were found. A high prevalence of Salmonella spp. and a relatively high resistance rate to multiple antimicrobials was found. This data indicates that chicken and pork meat could constitute a source of human exposure to multidrug-resistant Salmonella and therefore could be considered a potential vehicle of resistant Salmonella foodborne diseases. Further actions are needed to succesfully implement a national surveillance program for better monitoring of these resistant pathogens.


[Management of iron overload during pregnancy and childbirth in a patient with ferroportin disease].
An asymptomatic woman in her early 40s with a history of hyperferritinemia (5,412 ng/ml) was referred to our hospital after repeated phlebotomy for hemosiderosis. She had unexplained hyperferritinemia, low-normal transferrin saturation, and high hepcidin levels, in the absence of iron overload-induced organ injury. She was diagnosed with ferroportin disease based on detection of the SLC40A1 variant SLC40A1 c.485_487del (p.Val162del) on genetic analysis. Her ferritin levels remained stable during pregnancy, and postpartum anemia was successfully treated with 2-week oral iron therapy. Ferroportin disease is characterized by impaired iron export and preferential iron trapping in tissue macrophages. To reduce risk of anemia, a non-aggressive phlebotomy regimen is recommended in patients with ferroportin disease, which shows a milder clinical course compared with other classical hemochromatosis subtypes.


Surface Analysis by Secondary Ion Mass Spectrometry (SIMS): Principles and Applications from Swiss laboratories.
Secondary Ion Mass Spectrometry (SIMS) extracts chemical, elemental, or isotopic information about a localized area of a solid target by performing mass spectrometry on secondary ions sputtered from its surface by the impact of a beam of charged particles. This primary beam sputters ionized atoms and small molecules (as well as many neutral particles) from the upper few nanometers of the sample surface. The physical basis of SIMS has been applied to a large range of applications utilizing instruments optimized with different types of mass analyzer, either dynamic SIMS with a double focusing mass spectrometer or static SIMS with a Time of Flight (TOF) analyzer. Here, we present a short review of the principles and major applications of three different SIMS instruments located in Switzerland.


Wearable technology for compensatory reserve to sense hypovolemia.
Traditional monitoring technologies fail to provide accurate or early indications of hypovolemia-mediated extremis because physiological systems (as measured by vital signs) effectively compensate until circulatory failure occurs. Hypovolemia is the most life-threatening physiological condition associated with circulatory shock in hemorrhage or sepsis, and it impairs one's ability to sustain physical exertion during heat stress. This review focuses on the physiology underlying the development of a novel noninvasive wearable technology that allows for real-time evaluation of the cardiovascular system's ability to compensate to hypovolemia, or its compensatory reserve, which provides an individualized estimate of impending circulatory collapse. Compensatory reserve is assessed by real-time changes (sampled millions of times per second) in specific features (hundreds of features) of arterial waveform analog signals that can be obtained from photoplethysmography using machine learning and feature extraction techniques. Extensive experimental evidence employing acute reductions in central blood volume (using lower-body negative pressure, blood withdrawal, heat stress, dehydration) demonstrate that compensatory reserve provides the best indicator for early and accurate assessment for compromises in blood pressure, tissue perfusion, and oxygenation in resting human subjects. Engineering challenges exist for the development of a ruggedized wearable system that can measure signals from multiple sites, improve signal-to-noise ratios, be customized for use in austere conditions (e.g., battlefield, patient transport), and be worn during strenuous physical activity.


The treatment of traumatized permanent anterior teeth: case report & literature review. Part I--Management of intruded incisors.
A case is reported in which an intruded incisor was initially treated by an endodontic dressing with calcium hydroxide and then extruded using a removable orthodontic appliance. A follow-up examination seven years after completion of endodontic therapy and bleaching showed a favourable response. A review of the relevant literature indicates that intrusion occurs in five to twelve per cent of luxation cases. In this type of injury maximum damage occurs to the pulp and all supporting structures because the tooth is driven into the alveolar process. Complications which have been reported include: pulp necrosis, apical radiolucencies, partial or total pulp calcification, root resorption (surface, inflammatory or replacement), marginal periodontal bone breakdown, and arrested or disturbed root development. The prognosis for pulp survival after intrusion is much more favourable for teeth with incomplete root formation than for teeth with complete root formation. Treatment options available to bring an intruded tooth into alignment are: to await spontaneous re-eruption which may occur if root formation is incomplete, uncovering of the intruded crown, orthodontic extrusion which is allied with gentle luxation if the tooth does not move, and immediate surgical repositioning.


Management of coeliac disease: a changing diagnostic approach but what value in follow up?
To assess the management of patients with coeliac disease in relation to a change in diagnostic method from jejunal suction biopsy to endoscopic biopsy. 16 item questionnaire survey of consultant members of the British Society of Gastroenterology. 359 consultant physician and gastroenterologist members of the society. Type of routine biopsy; repeat biopsy after gluten withdrawal; gluten rechallenge; follow up measurements; screening for malignancy; and methods of follow up, including special clinics. 270(70%) members replied; 216(80%) diagnosed coeliac disease routinely by endoscopic duodenal biopsy, 30(11%) by jejunal capsule biopsy, and the remainder by either method. Only 156(58%) repeated the biopsy after gluten withdrawal, though more did so for duodenal than jejunal biopsies (134/216, 62% v 13/30, 43%; p < 0.02). Follow up biopsies featured more duodenal than jejunal biopsies (133/156, 82% v 23/156, 15%; p < 0.02). Regular follow up included assessments of weight (259, 96%) and full blood count (238, 88%) but limited assessment of serum B-12 and folate (120, 44%) and calcium (105, 39%) concentrations. Routine screening for malignancy is not performed, and there are few specialist clinics. 171(63%) respondents thought that patients should be followed up by a hospital specialist and 58(21%) by family doctors. The practice of diagnosing coeliac disease varies appreciably from that in many standard texts. Many patients could be effectively cared for by their family doctor. The British Society of Gastroenterology should support such management by family doctors by providing clear guidelines for them.


IL-1α Mediates Innate and Acquired Resistance to Immunotherapy in Melanoma.
Inflammation has long been associated with cancer initiation and progression; however, how inflammation causes immune suppression in the tumor microenvironment and resistance to immunotherapy is not well understood. In this study, we show that both innate proinflammatory cytokine IL-1α and immunotherapy-induced IL-1α make melanoma resistant to immunotherapy. In a mouse melanoma model, we found that tumor size was inversely correlated with response to immunotherapy. Large tumors had higher levels of IL-1α, Th2 cytokines, polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs), and regulatory T cells but lower levels of IL-12, Th1 cytokines, and activated T cells. We found that therapy with adenovirus-encoded CD40L (rAd.CD40L) increased tumor levels of IL-1α and PMN-MDSCs. Blocking the IL-1 signaling pathway significantly decreased rAd.CD40L-induced PMN-MDSCs and their associated PD-L1 expression in the tumor microenvironment and enhanced tumor-specific immunity. Similarly, blocking the IL-1 signaling pathway improved the antimelanoma activity of anti-PD-L1 Ab therapy. Our study suggests that blocking the IL-1α signaling pathway may increase the efficacy of immunotherapies against melanoma.


Association between a sequence variant in the IL-4 gene promoter and FEV(1) in asthma.
Recent family-based studies have revealed evidence for linkage of human chromosome 5q31 to the diagnosis of asthma, elevated serum IgE levels, and bronchial hyperresponsiveness. Among the candidate genes in this region is the gene encoding for human interleukin-4 (IL-4). We reasoned that this gene could also serve as a candidate gene with respect to asthma severity as indicated by the FEV(1) measured when bronchodilator treatment was withheld. To test this hypothesis, we examined a large population of patients with asthma (ascertained without respect to genetic characteristics), for associations between a genetic variant in the IL-4 promoter region (C-589T) and asthma severity, as indicated by FEV(1). We used amplification by the polymerase chain reaction followed by BsmF1 restriction digestion to assign genotypes at the IL-4 promoter C-589T locus. We compared genotypes at this locus in 772 Caucasian and African American patients with asthma of varying severity, and we used multiple regression analysis to relate genotypic findings to FEV(1). Among white individuals, the homozygous presence of the C-589T IL-4 promoter genotype (TT) was associated with a FEV(1) below 50% of predicted (p = 0.013; OR, 1.44; 95% CI: 1.09 to 1.90). Subjects with the TT genotype had mean FEV(1) (% predicted) values 4.5% lower than those of subjects with the wild-type (CC) genotype at this locus. FEV(1) values of white patients with a CC or CT genotype were broadly distributed, whereas the TT genotype was associated with a narrow distribution of low FEV(1) values. The frequency of the T allele was significantly greater (p = 1 x 10(-)(23)) among African American asthmatics (0.544) than among white asthmatics (0.183). These data provide the first evidence associating FEV(1) in patients with asthma and genetic determinants at any locus. Our data are consistent with the idea that the FEV(1) in asthma is the result of multiple factors; one of these factors is the genotype at the IL-4 C-589T locus. This locus is associated with a small but significant decrement in pulmonary function among white asthmatic subjects.


Effects of sealing the intramedullary femoral canal in total knee arthroplasty: A randomized study.
This study investigates the clinical effects of sealing the femoral canal by intramedullary alignment instrumentation in total knee arthroplasty (TKA). One hundred twenty consecutive patients with knee osteoarthritis, who underwent unilateral TKA, were enrolled in the study and equally randomized into 2 groups: the sealing group and the control group. In the sealing group, the femoral canal was sealed with autogenous bone and cement using intramedullary alignment instrumentation, while the femoral hole was left open for patients in the control group. Blood loss, hemoglobin (Hb) reduction, and other parameters were recorded, as well as the duration of hospital stay and complications. The Hospital for Special Surgery (HSS) knee score was used to assess knee function at the final follow-up appointment. The calculated blood loss, hidden blood loss, transfusion requirements, drainage volume, and Hb reduction measurements were significantly different (P < .05) between the 2 groups. There were no significant differences in the surgery time, intraoperative blood loss, length of hospital stay, HSS score or complications between the 2 groups (P > .05). Sealing the intramedullary canal with autologous bone and a cement plug is an effective method for reducing blood loss and decreasing blood transfusion requirements during TKA procedures that have increasing complication rates.


Influence of circadian rhythms on rat muscle glycogen metabolism during and after exercise.
Marked circadian fluctuations in skeletal muscle glycogen concentrations have previously been reported. The purpose of the present study was to estimate the influence of these rhythms on muscle glycogen metabolism during and after high-intensity treadmill exercise. Male Sprague-Dawley rats ran five 1-min sprints at 75 m min-1 interspersed by 1-3 min rest intervals either at 08.00 h (morning) or at 20.00 h (night). All muscles sampled lost significant amounts of glycogen during exercise at both time periods. There were no differences in rates of loss between morning and night, even though glycogen levels in several muscles (high-oxidative muscles) were significantly higher before exercise in the morning. Following exercise, glycogen restoration in muscle samples primarily composed of fast-twitch fibres was more rapid in the morning than at night. There was no difference in glycogen restoration rates between the two time periods in the muscle primarily composed of slow-twitch fibres. Although liver glycogen was lower after exercise at night than in the morning, there were no differences in post-exercise blood glucose levels between the two time periods. In conclusion, circadian rhythms do not appear to influence rates of glycogen loss during high-speed running. However, since glycogen loss is the same at all times of day, one would predict that circadian changes in pre-exercise muscle glycogen concentrations would affect muscular endurance. Muscle glycogen restoration after exercise does appear to be affected by circadian rhythms, although interpretation of these data is complicated by possible changes in patterns of muscle fibre contraction at different times of the day. These circadian influences should be considered in the design of exercise studies using laboratory rodents.


Highly specific alternative splicing of transcripts encoding BK channels in the chicken's cochlea is a minor determinant of the tonotopic gradient.
The frequency sensitivity of auditory hair cells in the inner ear varies with their longitudinal position in the sensory epithelium. Among the factors that determine the differential cellular response to sound is the resonance of a hair cell's transmembrane electrical potential, whose frequency correlates with the kinetic properties of the high-conductance Ca(2+)-activated K(+) (BK) channels encoded by a Slo (kcnma1) gene. It has been proposed that the inclusion of specific alternative axons in the Slo transcripts along the cochlea underlies the gradient of BK-channel kinetics. By analyzing the complete sequences of chicken Slo gene (cSlo) cDNAs from the chicken's cochlea, we show that most transcripts lack alternative exons. Transcripts with more than one alternative exon constitute only 10% of the total. Although the fraction of transcripts containing alternative exons increases from the cochlear base to the apex, the combination of alternative exons is not regulated. There is also a clear increase in the expression of BK transcripts with long carboxyl termini toward the apex. When long and short BK transcripts are expressed in HEK-293 cells, the kinetics of single-channel currents differ only slightly, but they are substantially slowed when the channels are coexpressed with the auxiliary beta subunit that occurs more widely at the apex. These results argue that the tonotopic gradient is not established by the selective inclusion of highly specific cSlo exons. Instead, a gradient in the expression of beta subunits slows BK channels toward the low-frequency apex of the cochlea.


Transmural variation in the relationship between myocardial infarct size and risk area.
To test the hypothesis that the relationship between infarct area (IA) and area at risk (AR) varies in different layers of the left ventricle (LV), we occluded the circumflex coronary artery for 48 h in 20 conscious dogs. AR was determined by postmortem coronary stereoarteriography, and infarct area by pathological examination. Both AR and IA were divided into four layers: posterior papillary muscle (PPM), subendocardium (Endo), midwall, and subepicardium (Epi) and quantified with planimetry. Hemodynamics and regional myocardial flow with tracer microspheres (7-10 micrometers diam) were measured before and after coronary occlusion. IA was closely correlated with AR for PPM (r = 0.96), Endo (r = 0.97), and Epi (r = 0.92). However, the slope of IA/AR for Endo (1.30 +/- 0.08) was significantly steeper (P less than 0.05) than that for Epi (0.89 +/- 0.11); furthermore, the intercepts at zero infarction for PPM (0.5 +/- 0.1% of LV), Endo (4.2 +/- 0.4%), and Epi (0.1 +/- 0.7%) were significantly different from each other. Regional blood flow measurements indicate that the differences in IA/AR in various layers reflected earlier and greater total collateral flow to the noninfarcted AR in the epicardium. Thus IA/AR for the entire LV is a composite representing separate IA/AR specific to various transmural layers of the LV. In addition, this study demonstrates that the lateral border zone between the IA and the AR is minimal (less than 3-5 mm) in the subendocardium and midwall layers of the left ventricle.


Patients with Orthostatic Intolerance: Relationship to Autonomic Function Tests results and Reproducibility of Symptoms on Tilt.
This study was designed to investigate the frequency and pattern of orthostatic symptoms during head-up tilt (HUT) in patients with orthostatic intolerance during daily life, and to identify the relationship between the orthostatic symptoms during HUT and autonomic parameters. We prospectively collected autonomic data from 464 patients with orthostatic symptoms. Adrenergic and cardiovagal function tests including HUT were performed. Based on HUT results, we divided patients into orthostatic hypotension (OH), postural tachycardia syndrome (POTS), or normal HUT groups. We also investigated orthostatic symptoms during HUT. Only 25% of the patients reported orthostatic symptoms during HUT and 75% were asymptomatic. Typical orthostatic symptoms such as orthostatic dizziness and blurred vision, and atypical symptoms like chest tightness and headache occurred in 86% and 66%, respectively. Patients with POTS had symptoms more frequently than patients with OH during HUT. There were no differences in degrees of BP or HR changes between symptomatic and asymptomatic groups within the OH and POTS groups. HUT fails to reproduce symptoms of orthostatic intolerance in the majority of patients. Clinicians need to be aware that most patients with OH are asymptomatic during HUT and patients with POTS are more likely to be symptomatic than patients with OH.


Complementation of deletion mutations in a cloned functional cluster of Erwinia chrysanthemi out genes with Erwinia carotovora out homologues reveals OutC and OutD as candidate gatekeepers of species-specific secretion of proteins via the type II pathway.
The type II or Sec-dependent secretion system is used by diverse Gram-negative bacteria for secretion of extracellular proteins. Of the 12-15 proteins involved in secretion, the requirement for many has not been demonstrated and little is known about their functions in the secretion process. The plant pathogens Erwinia chrysanthemi and Erwinia carotovora secrete extra-cellular pectate lyases (Pels) using the type II or Out pathway. However, these two bacteria cannot secrete Pels encoded by heterologously expressed genes from the other species, suggesting the presence of species-specific recognition factors in the Out systems of the two Erwinia species. We previously reported the isolation of a cosmid clone, pCPP2OO6, from E. chrysanthemi EC16, which enables Escherichia coil to secrete heterologously expressed E. chrysanthemi Pels. Sequencing in a region required for secretion revealed the presence of 12 genes, outC-M and outO. We report here the construction of functionally non-polar mutations in each gene in the outC-M operon and outS and outB using a polA(ts) strain of E. coli to facilitate homologous recombination between out genes carrying deletions and their wild-type copies on pCPP2006. By testing for complementation of each deletion with wild-type out genes from E. chrysanthemi EC16 and E. carotovora SCRI193 we have demonstrated that: (i) each out gene is required for secretion of E. chrysanthemi PelE from E. coli with the exception of outH; (ii) each mutation can be complemented by its homologue from E. carotovora, except for outC and outD; (iii) outC and outD from E. carotovora do not confer secretion of Pel1 on the E. chrysanthemi Out system; and (iv) Pel1 secretion can be conferred on the E. chrysanthemi Out system by the presence of outC-M, S and B from E. carotovora. The data suggest that OutC and OutD are gatekeepers of the Out system involved in recognition of Pels targeted for secretion but that OutC and OutD from E. carotovora cannot be successfully assembled into the E. chrysanthemi Out system.


Bisphenol A and bisphenol S disruptions of the mouse placenta and potential effects on the placenta-brain axis.
Placental trophoblast cells are potentially at risk from circulating endocrine-disrupting chemicals, such as bisphenol A (BPA). To understand how BPA and the reputedly more inert bisphenol S (BPS) affect the placenta, C57BL6J mouse dams were fed 200 μg/kg body weight BPA or BPS daily for 2 wk and then bred. They continued to receive these chemicals until embryonic day 12.5, whereupon placental samples were collected and compared with unexposed controls. BPA and BPS altered the expression of an identical set of 13 genes. Both exposures led to a decrease in the area occupied by spongiotrophoblast relative to trophoblast giant cells (GCs) within the junctional zone, markedly reduced placental serotonin (5-HT) concentrations, and lowered 5-HT GC immunoreactivity. Concentrations of dopamine and 5-hydroxyindoleacetic acid, the main metabolite of serotonin, were increased. GC dopamine immunoreactivity was increased in BPA- and BPS-exposed placentas. A strong positive correlation between 5-HT+ GCs and reductions in spongiotrophoblast to GC area suggests that this neurotransmitter is essential for maintaining cells within the junctional zone. In contrast, a negative correlation existed between dopamine+ GCs and reductions in spongiotrophoblast to GC area ratio. These outcomes lead to the following conclusions. First, BPS exposure causes almost identical placental effects as BPA. Second, a major target of BPA/BPS is either spongiotrophoblast or GCs within the junctional zone. Third, imbalances in neurotransmitter-positive GCs and an observed decrease in docosahexaenoic acid and estradiol, also occurring in response to BPA/BPS exposure, likely affect the placental-brain axis of the developing mouse fetus.


Anti-inflammatory activity of clove (Eugenia caryophyllata) essential oil in human dermal fibroblasts.
Clove (Eugenia caryophyllata Thunb. [Myrtaceae]) essential oil (CEO) has been shown to possess antimicrobial, antifungal, antiviral, antioxidant, anti-inflammatory and anticancer properties. However, few studies have focused on its topical use. We investigated the biological activity of a commercially available CEO in a human skin disease model. We evaluated the effect of CEO on 17 protein biomarkers that play critical roles in inflammation and tissue remodelling in a validated human dermal fibroblast system, which was designed to model chronic inflammation and fibrosis. Four concentrations of CEO (0.011, 0.0037, 0.0012, and 0.00041%, v/v) were studied. The effect of 0.011% CEO on genome-wide gene expression was also evaluated. CEO at a concentration of 0.011% showed robust antiproliferative effects on human dermal fibroblasts. It significantly inhibited the increased production of several proinflammatory biomarkers such as vascular cell adhesion molecule-1 (VCAM-1), interferon γ-induced protein 10 (IP-10), interferon-inducible T-cell α chemoattractant (I-TAC), and monokine induced by γ interferon (MIG). CEO also significantly inhibited tissue remodelling protein molecules, namely, collagen-I, collagen-III, macrophage colony-stimulating factor (M-CSF), and tissue inhibitor of metalloproteinase 2 (TIMP-2). Furthermore, it significantly modulated global gene expression and altered signalling pathways critical for inflammation, tissue remodelling, and cancer signalling processes. CEO significantly inhibited VCAM-1 and collagen III at both protein and gene expression levels. This study provides important evidence of CEO-induced anti-inflammatory and tissue remodelling activity in human dermal fibroblasts. This study also supports the anticancer properties of CEO and its major active component eugenol.


[The developmental dynamics of the erythroid stem cell in rat hematopoietic organs in ontogeny].
The cellular composition of peripheral blood, hematopoietic organs, and the content of erythroblastic islands of the red bone marrow revealed regular changes of concentrations of the RBCs, hemoglobin and reticulocytes thus corroborating the phenomenon of "early postnatal anemia" in rats. A considerable increase in number of the erythroblastic islands was revealed between the 10th and 45th days of life. Functional reconstruction of the islands in the postnatal ontogenesis was analysed in rats.


Dry Eye and Quantitative and Qualitative Changes of Meibomian Glands in Patients With Pemphigus.
To evaluate dry eye and quantitative and qualitative changes of meibomian glands with meibography in patients with pemphigus. Twenty left eyes of 20 patients with pemphigus (group 1) and 20 left eyes of 20 age- and sex-matched healthy volunteers (group 2) were enrolled. Dry eye was evaluated with Oxford scale scoring, tear film breakup time, Schirmer 1 tests, and Ocular Surface Disease Index score assessments. Morphological changes and dropout ratio of meibomian glands were evaluated by upper and lower eyelid meibography (SL-D701; Topcon, IJssel, The Netherlands). Meibomian gland dropout ratio was scored for each eyelid from grade 0 (no loss) through grade 3 (lost area >2/3 of the total meibomian gland area). The mean ages of group 1 and group 2 were 51.8 ± 11.1 (range: 32-73) and 50.4 ± 9.6 years (range: 32-70), respectively (P = 0.672). Schirmer 1 and tear film breakup time values were lower and Oxford and Ocular Surface Disease Index scores were higher in group 1 when compared with group 2, and the differences were statistically significant (P = 0.01, P < 0.001, P = 0.001, and P < 0.001, respectively). Upper eyelid, lower eyelid, and total eyelid meiboscore values were significantly higher in group 1 (P = 0.005, P = 0.018, and P = 0.002, respectively). Morphological changes in meibomian glands were detected in 16 eyes (80%) among group 1. Between patients with pemphigus and those who were healthy, there were significant differences in the results of dry eye tests and meibomian gland morphology. Patients with pemphigus should be monitored for dry eye and meibomian gland dysfunction and be promptly treated to prevent the serious consequences of dry eye.


A heavy metal-associated protein (AcHMA1) from the halophyte, Atriplex canescens (Pursh) Nutt., confers tolerance to iron and other abiotic stresses when expressed in Saccharomyces cerevisiae.
Many heavy metals are essential for metabolic processes, but are toxic at elevated levels. Metal tolerance proteins provide resistance to this toxicity. In this study, we identified and characterized a heavy metal-associated protein, AcHMA1, from the halophyte, Atriplex canescens. Sequence analysis has revealed that AcHMA1 contains two heavy metal binding domains. Treatments with metals (Fe, Cu, Ni, Cd or Pb), PEG6000 and NaHCO3 highly induced AcHMA1 expression in A. canescens, whereas NaCl and low temperature decreased its expression. The role of AcHMA1 in metal stress tolerance was examined using a yeast expression system. Expression of the AcHMA1 gene significantly increased the ability of yeast cells to adapt to and recover from exposure to excess iron. AcHMA1 expression also provided salt, alkaline, osmotic and oxidant stress tolerance in yeast cells. Finally, subcellular localization of an AcHMA1/GFP fusion protein expressed in tobacco cells showed that AcHMA1 was localized in the plasma membrane. Thus, our results suggest that AcHMA1 encodes a membrane-localized metal tolerance protein that mediates the detoxification of iron in eukaryotes. Furthermore, AcHMA1 also participates in the response to abiotic stress.


Pars plana Molteno implantation in complicated inflammatory glaucoma.
We retrospectively studied pars plana Molteno implantation in seven patients who had previously undergone multiple attempts, both medical and surgical, to control intraocular pressure (IOP), to no avail. In an effort to avoid further anterior chamber complications, a Molteno implant was inserted through the superior temporal vitrectomy port, following a complete pars plana vitrectomy. Acuity decreased in two of the seven patients due to preexisting conditions and one patient lost vision. Preoperative IOPs ranged from 29 to 57 mm Hg (mean 41 mm Hg) and postoperative IOPs ranged from 7 to 20 mm Hg (mean 12 mm Hg).


In vivo imaging of beta cells with radiotracers: state of the art, prospects and recommendations for development and use.
Radiotracer imaging is characterised by high in vivo sensitivity, with a detection limit in the lower picomolar range. Therefore, radiotracers represent a valuable tool for imaging pancreatic beta cells. High demands are made of radiotracers for in vivo imaging of beta cells. Beta cells represent only a small fraction of the volume of the pancreas (usually 1-3%) and are scattered in the tiny islets of Langerhans throughout the organ. In order to be able to measure a beta cell-specific signal, one has to rely on highly specific tracer molecules because current in vivo imaging technologies do not allow the resolution of single islets in humans non-invasively. Currently, a considerable amount of preclinical data are available for several radiotracers and three are under clinical evaluation. We summarise the current status of the evaluation of these tracer molecules and put forward recommendations for their further evaluation.


Comparative genomic hybridization selection of blastocysts for repeated implantation failure treatment: a pilot study.
The aim of this study is to determine if the use of preimplantation genetic screening (PGS) by array comparative genomic hybridization (array CGH) and transfer of a single euploid blastocyst in patients with repeated implantation failure (RIF) can improve clinical results. Three patient groups are compared: 43 couples with RIF for whom embryos were selected by array CGH (group RIF-PGS), 33 couples with the same history for whom array CGH was not performed (group RIF NO PGS), and 45 good prognosis infertile couples with array CGH selected embryos (group NO RIF PGS). A single euploid blastocyst was transferred in groups RIF-PGS and NO RIF PGS. Array CGH was not performed in group RIF NO PGS in which 1-2 blastocysts were transferred. One monoembryonic sac with heartbeat was found in 28 patients of group RIF PGS and 31 patients of group NO RIF PGS showing similar clinical pregnancy and implantation rates (68.3% and 70.5%, resp.). In contrast, an embryonic sac with heartbeat was only detected in 7 (21.2%) patients of group RIF NO PGS. In conclusion, PGS by array CGH with single euploid blastocyst transfer appears to be a successful strategy for patients with multiple failed IVF attempts.


Different Clinical Course and Complications in Interagency Registry for Mechanically Assisted Circulatory Support 1 (INTERMACS) Patients Managed With or Without Extracorporeal Membrane Oxygenation.
Extracorporeal membrane oxygenation (ECMO) as a bridge to left ventricular assist device (LVAD) implantation has shown promise in improving end-organ function and optimizing outcomes in some critically ill patients, but the practice remains controversial. Retrospective review of patients who received LVADs from May 2008 to September 2016 at a high-volume, tertiary care cardiovascular center was performed. Subjects were Interagency Registry for Mechanically Assisted Circulatory Support (INTERMACS) class 1 patients divided into ECMO bridge and non-ECMO bridge cohorts. Patient demographics, adverse events, and survival at immediate and 1 year postoperative time points were compared between groups. In total, 235 patients received a HeartMate II or HVAD during the study period. Among INTERMACS 1 patients, 18 were ECMO bridge and 17 were non-ECMO bridge. Age, gender and bridge-to-transplant proportions (50% vs. 53%) were similar between groups. The ECMO bridge group had lower hemoglobin (7.9 ± 1.1 vs. 10.2 ± 2.2; p < 0.01), platelet (101 [70] vs. 176 [115]; p < 0.05), and prealbumin levels (10.6 ± 4.3 vs. 17.3 ± 7.7; p < 0.01). Nearly half (n = 8; 44%) of the ECMO bridge patients required packed red blood cell transfusions before VAD and were more likely to be on an epinephrine drip (78% vs. 12%; p < 0.01). However, along with these adjunctive measures, the ECMO bridge did effectively improve hemodynamic profiles by the time of VAD implant resulting in lower central venous pressure (7.7 ± 2.5 vs. 10.4 ± 4.8; p < 0.01) and mean pulmonary arterial pressure (18 ± 9 vs. 32 ± 8; p < 0.01). It also allowed for restoration of end-organ function as noted by comparable creatinine (1.0 [1.2] vs. 1.4 [0.6]) and total bilirubin levels (1.6 ± 1 vs.1.5 ± 1.7) between the two groups. There was no difference in rates of adverse events. Survival at 30 days postoperative and at 1 year (77% vs. 88%; p = 0.6) was similar. This study demonstrates that ECMO bridge is a central component of a multifaceted strategy for stabilization of select patients with severe hemodynamic instability before LVAD implantation. Further studies to optimize patient selection should be further explored.


The Prognostic Value of FGFR3 Expression in Patients with T1 Non-Muscle Invasive Bladder Cancer.
Fibroblast growth factor receptor 3 (FGFR3) alterations are frequent in non-muscle-invasive bladder cancer (NMIBC), although current data regarding the prognostic and therapeutic relevance are inconsistent. We analyzed the prognostic role of FGFR3 mRNA expression in stage T1 NMIBC. The mRNA expression of FGFR3 and cyclin-dependent kinase inhibitor 2A (CDKN2A) was measured by RT-qPCR in 80 patients with stage T1 NMIBC treated with transurethral resection of the bladder and correlated with clinical data and KRT5 and KRT20 expression, used as surrogate markers for basal and luminal subtypes, respectively. FGFR3 and CDKN2A transcript levels were not correlated. FGFR3 expression was associated with the expression of KRT5 (p=0.002) and KRT20 (p < 0.001). CDKN2A expression was negatively correlated with KRT5 (p=0.030). In Kaplan-Meier analysis and univariable Cox regression analysis, high FGFR3 expression was associated with significantly reduced recurrence-free survival (RFS) (HR=3.78; p < 0.001) and improved overall survival (OS) (HR=0.50; p=0.043), while high CDKN2A expression was associated with reduced OS (HR=2.34; p=0.034). Patient age was the only clinicopathological parameter associated with reduced OS (HR=2.29; p=0.022). No parameter was an independent prognostic factor in multivariable analysis. Next, we stratified the patients depending on their lineage differentiation. In univariable analysis, the prognostic effect of FGFR3 and CDKN2A was observed primarily in patients demonstrating high expression of KRT5 or KRT20, whereas high FGFR3 expression was associated with significantly reduced RFS, irrespective of instillation therapy. Stage T1 NMIBC patients with high FGFR3 expression show shorter RFS but better OS than patients with low FGFR3 expression.