Supporting practice-based audit: a price to be paid for collecting data.
There has been considerable investment by health authorities in the funding of support staff whose job is to collect data for audit purposes. It is important to understand what costs are involved in such a data collection exercise. The cost advantages of using existing practice staff or externally funded staff are not known. To assess the cost of transposing data on workload to computer software for audit purposes and retrieving data on five chronic diseases from case records. Four audit support staff monitored the time taken to collect specific data as part of a broad audit programme in 12 training practices within one health board area in the West of Scotland in 1997. The time taken was used to estimate comparative costs for using a receptionist or practice nurse for carrying out a similar exercise. Average costs for collecting data per 1000 patients for waiting time, appointments, recall, and telephone audits were 5.24 Pounds for reception staff, 5.64 Pounds for audit support staff, and 9.68 Pounds for a practice nurse. The average cost for collecting data per patient with diabetes, asthma, epilepsy, hypertension, or rheumatoid arthritis was 1.48 Pounds for reception staff, 1.60 Pounds for audit support staff, and 2.74 Pounds for a practice nurse. The cost of collecting data varies considerably depending on which staff are chosen for the purpose. Practices should consider carefully how best to collect data for audit in terms of cost.


[Change in pulmonary artery and aorta reactions of isobaric hypoxic pulmonary hypertension in guinea-pig and milrinone's effect].
The pulmonary hypertensive model was built on guinea-pig after 1 week of isobaric hypoxia. Meanwhile, the response of pulmonary artery ring (PAR) to noradrenaline (NA) increased significantly as compared with control, but it returned to normal level after 2 week of hypoxia. The similar change was not found in aorta ring (AR). The cumulative concentration response curves of PAR, AR to NA shifted to right with dose dependent relationship after Mil pretreated either control or hypoxic groups. Hypoxia decreased the sensitivity of AR to Mil significantly but not much on PAR.


Gammaherpesvirus small noncoding RNAs are bifunctional elements that regulate infection and contribute to virulence in vivo.
Many viruses express noncoding RNAs (ncRNAs). The gammaherpesviruses (γHVs), including Epstein-Barr virus, Kaposi's sarcoma-associated herpesvirus, and murine γHV68, each contain multiple ncRNA genes, including microRNAs (miRNAs). While these ncRNAs can regulate multiple host and viral processes in vitro, the genetic contribution of these RNAs to infection and pathogenesis remains largely unknown. To study the functional contribution of these RNAs to γHV infection, we have used γHV68, a small-animal model of γHV pathogenesis. γHV68 encodes eight small hybrid ncRNAs that contain both tRNA-like elements and functional miRNAs. These genes are transcribed by RNA polymerase III and are referred to as the γHV68 TMERs (tRNA-miRNA-encoded RNAs). To determine the total concerted genetic contribution of these ncRNAs to γHV acute infection and pathogenesis, we generated and characterized a recombinant γHV68 strain devoid of all eight TMERs. TMER-deficient γHV68 has wild-type levels of lytic replication in vitro and normal establishment of latency in B cells early following acute infection in vivo. In contrast, during acute infection of immunodeficient mice, TMER-deficient γHV68 has reduced virulence in a model of viral pneumonia, despite having an enhanced frequency of virus-infected cells. Strikingly, expression of a single viral tRNA-like molecule, in the absence of all other virus-encoded TMERs and miRNAs, reverses both attenuation in virulence and enhanced frequency of infected cells. These data show that γHV ncRNAs play critical roles in acute infection and virulence in immunocompromised hosts and identify these RNAs as a new potential target to modulate γHV-induced infection and pathogenesis. The gammaherpesviruses (γHVs) are a subfamily of viruses associated with chronic inflammatory diseases and cancer, particularly in immunocompromised individuals. These viruses uniformly encode multiple types of noncoding RNAs (ncRNAs) that are not translated into proteins. It remains unclear how virus-expressed ncRNAs influence the course and outcome of infection in vivo. Here, we generated a mouse γHV that lacks the expression of multiple ncRNAs. Notably, this mutant virus is critically impaired in the ability to cause disease in immunocompromised hosts yet shows a paradoxical increase in infected cells early during infection in these hosts. While the original mouse virus encodes multiple ncRNAs, the expression of a single domain of one ncRNA can partially reverse the defects of the mutant virus. These studies demonstrate that γHV ncRNAs can directly contribute to virus-induced disease in vivo and that these RNAs may be multifunctional, allowing the opportunity to specifically interfere with different functional domains of these RNAs.


Infection dynamics of the monogenean parasite Gyrodactylus gasterostei on sympatric and allopatric populations of the three-spined stickleback Gasterosteus aculeatus.
Parasites with high host specificity maximally depend on their hosts, which should increase the likelihood of coevolution. However, coevolution requires reciprocal selection exerted by the host and the parasite, and thus a considerable level of parasite virulence. In species of the monogenean ectoparasite genus Gyrodactylus consecutive generations are confronted with a single host, which may constrain the evolution of virulence. Transmission, which is often important in the ecology of Gyrodactylus species, may have the opposite effect, but may also lead to the avoidance of coevolutionary arms races. We investigated the potential outcome of coevolution between Gyrodactylus gasterostei Gläser, 1974 and its host, the three-spined stickleback (Gasterosteus aculeatus L.) by determining the strength of genotype by genotype (GxG) interactions on two levels: within and between sympatric and allopatric host populations. To do so, we compared the parasite's infection dynamics on laboratory-reared sympatric (Belgian) and allopatric (German) hosts. We found that a parasite line successfully infected a range of sympatric host genotypes (represented by families), while it failed to establish on allopatric hosts. Phylogeographic studies suggest that neutral genetic divergence between the host populations cannot explain this dramatic difference. Provided that this result can be generalised towards other parasite lines, we conclude that coevolution in this host-parasite system is more likely to lead to local adaptation on the population level than to GxG interactions within populations.


Metabolism of peptides by rumen microorganisms.
Rumen microorganisms utilize tryptic peptides from Chlorella protein, forming carbon dioxide, volatile fatty acids, and bacterial protein. Peptide carbon is more efficiently converted into bacterial protein than is amino acid carbon. A progressive degradation of the peptides was demonstrated by use of columns of Sephadex G-25.


Treatment of osteoarthritis using a helper-dependent adenoviral vector retargeted to chondrocytes.
Osteoarthritis (OA) is a joint disease characterized by degeneration of the articular cartilage, subchondral bone remodeling, and secondary inflammation. It is among the top three causes of chronic disability, and currently there are no treatment options to prevent disease progression. The localized nature of OA makes it an ideal candidate for gene and cell therapy. However, gene and cell therapy of OA is impeded by inefficient gene transduction of chondrocytes. In this study, we developed a broadly applicable system that retargets cell surface receptors by conjugating antibodies to the capsid of helper-dependent adenoviral vectors (HDVs). Specifically, we applied this system to retarget chondrocytes by conjugating an HDV to an α-10 integrin monoclonal antibody (a10mab). We show that a10mab-conjugated HDV (a10mabHDV)-infected chondrocytes efficiently in vitro and in vivo while detargeting other cell types. The therapeutic index of an intra-articular injection of 10mabHDV-expressing proteoglycan 4 (PRG4) into a murine model of post-traumatic OA was 10-fold higher than with standard HDV. Moreover, we show that PRG4 overexpression from articular, superficial zone chondrocytes is effective for chondroprotection in postinjury OA and that α-10 integrin is an effective protein for chondrocyte targeting.


A Comparison of Life Stress and Depressive Symptoms in Pregnant Taiwanese and Immigrant Women.
An increasing number of women from other countries, mostly Mainland China and Southeast Asia, are marrying Taiwanese husbands and settling in Taiwan. Immigration, marriage abroad, and pregnancy may be stressful and adversely affect maternal health. Relatively little research has compared the life stress and depressive symptoms of pregnant women of different ethnic groups living in nonmetropolitan areas in Taiwan. This study investigates the levels of life stress and depressive symptoms in pregnant Taiwanese women and Vietnamese "foreign brides" currently living in southern Taiwan. Eligible women in their last trimester of pregnancy who attended their local antenatal clinic were recruited for the study. Participants completed standardized measures, including the Difficult Life Circumstances Scale, Social Support APGAR Scale, and Edinburgh Postnatal Depression Scale. Two hundred thirty-six Taiwanese women and 44 Vietnamese women participated. Major life difficulties for both groups of women were related to their marital relationship, housing, or health problems. Taiwanese participants reported perceiving financial strain more often than their Vietnamese peers, whereas Vietnamese participants reported perceiving greater concerns regarding their children's development and about recent physical abuse than their Taiwanese peers. Furthermore, the Vietnamese participants reported less social support and higher rates of antenatal depression than Taiwanese participants. Clinical nurses and midwives should be sensitive to the particular difficulties and insufficient social support faced by pregnant women from different backgrounds in Taiwan. Women from foreign countries or those under unique challenging circumstances may face a particular risk of adverse outcomes. Identifying stresses informs the development of effective nursing interventions and support activities for new mothers and their families.


Long-term wildlife mortality surveillance in northern Congo: a model for the detection of Ebola virus disease epizootics.
Ebolavirus (EBOV) has caused disease outbreaks taking thousands of lives, costing billions of dollars in control efforts and threatening great ape populations. EBOV ecology is not fully understood but infected wildlife and consumption of animal carcasses have been linked to human outbreaks, especially in the Congo Basin. Partnering with the Congolese Ministry of Health, we conducted wildlife mortality surveillance and educational outreach in the northern Republic of Congo (RoC). Designed for EBOV detection and to alert public health authorities, we established a low-cost wildlife mortality reporting network covering 50 000 km2. Simultaneously, we delivered educational outreach promoting behavioural change to over 6600 people in rural northern RoC. We achieved specimen collection by training project staff on a safe sampling protocol and equipping geographically distributed bases with sampling kits. We established in-country diagnostics for EBOV testing, reducing diagnostic turnaround time to 3 days and demonstrated the absence of EBOV in 58 carcasses. Central Africa remains a high-risk EBOV region, but RoC, home to the largest remaining populations of great apes, has not had an epidemic since 2005. This effort continues to function as an untested early warning system in RoC, where people and great apes have died from past Ebola virus disease outbreaks. This article is part of the theme issue 'Dynamic and integrative approaches to understanding pathogen spillover'.


Chronic cough due to angiotensin-converting enzyme inhibitors.
The ACE-inhibiting drugs enalapril and captopril may result in a chronic and sometimes severe cough for which no pathologic cause can be found. Drug-induced cough should therefore be considered in any symptomatic patient taking these medications. In such cases, prompt withdrawal of the drug and substitution of a non-ACE inhibitor is curative and conserves the time and resources of the patient and the physician by avoiding unnecessary diagnostic and therapeutic measures.


Phenomenography: An emerging qualitative research design for nursing.
Phenomenography emerged from pedagogy to examine the qualitatively different ways that individuals experience and perceive the same phenomenon. Despite its uniqueness, the uptake of phenomenography in nursing research is still limited. Potentially, this may be related to confusion regarding what the design is about, its philosophical underpinnings and how distinct it is from other qualitative designs. To offer a better understanding of phenomenography by comparing it with other established qualitative research designs, examining its theoretical foundations, highlighting some studies that have employed the approach in nursing and offering methodological guidance to improve its uptake in nursing. Discussion paper. Compared to the traditional qualitative designs employed in nursing, phenomenography has been utilized in fewer studies. The ontological, epistemological and methodological basis of phenomenography highlights it as a distinct design. The strength of phenomenography lies in its emphasis on understanding the collective variations between participants and presenting these holistically as an 'outcome space'. Phenomenography is a distinct qualitative research approach that presents a unique opportunity for nursing to further its use. Issues regarding bracketing, the inclusion of phenomenography studies in qualitative meta-synthesis and employing a hermeneutic approach to phenomenography are avenues for further work in nursing. No patient or public contribution.


Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli.
A biosynthetic antibody binding site, which incorporated the variable domains of anti-digoxin monoclonal antibody 26-10 in a single polypeptide chain (Mr = 26,354), was produced in Escherichia coli by protein engineering. This variable region fragment (Fv) analogue comprised the 26-10 heavy- and light-chain variable regions (VH and VL) connected by a 15-amino acid linker to form a single-chain Fv (sFv). The sFv was designed as a prolyl-VH-(linker)-VL sequence of 248 amino acids. A 744-base-pair DNA sequence corresponding to this sFv protein was derived by using an E. coli codon preference, and the sFv gene was assembled starting from synthetic oligonucleotides. The sFv polypeptide was expressed as a fusion protein in E. coli, using a leader derived from the trp LE sequence. The sFv protein was obtained by acid cleavage of the unique Asp-Pro peptide bond engineered at the junction of leader and sFv in the fusion protein [(leader)-Asp-Pro-VH-(linker)-VL]. After isolation and renaturation, folded sFv displayed specificity for digoxin and related cardiac glycosides similar to that of natural 26-10 Fab fragments. Binding between affinity-purified sFv and digoxin exhibited an association constant [Ka = (3.2 +/- 0.9) x 10(7) M-1] that was about a factor of 6 smaller than that found for 26-10 Fab fragments [Ka = (1.9 +/- 0.2) x 10(8) M-1] under the same buffer conditions, consisting of 0.01 M sodium acetate, pH 5.5/0.25 M urea.


A novel role for C-C motif chemokine receptor 2 during infection with hypervirulent Mycobacterium tuberculosis.
C-C motif chemokine receptor 2 (CCR2) is a major chemokine axis that recruits myeloid cells including monocytes and macrophages. Thus far, CCR2-/- mice have not been found to be susceptible to infection with Mycobacterium tuberculosis (Mtb). Here, using a prototype W-Beijing family lineage 2 Mtb strain, HN878, we show that CCR2-/- mice exhibit increased susceptibility to tuberculosis (TB). Following exposure to Mtb HN878, alveolar macrophages (AMs) are amongst the earliest cells infected. We show that AMs accumulate early in the airways following infection and express CCR2. During disease progression, CCR2-expressing AMs exit the airways and localize within the TB granulomas. RNA-sequencing of sorted airway and non-airway AMs from infected mice show distinct gene expression profiles, suggesting that upon exit from airways and localization within granulomas, AMs become classically activated. The absence of CCR2+ cells specifically at the time of AM egress from the airways resulted in enhanced susceptibility to Mtb infection. Furthermore, infection with an Mtb HN878 mutant lacking phenolic glycolipid (PGL) expression still resulted in increased susceptibility in CCR2-/- mice. Together, these data show a novel role for CCR2 in protective immunity against clinically relevant Mtb infections.


Biosimilar Insulins: Basic Considerations.
Until now most of the insulin used in developed countries has been manufactured and distributed by a small number of multinational companies. Beyond the established insulin manufacturers, a number of new players have developed insulin manufacturing capacities based on modern biotechnological methods. Because the patents for many of the approved insulin formulations have expired or are going to expire soon, these not yet established companies are increasingly interested in seeking market approval for their insulin products as biosimilar insulins (BI) in highly regulated markets like the EU and the United States. Differences in the manufacturing process (none of the insulin manufacturing procedures are 100% identical) can lead to insulins that to some extent may differ from the originator insulin. The key questions are if subtle differences in the structure of the insulins, purity, and so on are clinically relevant and may result in different biological effects. The aim of this article is to introduce and discuss basic aspects that may be of relevance with regard to BI.


A substantial level of donor hematopoietic chimerism is required to protect donor-specific islet grafts in diabetic NOD mice.
Mixed chimerism can induce tolerance to alloantigens and restore self-tolerance to autoantigens, thereby permitting islet transplantation. However, the minimal level of donor chimerism that is required to prevent islet allograft rejection and recurrence of autoimmune diabetes has not been established. We investigated whether allogeneic Balb/c donor chimerism can be induced in C57BL/6 mice, in prediabetic NOD mice, and in diabetic NOD mice after transplantation of a modest dose of bone marrow by using purine nucleoside analogue, fludarabine phosphate and cyclophosphamide conditioning therapy, followed by short-term anti-CD40 ligand monoclonal antibody and rapamycin posttransplant treatment. We also investigated whether the induced donor chimerism is sufficient to prevent the onset of diabetes in prediabetic NOD mice and protect donor islet grafts in diabetic NOD mice. Allogeneic donor chimerism could be induced under the authors' approach that is nonmyeloablative and radiation-free. Diabetes onset was prevented in chimeric prediabetic NOD mice. The induction of mixed chimerism protected donor-specific islet grafts in diabetic NOD mice. At 60 days after islet transplantation, all donor Balb/c islet grafts survived in diabetic NOD mice whose level of donor-derived lymphocytes was higher than 30% at the time of islet transplantation (n=8). In contrast, Balb/c islet grafts were rejected in five of seven diabetic NOD mice whose level was lower than 30%. Our data demonstrate that a donor lymphocyte chimerism (>30%) at the time of islet transplantation is required to protect donor-specific islet grafts, and indicate that a strictly non-irradiation-based protocol can be used to achieve this goal.


Longitudinal changes in diary- and laboratory-based sleep measures in healthy "old old" and "young old" subjects: a three-year follow-up.
We report a longitudinal study of diary- and laboratory-based sleep measures in 50 healthy elderly subjects followed prospectively over a 3-year interval. Our hypothesis was that "old old" subjects (aged 75 to 87; n = 27) would show decline over time in measures of sleep quality, continuity, and depth, whereas "young old" subjects (aged 61 to 74; n = 23) were expected to show stability of outcome measures. Using analysis of variance-based planned contrast procedures, we found that this hypothesis was strongly supported for subjective sleep quality and laboratory measures of sleep latency, sleep efficiency, wakefulness after sleep onset, and slow-wave sleep percent. These changes were accompanied by increased napping in the old old. However, there was no change of habitual time in bed (total time or temporal placement of nighttime sleep), daily social rhythms, or sleep apnea. Change in medical burden scores did not correlate significantly with change in sleep efficiency or other outcome variables in the old old. Intervention designed to slow age-dependent decreases in sleep quality, continuity, and depth is discussed. The current results are representative of healthy elderly; sleep would probably deteriorate earlier and more quickly in elderly with more serious health problems and heavier medication use.


Epigenetic regulation of pancreatic islets.
Epigenetic mechanisms, including DNA methylation, histone modifications, and noncoding RNA expression, contribute to regulate islet cell development and function. Indeed, epigenetic mechanisms were recently shown to be involved in the control of endocrine cell fate decision, islet differentiation, β-cell identity, proliferation, and mature function. Epigenetic mechanisms can also contribute to the pathogenesis of complex diseases. Emerging knowledge regarding epigenetic mechanisms suggest that they may be involved in β-cell dysfunction and pathogenesis of diabetes. Epigenetic mechanisms could predispose to the diabetic phenotype such as decline of β-cell proliferation ability and β-cell failure, and account for complications associated with diabetes. Better understanding of epigenetic landscapes of islet differentiation and function may be useful to improve β-cell differentiation protocols and discover novel therapeutic targets for prevention and treatment of diabetes.


Y-Site Compatibility Studies of Ketoprofen with Parenteral Nutrition Admixtures for Central and Peripheral Administration.
Ketoprofen (KTF) is often used in hospital wards, especially in its intravenous form. According to the literature review, the compatibility of ketoprofen with parenteral nutrition (PN) admixtures has not yet been investigated. For this reason, we aimed to provide data contributing to physical compatibility to ensure the safe co-administration of these medications. In this study, we examined the compatibility of KTF with eight selected commercial PN admixtures intended for central (Lipoflex Special, Omegaflex Special, Kabiven, SmofKabiven) and peripheral (Lipoflex peri, Omegaflex peri, Kabiven Peripheral, Olimel Peri N4E) administration. The KTF solution for infusion was combined in three different volume ratios with studied PN admixtures reflecting the conditions in clinical practice. The evaluation of undesirable physical destabilization of oil-in-water system or precipitate formation involved the visual inspection and the determination of mean droplet diameter, zeta potential, pH, and turbidity changes. The results of compatibility of KTF with eight commercial PN admixtures showed that three of them: Kabiven, SmofKabiven, and Kabiven Peripheral, are incompatible with KTF and should not be concomitantly administered.


CTLA-4 Expression Is a Promising Biomarker of Idiopathic Pulmonary Arterial Hypertension and Allows Differentiation of the Type of Pulmonary Hypertension.
Pulmonary arterial hypertension (PAH) is an increasingly frequently diagnosed disease, the molecular mechanisms of which have not been thoroughly investigated. The aim of our study was to investigate subpopulations of lymphocytes to better understand their role in the molecular pathomechanisms of various types of PAH and to find a suitable biomarker that could be useful in the differential diagnosis of PAH. Using flow cytometry, we measured the frequencies of lymphocyte subpopulations CD4+CTLA-4+, CD8+ CTLA-4+ and CD19+ CTLA-4+ in patients with different types of PAH, namely pulmonary arterial hypertension associated with congenital heart disease (CHD-PAH), pulmonary arterial hypertension associated with connective tissue disorders (CTD-PAH), chronic thromboembolic pulmonary hypertension (CTEPH) and idiopathic pulmonary arterial hypertension (iPAH), and in an age- and sex-matched control group in relation to selected clinical parameters. Patients in the iPAH group had the significantly highest percentage of CD4+CTLA-4+ T lymphocytes among all PAH groups, as compared to those in the control group (p < 0.001), patients with CTEPH (p < 0.001), CTD-PAH (p < 0.001) and CHD-PAH (p < 0.01). In iPAH patients, the percentages of CD4+CTLA-4+ T cells correlated strongly positively with the severity of heart failure New York Heart Association (NYHA) Functional Classification (r = 0.7077, p < 0.001). Moreover, the percentage of B CD19+CTLA-4+ cells strongly positively correlated with the concentration of NT-proBNP (r = 0.8498, p < 0.001). We have shown that statistically significantly higher percentages of CD4+CTLA-4+ (p ≤ 0.01) and CD8+ CTLA-4+ (p ≤ 0.001) T cells, measured at the time of iPAH diagnosis, were found in patients who died within 5 years of the diagnosis, which allows us to consider both of the above lymphocyte subpopulations as a negative prognostic/predictive factor in iPAH. CTLA-4 may be a promising biomarker of noninvasive detection of iPAH, but its role in planning the treatment strategy of PAH remains unclear. Further studies on T and B lymphocyte subsets are needed in different types of PAH to ascertain the relationships that exist between them and the disease.


Medicine's inconvenient truth: the placebo and nocebo effect.
Placebo and nocebo effects are often regarded by clinicians as either a quaint reminiscence from the pre-therapeutic era, or simply as a technique for establishing the efficacy of therapeutic interventions within the locus of evidence-based practice. However, neither of these explanations sufficiently account for their complexity or their persistence and impact in clinical medicine. Placebo and nocebo effects are embedded in the very fabric of therapeutic relationships and are both a manifestation and outcome of the rituals that characterise clinical practice. They are also a stark reminder of the many personal and environmental factors, including the attitudes, beliefs and expectations of both doctor and patient, that shape the outcomes of health professional-patient interactions. We describe how recent biological and neuropsychiatric data have clarified the operation of placebo and nocebo effects in clinical practice - demonstrating the ability of the therapeutic context to modulate endogenous biological processes in a targeted manner. This, in turn, illustrates the potent philosophical and sociocultural aspects of medical praxis.


Genetic reduction of vascular endothelial growth factor receptor 2 rescues aberrant angiogenesis caused by epsin deficiency.
We previously showed that endothelial epsin deficiency caused elevated vascular endothelial growth factor receptor 2 (VEGFR2) and enhanced VEGF signaling, resulting in aberrant tumor angiogenesis and reduced tumor growth in adult mice. However, direct evidence demonstrating that endothelial epsins regulate angiogenesis specifically through VEGFR2 downregulation is still lacking. In addition, whether the lack of epsins causes abnormal angiogenesis during embryonic development remains unclear. A novel strain of endothelial epsin-deleted mice that are heterozygous for VEGFR2 (Epn1(fl/fl); Epn2(-/-); Flk(fl/+); iCDH5 Cre mice) was created. Analysis of embryos at different developmental stages showed that deletion of epsins caused defective embryonic angiogenesis and retarded embryo development. In vitro angiogenesis assays using isolated primary endothelial cells (ECs) from Epn1(fl/fl); Epn2(-/-); iCDH5 Cre (EC-iDKO) and Epn1(fl/fl); Epn2(-/-); Flk(fl/+); iCDH5 Cre (EC-iDKO-Flk(fl/+)) mice demonstrated that VEGFR2 reduction in epsin-depleted cells was sufficient to restore normal VEGF signaling, EC proliferation, EC migration, and EC network formation. These findings were complemented by in vivo wound healing, inflammatory angiogenesis, and tumor angiogenesis assays in which reduction of VEGFR2 was sufficient to rescue abnormal angiogenesis in endothelial epsin-deleted mice. Our results provide the first genetic demonstration that epsins function specifically to downregulate VEGFR2 by mediating activated VEGFR2 internalization and degradation and that genetic reduction of VEGFR2 level protects against excessive angiogenesis caused by epsin loss. Our findings indicate that epsins may be a potential therapeutic target in conditions in which tightly regulated angiogenesis is crucial, such as in diabetic wound healing and tumors.


Comparison of absorbable and permanent sutures for laparoscopic sacrocervicopexy: A randomized controlled trial.
Pelvic organ prolapse is a common cause of morbidity and decreased quality of life among women and is treatable by laparoscopic sacrocolpopexy. Recent data suggest that absorbable sutures are a feasible and appealing option for mesh attachment given a potential decreased risk of complications related to mesh erosion. The aim of the present study was to demonstrate the non-inferiority of absorbable sutures to permanent sutures for laparoscopic sacrocervicopexy. We performed a randomized, single-blinded, non-inferiority trial comparing late-absorbable sutures (group A) to non-absorbable sutures (group B) for anterior and posterior vaginal mesh fixation during laparoscopic sacrocervicopexy at a single center in Italy. The primary outcome was prolapse correction at 12 months after surgery, defined as the absence of a pelvic organ prolapse leading edge reaching or extending below the level of the hymen and the absence of bulge symptoms. Secondary outcomes included intraoperative parameters, postoperative characteristics, and long-term morbidity. Statistical analyses were performed using STATA version 16. A total of 150 patients with pelvic organ prolapse were prospectively randomized 1:1 into two groups (A or B). Baseline characteristics and intraoperative parameters including blood loss, operation time, and intraoperative complications were comparable between groups. The success rate was 100% in both groups and no differences in prolapse correction were observed. The rates of de novo urinary incontinence and persistent urinary incontinence were also similar between groups. The rate of mesh erosion at 12 months was 0% in group A and 4% in group B (P = .24). Late absorbable sutures are non-inferior to non-absorbable sutures for laparoscopic sacrocervicopexy in terms of procedural success. Moreover we did not see any differences in terms of operative parameters, or intraoperative and postoperative characteristics, although the study was not powered to these outcomes.


Nitrosylation mechanisms of Mycobacterium tuberculosis and Campylobacter jejuni truncated hemoglobins N, O, and P.
Truncated hemoglobins (trHbs) are widely distributed in bacteria and plants and have been found in some unicellular eukaryotes. Phylogenetic analysis based on protein sequences shows that trHbs branch into three groups, designated N (or I), O (or II), and P (or III). Most trHbs are involved in the O2/NO chemistry and/or oxidation/reduction function, permitting the survival of the microorganism in the host. Here, a detailed comparative analysis of kinetics and/or thermodynamics of (i) ferrous Mycobacterium tuberculosis trHbs N and O (Mt-trHbN and Mt-trHbO, respectively), and Campylobacter jejuni trHb (Cj-trHbP) nitrosylation, (ii) nitrite-mediated nitrosylation of ferrous Mt-trHbN, Mt-trHbO, and Cj-trHbP, and (iii) NO-based reductive nitrosylation of ferric Mt-trHbN, Mt-trHbO, and Cj-trHbP is reported. Ferrous and ferric Mt-trHbN and Cj-trHbP display a very high reactivity towards NO; however, the conversion of nitrite to NO is facilitated primarily by ferrous Mt-trHbN. Values of kinetic and/or thermodynamic parameters reflect specific trHb structural features, such as the ligand diffusion pathways to/from the heme, the heme distal pocket structure and polarity, and the ligand stabilization mechanisms. In particular, the high reactivity of Mt-trHbN and Cj-trHbP reflects the great ligand accessibility to the heme center by two protein matrix tunnels and the E7-path, respectively, and the penta-coordination of the heme-Fe atom. In contrast, the heme-Fe atom of Mt-trHbO the ligand accessibility to the heme center of Mt-trHbO needs large conformational readjustments, thus limiting the heme-based reactivity. These results agree with different roles of Mt-trHbN, Mt-trHbO, and Cj-trHbP in vivo.


Self-determination and first-episode psychosis: associations with symptomatology, social and vocational functioning, and quality of life.
Self-determination theory (SDT) postulates that satisfaction of three basic psychological needs (i.e., autonomy, competence, and well-being) promotes motivation, well-being, and growth across domains of functioning. Thus, per SDT, we examined satisfaction of basic psychological needs among individuals with first-episode psychosis. First, we quantified the level of need satisfaction among a sample of individuals with first-episode psychosis and compared their level of need satisfaction to that of individuals without psychosis. Second, we examined the association between need satisfaction and several domains of well-being among individuals with first-episode psychosis (i.e., symptomatology, social/vocational functioning, and quality of life). Our results indicated that individuals with first-episode psychosis experience less satisfaction of basic psychological needs as compared to their same-aged counterparts. There was a modest association between need satisfaction and well-being among individuals with first-episode, with the need of relatedness being the need most frequently associated with indices of well-being. Although modest in scope, the results of the current study raise the possibility that further investigation of SDT among individuals with first-episode psychosis may reveal important strategies through which early intervention services can better promote well-being and recovery.


Abortion in Brazil: what do the official data say?
According to the World Health Organization, from 2010 to 2014, there were around 55 million abortions worldwide, 45% of which were unsafe. In Brazil, data on abortion and its complications are incomplete. Health care data are only available for the public sector and mortality data depend on investigations of deaths. This study sought to describe the situation of abortion in the country using public data available in the different Information Systems - SIM (mortality), SINASC (live births) and SIH (hospitalization). From 2008 to 2015, there were around 200,000 hospitalizations/year for procedures related to abortion, 1,600 of which for medical and legal reasons. From 2006 to 2015, we found 770 maternal deaths in SIM whose underlying cause was abortion. There was a discreet reduction in the number of deaths from abortion in the period, with regional variation. This number could be increased by around 29% per year if deaths with mentions of abortion and declared with a different underlying cause were considered. Among the deaths reported as resulting from abortion, 1% were abortions due to medical and legal reasons and 56.5% were non-specified abortions. The proportion of deaths from abortion identified in SIH, in relation to the total number of deaths from abortion identified in SIM, varied between 47.4% in 2008 and 72.2% in 2015. Although official health data do not allow us to estimate the number of abortions in Brazil, we were able to establish the profile of women at higher risk for death from abortion: black and indigenous women, with low educational levels, under 14 and over 40 years of age, living in the North Northeast and Central regions, without a partner.


A Cluster-Randomized Trial of Hydroxychloroquine for Prevention of Covid-19.
Current strategies for preventing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection are limited to nonpharmacologic interventions. Hydroxychloroquine has been proposed as a postexposure therapy to prevent coronavirus disease 2019 (Covid-19), but definitive evidence is lacking. We conducted an open-label, cluster-randomized trial involving asymptomatic contacts of patients with polymerase-chain-reaction (PCR)-confirmed Covid-19 in Catalonia, Spain. We randomly assigned clusters of contacts to the hydroxychloroquine group (which received the drug at a dose of 800 mg once, followed by 400 mg daily for 6 days) or to the usual-care group (which received no specific therapy). The primary outcome was PCR-confirmed, symptomatic Covid-19 within 14 days. The secondary outcome was SARS-CoV-2 infection, defined by symptoms compatible with Covid-19 or a positive PCR test regardless of symptoms. Adverse events were assessed for up to 28 days. The analysis included 2314 healthy contacts of 672 index case patients with Covid-19 who were identified between March 17 and April 28, 2020. A total of 1116 contacts were randomly assigned to receive hydroxychloroquine and 1198 to receive usual care. Results were similar in the hydroxychloroquine and usual-care groups with respect to the incidence of PCR-confirmed, symptomatic Covid-19 (5.7% and 6.2%, respectively; risk ratio, 0.86 [95% confidence interval, 0.52 to 1.42]). In addition, hydroxychloroquine was not associated with a lower incidence of SARS-CoV-2 transmission than usual care (18.7% and 17.8%, respectively). The incidence of adverse events was higher in the hydroxychloroquine group than in the usual-care group (56.1% vs. 5.9%), but no treatment-related serious adverse events were reported. Postexposure therapy with hydroxychloroquine did not prevent SARS-CoV-2 infection or symptomatic Covid-19 in healthy persons exposed to a PCR-positive case patient. (Funded by the crowdfunding campaign YoMeCorono and others; BCN-PEP-CoV2 ClinicalTrials.gov number, NCT04304053.).


Biofunctional porous anodized titanium implants for enhanced bone regeneration.
Efficient osseointegration is a key factor in dental implants to reduce the total time-course of therapy. Titanium implants with anodized surface gained much interest for their enhanced osseointegration. Anodized implant combined with bioactive drugs is an ideal candidate for enhance bone regeneration. Previously delivery of drugs from the metal implants has been attempted by utilizing a polymeric dip-coating method. However, the entire surface coating with polymer can diminish the advantageous surface roughness of anodized implants and cause contact inhibition between bone and implant surface. In this study, fibroblast growth factor-2 (FGF-2) loaded poly(lactide-co-glycolide) nanoparticles were partially coated on anodized Ti discs by an electrospray deposition. Nanoparticle coated anodized discs maintained their native porous structure and provided a sustained release of FGF-2 for more than 2 weeks with 40% initial burst. In vitro study confirmed the influence of polymeric nanoparticles and the release of growth factors from the Ti disc. Nanoparticle-coated groups significantly enhanced cell spreading and differentiation. For in vivo evaluation, the anodized titanium implants were applied to rabbit tibia model. The osseointegration was estimated by bone to implant contact of best three consecutive threads at the border of the implant. The mean osteointegration value of FGF-2 releasing implant groups (70.1%) was significantly higher than that of untreated implants (47.1%). We believe that the electrospray deposition technique is a particularly attractive approach for the coating of medical devices with porous surface to maintain their surface topography while allowing a sustained delivery of growth factors for bone regeneration. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 3639-3648, 2014.


Individual, interpersonal, and organisational factors associated with discrimination in medical decisions affecting people with a migration background with mental health problems: the case of general practice.
Although people with a migration background (MB) have more unmet mental health needs than the general population, patients with a MB are still underrepresented in mental health care services. Provider bias towards these patients has been evidenced repeatedly but its driving factors remain elusive. We assessed the moderating effect of the individual (e.g. age and ethnicity), interpersonal (e.g. healthcare provider trust), and organisational (e.g. perceived workload) factors on general practitioners (GPs) differential decision-making regarding diagnosis, treatment, and referral for a depressed patient with or without a MB. An experimental study was carried out in which GPs were shown one of two video vignettes featuring adult male depressed patients, one with a MB and the other without. Belgian GPs (n = 797, response rate was 13%) had to decide on their diagnosis, treatment, and referral. Analysis of variance and logistic regression were used to analyse the effect of a MB, adding interaction terms for the explanatory variables. Overall, we found that there were ethnic differences in GPs' decisions regarding diagnosis and treatment recommendations. GPs perceived the symptoms of the patient with a MB as less severe (F = 7.68, p < 0.01) and demonstrated a reduced likelihood to prescribe a combination of medical and non-medical treatments (F = 11.55, p < 0.001). Those differences increased in accordance with the GP's age and perceived workload; at an interpersonal level, we found that differences increased when the GP thought the patient was exaggerating his distress. This paper showed that lower levels of trust among GPs' towards their migrant patients and high GP workloads contribute to an increased ethnic bias in medical decision-making. This may perpetuate ethnic inequalities in mental health care. Future researchers should develop an intervention to decrease the ethnic inequities in mental health care by addressing GPs' trust in their migrant and ethnic minority patients.


[Risk factor analysis of systemic inflammatory response syndrome in type 2 diabetics after percutaneous nephrolithotomy].
To evaluate the risk factors of systemic inflammatory response syndrome (SIRS) in the patients with type 2 diabetes diagnosed with kidney stone after percutaneous nephrolithotomy (PCNL). In this study, 461 patients with type 2 diabetes who received PCNL in Peking University People's Hospital from June 2006 to December 2015 were reviewed. There were 281 males and 180 females with an average age of 57 years were included, of whom, 137 were diagnosed with SIRS after PCNL. The demographic data, clinical features, and test results were compared between the patients with SIRS and without SIRS, trying to identify the correlation between their clinical characters and the occurrence of SIRS. The SIRS was significantly correlated with the patients' preoperative white blood cell counting (×109/L) [7.76 (4.00-17.96) vs. 6.31 (2.00-17.40), P<0.001], preoperational blood glucose level (mmol/L) [7.30 (3.08-19.90) vs. 6.40 (3.42-16.78), P<0.001], operative time (min) [75 (20-270) vs. 60 (20-200), P<0.001], length of stay (d) [12 (2-46) vs. 11 (3-29), P=0.019], staghorn stones [38.8% (33/85) vs. 27.7% (104/376), P=0.042], and preoperational urinary tract infection [36.8% (50/136) vs. 26.6% (81/304), P=0.032]. There was no significant correlation between the SIRS and the patients' age, body mass index, preoperative hemoglobin level, preoperative serum creatinine, and transfusion. In multivariate analysis, abnormal preoperative white blood cell counting (OR=3.194, 95% CI: 1.531-6.666, P=0.002), operative time longer than 60 min (OR=1.635, 95% CI: 1.088-2.456, P=0.018), and preoperational blood glucose level higher than normal 7.1 mmol/L were significantly correlated with the presence of SIRS. The high level of preoperational blood glucose, abnormal preoperative white blood cell counting, and long operative time were significantly correlated with the presence of SIRS in patients with type 2 diabetes after PCNL.


A phase II study of V-BEAM as conditioning regimen before second auto-SCT for multiple myeloma.
High-dose melphalan has been the standard conditioning regimen for auto-SCT in multiple myeloma (MM) for decades. A more effective conditioning regimen may induce deeper responses and longer remission duration. It is especially needed in the setting of second auto-SCT, which rarely achieves comparable results with the first auto-SCT using the same conditioning regimen. Here we conducted a phase II study to investigate the efficacy and safety of a conditioning regimen V-BEAM (bortezomib-BEAM) before second auto-SCT for multiple myeloma. Ten patients were enrolled from September 2012 to May 2013. The CR rate at day +100 after auto-SCT was 75%; all except for one patient remained in remission after a median follow-up of 6 months. Three patients developed Clostridium difficile infection. Two patients died within the first 30 days of auto-SCT from neutropenic colitis and overwhelming sepsis, respectively. Due to the high rate of morbidity and mortality, the study was terminated after 10 patients. In summary, although the conditioning regimen V-BEAM before second auto-SCT for MM provided promising responses, it was associated with unexpected treatment-related toxicity and should not be investigated further without modifications.


LIMK-1 and LIMK-2, two members of a LIM motif-containing protein kinase family.
We previously isolated human cDNA encoding LIM-kinase (LIMK), a putative protein kinase which contains two repeats of the LIM motif at the N-terminus and a protein kinase consensus sequence at the C-terminus. Using as a probe a cDNA fragment of human LIMK, we isolated from a rat brain cDNA library cDNA clones encoding two distinct protein kinases (termed LIMK-1 and LIMK-2) related to human LIMK. LIMK-1 shares with human LIMK 95% of the total 647 amino acids and is probably a rat equivalent of human LIMK. LIMK-2 has an overall sequence and a domain structure similar to that of human LIMK and rat LIMK-1, but overall identity is 50-51% at the amino acid level. Like human LIMK, the protein kinase domains of rat LIMK-1 and -2 contain a characteristic sequence DLNSHN in subdomain VIB and a highly basic insert between subdomain VII and VIII. LIMK-1 and -2 are therefore closely related but distinct members of a novel LIM-containing protein kinase subfamily. Several forms of LIMK-2 transcripts encoding proteins that are N-terminally modified and/or C-terminally truncated are generated by alternative splicing or alternative initiation. Northern blot analysis revealed the expression of LIMK-1 mRNA predominantly in the brain and the expression of LIMK-2 mRNA in various tissues in the rat. Antibody raised against LIMK-1 specifically immunoprecipitated and identified in Rat2 fibroblast cells a 72 kDa protein, which has no detectable autophosphorylating activity but is capable of phosphorylating serine and threonine residues of myelin basic protein, by in vitro kinase reaction. As the LIMK family kinases have unique structural features, they are likely to have specific functions in previously uncharacterized signaling pathways.


Reflections on the public interpretation of regional environmental history in western Pennsylvania.
This essay attempts to counter the scarcity of efforts to address issues of natural resource extraction and environmental exploitation in public history forums. Focused on western Pennsylvania, it argues that the history of industrial development and its deleterious environmental impacts demands a regional vision that not only frames these stories within the ideological and economic context of the past, but also challenges residents and visitors to consider this history in light of the related environmental concerns of our own time. The essay explores some of the difficult issues faced by public historians and practitioners as they seek to produce public environmental histories that do not elude opportunities to link past and present in meaningful ways.


Pharmacological characterization of the nociceptin receptor, ORL1. Insight from the inward rectifier activation in the periaqueductal gray.
A novel opioid receptor-like orphan receptor (ORL1) was cloned and identified to be homologous to classical opioid receptors but insensitive to traditional opioids. A heptadecapeptide, termed orphanin FQ or nociceptin (OFQ/N), was identified as its endogenous ligand. OFQ/N shares overlapping distribution sites in pain-processing areas and common cellular mechanisms with opioids but exerts diverse effects on nociceptive responses. Of the two reported ORL1 antagonists, [Phe(1)psi(CH(2)-NH)- Gly(2)] nociceptin-(1-13)-NH(2) (Phepsi) and naloxone benzoylhydrazone (NBZ), antagonisms were validated in the activation of inward rectifying K channels induced by OFQ/N, using the patch clamp technique in ventrolateral periaqueductal gray slices. Results showed that Phepsi acted as a partial agonist and NBZ was a weak nonselective antagonist of ORL1. It is comparable with most but not all of the findings from other tissues. Comparing all the reports supports the above inference for these two antagonists. The possible causes for the discrepancy were discussed. A brief review on the putative ORL1 antagonists, acetyl-RYYRIK-NH2, some sigma-ligands and the functional antagonist, nocistatin, is also included. It indicates that a potent and selective ORL1 antagonist is expecting to elucidate the physiological role of OFQ/N.


Is endoscopic resection a useful technique for a cavernous sinus sellar cavernoma? A case report and literature review.
Cavernous sinus cavernous malformations (CSCMs) is a vascular malformation of the cavernous sinus. Nowadays, there is an increasing preference to withhold using the terms cavernoma or cavernous hemangioma in order to stop considering these lesions as vascular neoplasms. These lesions are highly vascularized making surgical resection a challenge, mainly in endoscopic approaches. We present a case of this tumor treated in our institution with an endoscopic endonasal approach and incomplete resection. Because of the strenuous resection through this approach, we systematically reviewed the reported endoscopic cases of CSCMs to determine their intraoperative complications, results and tumor features. Using the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines, one database (PubMed) and crossed references were queried for CSCMs from 1948 to 2020. Data regarding demographic features, clinical presentation, MRI features, surgical results and overall pathology features extracted. Eighteen patients were selected (including our case). The mean age was 50.4 ± 14 years. Pituitary dysfunction and cavernous sinus nerve compression were the most reported symptoms. Only five cases (27%) reported a gross total resection (GTR) through endoscopic endonasal approach. Intraoperative bleeding was the most frequent intraoperative complication. We present a comprehensive analysis of every reported CSCM treated through endoscopic approach. Partial or subtotal resection are the most used techniques because of the intraoperative bleeding and the adherence to surrounding structures. Radiotherapy is a very good option for patients with incomplete resections.


Supporting Weight Management during COVID-19: A Randomized Controlled Trial of a Web-Based, ACT-Based, Guided Self-Help Intervention.
Adults with overweight and obesity are vulnerable to weight gain and mental health deterioration during the COVID-19 pandemic. We developed a web-based, guided self-help intervention based on Acceptance and Commitment Therapy (ACT) that aims to support adults with overweight and obesity to prevent weight gain by helping them to manage their eating behaviours, be more physically active, and protect their emotional wellbeing ("SWiM-C"). SWiM-C is a guided self-help programme using non-specialist guides to enhance scalability and population reach while minimizing cost. This study evaluated the effect of SWiM-C on bodyweight, eating behaviour, physical activity, and mental wellbeing in adults with overweight and obesity over 4 months during the COVID-19 pandemic in the UK. We randomized adults (BMI ≥25 kg/m2) to SWiM-C or to a wait-list standard advice group. Participants completed outcome assessments online at baseline and 4 months. The primary outcome was self-measured weight; secondary outcomes were eating behaviour, physical activity, experiential avoidance/psychological flexibility, depression, anxiety, stress, and wellbeing. We estimated differences between study groups in change in outcomes from baseline to 4 months using linear regression, adjusted for outcome at baseline and the randomization stratifiers (BMI, sex). The trial was pre-registered (ISRCTN12107048). 486 participants were assessed for eligibility; 388 participants were randomized (196 standard advice, 192 SWiM-C), and 324 were analysed. The adjusted difference in weight between SWiM-C and standard advice was -0.60 kg (-1.67 to 0.47, p = 0.27). SWiM-C led to improvements in uncontrolled eating (-3.61 [-5.94 to -1.28]), cognitive restraint (5.28 [2.81-7.75]), experiential avoidance (-3.39 [-5.55 to -1.23]), and wellbeing (0.13 [0.07-0.18]). SWiM-C improved several psychological determinants of successful weight management and had a protective effect on wellbeing during the pandemic. However, differences in weight and some other outcomes were compatible with no effect of the intervention, suggesting further refinement of the intervention is needed.


Acute and sub-acute metabolic change in different brain regions induced by moderate blunt traumatic brain injury.
The objective of the study was to observe the effect of moderate closed-head injury on hippocampal, thalamic, and striatal tissue metabolism with time. Closed head injury is responsible for metabolic changes. These changes can be permanent or temporary, depending on the injury's impact. For the experiment, 20 rats were randomly divided into four groups, each containing five animals. Animals were subjected to injury using a modified Marmarou's weight drop device; hippocampal, thalamic, and striatal tissue samples were collected after 1 day, 3 days, and 7 days of injury. NMR spectra were acquired following sample processing. Changes in myo-inositol, creatine, glutamate, succinate, lactate, and N-acetyl aspartic acid in hippocampal tissues were observed at day 3 PI. The tyrosine level in the hippocampus was altered at day 7 PI. While thalamic and striatal tissue samples showed altered levels of branched-chain amino acids and myo-inositol at day 1PI. Taurine, gamma amino butyric acid (GABA), choline, and alpha keto-glutarate levels were found to be significantly altered in striatal tissues at days 1 and 3PI. Acetate and GABA levels were altered in the thalamus on day 1 PI. The choline level in the thalamus was found to alter at all-time points after injury. The alteration in these metabolites may be due to the alteration in their respective pathways. Neurotransmitter and energy metabolism pathways were found to be altered in all three brain regions after TBI. This study may help better understand the effect of injury on the metabolic balance of a specific brain region and recovery.


Parkin facilitates the elimination of expanded polyglutamine proteins and leads to preservation of proteasome function.
Parkin, the most commonly mutated gene in familial Parkinson's disease, encodes an E3 ubiquitin ligase. A number of candidate substrates have been identified for parkin ubiquitin ligase action including CDCrel-1, o-glycosylated alpha-synuclein, Pael-R, and synphilin-1. We now show that parkin promotes the ubiquitination and degradation of an expanded polyglutamine protein. Overexpression of parkin reduces aggregation and cytotoxicity of an expanded polyglutamine ataxin-3 fragment. Using a cellular proteasome indicator system based on a destabilized form of green fluorescent protein, we demonstrate that parkin reduces proteasome impairment and caspase-12 activation induced by an expanded polyglutamine protein. Parkin forms a complex with the expanded polyglutamine protein, heat shock protein 70 (Hsp70) and the proteasome, which may be important for the elimination of the expanded polyglutamine protein. Hsp70 enhances parkin binding and ubiquitination of expanded polyglutamine protein in vitro suggesting that Hsp70 may help to recruit misfolded proteins as substrates for parkin E3 ubiquitin ligase activity. We speculate that parkin may function to relieve endoplasmic reticulum stress by preserving proteasome activity in the presence of misfolded proteins. Loss of parkin function and the resulting proteasomal impairment may contribute to the accumulation of toxic aberrant proteins in neurodegenerative diseases including Parkinson's disease.


Myostatin inhibits proliferation and insulin-stimulated glucose uptake in mouse liver cells.
Although myostatin functions primarily as a negative regulator of skeletal muscle growth and development, accumulating biological and epidemiological evidence indicates an important contributing role in liver disease. In this study, we demonstrate that myostatin suppresses the proliferation of mouse Hepa-1c1c7 murine-derived liver cells (50%; p < 0.001) in part by reducing the expression of the cyclins and cyclin-dependent kinases that elicit G1-S phase transition of the cell cycle (p < 0.001). Furthermore, real-time PCR-based quantification of the long noncoding RNA metastasis associated lung adenocarcinoma transcript 1 (Malat1), recently identified as a myostatin-responsive transcript in skeletal muscle, revealed a significant downregulation (25% and 50%, respectively; p < 0.05) in the livers of myostatin-treated mice and liver cells. The importance of Malat1 in liver cell proliferation was confirmed via arrested liver cell proliferation (p < 0.05) in response to partial Malat1 siRNA-mediated knockdown. Myostatin also significantly blunted insulin-stimulated glucose uptake and Akt phosphorylation in liver cells while increasing the phosphorylation of myristoylated alanine-rich C-kinase substrate (MARCKS), a protein that is essential for cancer cell proliferation and insulin-stimulated glucose transport. Together, these findings reveal a plausible mechanism by which circulating myostatin contributes to the diminished regenerative capacity of the liver and diseases characterized by liver insulin resistance.


Regulatory polymorphisms in the interleukin-18 promoter are associated with hepatitis C virus clearance.
The immune response is critical in determining the outcome of hepatitis C virus (HCV) infection. Interleukin (IL)-18 is a pivotal mediator of the Th1/Th2-driven immune response. Two IL-18 promoter polymorphisms (-607C/A and -137G/C) and their haplotypes were known to affect IL-18 expression. We examined the role played by these polymorphisms in determining HCV clearance or persistence. Genotyping was performed among African American injection drug users with HCV clearance (n = 91) or HCV persistence (n = 182) and among European Americans with hemophilia who were mainly infected through plasma transfusion. Among injection drug users, IL18 -607A (odds ratio [OR], 3.68 [95% confidence interval {CI},1.85-7.34]) and IL18 -137C (OR, 2.33 [95% CI, 1.24-4.36]) were significantly associated with HCV clearance. A haplotype carrying -607A and -137C (OR, 4.53 [95% CI, 1.77-11.6]) was also strongly associated with viral clearance. No association was found among those with hemophilia. These results suggest that IL18 promoter polymorphism may affect the outcome of HCV infection in certain groups.


Comparing age at onset of major depression and other psychiatric disorders by birth cohorts in five US community populations.
Using data collected in the National Institute of Mental Health (Rockville, Md) Epidemiologic Catchment Area Program, we examined the proposed hypothesis that there has been a shift in major depression to younger ages at onset, or increased prevalence in younger age periods, for recent birth cohorts. Life-table survival methods were used to examine the hazard rates for major depression as well as for other specific mental disorders. The findings are consistent with a gradual shift to increased rates for major depression between the ages of 15 and 19 years for Epidemiologic Catchment Area respondents born more recently. The findings also suggest a similar shift for drug abuse/dependence; similar but less pronounced changes were found for alcohol abuse/dependence and obsessive-compulsive disorder. However, in this study, bipolar disorder, panic disorder, and phobias did not exhibit a consistent increase in onset at younger ages. Further research is required to determine if the shifts in major depression, drug abuse/dependence, and possibly alcohol abuse/dependence are linked. It is important to note that these shifts to adolescent onset are occurring when nearly half the 31 million Americans without health insurance are aged 24 years or younger.


Removal of reflections in LWIR image with polarization characteristics.
Long-wave infrared (LWIR) imaging has been successfully used in surveillance applications in low illumination conditions. However, infrared energy reflected from smooth surfaces such as floors and metallic objects may reduce object detection and tracking accuracies. In this paper, we present a novel reflection removal method using polarization properties of the reflection in LWIR imagery. Reflection can be distinguished from the scene by two unique characteristics of polarization: the difference of two orthogonal polarized components (OPC) and the uniformity of angle of polarization (AoP). The OPC difference helps locate the regions of reflection. The uniformity of AoP in the reflection region pose a strong constraint for reflection detection. The proposed joint reflection detection method combines the OPC difference and the uniformity of AoP can detect actual reflection region. Then the closed-form matting method improves the robustness of the method and removes the reflection from the scene. Experiment results demonstrate that the proposed scheme effectively removes the reflection in challenging situations where many existing techniques may fail.


Compaction of the Escherichia coli nucleoid caused by Cyt1Aa.
Compaction of the Escherichia coli nucleoid in the cell's centre was associated with the loss of colony-forming ability; these effects were caused by induction of Cyt1Aa, the cytotoxic 27 kDa protein from Bacillus thuringiensis subsp. israelensis. Cyt1Aa-affected compaction of the nucleoids was delayed but eventually more intense than compaction caused by chloramphenicol. The possibility that small, compact nucleoids in Cyt1Aa-expressing cells resulted in DNA replication run-out and segregation following cell division was ruled out by measuring relative nucleoid length. Treatments with membrane-perforating substances other than Cyt1Aa did not cause such compaction of the nucleoids, but rather the nucleoids overexpanded to occupy nearly all of the cell volume. These findings support the suggestion that, in addition to its perforating ability, Cyt1Aa causes specific disruption of nucleoid associations with the cytoplasmic membrane. In situ immunofluorescence labelling with Alexa did not demonstrate a great amount of Cyt1Aa associated with the membrane. Clear separation between Alexa-labelled Cyt1Aa and 4',6-diamidino-2-phenylindole (DAPI)-stained DNA indicates that the nucleoid does not bind Cyt1Aa. Around 2 h after induction, nucleoids in Cyt1Aa-expressing cells started to decompact and expanded to fill the whole cell volume, most likely due to partial cell lysis without massive peptidoglycan destruction.


Advances in adenovirus-mediated p53 cancer gene therapy.
The tumor suppressor p53 gene regulates diverse cellular processes, such as cell-cycle arrest, senescence, apoptosis and autophagy, and it is frequently inactivated by genetic alterations in ∼ 50% of all types of human cancers. To restore wild-type p53 function in p53-inactivated tumors, adenovirus-mediated p53 gene therapy has been developed as a promising antitumor strategy in preclinical experiments and clinical studies. This review focuses on the clinical relevance of replication-deficient adenovirus vectors that carry the wild-type p53 gene (Ad-p53; Advexin, Gendicine and SCH-58500) in clinical studies of patients with various cancers and the future perspectives regarding conditionally replicating adenovirus vectors expressing the wild-type p53 gene (CRAd-p53; AdDelta24-p53, SG600-p53, OBP-702) in preclinical experiments. Moreover, the recent advances in our understanding of the molecular basis for the p53-mediated tumor suppression network induced by Ad-p53 and CRAd-p53 vectors and the combination therapies for promoting the therapeutic potential of adenovirus-mediated p53 gene therapy are discussed. Exploration of the molecular mechanism underlying the p53-mediated tumor suppression network and the effective strategy for enhancing the p53-mediated cell death signaling pathway would provide novel insights into the improvement of clinical outcome in p53-based cancer gene therapy.


Ontogenetic changes in leaf phenology of Ulmus davidiana var. japonica, a deciduous broad-leaved tree.
To determine how plants control leaf phenology to maximize annual carbon gain, I examined ontogenetic changes in leaf phenology of Japanese elm, Ulmus davidiana var. japonica Nakai plants of different ages growing in contrasting light environments. Leaf emergence occurred earlier in 1- and 2-year-old seedlings than in current-year seedlings. Although leaf emergence was not affected by light conditions at the sites, it was influenced by plant height. The delay in leaf emergence increased with increasing plant height. These traits indicate that seedlings that received the least light during the summer intercepted light for a long period during the spring; however, the advantage of earlier leaf emergence decreased with increasing plant height. At each site, 1-year-old seedlings had a longer duration of leaf emergence than adults, because of a longer period of favorable light conditions even in the forest understory. Duration of leaf emergence, leaf duration and leaf longevity were usually longer in sun than in shade for both seedlings and adults; however, flexibility in the response to light was greater in seedlings than in adults. The plastic response in leaf phenology during the juvenile stages may contribute to the optimization of light acquisition in habitats with differing light conditions, thereby enhancing seedling survival.


Ocular dimensions by three-dimensional magnetic resonance imaging in emmetropic versus myopic school children.
Magnetic resonance imaging (MRI) has been used to investigate eye shapes; however, reports involving children are scarce. This study aimed to determine ocular dimensions, and their correlations with refractive error, using three-dimensional MRI in emmetropic versus myopic children. Healthy school children aged < 10 years were invited to take part in this cross-sectional study. Refraction and best-corrected distance visual acuity (BCDVA) were determined using cycloplegic refraction and a logarithm of the minimum angle of resolution (logMAR) chart, respectively. All children underwent MRI using a 3-Tesla whole-body scanner. Quantitative eyeball measurements included the longitudinal axial length (LAL), horizontal width (HW), and vertical height (VH) along the cardinal axes. Correlation analysis was used to determine the association between the level of refractive error and the eyeball dimensions. A total of 70 eyes from 70 children (35 male, 35 female) with a mean (standard deviation [SD]) age of 8.38 (0.49) years were included and analyzed. Mean (SD) refraction (spherical equivalent, SEQ) and BCDVA were -2.55 (1.45) D and -0.01 (0.06) logMAR, respectively. Ocular dimensions were greater in myopes than in emmetropes (all P < 0.05), with no significant differences according to sex. Mean (SD) ocular dimensions were LAL 24.07 (0.91) mm, HW 23.41 (0.82) mm, and VH 23.70 (0.88) mm for myopes, and LAL 22.69 (0.55) mm, HW 22.65 (0.63) mm, and VH 22.94 (0.69) mm for emmetropes. Significant correlations were noted between SEQ and ocular dimensions, with a greater change in LAL (0.46 mm/D, P < 0.001) than in VH (0.27 mm/D, P < 0.001) and HW (0.22 mm/D, P = 0.001). Myopic eyeballs are larger than those with emmetropia. The eyeball elongates as myopia increases, with the greatest change in LAL, the least in HW, and an intermediate change in VH. These changes manifest in both sexes at a young age and low level of myopia. These data may serve as a reference for monitoring the development of refractive error in young Malaysian children of Chinese origin.


Xanthan Gum Ameliorates Osteoarthritis and Mitigates Cartilage Degradation via Regulation of the Wnt3a/β-Catenin Signaling Pathway.
BACKGROUND Osteoarthritis (OA) is a joint disease characterized by articular cartilage degeneration and inflammation. We have previously clarified that a xanthan gum (XG) preparation exerts ameliorating effect on a rabbit OA model by regulating matrix metalloproteinase (MMP)-1 and MMP-3, which are critical proteins in the Wnt3a/ß-catenin pathway. Thus, it is reasonable to predict that the Wnt3a/ß-catenin pathway is involved in the treatment of OA with XG. MATERIAL AND METHODS The effect of XG in OA model animals were observed by hematoxylin and eosin staining (HE), Safranin O staining, and Fast Green staining. Articular cartilage degradation on the medial plateau sides was quantified using the modified Pritzker OARSI score. The levels of IL-6, TNF-alpha, and IL-1ß in synovial fluid were determined with ELISA. The protective effect of XG in rat chondrocytes was assessed by CCK8 assay. Moreover, activation of the Wnt3a/ß-catenin pathway and the expression of MMP13, ADAMTS5, aggrecan, and collagen II under the influence of XG was measured by Western blot and qRT-PCR. RESULTS Our results showed that XG reduced the OARSI score and the concentration of inflammatory cytokines in OA after intra-articular injection. XG acted on Wnt3a/ß-catenin in ATDC5 cells in a dose-dependent manner and exhibited a protective effect. XG also decreased the expression of MMP13 and ADAMTS5 and rescued the inhibition of aggrecan and collagen II expression in SNP-stimulated chondrocytes. CONCLUSIONS These results indicate that the effects of XG are related to the Wnt3a/ß-catenin pathway and XG suppresses matrix degradation by inhibiting the expression of MMPs and ADAMTS and promotes aggrecan and collagen II content in the ECM, indicating its favorable potential for use in OA therapy.


A Low Cost Sensors Approach for Accurate Vehicle Localization and Autonomous Driving Application.
Autonomous driving in public roads requires precise localization within the range of few centimeters. Even the best current precise localization system based on the Global Navigation Satellite System (GNSS) can not always reach this level of precision, especially in an urban environment, where the signal is disturbed by surrounding buildings and artifacts. Laser range finder and stereo vision have been successfully used for obstacle detection, mapping and localization to solve the autonomous driving problem. Unfortunately, Light Detection and Ranging (LIDARs) are very expensive sensors and stereo vision requires powerful dedicated hardware to process the cameras information. In this context, this article presents a low-cost architecture of sensors and data fusion algorithm capable of autonomous driving in narrow two-way roads. Our approach exploits a combination of a short-range visual lane marking detector and a dead reckoning system to build a long and precise perception of the lane markings in the vehicle's backwards. This information is used to localize the vehicle in a map, that also contains the reference trajectory for autonomous driving. Experimental results show the successful application of the proposed system on a real autonomous driving situation.


Endothelial cell injury caused by Candida albicans is dependent on iron.
Although it is known that Candida albicans causes endothelial cell injury, in vitro and in vivo, the mechanism by which this process occurs remains unknown. Iron is critical for the induction of injury in many types of host cells. Therefore, we investigated the role of iron in Candida-induced endothelial cell injury. We found that pretreatment of endothelial cells with the iron chelators phenanthroline and deferoxamine protected them from candidal injury, even though the organisms germinated and grew normally. Loading endothelial cells with iron reversed the cytoprotective effects of iron chelation. Moreover, chelation of endothelial cell iron significantly reduced phagocytosis of C. albicans by these cells, while candidal adherence to chelator-treated endothelial cells was slightly enhanced. Since endothelial cell phagocytosis of C. albicans is required for endothelial cell injury to occur, inhibition of phagocytosis is likely the principal mechanism of the cytoprotective effects of iron chelation. The production of toxic reactive oxygen intermediates by host cells is known to be inhibited by iron chelation. Therefore, we investigated whether treating endothelial cells with antioxidants could mimic the cytoprotective effects of iron chelation. Neither extracellular nor membrane-permeative antioxidants reduced candidal injury of endothelial cells. Furthermore, depleting endothelial cells of the endogenous antioxidant glutathione did not render them more susceptible to damage by C. albicans. These results suggest that candidal injury of endothelial cells is independent of the production of reactive oxygen intermediates and that the cytoprotective effects of iron chelation are not due to inhibition of the synthesis of these toxic intermediates.


Evaluating the endometabolic and bone health effects of Tyrosine Kinase Inhibitors in Chronic Myeloid Leukaemia: a systematic review protocol.
Chronic Myeloid Leukaemia (CML) constitutes 15% of new adult leukaemia cases as well as 2%-3% of leukaemia in children under 15% and 9% of leukaemias in adolescents 15-19 years of age annually. The introduction of Tyrosine Kinase Inhibitors (TKI) therapy has dramatically improved survival in these patients, yet the off-target effects of this treatment may have long-term health impacts on CML survivors. The risk of adverse health outcomes is especially important in children, where TKI exposure may occur during critical windows of growth and puberty, and patients require treatment for prolonged periods of time. The aim of this systematic review protocol is to report on the methods used to conduct a systematic review to investigate the endometabolic and bone health effects of TKI therapy in CML. Searches will be conducted in the Cochrane Central Register of Controlled Trials, EMBASE and MEDLINE from inception on August 1st, 2019. Searches may be updated while performing the systematic review to ensure new evidence is included if applicable. Grey literature search will include ClinicalTrials.gov and ProQuest Dissertations and Theses A&I. We will perform a meta-analysis if there are at least two studies reporting similar populations, interventions, methods and tracking the same outcome measures. The studies should also have similar age and sex distributions. As this is a systematic review protocol, it does not include patient data; therefore, Research Ethics Board approval is not indicated. The systematic review will be published in a peer-reviewed journal and presented at international conferences. CRD42018091175.


RrGT2, A Key Gene Associated with Anthocyanin Biosynthesis in Rosa rugosa, Was Identified Via Virus-Induced Gene Silencing and Overexpression.
In this study, a gene with a full-length cDNA of 1422 bp encoding 473 amino acids, designated RrGT2, was isolated from R. rugosa 'Zizhi' and then functionally characterized. RrGT2 transcripts were detected in various tissues and were proved that their expression patterns corresponded with anthocyanins accumulation. Functional verification of RrGT2 in R. rugosa was performed via VIGS. When RrGT2 was silenced, the Rosa plants displayed a pale petal color phenotype. The detection results showed that the expression of RrGT2 was significantly downregulated, which was consistent with the decrease of all anthocyanins; while the expression of six key upstream structural genes was normal. Additionally, the in vivo function of RrGT2 was investigated via its overexpression in tobacco. In transgenic tobacco plants expressing RrGT2, anthocyanin accumulation was induced in the flowers, indicating that RrGT2 could encode a functional GT protein for anthocyanin biosynthesis and could function in other species. The application of VIGS in transgenic tobacco resulted in the treated tobacco plants presenting flowers whose phenotypes were lighter in color than those of normal plants. These results also validated and affirmed previous conclusions. Therefore, we speculated that glycosylation of RrGT2 plays a crucial role in anthocyanin biosynthesis in R. rugosa.


Combining radiotherapy with AZD2171, a potent inhibitor of vascular endothelial growth factor signaling: pathophysiologic effects and therapeutic benefit.
AZD2171 is a highly potent, orally active inhibitor of vascular endothelial growth factor receptor signaling. The potential for AZD2171 to enhance the antitumor effects of radiotherapy was investigated in lung (Calu-6) and colon (LoVo) human tumor xenograft models. Combined treatment resulted in a significantly enhanced growth delay compared with either modality alone. The enhancement was independent of whether chronic once daily AZD2171 treatment was given 2 h prior to each radiation fraction (2 Gy daily for 3 or 5 consecutive days), and daily thereafter, or commenced immediately following the course of radiotherapy. Histologic assessments revealed that 5 days of radiation (2 Gy) or AZD2171 (3 or 6 mg/kg/d) reduced vessel density and perfusion. Concomitant AZD2171 and radiation enhanced this effect and produced a significant increase in tumor hypoxia. Concomitant AZD2171 (6 mg/kg/d) was also found to reduce tumor growth significantly during the course of radiotherapy (5 x 2 Gy). However, the extent and duration of tumor regression observed postradiotherapy was similar to sequentially treated tumors, suggesting that preirradiated tumors were sensitized to AZD2171 treatment. An enhanced antivascular effect of administering AZD2171 postradiotherapy was observed in real-time in Calu-6 tumors grown in dorsal window chambers. Collectively, these data support the clinical development of AZD2171 in combination with radiotherapy.


Alterations in brain activation patterns in women with functional defecatory disorder: A novel fMRI rectal balloon expulsion study.
Functional defecatory disorders (FDD) are common among women. Despite the extensive research on peripheral mechanisms involved in FDD, the central-neural contribution to its pathophysiology remains poorly understood. We aimed to delineate specific supra-spinal regions involved in defecation and examine whether their activity, as measured by blood-oxygen-level-dependent (BOLD) signals, is different in FDD. We performed functional MRI (fMRI) with concurrent rectal manometry in 15 controls and 18 females with ROME III diagnosis of FDD. A block design was used and brain activation maps based on BOLD effect employing the generalized linear model were calculated for each subject. Statistical significance between groups was assessed by a Student t-test with cluster-based multiple comparisons correction (corrected p < 0.01). Simulated defecation was associated with activation of regions of primary and supplementary motor (SMA) and somatosensory cortices, homeostatic afferent (thalamus, mid-cingulate cortex, and insula), and emotional arousal networks (hippocampus and prefrontal cortex), occipital and cerebellum along with deactivation of right anterior cingulate cortex (ACC) in controls. Women with FDD had fewer regions engaged in defecation and BOLD activation was much decreased is areas related to executive-cognitive function (insula, parietal, and prefrontal cortices). Patients unlike controls showed activation in right ACC and otherwise had similar brain activation patterns during anal squeeze. Our results provide evidence that distinct differences exist in supra-spinal control of defecation in key regions of motivational-affective regulation and executive-cognitive function, in patients with FDD as compared to controls.


Robot-assisted Transplanted Ureteral Stricture Management.
To assess the feasibility of robot-assisted transplanted ureteral reimplantation as a minimally invasive alternative to open surgery. Between August 2015 and March 2016, 5 patients presented with transplanted ureteral strictures after failure of a previous endoscopic management. All patients underwent robot-assisted ureteral reimplantation. Patients' demographics, perioperative outcomes, and complications are reported. All patients presented with deterioration of kidney function with or without recurrent urinary tract infection. Two patients had short strictures (<1 cm) and 2 had long strictures (>1 cm), whereas 1 patient had a nitinol ureteral stent in situ. The location of the stricture varied among these patients with 3 distal and 1 proximal. Intraoperatively, 3 patients had a modified Lich-Gregoir reimplantation and 2 patients had a pyelovesicostomy. The mean operative time was 164 (±52) minutes. There were no intraoperative complications, conversion to open surgery, or significant blood loss necessitating blood transfusion. There were no urine leaks in the immediate or late postoperative period. One patient developed a Clavien grade IVa complication (sepsis). The median length of stay, the duration of catheterization, and the duration of stenting were 1 day (range 1-5 days), 7 days (range 6-14 days), and 39 days (range 25-51 days), respectively. After a median follow-up of 79 days (range 40-139 days), no strictures or delayed leakages were identified. Robot-assisted transplanted ureteral reimplantation is technically feasible. With a larger number of cases and a longer follow-up, robot-assisted transplanted ureteral reimplantation may provide a new and effective, minimally invasive alternative for the treatment of this complex surgical problem.


Safety of long-term use of daily and long-acting growth hormone in growth hormone-deficient adults on cancer risk.
Daily injections of recombinant human growth hormone (rhGH) have been used in clinical practice for almost four decades as a replacement therapy in adult patients with GH deficiency (GHD). Long-term adherence to daily injections of rhGH is a clinical concern that may result in reduced therapeutic efficacy, and long-acting GH (LAGH) formulations have been developed in an attempt of overcoming this problem. Long-term safety issues of rhGH are the other side of the coin that has been carefully monitored over the years, particularly related to the proliferative actions of GH that could increase the risk of tumor recurrence or induce the development of new benign and malignant tumors. In this review, we present what is currently known about the cancer risk in GHD adults treated with daily rhGH injections and we discuss the major concerns and responses needed from future surveillance studies regarding the safety of LAGH preparations.


No Time for Statistics: Joseph Lister's Antisepsis and Types of Knowledge in Nineteenth-Century British Surgery.
This article documents Joseph Lister's reluctance to publish numerical material and aims at explaining his skeptical view about statistics through an investigation of his approach in its historical context. In this context, statistics was only one kind of evidence used in surgery, along with case histories and experimental results from the laboratory. They represent different "ways of knowing," anchored in different social, conceptual, and practical contexts. The account looks at Lister's approach to wound disease and analyzes how this relates to his attitude toward different types of evidence about surgical outcomes. For this, it also examines his contemporaries' approaches to fighting wound disease as well as their evaluation of different kinds of evidence. This article is a contribution to the history of Lister's antisepsis, but also to the history of the production and use of therapeutic knowledge in nineteenth- and early twentieth-century surgery more generally.


Growth factors as potential drugs for the sensory epithelia of the ear.
The highly ordered structures of the hearing and balance organs of vertebrate ears go through a coordinated sequence of cellular and morphogenetic events. It is to be expected that protein growth factors and other extracellular signals will regulate many events during embryonic development of the ear, including the induction of the ear, the specific induction of sensory epithelia, the proliferation of the cells that form the sensory epithelia, the differentiation of the sensory and supporting cells, and the attraction and maintenance of innervation. After embryonic development, growth factors will support cell survival and innervation of new sensory cells. In damaged sensory epithelia, supplementation of the normal growth factors in these tissues has the potential to influence cellular responses to trauma, to reduce cell death and to promote the replacement of dead cells through renewed proliferation and differentiation, so as to improve hearing and balance health via preventive and restorative treatments. Assessment of the influences of specific growth factors on the sensory epithelia of vertebrate ears is at an early stage: this paper provides a brief account of what we know from studies of normal and experimentally manipulated epithelia, discusses the current questions and suggests directions for future studies.


Halothane hepatitis. Detection of a constitutional susceptibility factor.
We studied susceptibility to halothane hepatitis with an in vitro test that detects cell damage from electrophilic drug intermediates. Metabolites of phenytoin were generated by incubation of phenytoin with rat hepatic microsomes in the presence of the epoxide hydrolase inhibitor 1,1,1-trichloropropene oxide (TCPO), which prevents the further metabolism of phenytoin to an inert metabolite. In lymphocytes exposed to this system, cytotoxicity was measured by trypan blue dye exclusion and was expressed as the percentage increase in trypan blue-positive cells after the addition of TCPO. In the presence of TCPO, lymphocytes from 11 patients with halothane hepatitis exhibited an increase in cytotoxicity at 0.06 mM phenytoin that was eight times greater than the increase in healthy controls (54 +/- 10 per cent [mean +/- S.E.M.] vs. 7.1 +/- 2.2 per cent, P less than 0.0001). Patients with other liver diseases and persons recently exposed to halothane without adverse effects did not differ from healthy controls. In three patients with halothane hepatitis who were studied serially, the lymphocyte abnormality was still present after 13 months. Family studies revealed abnormal results on 10 cytotoxicity tests among 19 members of four families. We propose that there is a familial, constitutional susceptibility factor that predisposes persons to halothane hepatitis.


Maintenance of differentiated rat hepatocytes in primary culture.
Normal adult rat hepatocytes remained viable and functional for at least 43 days when plated on collagen-coated dishes and fed chemically defined medium supplemented with dimethyl sulfoxide (Me2SO). Hepatocytes isolated by collagenase perfusion and cultured in the presence or absence of Me2SO were (i) examined by light and electron microscopy for morphological changes; (ii) analyzed for the production of albumin and other plasma proteins; and (iii) tested by autoradiography for DNA synthesis. Me2SO-treated cells continued to produce specific plasma proteins during the entire culture period; albumin production was consistently high (11-19 micrograms/ml of culture medium per 24 hr) from day 2 to at least day 43 after plating. Ultrastructural analyses demonstrated that Me2SO-treated hepatocytes resembled those from intact liver in organization of cytoplasmic organelles and cellular junctions. The optimal concentration for observing the morphological and biochemical effects of Me2SO was 2% (vol/vol). We conclude that supplementation of chemically defined medium with Me2SO enables maintenance of differentiated hepatocytes in culture for extended periods of time.


Dementia and mortality in older adults: A twin study.
Dementia predicts increased mortality. We used case-control and co-twin control models to investigate genetic and shared environmental influences on this association. Case-control design, including 987 twins with dementia and 2938 age- and sex-matched controls in the Swedish Twin Registry. Co-twin control design, including 90 monozygotic (MZ) and 288 dizygotic (DZ) twin pairs discordant for dementia. To test for genetic and environmental confounding, differences were examined in mortality risk between twins with dementia and their matched or co-twin controls. Twins with dementia showed greater mortality risk than age- and sex-matched controls (HR = 2.02 [1.86, 2.18]). Mortality risk is significantly elevated but attenuated substantially in discordant twin pairs, for example, comparing MZ twins with dementia to their co-twin controls (HR = 1.48 [1.08, 2.04]). Findings suggest that genetic factors partially confound the association between dementia and mortality and provide an alternative hypothesis to increased mortality due to dementia itself. Highlights We studied dementia and mortality in twin pairs discordant for dementia. People without dementia outlived people with dementia. Identical twins with dementia and their co-twin controls had similar survival time. Findings suggest genotype may explain the link between dementia and mortality.


[Tobacco control in France: what's new ?]
French governments only very recently introduced comprehensive tobacco control policies including several measures recommended by WHO Framework Convention on Tobacco Control, leading to a speeding-up of tobacco smoking decrease, but still with a high current level of use. In the meantime, research confirms that nicotine is highly addictive, that smoking just a cigarette a day is highly detrimental for health, and that health professionals are efficient in helping smokers to give up. These measures are such diverse as tobacco taxes increases, introducing plain packs, promoting November as month without tobacco, getting tobacco cessation medications paid for by health insurances, implementing a comprehensive advertising ban of tobacco products in retailor shops, and increasing protection of public health policies from tobacco industry intrusion. This industry via frenzied marketing reacts by promoting "heat not burn tobacco" as a harm reduction mean, which it is not, and invests in e-cigarettes, whose most recent ones are as addictive as classic tobacco cigarettes.


[Partial characteristics of the basic phenylmethylsulfonylfluoride-inhibited carboxypeptidase from cat brain].
Cat brain carboxypeptidase releasing C-terminal arginine from the synthetic substrate, dansyl-Phe-Leu-Arg, was partially characterized. The enzyme has a molecular weight of 100-120 kDa, displays the maximal activity at pH 6.0-6.5 and is strongly inhibited by phenylmethanesulfonylfluoride and 4-chloromercuribenzoate, less strongly (by 40%) by iodoacetamide and is not inhibited by N-ethylmaleimide, 2-mercaptoethanol, EDTA, Co2+ and guanidinoethylmercaptosuccinic acid. The Km values for the hydrolysis of dansyl-Phe-Leu-Arg and dansyl-Phe-Ala-Arg by soluble carboxypeptidase are 48 and 96 microM, respectively. By all properties, this carboxypeptidase differs from other known carboxypeptidases. Possible participation of the enzyme in neuropeptide metabolism is discussed.


[Immunoenzyme method of determination of m5Cyd level; characteristics of the epitope and estimation of m5Cyd levels in the blood of leukemic mice].
Antibodies specific to m5Cyd were obtained and a sensitive enzyme-linked immunosorbent assay (ELISA) for detecting m5Cyd was developed. This method was used for the characterization of m5Cyd epitope. It was shown that the 4-NH2 and 5-CH3-groups of the pyrimidine ring are of primary importance for the antigenic determinant assembly, whereas the ribosyl residue and its structure are less important. It was found also that the amount of m5Cyd in leukemic La murine serum after gamma-irradiation is markedly increased.


Pretreatment-Etidronate Alleviates CoCl2 Induced-SH-SY5Y Cell Apoptosis via Decreased HIF-1α and TRPC5 Channel Proteins.
Chronic hypoxic damage is one of the most common pathogenic factors that can cause neurodegenerative disorder in most cases. Etidronate (Eti) is one of the best-known earlier-generations of bisphosphonate derivatives for the treatment of bone-loss related diseases. Building on the preceding study of our laboratory, we found that Eti showed neuroprotective effects against 2-vessel occlusion induced vascular dementia (VD) in rats. Therefore, in this study, we attempted to elucidate the mechanism of action, which Eti protected cells from chronic hypoxic damage caused by CoCl2 in SH-SY5Y cells in vitro. Our data showed that the pretreatment with 100 µM Eti partially improved hypoxic damage in cell viability and reduced the hypoxia-inducible factor-1α (HIF-1α) expression, which indicated chronic hypoxic level. Furthermore, the protein expression of TRPC5 channel and its mediated intracellular calcium ion concentration ([Ca2+]i) were decreased. In addition, the apoptosis-related proteins caspase-9, and caspase-3 as well as calcium/calmodulin-dependent protein kinase II (CaMK-II) were down-regulated after treatment with Eti. In conclusion, Eti shows neuroprotective effects on SH-SY5Y cells injured by CoCl2 through resisting apoptosis caused by calcium influx, which may be related to the inhibition of HIF-1α protein and the decreased TRPC5 channel protein.


Central adiposity and the functional magnetic resonance imaging response to cognitive challenge.
Excessive adipose tissue, particularly with a centralized distribution, propagates hormonal and metabolic disturbance. The detrimental effects of adiposity may extend beyond the periphery and target the central nervous system, increasing vulnerability to cognitive decline. The aim of the current study was to determine how central adiposity impacts the brain at midlife by examining the blood oxygen level-dependent (BOLD) response to a challenging cognitive task. Seventy-three adults, aged 40-60 years, completed a 2-back verbal working memory task during functional magnetic resonance imaging. Central adiposity was assessed with waist circumference. The association between waist circumference and task-related activation in a priori regions of interest was modeled using bootstrapping regression models corrected for multiple-comparisons. Larger waist circumference was associated with diminished working-memory-related BOLD response in the right superior frontal gyrus (β=-0.008, P=0.001, 95% CI: -0.012 to -0.004) and left middle frontal gyrus (β=-0.009, P=0.002, 95% CI: -0.015 to -0.003), statistically adjusting for age, sex, systolic blood pressure and total cholesterol. Reduced task-related activation in the right superior frontal gyrus (r=-0.369, P=0.002) and left middle frontal gyrus (r=-0.266, P=0.025) were related to slower reaction time on the task, controlling for age and education. Larger waist circumference predicted alterations in the BOLD response that coupled with decrements in task performance. While future studies are necessary, the results suggest that similar to its role in the periphery, central adiposity may be a robust predictor of metabolic and hormonal alterations that impinge upon central nervous system functioning.


Comparison of stress and coping in male and female dentists.
This study was designed to examine how female dentists withstand the pressures of dental practice in comparison to male dentists. Two-hundred thirty-eight dentists responded to a national survey that collected demographic data, information about the kinds and degree of stress experienced, coping strategies used and coping effectiveness. The greatest amount of stress experienced was attributed to interactions with patients. The business aspects of dentistry were also associated with significant amounts of stress. In general, male and female dentists' responses to stress were similar, although female dentists experienced more personal/domestic stress. The coping styles of males and females were similar in most respects, except that females were more inclined to discuss their problems. Coping strategies were seen as moderately effective by both sexes. It was suggested that dental students receive training in interpersonal skills, stress management and practice management along with technical training to better prepare them to cope with the realities of dental practice. An alternative suggestion was to address these nontechnical topics through continuing education programs designed for dentists beginning their practice.