[Directional osteotomy of the femur in the bedridden patient with cerebral palsy].
Wind-blown or scissor deformity of the hip in severely spastic bed-ridden patients with cerebral palsy requires surgical correction. The methods generally used involve a number of disadvantages which are analysed. Corrective sub-trochanteric osteotomy following extensive soft tissue release is the method of choice in this exceptional situation. The treatment of six patients since 1982 has emphasised the good tolerance of this operation by patients who are frail and suffer from multiple handicaps and the reliability of the results obtained in the relief of pain, the improved ease of nursing management and attention to toilet and, in certain cases, the ability to sit.


CALIBER: a phase II randomized feasibility trial of chemoablation with mitomycin-C vs surgical management in low-risk non-muscle-invasive bladder cancer.
To evaluate the activity of intravesical mitomycin-C (MMC) to ablate recurrent low-risk non-muscle-invasive bladder cancer (NMIBC) and assess whether it may enable patients to avoid surgical intervention for treatment of recurrence. CALIBER is a phase II feasibility study. Participants were randomized (2:1) to treatment with four once-weekly MMC 40-mg intravesical instillations (chemoablation arm) or to surgical management. The surgical group was included to assess the feasibility of randomization. The primary endpoint was complete response to intravesical MMC in the chemoablation arm at 3 months, reported with exact 95% confidence intervals (CIs). Secondary endpoints included time to subsequent recurrence, summarized by Kaplan-Meier methods. Between February 2015 and August 2017, 82 patients with visual diagnosis of recurrent low-risk NMIBC were enrolled from 24 UK hospitals (chemoablation, n = 54; surgical management, n =28). The median follow-up was 24 months. Complete response at 3 months was 37.0% (20/54; 95% CI 24.3-51.3) with chemoablation and 80.8% (21/26; 95% CI 60.6-93.4) with surgical management. Amongst patients with complete response at 3 months, a similar proportion was recurrence-free by 12 months in both groups (84%). Amongst those with residual disease at 3 months, the 12-month recurrence-free proportion was lower in the surgical management group (40.0%) than in the chemoablation group (84%). Recruitment stopped early as chemoablation did not meet the prespecified threshold of 45% complete responses at 3 months. Intravesical chemoablation in low-risk NMIBC is feasible and safe, but did not demonstrate sufficient response in the present trial. After chemoablation there may be a reduction in recurrence rate, even in non-responders, that is greater than with surgery alone. Further research is required to investigate the role and optimal schedule of neoadjuvant intravesical chemotherapy prior to surgery for NMIBC.


Validated Growth Rate-Dependent Regulation of Lipid Metabolism in Yarrowia lipolytica.
Given the strong potential of Yarrowia lipolytica to produce lipids for use as renewable fuels and oleochemicals, it is important to gain in-depth understanding of the molecular mechanism underlying its lipid accumulation. As cellular growth rate affects biomass lipid content, we performed a comparative proteomic analysis of Y. lipolytica grown in nitrogen-limited chemostat cultures at different dilution rates. After confirming the correlation between growth rate and lipid accumulation, we were able to identify various cellular functions and biological mechanisms involved in oleaginousness. Inspection of significantly up- and downregulated proteins revealed nonintuitive processes associated with lipid accumulation in this yeast. This included proteins related to endoplasmic reticulum (ER) stress, ER-plasma membrane tether proteins, and arginase. Genetic engineering of selected targets validated that some genes indeed affected lipid accumulation. They were able to increase lipid content and were complementary to other genetic engineering strategies to optimize lipid yield.


Pleiotropic Properties of Amphiphilic Dihydropyridines, Dihydropyridones, and Aminovinylcarbonyl Compounds.
Three groups of synthetic lipids are chosen for studies: (1) 1,4-dihydropyridines (1,4-DHPs) containing two cationic moieties and their analogues; (2) 3,4-dihydro-2(1H)-pyridones containing a cationic moiety; and (3) acyclic, open-chain analogues, i.e., 2-amino-3-alkoxycarbonylalkylammonium derivatives. 1,4-DHPs possessing dodecyl alkyl chains in the ester groups in positions 3 and 5 and cationic nitrogen-containing groups in positions 2 and 6 have high cytotoxicity in cancer cells HT-1080 (human lung fibrosarcoma) and MH-22A (mouse hepatoma), but low cytotoxicity in the noncancerous NIH3T3 cells (mouse embryonic fibroblast). On the contrary, similar compounds having short (methyl, ethyl, or propoxyethyl) chains in the ester groups in positions 3 and 5 lack cytotoxicity in the cancer cells HT-1080 and MH-22A even at high doses. Inclusion of fluorine atoms in the alkyl chains in positions 3 and 5 of the DHP cycle decreases the cytotoxicity of the mentioned compounds. Structurally related dihydropyridones with a polar head group are substantially more toxic to normal and cancerous cells than the DHP analogues. Open-chain analogues of DHP lipids comprise the same conjugated aminovinylcarbonyl moiety and possess anticancer activity, but they also have high basal cytotoxicity. Electrochemical oxidation data demonstrate that oxidation potentials of selected compounds are in the range of 1.6-1.7 V for cationic 1,4-DHP, 2.0-2.4 V for cationic 3,4-dihydropyridones, and 1.2-1.5 V for 2-amino-3-alkoxycarbonylalkylammonium derivatives. Furthermore, the tested cationic 1,4-DHP amphiphiles possess antiradical activity. Molecular topological polar surface area values for the tested compounds were defined in accordance with the main fragments of compound structures. The determined logP values were highest for dodecyl ester groups in positions 3 and 5 of the 1,4-DHP and lowest for short alkyl chain-containing amphiphiles.


Outcomes of electroconvulsive therapy in patients with depressive symptoms with versus without comorbid personality disorders/traits: A systematic review and meta-analysis.
To assess electroconvulsive therapy (ECT) outcomes in patients affected by depressive symptoms with versus without additional comorbid personality disorders/traits. We identified observational studies investigating ECT clinical outcomes in patients affected by depressive symptoms with versus without comorbid personality disorders/traits in Embase/Medline in 11/2022. Our protocol was registered with PROSPERO (CRD42023390833). Study quality was evaluated using the Newcastle-Ottawa-Scale. Our primary outcomes were ECT response and remission rates. Meta-regression analyses included effects of in/outpatient percentages, age, number of ECT sessions, and electrode placement; subgroup analyses included the assessment methods for personality disorders/traits. We performed sensitivity analyses after excluding poor-quality studies. A total of 20 studies (n = 11,390) were included in our analysis. Patients with comorbid personality disorders/traits had lower remission rates (OR = 0.42, 95% CI = 0.31, 0.58, p < 0.001) with substantial heterogeneity (I2 = 93.0%) as well as lower response rates (OR = 0.35, 95% CI = 0.24, 0.51, n = 5129, p < 0.001) with substantial heterogeneity (I2 = 93.0%) compared with patients without comorbid personality disorders/traits. Relapse rates were higher in patients with versus without comorbid personality disorders/traits (OR = 3.23, 95% CI = 1.40, 7.45, k = 4, n = 239, p = 0.006) with moderate heterogeneity (I2 = 75.0%) and post-ECT memory impairment was more frequent in patients with versus without comorbid personality disorders/traits (OR = 1.41, 95% CI = 1.36, 1.46, k = 4, n = 471, p < 0.001) with minimal heterogeneity (I2 = 0.0%). Dropout rates were higher in patients with versus without comorbid personality disorders/traits (OR = 1.58, 95% CI = 1.13, 2.21, k = 3, n = 6145, p = 0.008). Patients with comorbid personality disorders/traits treated with ECT are reported to have lower response and remission rates and higher rates of side effects and relapse rates compared with patients without personality disorders/traits.


Peroxiredoxin 2 and peroxidase enzymatic activity of mammalian spermatozoa.
Peroxiredoxin 2 (PRDX2) is a highly efficient redox protein that neutralizes hydrogen peroxide, resulting in protection of cells from oxidative damage and in regulation of peroxide-mediated signal transduction events. The oxidized form of PRDX2 is reverted back to the reduced form by the thioredoxin system. In the present study, we investigated the presence of PRDX2 in mouse and boar spermatozoa and in mouse spermatids using proteomic techniques and immunocytochemistry. Sperm and spermatid extracts displayed a 20-kDa PRDX2 band on Western blotting. PRDX2 occurred as a Triton-soluble form in spermatids and as a Triton-insoluble form in mature spermatozoa. Boar seminiferous tubule extracts were immunoprecipitated with PRDX2 antibody and separated by SDS-PAGE. Peptide mass fingerprinting by matrix-assisted laser desorption ionization-time of flight (TOF) and microsequencing by nanospray quadrupole-quadrupole TOF tandem mass spectrometry revealed the presence of PRDX2 ions in the immunoprecipitated band, along with sperm mitochondria-associated cysteine-rich protein, cellular nucleic acid-binding protein, and glutathione peroxidase 4. In mouse spermatocytes and spermatids, diffuse labeling of PRDX2 was observed in the cytoplasm and residual bodies. After spermiation, PRDX2 localization became confined to the mitochondrial sheath of the sperm tail midpiece. Boar spermatozoa displayed similar PRDX2 localization as in mouse spermatozoa. Boar spermatozoa with disrupted acrosomes expressed PRDX2 in the postacrosomal sheath region. Peroxidase enzyme activity of boar sperm extracts was evaluated by estimating the rate of NADPH oxidation in the presence or absence of a glutathione depletor (diethyl maleate) or a glutathione reductase inhibitor (carmustine). Diethyl maleate partially inhibited peroxidase activity, whereas carmustine showed an insignificant effect. These observations suggest that glutathione and glutathione reductase activity contribute only partially to the total peroxidase activity of the sperm extract. While the specific role of PRDX2 in the total peroxidase activity of sperm extract is still an open question, the present study for the first time (to our knowledge) shows the presence of PRDX2 in mammalian spermatozoa. Peroxidase activity in sperm extracts is not due to the glutathione system and therefore possibly involves PRDX2 and other peroxiredoxins.


Interferon lambda alleles predict innate antiviral immune responses and hepatitis C virus permissiveness.
Hepatitis C virus (HCV) infection can result in viral chronicity or clearance. Although host genetics and particularly genetic variation in the interferon lambda (IFNL) locus are associated with spontaneous HCV clearance and treatment success, the mechanisms guiding these clinical outcomes remain unknown. Using a laser capture microdissection-driven unbiased systems virology approach, we isolated and transcriptionally profiled HCV-infected and adjacent primary human hepatocytes (PHHs) approaching single-cell resolution. An innate antiviral immune signature dominated the transcriptional response but differed in magnitude and diversity between HCV-infected and adjacent cells. Molecular signatures associated with more effective antiviral control were determined by comparing donors with high and low infection frequencies. Cells from donors with clinically unfavorable IFNL genotypes were infected at a greater frequency and exhibited dampened antiviral and cell death responses. These data suggest that early virus-host interactions, particularly host genetics and induction of innate immunity, critically determine the outcome of HCV infection.


Hospital clinical career advancement programs: comparing perceptions of nurse participants and nonparticipants.
The study reported in this article describes nurse participants and nonparticipants in hospital clinical career advancement programs (clinical ladders) and compares perceptions in relation to program intrinsic outcomes, needs, and criteria for level advancement. A stratified random sample consisted of 120 participants and 480 nonparticipants drawn from 1,769 clinical nurses employed full time at five regional general hospitals. The findings suggest that nurses' demographic characteristics do not influence program participation status and that clinical advancement programs have limited impact upon job enrichment designed to recruit and retain clinical nurses providing direct patient care. Program participants (106; 88.3 percent) differed significantly at the .05 level from nonparticipants (385; 80.2 percent) on each factor area measuring perceptions of clinical advancement programs.


Supracondylar humeral fractures in children.
Operative fixation is indicated for most type-II and III supracondylar humeral fractures in order to prevent malunion. Medial comminution is a subtle finding that, if treated nonoperatively, is likely to lead to unacceptable varus malunion. Angiography is not indicated for a pulseless limb, as it delays fracture reduction, which usually corrects the vascular problem. A high index of suspicion is necessary to avoid missing an impending compartment syndrome, especially when there is a concomitant forearm fracture or when there is a median nerve injury, which may mask symptoms of compartment syndrome. Lateral entry pins have been shown, in biomechanical and clinical studies, to be as stable as cross pinning if they are well spaced at the fracture line, and they are not associated with the risk of iatrogenic ulnar nerve injury.


Geostatistical integration and uncertainty in pollutant concentration surface under preferential sampling.
In this paper the focus is on environmental statistics, with the aim of estimating the concentration surface and related uncertainty of an air pollutant. We used air quality data recorded by a network of monitoring stations within a Bayesian framework to overcome difficulties in accounting for prediction uncertainty and to integrate information provided by deterministic models based on emissions meteorology and chemico-physical characteristics of the atmosphere. Several authors have proposed such integration, but all the proposed approaches rely on representativeness and completeness of existing air pollution monitoring networks. We considered the situation in which the spatial process of interest and the sampling locations are not independent. This is known in the literature as the preferential sampling problem, which if ignored in the analysis, can bias geostatistical inferences. We developed a Bayesian geostatistical model to account for preferential sampling with the main interest in statistical integration and uncertainty. We used PM10 data arising from the air quality network of the Environmental Protection Agency of Lombardy Region (Italy) and numerical outputs from the deterministic model. We specified an inhomogeneous Poisson process for the sampling locations intensities and a shared spatial random component model for the dependence between the spatial location of monitors and the pollution surface. We found greater predicted standard deviation differences in areas not properly covered by the air quality network. In conclusion, in this context inferences on prediction uncertainty may be misleading when geostatistical modelling does not take into account preferential sampling.


Electrogenesis in the lower Metazoa and implications for neuronal integration.
Electrogenic communication appears to have evolved independently in a variety of animal and plant lineages. Considered here are metazoan cells as disparate as the loose three-dimensional parenchyma of glass sponges, the two-dimensional epithelial sheets of hydrozoan jellyfish and the egg cell membranes of the ctenophore Beroe ovata, all of which are capable of generating electrical impulses. Neuronal electrogenesis may have evolved independently in ctenophores and cnidarians but the dearth of electrophysiological data relating to ctenophore nerves means that our attention is focused on the Cnidaria, whose nervous systems have been the subject of extensive study. The aim here is to show how their active and passive neuronal properties interact to give integrated behaviour. Neuronal electrogenesis, goes beyond simply relaying 'states of excitement' and utilizes the equivalent of a set of basic electrical 'apps' to integrate incoming sensory information with internally generated pacemaker activity. A small number of membrane-based processes make up these analogue applications. Passive components include the decremental spread of current determined by cellular anatomy; active components include ion channels specified by their selectivity and voltage dependence. A recurring theme is the role of inactivating potassium channels in regulating performance. Although different aspects of cnidarian behaviour are controlled by separate neuronal systems, integrated responses and coordinated movements depend on interactions between them. Integrative interactions discussed here include those between feeding and swimming, between tentacle contraction and swimming and between slow and fast swimming in the hydrozoan jellyfish Aglantha digitale.


Uncertainty quantification of the effects of biotic interactions on community dynamics from nonlinear time-series data.
Biotic interactions are expected to play a major role in shaping the dynamics of ecological systems. Yet, quantifying the effects of biotic interactions has been challenging due to a lack of appropriate methods to extract accurate measurements of interaction parameters from experimental data. One of the main limitations of existing methods is that the parameters inferred from noisy, sparsely sampled, nonlinear data are seldom uniquely identifiable. That is, many different parameters can be compatible with the same dataset and can generalize to independent data equally well. Hence, it is difficult to justify conclusive assertions about the effect of biotic interactions without information about their associated uncertainty. Here, we develop an ensemble method based on model averaging to quantify the uncertainty associated with the effect of biotic interactions on community dynamics from non-equilibrium ecological time-series data. Our method is able to detect the most informative time intervals for each biotic interaction within a multivariate time series and can be easily adapted to different regression schemes. Overall, this novel approach can be used to associate a time-dependent uncertainty with the effect of biotic interactions. Moreover, because we quantify uncertainty with minimal assumptions about the data-generating process, our approach can be applied to any data for which interactions among variables strongly affect the overall dynamics of the system.


Immunotherapy and chimeric antigen receptor T-cell therapy in hepatocellular carcinoma.
Advanced hepatocellular carcinoma (HCC) is a deadly disease. With increasing incidence of new cases over the last years multiple efforts have been made to ameliorate survival and quality of life. Recent advances in understanding the tumor microenvironment and cancer immune evasion led to development of potent immune therapies targeting programmed death-ligand-1 (PD-L1), programmed cell death protein 1 (PD-1) and anti-cytotoxic T-lymphocyte-associated protein-4 (CTLA-4). Early clinical studies highlighted the activity and synergism of checkpoint inhibitors with antiangiogenic drugs, including anti-vascular endothelial growth factor (VEGF) antibodies and multi-tyrosine kinase inhibitors. Most recently, the combination of bevacizumab and atezolizumab improved survival compared to sorafenib in the treatment of advanced HCC on first-line therapy in a randomized phase III trial and now is considered the standard of care. Multiple options are available for the treatment of metastatic HCC including atezolizumab, bevacizumab, nivolumab, pembrolizumab, ipilimumab, sorafenib, lenvatinib, regorafenib, cabozantinib and ramucirumab. Furthermore, other checkpoint inhibitors are being evaluated including durvalumab, tremelimumab, CS1003, sintilimab and camrelizumab. In this review article, we focus on the landscape of different immunotherapy strategies in the management of HCC and the combination of checkpoint inhibitors antibodies with antiangiogenics. In addition, we will address the limitation of cell therapies in advanced HCC and current strategies to improve efficacy.


Ethical considerations for children's participation in data collection activities during humanitarian emergencies: A Delphi review.
Children's right to participate in data collection during emergencies has been widely recognized by humanitarian actors. However, participation in such activities can expose children to risk. Tensions have been noted between the right to participate and other principles, such as the imperative to 'do no harm.' With little evidence to inform guidance on addressing this tension, our study sought to identify expert consensus on whether and how children participate in emergency-related data collection activities. We employed a three-round Delphi technique with a purposive sample of 52 child protection specialists. Respondents answered two open-ended questions in round one. A thematic analysis of responses generated a set of unique statements addressing the study questions. In the second round, respondents rated each statement on a five-point scale. In the final round, respondents reviewed the group's average ratings for each statement with the option to revise their own ratings. A statement was said to have reached clear consensus when at least 90% of respondents agreed or strongly agreed with the statement. A total of 124 statements and 14 themes emerged from the thematic analysis, with 46.0% of statements reaching clear consensus in the third round. Respondents strongly supported children's right to participate in data collection in humanitarian settings, while also recognizing that protecting children from harm may "over-ride" the participation principle in some contexts. Respondents identified capacity and contextual considerations as important factors influencing participation decisions, though they sometimes disagreed about how these factors should determine participation. Respondents also considered the role of individual child factors and the presence of caregivers in selecting child participants, and proposed best practice approaches for securing children's safe and meaningful participation. With almost half of statements reaching clear consensus, these findings reflect broad agreement within the sector about engaging children in data collection in emergencies. At the same time, points of ongoing debate around how to factor different risks into child participation decisions may indicate discordant practice. Further reflection is needed around how factors such as the phase of emergency, the existence of basic services, and cultural beliefs should influence whether and how children participate.


Risk factors for febrile urinary tract infection in boys with posterior urethral valves.
Boys with posterior urethral valves (PUV) present an increased risk of febrile urinary tract infection (fUTI). Identifying specific risk factors could allow for tailoring UTI prevention. The aim of this study was to use the data from the CIRCUP randomized controlled trial data to identify patient characteristics associated with a higher risk of fUTI. We performed a secondary analysis of the data from the CIRCUP randomized trial which included boys with PUV, randomized to circumcision and antibiotic prophylaxis vs. antibiotic prophylaxis alone and followed for 2 years. There was only 1 episode of fUTI in the circumcision group vs. 17 in the uncircumcised group. We therefore only studied the antibiotic prophylaxis alone group and compared age at prenatal diagnosis, size and weight at birth, presence of dilating VUR at diagnosis, abnormal DMSA scan at 2 months, and nadir creatinine between children who presented a fUTI and those who did not, as well as age at first episode of fUTI. The study group consisted of 42 patients of which 17 presented at least on fUTI. Presence of dilating VUR was significantly associated with risk of fUTI (p = 0.03), OR: 6 [CI 95% = (1.13-27.52)]. None of the other parameters were associated with increased risk of fUTI. We observed three distinct time periods for presenting a fUTI with a decrease in infection rate after the first 40 days of life, then at 240 days of life. In boys with PUV, presence of high-grade VUR is associated with a higher risk of presenting a fUTI. The rate of febrile UTIs seems to decrease after 9 months.


Smokers' use of nicotine replacement therapy for reasons other than stopping smoking: findings from the ITC Four Country Survey.
To measure the prevalence and correlates of nicotine replacement therapy (NRT) use for reasons other than quitting smoking among smokers in four countries. Population-based, cross-sectional telephone survey with nationally representative samples of adult smokers in Canada, the United States, the United Kingdom and Australia, conducted in 2005. A total of 6532 adult daily smokers in Canada (n = 1660), the United States (n = 1664), the United Kingdom (n = 1617) and Australia (n = 1591). Survey questions included demographics, smoking behaviour, use of NRT and reasons for NRT use, as well as access and availability of NRT. Approximately 17% of smokers surveyed had used NRT in the past year. Among NRT users, approximately one-third used NRT for a reason other than quitting smoking, including temporary abstinence or reducing the number of cigarettes smoked. The prevalence of non-standard NRT use was remarkably consistent across countries. Using NRT for reasons other than quitting was associated with higher education level, heavier smoking, having no quit intentions, having no past-year quit attempts, the type of NRT product used and accessing NRT without a prescription. The use of NRT for purposes other than quitting smoking is fairly common and may help to explain the difficulty in detecting significant quitting benefits associated with NRT use in population studies. Tobacco control policies, including the accessibility of NRT, may have important implications for patterns of NRT use.


Dual effects of extracellular Ca2+ on cardiotoxin-induced cytotoxicity and cytosolic Ca2+ changes in cultured single cells of rabbit aortic endothelium.
The effects of extracellular Ca2+ on cytotoxicity induced by cardiotoxin (CTX), isolated from Chinese cobra venom, were investigated in cultured rabbit aortic endothelial cells (RAECs). In Hank's buffered saline solution (HBSS) containing 1.2 mM Ca2+, CTX (1-30 microM) caused cell necrosis and cell death in a concentration-dependent manner, as determined by trypan blue exclusion test performed after a 20-min CTX treatment. The concentration of CTX that caused 50% cell death was about 6.5 microM. CTX (10 microM)-induced RAEC damage was also evident but less prominent in Ca2+-free medium and almost completely prevented in medium containing 7-10 mM Ca2+. Therefore, Ca2+ appears to provoke CTX-induced injury at physiological concentrations, but protects against it at high concentrations. The protection of RAECs from CTX-induced injury could also be achieved by high concentrations of Ni2+ and Mg2+. Using the fura-2 fluorescence technique to measure the cytosolic free Ca2+ concentration ([Ca2+]i) of single RAEC, it was shown that in 1.2 mM Ca2+-containing HBSS, treatment of RAECs with 10 microM CTX for 7-35 min resulted in a tremendous and irreversible [Ca2+]i elevation, suggestive of cell membrane damage and extracellular Ca2+ entry. Ni2+ could also enter the cytosol of these damaged RAECs. However, there was no [Ca2+]i elevation or Ni2+ entry in RAECs that were preincubated in HBSS containing 7 mM Ca2+ or Ni2+ before CTX exposure. In RAECs protected with 7 mM Ca2+, the intracellular Ca2+ signals triggered by 100 microM extracellular ATP or 10 microM bradykinin in CTX-treated groups were similar to those in the untreated control groups. Taken together, the results indicate that high extracellular Ca2+ concentrations protected RAECs from CTX-induced injury, and preserved the ability of CTX-treated RAECs to generate Ca2+ signals in response to physiological stimuli.


Effectiveness of Valganciclovir 900mg Versus 450mg for Cytomegalovirus Prophylaxis in Renal Transplantation: A Systematic Review and Meta-Analysis.
Valganciclovir 900 mg/day is approved for cytomegalovirus (CMV) prophylaxis, but 450 mg/day is seems also effective. We systematically reviewed the efficacy and safety of low-dose versus high-dose valganciclovir prophylaxis in renal transplantation recipients. An electronic search was conducted up to November 29, 2016. The primary outcomes were incidences of CMV, CMV disease, mortality and opportunistic infection. The second outcomes were acute rejection, allograft loss, adverse drug reaction (ADR). 7 cohort studies, all with high quality involving (1431 patients) were included. There was no significant difference of the incidence of following CMV disease (1271 patients, odds ratio [OR] 0.74, 95% confidence interval [CI], 0.38-1.43, p=0.36), acute rejection (1343 patients, OR 0.77, 95%CI 0.53-1.14, p=0.19), allograft loss (1271 patients, OR 0.64, 95%CI 0.31-1.35, p=0.24), mortality (1271 patients, OR 0.55, 95%CI 0.20-1.47, p=0.23) and opportunistic infections (OI) (985 patients, OR 0.76, 95%CI 0.52-1.10, p=0.14) between the low-dose and the high-dose valganciclovir prophylaxis. And no significant difference was observed for premature valganciclovir discontinuation (1010 patients, OR 0.81, 95%CI 0.52-1.25, p=0.33) and the incidence of leukopenia (1082 patients, OR 0.65, 95%CI 0.34-1.22, p=0.18) between the two regimens. 450 mg and 900 mg doses of valganciclovir are equipotent for CMV universal prophylaxis. CMV 450 mg prophylaxis should be used for renal transplant recipients. This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page.


A novel human arterial wall-on-a-chip to study endothelial inflammation and vascular smooth muscle cell migration in early atherosclerosis.
Mechanistic understanding of atherosclerosis is largely hampered by the lack of a suitable in vitro human arterial model that recapitulates the arterial wall structure, and the interplay between different cell types and the surrounding extracellular matrix (ECM). This work introduces a novel microfluidic endothelial cell (EC)-smooth muscle cell (SMC) 3D co-culture platform that replicates the structural and biological aspects of the human arterial wall for modeling early atherosclerosis. Using a modified surface tension-based ECM patterning method, we established a well-defined intima-media-like structure, and identified an ECM composition (collagen I and Matrigel mixture) that retains the SMCs in a quiescent and aligned state, characteristic of a healthy artery. Endothelial stimulation with cytokines (IL-1β and TNFα) and oxidized low-density lipoprotein (oxLDL) was performed on-chip to study various early atherogenic events including endothelial inflammation (ICAM-1 expression), EC/SMC oxLDL uptake, SMC migration, and monocyte-EC adhesion. As a proof-of-concept for drug screening applications, we demonstrated the atheroprotective effects of vitamin D (1,25(OH)2D3) and metformin in mitigating cytokine-induced monocyte-EC adhesion and SMC migration. Overall, the developed arterial wall model facilitates quantitative and multi-factorial studies of EC and SMC phenotype in an atherogenic environment, and can be readily used as a platform technology to reconstitute multi-layered ECM tissue biointerfaces.


An RNAi screen identifies Msi2 and Prox1 as having opposite roles in the regulation of hematopoietic stem cell activity.
In this study, we describe an in vivo RNA interference functional genetics approach to evaluate the role of 20 different conserved polarity factors and fate determinants in mouse hematopoietic stem cell (HSC) activity. In total, this screen revealed three enhancers and one suppressor of HSC-derived reconstitution. Pard6a, Prkcz, and Msi2 shRNA-mediated depletion significantly impaired HSC repopulation. An in vitro promotion of differentiation was observed after the silencing of these genes, consistent with their function in regulating HSC self-renewal. Conversely, Prox1 knockdown led to in vivo accumulation of primitive and differentiated cells. HSC activity was also enhanced in vitro when Prox1 levels were experimentally reduced, identifying it as a potential antagonist of self-renewal. HSC engineered to overexpress Msi2 or Prox1 showed the reverse phenotype to those transduced with corresponding shRNA vectors. Gene expression profiling studies identified a number of known HSC and cell cycle regulators as potential downstream targets to Msi2 and Prox1.


Under-five mortality pattern and associated risk factors: a case-control study at the Princess Marie Louise Children's Hospital in Accra, Ghana.
Millions of children under the age of 5 years die every year. Some of these deaths occur in hospitals and are related to both clinical characteristics and modifiable risk factors. This study investigates the association between malnutrition and mortality and profiles the presenting features in a case-control study of children under 5 years of age who attended Princess Marie Louise Children's Hospital (PML) in 2011. A total of 120 cases of children under the age of 5 years who were admitted to hospital and died there were matched by sex and age to 120 controls who were children who survived on 1:1 basis from a record of patients admitted to PML in 2011. Data on socio-demographic and clinical characteristics were extracted from the medical records of the study participants. The association between malnutrition and mortality was determined by conditional logistic regression reported as odds ratios (OR) and their 95 % confidence interval (95 % CI). P < 0.05 was considered significant in all analyses. Malnutrition was significantly associated with mortality in children under-5 years of age attending PML. In the adjusted analysis, the odds of dying was significantly higher in malnourished children compared with well-nourished children (adjusted OR = 4.32 [95 % CI, 1.33-13.92], p = 0.014]). In addition, a previous episode of diarrhoea within the last year was associated with mortality (adjusted OR = 7.25 [95 % CI, 1.68-31.22], p = 0.008). The proportion of patients with noisy or difficulty breathing, pallor, lethargic appearance, ill-looking appearance, febrile convulsion, altered sensorium, skin lesions, hepatomegaly or oedema was significantly higher among cases than in controls (p < 0.05). Malnutrition and a previous episode of diarrhoea within the last year were the main risk factors for mortality. Efforts to prevent malnutrition and diarrhoea must be intensified and a protocol to follow-up diarrhoea patients may be beneficial. Six out of the nine clinical features that were proportionally higher in children who died than those who survived, are captured by the Emergency Triage Assessment and Treatment (ETAT) screening protocol as emergency or priority signs, giving credence to the use of ETAT in this setting. Thus education of health professionals on the use of the tool to triage patients should be on-going. However, further studies are needed to establish whether the other clinical signs are consistently associated with mortality and if so, whether they can be included among triage criteria, danger signs or in a prognostic scoring system for this setting.


Co-Occurrence of ANCA-Associated Vasculitis and Sjögren's Syndrome in a Patient With Acromegaly: A Case Report and Retrospective Single-Center Review of Acromegaly Patients.
ANCA-associated vasculitis (AAV) and Sjögren's syndrome (SS) are uncommon autoimmune diseases. The co-occurrence in the same patient has been rarely described. Acromegaly has been associated with autoimmune thyroiditis, but the prevalence of other autoimmune disorders such as AAV and SS has not been evaluated in acromegaly. Characterization of a patient with acromegaly and two rare autoimmune diseases-SS and AAV (microscopic polyangiitis (MPA))-by autoantibody-array and whole exome sequencing (WES). Single-center retrospective review of medical records of acromegaly patients to explore the prevalence of diagnosed autoimmune diseases. We report a Caucasian woman in her 50's with a serologically (anti-SSA/Ro, anti-MPO-ANCA antibodies) and histologically confirmed diagnosis of symptomatic SS and MPA. SS with MPO-ANCA positivity preceded MPA. An exploratory autoantigen array detected a broad spectrum of autoantibodies. WES revealed heterozygous carrier status of the PTPN22 mutation R620W, which is associated with an increased risk for autoimmunity. A similar combination of positive anti-SSA/Ro autoantibodies and ANCA was only present in 5/1184 (0.42%) other patients tested for both antibodies in our clinic over six years. Amongst 85 acromegaly patients seen at our clinic in a 20-year period, 12% had a clinically relevant associated immunological disease. We present a rare case of SS and AAV in a patient with acromegaly and multiple autoantibody specificities. Patients with SS and ANCA should be closely monitored for the development of (subclinical) AAV. Whether acromegaly represents a risk for autoimmunity should be further investigated in prospective acromegaly cohorts.


Antiviral Properties of Chemical Inhibitors of Cellular Anti-Apoptotic Bcl-2 Proteins.
Viral diseases remain serious threats to public health because of the shortage of effective means of control. To combat the surge of viral diseases, new treatments are urgently needed. Here we show that small-molecules, which inhibit cellular anti-apoptotic Bcl-2 proteins (Bcl-2i), induced the premature death of cells infected with different RNA or DNA viruses, whereas, at the same concentrations, no toxicity was observed in mock-infected cells. Moreover, these compounds limited viral replication and spread. Surprisingly, Bcl-2i also induced the premature apoptosis of cells transfected with viral RNA or plasmid DNA but not of mock-transfected cells. These results suggest that Bcl-2i sensitizes cells containing foreign RNA or DNA to apoptosis. A comparison of the toxicity, antiviral activity, and side effects of six Bcl-2i allowed us to select A-1155463 as an antiviral lead candidate. Thus, our results pave the way for the further development of Bcl-2i for the prevention and treatment of viral diseases.


The rise of the general practitioner in the nineteenth century.
It was the Apothecaries Act of 1815 which led to the emergence of general practice as we know it today and it was this one Act which produced a flood of changes on the medical scene that are without parallel in our history. Students were soon to undergo new forms of training in new medical schools and hospitals, and many medical associations and journals were founded. The term ;general practitioner' was soon in use. The driving force behind all these changes was the Society of Apothecaries and the new general practitioners, and all too often they were opposed by the two Royal Colleges. It was only at the beginning of the twentieth century that these new practitioners were allowed to call themselves ;doctors'.


A protein functional leap: how a single mutation reverses the function of the transcription regulator TetR.
Today's proteome is the result of innumerous gene duplication, mutagenesis, drift and selection processes. Whereas random mutagenesis introduces predominantly only gradual changes in protein function, a case can be made that an abrupt switch in function caused by single amino acid substitutions will not only considerably further evolution but might constitute a prerequisite for the appearance of novel functionalities for which no promiscuous protein intermediates can be envisaged. Recently, tetracycline repressor (TetR) variants were identified in which binding of tetracycline triggers the repressor to associate with and not to dissociate from the operator DNA as in wild-type TetR. We investigated the origin of this activity reversal by limited proteolysis, CD spectroscopy and X-ray crystallography. We show that the TetR mutant Leu17Gly switches its function via a disorder-order mechanism that differs completely from the allosteric mechanism of wild-type TetR. Our study emphasizes how single point mutations can engender unexpected leaps in protein function thus enabling the appearance of new functionalities in proteins without the need for promiscuous intermediates.


Tolvaptan Improves the Long-Term Prognosis in Patients With Congestive Heart Failure With Preserved Ejection Fraction as Well as in Those With Reduced Ejection Fraction.
Tolvaptan (TLV), an arginine vasopressin type 2 antagonist, has been shown to play a role in ameliorating symptomatic congestion and normalizing diluted hyponatremia in patients with congestive heart failure (HF). However, most evidence was derived from patients with HF with reduced ejection fraction (HFrEF), and the clinical efficacy of TLV in patients with HF with preserved ejection fraction (HFpEF) remains uncertain. In this study, we retrospectively enrolled 60 in-hospital patients with stage D HF, who had received TLV to treat symptomatic congestion at our institute between 2011 and 2013. As a control group, we also enrolled 60 background-matched HF patients who did not receive TLV therapy. Patients with HFpEF (n = 29), whose left ventricular ejection fraction was > 45%, had higher age and a lower urine aquaporin-2 level relative to the plasma arginine vasopressin concentration compared with those with HFrEF (n = 91). TLV therapy significantly reduced the 2-year readmission rates in both the HFrEF and HFpEF populations (P < 0.05 for both), indicating that TLV therapy may improve the long-term prognosis not only in patients with HFrEF but also in those with HFpEF.


Salicylic acid is part of the Mi-1-mediated defense response to root-knot nematode in tomato.
The Mi-1 gene of tomato confers resistance against three species of root-knot nematode in tomato (Lycopersicon esculentum). Transformation of tomato carrying Mi-1 with a construct expressing NahG, which encodes salicylate hydroxylase, a bacterial enzyme that degrades salicylic acid (SA) to catechol, results in partial loss of resistance to root-knot nematodes. Exogenous SA was toxic to roots expressing NahG but not to control roots. This toxicity is most likely due to the production of catechol from SA, and we report here that 100 microM catechol is toxic to tomato roots. Benzothiadiazole, a SA analog, completely restores nematode resistance in Mi-1 roots transformed with NahG but does not confer resistance to susceptible tomato roots. The localized cell death produced by transient expression in Nicotiana benthamiana of Mi-DS4, a constitutively lethal chimera of Mi-1 with one of its homologs, was prevented by coexpression of NahG. These results indicate that SA is an important component of the signaling that leads to nematode resistance and the associated hypersensitive response.


Groucho acts as a corepressor for a subset of negative regulators, including Hairy and Engrailed.
Relatively little is known about the molecular mechanisms involved in transcriptional repression, despite its importance in development and differentiation. Recent evidence suggests that some transcriptional repressors act by way of adaptor molecules known as corepressors. Here, we use in vivo functional assays to test whether different repressor activities are mediated by the Groucho (Gro) corepressor in the Drosophila embryo. Previously, Gro was proposed to mediate repression by the Hairy-related family of basic helix-loop-helix proteins. Our results indicate not only that repression by Hairy requires Gro, but that a repressor domain from the Engrailed (En) homeodomain protein is also Gro dependent. The latter result correlates with an ability of this En domain to bind to Gro in vitro. In contrast, repressor regions from the Even-skipped, Snail, Krüppel, and Knirps transcription factors are effective in the absence of Gro. These results show that Gro is not generally required for repression, but acts as a specific corepressor for a fraction of negative regulators, including Hairy and En.


The "sphere of care": A qualitative study of colorectal cancer patient and caregiver experiences of support within the cancer treatment setting.
Colorectal cancer is associated with considerable physical and psychosocial burden. Whilst social support is known to facilitate psychological adjustment to cancer, patients' and caregivers' experiences of social support within a treatment setting and their perceptions of the role of the treating team in providing this support is unknown. Specifically, there is a gap in the research that explores in detail who people affected by colorectal cancer consider to be supportive, and the function, timing and nature of this support, whilst receiving treatment. This study explored both patients' and caregivers' a) experiences of social support and how this relates to their experience of treatment; and b) what facilitates support in the treatment setting. Individual interviews (N = 20) were conducted with patients diagnosed with colorectal cancer and caregivers of such patients. Audiotaped interviews were transcribed verbatim and analysed using the framework method. Three major themes emerged from the data: a) treating team as a source of support, highlighting the importance of connection with the treating team; b) changes in existing social supports, encompassing issues regarding distance in interpersonal relationships as a consequence of cancer; and c) differing dimensions of support, exploring the significance of shared experience, practical, financial, and emotional support. Patients and caregivers perceived the treating team as a major source of support. Support from the treating team was particularly important in the context of the changes that occur as a result of a diagnosis of colorectal cancer and the effects of subsequent treatment. Incidental support from others encountered in the treatment setting was also experienced and was equally important to both patients and caregivers. This has implications for the way health care professionals respond to both patients and caregivers in the treatment setting in terms of communication, interventions and environment.


Laboratory evaluation of pathogenicity of entomogenous nematodes to African tick species.
Five strains of entomogenous nematodes, Steinernema carpocapsae strains DD, Mexican, SR, and Heterorhabditis bacteriophora strains 1S5 and HP88, were tested for their pathogenicity to various developmental stages of five African tick species namely; Rhipicephalus appendiculatus, R. evertsi, Amblyomma variegatum, A. gemma, and Boophilus decoloratus. In engorged female R. appendiculatus, all nematodes at a concentration of 1,000 infective juveniles (IJ)/dish, except S. carpocapsae Mexican strain, induced high mortalities (56-100%), whereas in engorged female R. evertsi, only S. carpocapsae DD and H. bacteriophora HP88 induced high mortalities (78% and 56%, respectively). In engorged B. decoloratus, S. carpocapsae DD, Mexican, SR and H. bacteriophora HP88 (100 IJ/dish) induced mortalities of 85%, 65%, 80%, and 100%, respectively. In all cases, except for S. carpocapsae Mexican strain, a higher concentration (5,000 IJ/dish) did not result in higher mortality than occurred with 1,000 IJ/dish. Unfed females and immature stages of ticks were found to be generally resistant to the nematodes. The feasibility of using entomogenous nematodes for biological control of African tick species are briefly discussed.


Relationships Among Retinal Nerve Fiber Layer Thickness, Vascular Endothelial Growth Factor, and Cognitive Impairment in Patients with Schizophrenia.
Studies have suggested retinal nerve fiber layer (RNFL) involvement in the pathogenesis of schizophrenia. Additionally, research has shown that vascular endothelial growth factor (VEGF) potentially contributes to the pathophysiology of psychiatric disorders. Therefore, this study aimed to investigate VEGF, RNFL, and correlations with cognitive impairments in schizophrenia patients. Patients with schizophrenia (n = 138) were compared to healthy controls (n = 160). RNFLs were measured with optical coherence tomography (OCT). The Stroop color and word test (SCWT) was used to evaluate neurocognition. Blood samples were collected to measure VEGF. SPSS 20.0 was used to perform analysis of covariance, t-tests, partial correlation analysis, and linear regression. Thinner RNFLs were found in schizophrenia patients (p < 0.001). RNFL showed a significant correlation with SCWT scores (all p < 0.05). Serum level of VEGF was lower in patients with schizophrenia (p = 0.010). Total and inferior RNFL thicknesses of right eyes were positively correlated to VEGF level (RNFL total thickness p = 0.032, inferior thickness p = 0.014).Total RNFL thicknesses were shown to be reduced following a prolonged duration of illness (both p < 0.01). These findings suggest that patients with schizophrenia have degeneration with RNFL thickness following illness duration, which may contribute to neurocognitive impairments observed in schizophrenia. VEGF is speculated to play some important role on RNFL degeneration with schizophrenia patients.


Higher HTLV-1c proviral loads are associated with blood stream infections in an Indigenous Australian population.
An association between blood stream infections (BSI) and HTLV-1 seropositivity in Indigenous Australians might result from HTLV-1 mediated inflammation and parasite coinfections that provide portals of entry for bacteria. To determine whether BSI risk increases with HTLV-1c proviral load (PVL) and to identify the pathogens responsible in the context of HTLV-1 related conditions. Indigenous adults admitted to Alice Springs Hospital, central Australia, were recruited as cases or controls according to whether they had a BSI. Clinical data were extracted from case notes and the hospital pathology database. HTLV-1 serology and PVL assays were then performed and risk factors for BSI were determined for HTLV-1 infected subjects. Risk factors were compared between 44 cases and 30 controls who were HTLV-1 Western blot positive. In a multivariable model, high HTLV-1c PVL was predictive of BSI during the study period (OR, 9.10; 95% CI, 2.40-34.4). Median (IQR) HTLV-1c PVL (copies per 100 PBL) was 39-fold higher for patients recording any BSI (0.116 (0.009, 0.277)) relative to those who had no BSI (0.003 (0.000, 0.067))(p=0.0007). Mean (SD) PVL for subjects with no BSI (n=27), 1 BSI (n=37) and ≥2 BSI (n=10) during the study period were 0.120 (0.301), 0.271 (0.622) and 0.500 (0.654) copies per 100 PBL, respectively (p=0.0007). Five BSI were associated with possible complications of HTLV-1; strongyloidiasis (3), infective dermatitis (1), HTLV-1 associated bronchiectasis (1). Higher HTLV-1c PVL predicts BSI risk; however; most BSI were not associated with recognised complications of HTLV-1 infection.


Levothyroxine Dosing after Delivery in Women Diagnosed with Hypothyroidism During Pregnancy-A Retrospective, Observational Study.
Pregnancy leads to profound alteration in thyroid function and dysthyroidism contributes to adverse pregnancy outcomes. Though the management of hypothyroidism during pregnancy is highlighted, the same is often neglected during postpartum. We have evaluated the postpartum levothyroxine (LT4) dose change in patients with new onset hypothyroidism. We conducted this retrospective, observational study between 2014 and 2016 using the medical records of patients with new onset hypothyroidism during pregnancy. We included patients who continued with LT4 after delivery (as per predetermined protocol) and the availability of 2-year follow up record. We excluded patients who stopped LT4 and use of other drugs that affect the thyroid function tests (TFT) after delivery. The patients were divided into 2 groups for comparison [Group 1-Overt hypothyroidism (OH) and Group 2-Subclinical hypothyroidism (SCH)] based on the initial TFT reports. The data were analyzed using appropriate statistical methods and a P value of less than 0.05 was considered significant. A total of 159 women continued using LT4 after delivery and the final follow up data were available for 130 patients only. LT4 dose up titration was observed more in group 1 than in group 2 (P = 0.0336). In both the groups, the presence of goitre, thyroid autoimmunity and a repeat pregnancy are associated with increasing LT4 requirement. Majority of patients with OH during pregnancy require more than half of the final dose after delivery. Goitre and autoimmunity are associated with higher LT4 dose after delivery.


Minimizing the impact of drug-resistant Streptococcus pneumoniae (DRSP). A strategy from the DRSP Working Group.
Emergence of drug-resistant Streptococcus pneumoniae (DRSP) presents a challenge to the medical and public health communities since the magnitude of the problem is not known, the clinical impact of DRSP infections is not well described, national vaccination rates are low, and antimicrobial drugs are often used excessively and inappropriately. To address the problem of DRSP, a working group by Centers for Disease Control and Prevention was formed in June 1994 consisting of public health practitioners, health care providers, and clinical laboratorians representing state and federal agencies and various professional organizations. Through periodic open meetings, the working group has developed a strategy for surveillance, investigation, prevention, and control of infections due to DRSP. The strategy focuses on (1) implementing an electronic laboratory-based surveillance (ELBS) system for reporting invasive DRSP infections and providing clinically relevant feedback to clinicians, (2) identifying risk factors and outcomes of DRSP infection, (3) increasing pneumococcal vaccination, and (4) promoting judicious antimicrobial drug use. Data received through ELBS will be used to make timely estimates of the community-specific prevalence of drug-resistant pneumococci. National, regional, and local trends will be made available to health care providers and clinicians to promote optimal antimicrobial drug use and increased vaccination in targeted areas. Once in operation, the ELBS network will be adaptable to other diseases, improving the comprehensiveness and timeliness of public health surveillance. The intended outcome of the strategy is to reduce complications of DRSP infection, such as long-term sequelae of infection, health care expenditures, morbidity, and mortality.


Enantiomer-specific accumulation of PCB atropisomers in the bowhead whale (Balaena mysticetus).
Blubber (n = 40) and liver (n = 20) samples from the bowhead whale (Balaena mysticetus) were collected during the 1997-1998 Native (Inuit) subsistence harvests in Barrow, AK. Bowhead tissues and zooplankton were analyzed for polychlorinated biphenyl (PCB) concentrations and the enantiomeric fractions (EFs) of eight chiral PCB congeners (PCB-91, 95, 135, 136, 149, 174, 176, and 183) to quantify the enantiomer-specific accumulation of PCBs in this cetacean. PCB concentrations in bowhead blubber were low (mean +/- 1 SE: 610 +/- 54 ng g(-1) lipid) relative to other cetaceans. The accumulation of several chiral PCBs (PCB-91, 135, 149, 174, 176, and 183) in bowhead blubber was enantiomer-specific relative to bowhead liver and zooplankton, suggesting that biotransformation processes within the bowhead whale are enantioselective. The EFs for PCB-95 and 149 were significantly correlated with body length in male and female whales, while EFs for PCB-91 correlated with length in males only. Despite evidence for enantioselective biotransformation, all three congeners bioaccumulated in the bowhead relative to PCB-153. Results suggest that enantioselective accumulation of PCB-91, 95, and 149 is influenced by PCB concentrations, age, and/or the modification of an uncharacterized stereoselective process (or processes) during sexual maturity.


Plasma Cell Myeloma in Children and Young Adults: A Report of 4 Cases From a Single Institution and a Review of the Literature.
Plasma cell myeloma (PCM) is rare in children and young adults and therefore may be difficult to diagnose. Here we report the clinicopathologic findings of 4 patients under the age of 30 diagnosed with PCM at our institution and summarize the literature about 48 other cases of PCM in this age group. The male:female ratio was 1.2:1 and the number of cases increased with age. Children and young adults with PCM often present with a plasmacytoma and are less likely to have asymptomatic PCM than their adult counterparts. From the cases that reported ethnicity, the majority (55%) were non-white suggesting a possible ethnic predisposition to PCM in this age group. PCM should be included in the differential diagnosis of mass lesions, especially a destructive bony lesion, after more common causes have been ruled out in this age group. The optimal treatment for PCM in this patient population is unclear and conclusions into this are hampered by the paucity of cases and the lack of standardized follow-up.


Developmental Changes in Sexual Risk and Substance Use Among African American Females: an Integrated Data Analysis Approach Using Time-varying Effect Models.
The aim of this study was to describe age-related changes in sexual risk and substance use using existing data from three HIV/sexually transmitted infection (STI) prevention trials conducted in Atlanta, Georgia, that enrolled young African American women. We used two novel analysis methods: integrative data analysis (IDA) and time-varying effect models (TVEM). Each trial collected self-reported behavioral data and vaginal swab specimens assayed for chlamydia, gonorrhea, and trichomonas. Baseline data from all participants and follow-up data from participants not randomized to an active intervention arm were integrated in a pooled dataset using an IDA approach. The pooled dataset included observations for 1974 individuals, aged 14-25 years for behavioral outcomes and 16-25 years for STI outcomes. We used TVEM to model age-related changes in sexual risk and substance use behaviors and STI acquisition, adjusting for trial as a fixed effect. Coital frequency and condomless sex increased through the early 20s while multiple partnerships remained fairly steady. Alcohol use and cigarette smoking increased through about age 24 years before declining. Marijuana use peaked at age 16-17 years and thereafter generally declined. STI acquisition was highest at age 16 years. This study demonstrates the feasibility and utility of innovative methodological techniques to address novel questions related to adolescent development using existing data from multiple trials. The results suggest that mid-adolescence and the early 20s may be periods of particular risk. The findings may be useful for timing culturally and developmentally relevant prevention interventions for young African American women.


Resveratrol inhibits matrix metalloproteinases to attenuate neuronal damage in cerebral ischemia: a molecular docking study exploring possible neuroprotection.
The main pathophysiology of cerebral ischemia is the structural alteration in the neurovascular unit, coinciding with neurovascular matrix degradation. Resveratrol has been reported to be one of the most potent chemopreventive agents that can inhibit cellular processes associated with ischemic stroke. Matrix metalloproteinases (MMPs) has been considered as a potential drug target for the treatment of cerebral ischemia. To explore this, we tried to investigate the interaction of resveratrol with MMPs through molecular docking studies. At 30 minutes before and 2 hours after cerebral ischemia/reperfusion induced by occlusion of the middle cerebral artery, 40 mg/kg resveratrol was intraperitoneally administered. After resveratrol administration, neurological function and brain edema were significantly alleviated, cerebral infarct volume was significantly reduced, and nitrite and malondialdehyde levels in the cortical and striatal regions were significantly decreased. The molecular docking study of resveratrol and MMPs revealed that resveratrol occupied the active site of MMP-2 and MMP-9. The binding energy of the complexes was -37.848672 kJ/mol and -36.6345 kJ/mol for MMP-2 and MMP-9, respectively. In case of MMP-2, Leu 164, Ala 165 and Thr 227 were engaged in H-Bonding with resveratrol and in case of MMP-9, H-bonding was found with Glu 402, Ala 417 and Arg 424 residues. These findings collectively reveal that resveratrol exhibits neuroprotective effects on cerebral ischemia through inhibiting MMP-2 and MMP-9 activity.


Incidence, clinical spectrum, and outcomes of primary sclerosing cholangitis in a United States community.
The epidemiology of primary sclerosing cholangitis (PSC) in the United States is unknown. We report the incidence, clinical spectrum, and outcomes of PSC in Olmsted County, Minnesota. Using the Rochester Epidemiology Project, a medical records linkage system in Olmsted County, Minnesota, we identified county residents with PSC, and the diagnosis was confirmed according to clinical, biochemical, radiographic, and histologic criteria. Twenty-two patients met diagnostic criteria for PSC in 1976-2000. The age-adjusted (to 2000 U.S. whites) incidence of PSC in men was 1.25 per 100,000 person-years (95% CI, 0.70 to 2.06) compared with 0.54 per 100,000 person-years (95% CI, 0.22 to 1.12) in women. The prevalence of PSC in 2000 was 20.9 per 100,000 men (95% CI, 9.5 to 32.4) and only 6.3 per 100,000 women (95% CI, 0.1 to 12.5). Seventy-three percent of cases had inflammatory bowel disease, the majority with ulcerative colitis. Survival among PSC patients was significantly less than expected for the Minnesota white population of similar age and gender (P < 0.001). These data represent the first population-based estimates of the incidence and prevalence of PSC in the United States. The incidence and prevalence of PSC were approximately one third of those previously described for primary biliary cirrhosis in the same population. Our data suggest that the prevalence of PSC in the United States, with its attendant medical burdens, is significantly greater than previously estimated.


[Lung cancer mortality trends in Colombia, 1985-2018Tendências na mortalidade por câncer de pulmão na Colômbia de 1985 a 2018].
To determine lung cancer mortality trends in Colombia during the period 1985-2018 in the population aged 35 years and over and identify changes in the trend. Analysis of mortality time series. The specific standardized rates by sex and age group were calculated. Using joinpoint regression, the annual percentage change in the rates was estimated and points of change were identified. During the period 1985-2018, 105 553 deaths from lung cancer were reported in the population aged 35 and over. The standardized rates exhibit a downward trend during the period 1985-2005, except in people over the age of 64. Lung cancer death rates in Colombia are trending downward. Primary and secondary prevention measures with respect to tobacco use need to be enhanced and other risk factors, such as residential radon or occupation, monitored.


Water drinking in rats resulting from intravenous relaxin and its modification by other dipsogenic factors.
The purpose of the study was to determine whether iv infusion of relaxin would acutely stimulate water drinking in rats and, if it did, whether such drinking is affected by other dipsogenic stimuli or is blocked by centrally administered losartan. iv infusions of human gene 2 relaxin at doses of 25, 40, 55, or 80 microg/kg x h for 1 h induced dose-dependent water drinking in both male and female rats within 15-30 min of commencement of infusions. iv infusion of a nondipsogenic dose of angiotensin II (0.5 microg/h), combined with relaxin (40 microg/kg x h), almost tripled the relaxin-induced water intake. iv infusion of hypertonic (1 M) NaCl did not potentiate relaxin-induced drinking. Intracerebroventricular injection of the angiotensin AT1 antagonist losartan (10 microg) reduced water drinking induced by iv infusion of relaxin. The water drinking induced by iv infusion of relaxin in the rat suggests that blood-borne relaxin may be a dipsogenic hormone. Potentiation of this relaxin-induced drinking by moderate levels of circulating angiotensin II is additional evidence in support of this view. The results also indicate that a central angiotensinergic neural pathway, utilizing AT1 receptors, subserves relaxin-induced drinking.


New substrate analogue inhibitors of factor Xa containing 4-amidinobenzylamide as P1 residue: part 1.
The trypsin-like serine protease factor Xa (fXa) is located at the convergence point of the intrinsic and extrinsic coagulation cascade, and therefore has emerged as an attractive target for the design of novel anticoagulants. During the development of substrate-analogue urokinase inhibitors we have found that the protection of the P3-dSer side chain leads to a scaffold of potent fXa inhibitors with the general structure R1-SO2-dSer(R2)-Gly-4-amidinobenzylamide. The first lead (3) with an N-terminal benzylsulfonyl group and dSer(tBu) as P3 residue inhibits human fXa with a Ki of 14 nM. A variety of derivatives with modified P4, P3, and P2 residues have been investigated in terms of inhibition of fXa and related proteases and for their anticoagulant potency and elimination behaviour. Most inhibitors were rapidly cleared from the circulation of rats. However, compound 48 (Ki= 3.5 nM), one of the most potent and selective inhibitors containing a dArg as P3 residue was relatively slowly eliminated (t1/2 approximately 1 h). Inhibitor 48 doubled clotting times in human plasma at 0.32 microM (aPTT) and 0.28 microM (PT), and is approximately 10-fold more potent than the reference fXa inhibitor DX-9065a in the inhibition of the prothrombinase complex. The structures of two inhibitors in complex with human fXa were solved by X-ray crystallography.


Lidar Observation of Cloud.
Lidar (from "light detection and ranging") is the optical counterpart of meteorological radar. At optical wavelengths, very much smaller atmospheric particles can be detected than at microwave wavelengths. With a laser power source, a transmitter uses a lens system to beam very intense pulses of monochromatic light of extremely short duration. Light backscattered by the atmosphere is collected in a receiver system that is essentially a telescope coaligned with the transmitter, and a narrow-pass filter allows only light of the transmitted frequency to be detected by a photomultiplier. Data are presented on an oscilloscope as a trace of signal intensity versus range (the A-scope of radar practice) and photographed.


Predictive factors for the outcome of anterior cervical decompression and fusion.
In a prospective study, 103 patients were randomised to anterior cervical decompression and fusion (ACDF) with a cervical carbon-fibre intervertebral fusion cage or the Cloward procedure. Preoperative background variables, active range of neck motion, handgrip strength, radiological evaluation and subjective variables were used in a multiple regression model to find the strongest predictors of postoperative outcome as measured by current pain intensity and the Neck Disability Index (NDI). Male sex, greater kyphosis at the level operated on, non-smoking, a greater neck mobility in right rotation, low disability on NDI, and older age were predictors of pain reduction and explained 30% of current pain intensity at follow-up. Higher educational level, non-smoking, greater kyphosis at the level operated on, a greater flexion mobility, greater right handgrip strength and lower current pain intensity were predictors of improvement, and explained 28% of the postoperative outcome on NDI. The most important predictor for postoperative pain intensity was the magnitude of the preoperative kyphosis. Preoperative pain intensity was the most important predictor for improved NDI. At follow-up about 70% of the patients still had deficit based on current pain intensity and NDI, and 44% had remaining dysfunction based on Odom's criteria. In conclusion, the multivariate analysis shows that male sex, non-smoking, greater segmental kyphosis and a low pain and disability level are preoperative predictors of a good outcome in ACDF. In addition, the study suggests the importance of other predictive variables than those studied for the outcome of ACDF.


Gene expression and distribution of Swi6 in partial aneuploids of the fission yeast Schizosaccharomyces pombe.
Imbalances of gene expression in aneuploids, which contain an abnormal number of chromosomes, cause a variety of growth and developmental defects. Aneuploid cells of the fission yeast Schizosaccharomyces pombe are inviable, or very unstable, during mitotic growth. However, S. pombe haploid cells bearing minichromosomes derived from the chromosome 3 can grow stably as a partial aneuploid. To address biological consequences of aneuploidy, we examined the gene expression profiles of partial aneuploid strains using DNA microarray analysis. The expression of genes in disomic or trisomic cells was found to increase approximately in proportion to their copy number. We also found that some genes in the monosomic regions of partial aneuploid strains increased their expression level despite there being no change in copy number. This change in gene expression can be attributed to increased expression of the genes in the disomic or trisomic regions. However, even in an aneuploid strain that bears a minichromosome containing no protein coding genes, genes located within about 50 kb of the telomere showed similar increases in expression, indicating that these changes are not a secondary effect of the increased gene dosage. Examining the distribution of the heterochromoatin protein Swi6 using DNA microarray analysis, we found that binding of Swi6 within ~50 kb from the telomere occurred less in partial aneuploid strains compared to euploid strains. These results suggest that additional chromosomes in aneuploids could lead to imbalances in gene expression through changes in distribution of heterochromatin as well as in gene dosage.


The effect of blood volume and volume loading on left ventricular diastolic function in individuals with spinal cord injury.
Cohort Study (Prospective Observational Study). The objectives of the study were (i) to examine left ventricular (LV) diastolic function at rest and during rapid saline infusion in those with spinal cord injury (SCI) and (ii) to determine the contribution of blood volume on the purported diastolic impairments in individuals with SCI. St Catharines, ON, Canada. Thirteen SCI (AIS:A-D; C4-T6; age: 41±8.5; 10 males, 3 females) and 12 able-bodied (AB) individuals (age: 40±8.5; 9 males, 3 females) without a history of cardiovascular disease participated in the study. LV structure and diastolic function were assessed via conventional echocardiography. The carbon monoxide rebreathe test was used to measure the blood volume. LV diastolic function was assessed once more following rapid saline infusion (dose: 15 ml kg-1; rate: 100 ml min-1). Compared with the AB group, individuals with SCI had a smaller LV internal diameter (SCI: 4.5±0.3 cm vs AB: 5.1±0.7 cm; P=0.01), lower blood volume (SCI: 3.9±0.6 l vs AB: 5.0±1.2 l; P=0.02) and end-diastolic volume (SCI: 97.2±29.4 ml vs AB: 128.6±38.3 ml; P=0.03). There were no between-group differences in baseline diastolic parameters; however, when LV internal diameter was adjusted for individuals with SCI demonstrated lower early to late transmitral velocity ratio (SCI: 1.9±0.5 vs AB: 2.2±0.7; P=0.03). There was no between-group difference in diastolic responses to the saline infusion, as both groups showed similar diastolic changes following volume loading. Individuals with SCI have preserved LV diastolic function despite having lower preload. Preserved diastolic function may be mediated by the cardiac atrophy that occurs following SCI. Individuals with SCI also demonstrate normal diastolic responses to increased volume loading, suggesting compliant ventricles.


The Dihydroxy Metabolite of the Teratogen Thalidomide Causes Oxidative DNA Damage.
Thalidomide [α-(N-phthalimido)glutarimide] (1) is a sedative and antiemetic drug originally introduced into the clinic in the 1950s for the treatment of morning sickness. Although marketed as entirely safe, more than 10 000 babies were born with severe birth defects. Thalidomide was banned and subsequently approved for the treatment of multiple myeloma and complications associated with leprosy. Although known for more than 5 decades, the mechanism of teratogenicity remains to be conclusively understood. Various theories have been proposed in the literature including DNA damage and ROS and inhibition of angiogenesis and cereblon. All of the theories have their merits and limitations. Although the recently proposed cereblon theory has gained wide acceptance, it fails to explain the metabolism and low-dose requirement reported by a number of groups. Recently, we have provided convincing structural evidence in support of the presence of arene oxide and the quinone-reactive intermediates. However, the ability of these reactive intermediates to impart toxicity/teratogenicity needs investigation. Herein we report that the oxidative metabolite of thalidomide, dihydroxythalidomide, is responsible for generating ROS and causing DNA damage. We show, using cell lines, the formation of comet (DNA damage) and ROS. Using DNA-cleavage assays, we also show that catalase, radical scavengers, and desferal are capable of inhibiting DNA damage. A mechanism of teratogenicity is proposed that not only explains the DNA-damaging property but also the metabolism, low concentration, and species-specificity requirements of thalidomide.


Coping with large litters: management effects on welfare and nursing capacity of the sow.
A number of management issues can be used as drivers for change in order to improve animal welfare and nursing capacity of the hyperprolific sow. Group housing of sows during gestation is a recommended practice from the perspective of animal welfare. Related health issues include reproductive health and the locomotor system. It appears that management of pregnant sows in groups is challenging for a producer and considerable skill is required. We explored the benefits and challenges of group housing, including feeding issues. Increasing litter size requires additional attention to the mammary gland and its ability to provide sufficient nursing for the growing litter. We discuss the fundamentals of mammary development and the specific challenges related to the hyperprolific sow. We also address challenges with the farrowing environment. It appears that the old-fashioned farrowing crate is not only outdated in terms of welfare from the public's perspective, but also fails to provide the environment that the sow needs to support her physiology of farrowing, nursing, and maternal behaviour. Studies from our group and others indicate that providing the sow with a loose housing system adequate in space and nesting material, along with reasonable chance for isolation, can be considered as fundamental for successful farrowing of the hyperprolific sow. It has also been shown that management strategies, such as split suckling and cross fostering, are necessary to ensure proper colostrum intake for all piglets born alive in a large litter. We thus conclude that welfare and nursing capacity of the sow can be improved by management. However, current megatrends such as the climate change may change sow management and force the industry to rethink goals of breeding and, for instance, breeding for better resilience may need to be included as goals for the future.


Increase in the population of duodenal immunoglobulin A plasmocytes in axenic mice associated with different living or dead bacterial strains of intestinal origin.
Various bacterial strains were tested for their ability to stimulate immunoglobulin A (IgA) plasmocytes to populate the duodenal lamina propria in axenic mice. The mice were associated with the strains for at least 4 weeks. The strains inhabiting the conventional mouse intestine and belonging to the genera Lactobacillus, Streptococcus, Eubacterium, Actinobacillus, Micrococcus, Corynebacterium, and Clostridium (including the extremely oxygen-sensitive ones) are only slightly or nonimmunogenic, whereas the strains belonging to the genera Bacteroides and Escherichia have an immunogenic effect. The same result was obtained with Bacteroides and Escherichia strains isolated from the digestive tract of other animal species. The kinetics of appearance of intestinal IgA plasmocytes are similar in axenic mice monoassociated with a stimulatory strain and in conventional mice. The association of two or more strains with axenic mice leads either to the same or a greater number of duodenal IgA plasmocytes as that obtained with the most stimulatory strain monoassociated with axenic mice. The maximum stimulation recorded in all of these trials represents about two-thirds of that observed in conventional mice and was obtained in the duodenum of gnotoxenic mice harboring four bacterial strains isolated from the conventional mouse microflora. The orally administered killed cells of two immunogenic strains, E. coli and Bacteroides sp., are as immunogenic as the living cells, provided that their concentration in the digestive tract is sufficient.


Bridging the gap: can impairment-based therapy for anomia have an impact at the psycho-social level?
Studies of therapy with people with aphasia tend to use impairment-based and functional measures of outcome. The views of participants are not formally evaluated. Current health and socialcare practice requires intervention to be explicitly client-centred and evidence-based. It is therefore important to investigate the broader effects of speech and language therapy. To explore the outcome of a therapy for anomia using the Communication Disability Profile (CDP), focusing particularly on participants' ratings of 'activity'. Overall eight people with aphasia and their conversation partners participated in the study. There was a range of severity and type of aphasia. Following two baselines (at least 8 weeks apart), there were two phases of therapy for anomia each lasting 8 weeks. This first involved the use of spoken and written cues to aid word finding. The second encouraged the use of targeted words in connected speech and conversation. Eight weeks later, after no further therapy, participants were reassessed. Participants' word finding in picture-naming improved significantly, as did their activity ratings. The relationship between the group's word-retrieval scores and CDP activity ratings over the course of the study tended towards significance, although there was considerable variation across individuals. Furthermore, all participants rated participation in activities requiring communication higher at the end than the start of the project. The findings suggest that therapy which targets word retrieval can have an impact on people with aphasia's views of their communicative activity and life participation. The findings support therapists' clinical insight that impairment-based interventions can effect change beyond scores on language tests.


Effect of occipitoatlantal decompression on cerebral blood flow dynamics as evaluated by Doppler ultrasonography.
Osteopathic manipulative treatment reduces symptoms in patients with headache disorders, but the underlying mechanisms are unclear. To evaluate blood flow in the intracranial and extracranial vasculature before and after occipitoatlantal decompression (OAD) using Doppler ultrasonography. Healthy, first-year osteopathic medical students from A.T. Still University's Kirksville College of Osteopathic Medicine participated in a randomized, single-blinded, two-period, two-treatment crossover study. The participants were randomly assigned to 1 of 2 treatment interventions: OAD or sham touch. After one week, participants returned to have the other intervention performed. Blood flow parameters-peak systolic velocity (PSV) and end-diastolic velocity (EDV)-in the middle cerebral artery (MCA), internal carotid artery (ICA), and vertebral artery (VA) were evaluated before, immediately after, 5 minutes after, and 10 minutes after treatment. Differences in PSV, EDV, heart rate (HR), and blood pressure (BP) for both interventions were analyzed for the four time points using mixed-effects models. Thirty healthy medical students (11 men, 19 women; mean age, 24 years) participated in this study. EDV increased after OAD in the MCA, ICA, and VA (all p<0.001); no change occurred after sham touch (all p>0.05). EDV was greater for all post-treatment timepoints after OAD in the MCA, ICA, and VA than after sham touch (all p<0.001). Although baseline PSV in the MCA measured before treatment was different between treatment interventions (p=0.01), no difference was found between interventions at any post-treatment time point (all p>0.59). Changes in PSV in the ICA and VA and for HR and BP did not depend on treatment intervention (p>0.06). Increases in EDV occurred in major cranial arteries after OAD but not after sham touch, indicating that OAD improves blood flow to the brain. The exact mechanism of this increase is unknown; however, it can be explained by either parasympathetic stimulation through the secretion of vasodilating neurotransmitters or by a decrease in external tissue pressure on ICA and VA, with the resulting flow causing further dilation in the MCA.


Metabolic signatures of GS-CHO cell clones associated with butyrate treatment and culture phase transition.
Chinese hamster ovary (CHO) cells are preferred hosts for the production of recombinant biopharmaceuticals. Efforts to optimize these bioprocesses have largely relied on empirical experience and our knowledge of cellular behavior in culture is incomplete. More recently, comprehensive investigations of metabolic network operation have started to be used to uncover traits associated with optimal growth and recombinant protein production. In this work, we used (1) H-nuclear magnetic resonance ((1) H-NMR) to analyze the supernatants of glutamine-synthetase (GS)-CHO cell clones expressing variable amounts of an IgG4 under control and butyrate-treated conditions. Exometabolomic data revealed accumulation of several metabolic by-products, indicating inefficiencies at different metabolic nodes. These data were contextualized in a detailed network and the cellular fluxomes estimated through metabolic flux analysis. This approach allowed comparing metabolic activity across different clones, growth phases and culture conditions, in particular the efficiency pertaining to carbon lost to glycerol and lactate accumulation and the characteristic nitrogen metabolism involving high asparagine and serine uptake rates. Importantly, this study shows that early butyrate treatment has a marked effect on sustaining high nutrient consumption along culture time, being more pronounced during the stationary phase when extra energy generation and biosynthetic activity is fueled to increase IgG formation. Collectively, the information generated contributes to deepening our understanding of CHO cells metabolism in culture, facilitating future design of improved bioprocesses.


Concentration and bioavailability of ciprofloxacin and teicoplanin in the cornea.
To investigate the concentration and bioavailability of ciprofloxacin and teicoplanin in the cornea. A biological assay was developed with corneal tissue used as a carrier for the antimicrobial. Concentration and biological activity were determined with a chemical assay and zone of inhibition (ZOI) around corneal samples with epithelial and endothelial surfaces in contact with the indicator organism. Patients undergoing penetrating keratoplasty received ciprofloxacin 0.3% or teicoplanin 1%. There were good correlations between antimicrobial concentration and ZOI, when either filter paper or corneal discs were used (R(2) > 92%). Of 33 patients, the mean (median) concentration of ciprofloxacin in the cornea was 1.37 mg/L (0.46 mg/L) and 1.89 mg/L (1.44 mg/L; bioassay) in the epithelial and endothelial orientations, respectively, and 14.87 mg/L (7.41) in the cornea and 0.51 mg/L (0.42) in the aqueous (chemical assay). For teicoplanin, the mean (median) concentration in the cornea was 9.58 mg/L (0 mg/L) in the epithelial and 4.78 mg/L (0 mg/L) in the endothelial orientations (bioassay). In the chemical assay, teicoplanin could not be detected in the cornea or aqueous at the lower limit of detection of 3.6 mg/L. The ZOI produced by corneal tissue provides a potential bioassay of antimicrobial activity and concentration. Although in contrast to teicoplanin ciprofloxacin shows good corneal penetration, with high endothelial-to-epithelial levels, only approximately 10% of measured levels in a chemical assay are available, according to a bioassay. Teicoplanin shows relatively poor corneal penetration through intact epithelium. These methods may be useful in evaluating the biological activity across the cornea of antimicrobials introduced into ophthalmic practice to deal with changing bacterial resistance.


(Im)Perfect robustness and adaptation of metabolic networks subject to metabolic and gene-expression regulation: marrying control engineering with metabolic control analysis.
Metabolic control analysis (MCA) and supply-demand theory have led to appreciable understanding of the systems properties of metabolic networks that are subject exclusively to metabolic regulation. Supply-demand theory has not yet considered gene-expression regulation explicitly whilst a variant of MCA, i.e. Hierarchical Control Analysis (HCA), has done so. Existing analyses based on control engineering approaches have not been very explicit about whether metabolic or gene-expression regulation would be involved, but designed different ways in which regulation could be organized, with the potential of causing adaptation to be perfect. This study integrates control engineering and classical MCA augmented with supply-demand theory and HCA. Because gene-expression regulation involves time integration, it is identified as a natural instantiation of the 'integral control' (or near integral control) known in control engineering. This study then focuses on robustness against and adaptation to perturbations of process activities in the network, which could result from environmental perturbations, mutations or slow noise. It is shown however that this type of 'integral control' should rarely be expected to lead to the 'perfect adaptation': although the gene-expression regulation increases the robustness of important metabolite concentrations, it rarely makes them infinitely robust. For perfect adaptation to occur, the protein degradation reactions should be zero order in the concentration of the protein, which may be rare biologically for cells growing steadily. A proposed new framework integrating the methodologies of control engineering and metabolic and hierarchical control analysis, improves the understanding of biological systems that are regulated both metabolically and by gene expression. In particular, the new approach enables one to address the issue whether the intracellular biochemical networks that have been and are being identified by genomics and systems biology, correspond to the 'perfect' regulatory structures designed by control engineering vis-à-vis optimal functions such as robustness. To the extent that they are not, the analyses suggest how they may become so and this in turn should facilitate synthetic biology and metabolic engineering.


Chinese hamster ovary cell mutants specifically affected in the phosphorylation of C-purine nucleosides.
In Chinese hamster ovary cells, two different types of mutants resistant to purine nucleoside analogs have been isolated. One type of mutants selected in presence of C-nucleosides formycin A and formycin B (FomR mutants) exhibited a high degree of cross resistance to different C-nucleosides but showed very slight to no cross resistance to various N-nucleosides. In contrast, mutants selected in presence of toyocamycin (Toyr mutants) exhibited a high degree of cross resistance to all C- and N-adenosine analogs. Studies on the cellular uptake and phosphorylation of [3H]adenosine, [3H]tubercidin, and [3H]formycin A show that, unlike the Toyr mutants which show reduced phosphorylation of all three compounds, the FomR mutants show reduced cellular phosphorylation of only [3H]formycin A. It is suggested that the genetic lesion in the FomR mutants affects adenosine kinase in a novel manner that specifically affects the phosphorylation of C-purine nucleosides.


Fifteen years of Servitude et Grandeur to the application of a biophysical technique in medicine: The tale of AFMBioMed.
AFMBioMed is the founding name under which international conferences and summer schools are organized around the application of atomic force microscopy in life sciences and nanomedicine. From its inception at the Atomic Energy Commission in Marcoule near 2004 to its creation in 2007 and to its 10th anniversary conference in Krakow, a brief narrative history of its birth and rise will demonstrate how and what such an organization brings to laboratories and the AFM community. With the current planning of the next AFMBioMed conference in Münster in 2019, it will be 15 years of commitment to these events.


The normal breast epithelium of women with breast cancer displays an aberrant response to estradiol.
Breast epithelial response to estradiol may play an important role in breast cancer etiology. We have examined the relationship between serum estradiol and progesterone levels and normal breast epithelial expression of estrogen receptor (ER) alpha, progesterone receptor (PgR), and epithelial proliferation (as reflected by the Ki-67 labeling index) in 121 women (50 newly diagnosed breast cancer cases and 71 benign breast disease controls). Simultaneous samples of grossly normal breast tissue and venous blood were obtained from women undergoing breast surgery. Serum estradiol and progesterone levels were measured by radioimmunoassay; breast epithelial ER, PgR, and Ki-67 expression was measured by immunohistochemistry. Linear regression, controlled for patient age and ductal and lobular composition of the tissue, showed that the breast epithelium of control women displayed an inverse correlation between serum estradiol and ER-alpha, which was not seen in case women (P for the difference in regression slopes = 0.001). PgR expression displayed a significant positive correlation with serum estradiol in cases, but not in controls. Epithelial proliferation had no relationship to either estradiol or progesterone in both cases and controls but showed an inverse relationship with ER in controls and a direct relationship in cases (P for the difference in regression slopes = 0.066). These results suggest a dysregulation of hormonal response in the normal breast epithelium of high-risk women, with lack of regulation of ER by estradiol, increased estrogen responsiveness as reflected by PgR expression, and a dissociation of ER expression and proliferative response.


Basal DNA repair machinery is subject to positive selection in ionizing-radiation-resistant bacteria.
Ionizing-radiation-resistant bacteria (IRRB) show a surprising capacity for adaptation to ionizing radiation and desiccation. Positive Darwinian selection is expected to play an important role in this trait, but no data are currently available regarding the role of positive adaptive selection in resistance to ionizing-radiation and tolerance of desiccation. We analyzed the four known genome sequences of IRRB (Deinococcus geothermalis, Deinococcus radiodurans, Kineococcus radiotolerans, and Rubrobacter xylanophilus) to determine the role of positive Darwinian selection in the evolution of resistance to ionizing radiation and tolerance of desiccation. We used the programs MultiParanoid and DnaSP to deduce the sets of orthologs that potentially evolved due to positive Darwinian selection in IRRB. We find that positive selection targets 689 ortholog sets of IRRB. Among these, 58 ortholog sets are absent in ionizing-radiation-sensitive bacteria (IRSB: Escherichia coli and Thermus thermophilus). The most striking finding is that all basal DNA repair genes in IRRB, unlike many of their orthologs in IRSB, are subject to positive selection. Our results provide the first in silico prediction of positively selected genes with potential roles in the molecular basis of resistance to gamma-radiation and tolerance of desiccation in IRRB. Identification of these genes provides a basis for future experimental work aimed at understanding the metabolic networks in which they participate.


Obstructive Sleep Apnea and Hallmarks of Aging.
Obstructive sleep apnea (OSA) is one of the most common sleep disorders. Since aging is a risk factor for OSA development, it is expected that its prevalence will increase with the current increase in life span. In recent years, several studies have shown that OSA potentially contributes to functional decline, mainly prompted by chronic intermittent hypoxia and sleep fragmentation. Here, we propose that OSA might anticipate/aggravate aging by inducing cellular and molecular impairments that characterize the aging process, such as stem cell exhaustion, telomere attrition and epigenetic changes. We suggest that further knowledge on the impact of OSA on aging mechanisms might contribute to a better understanding of how OSA might putatively accelerate aging and aging-related diseases.


Elucidation of an alternate isoleucine biosynthesis pathway in Geobacter sulfurreducens.
The central metabolic model for Geobacter sulfurreducens included a single pathway for the biosynthesis of isoleucine that was analogous to that of Escherichia coli, in which the isoleucine precursor 2-oxobutanoate is generated from threonine. 13C labeling studies performed in G. sulfurreducens indicated that this pathway accounted for a minor fraction of isoleucine biosynthesis and that the majority of isoleucine was instead derived from acetyl-coenzyme A and pyruvate, possibly via the citramalate pathway. Genes encoding citramalate synthase (GSU1798), which catalyzes the first dedicated step in the citramalate pathway, and threonine ammonia-lyase (GSU0486), which catalyzes the conversion of threonine to 2-oxobutanoate, were identified and knocked out. Mutants lacking both of these enzymes were auxotrophs for isoleucine, whereas single mutants were capable of growth in the absence of isoleucine. Biochemical characterization of the single mutants revealed deficiencies in citramalate synthase and threonine ammonia-lyase activity. Thus, in G. sulfurreducens, 2-oxobutanoate can be synthesized either from citramalate or threonine, with the former being the main pathway for isoleucine biosynthesis. The citramalate synthase of G. sulfurreducens constitutes the first characterized member of a phylogenetically distinct clade of citramalate synthases, which contains representatives from a wide variety of microorganisms.


Self-administration with UniJect of the once-a-month injectable contraceptive Cyclofem.
The objective of this study was to evaluate women's acceptance of and ability to self-administrate the injectable contraceptive Cyclofem using prefilled UniJect devices. A total of 102 women were invited to participate in the study. Fourteen women (13.7%) refused to participate. Of the remaining 88 women, 32 women (31.4%) consented to participate and were trained using oranges but were still afraid of the procedure and ultimately refused to self-administer the injections. Only 56 women (55%) ultimately self-injected Cyclofem with UniJect. They performed a total of 144 injections, all of them on the ventral side of the thigh. When nurses evaluated women's ability to activate the devices, they found that more than 80% were successful in both the group of women that later self-administered the injections and the group that did not. The evaluation of the self-administered injection technique showed that more than 90% of the women correctly self-administered the contraceptive using UniJect. With respect to the opinion of the women about the self-administration of the contraceptive, more than 50% (32 of 56) of women who self-injected preferred to self-administer the injection and said that they wished to continue with the self-administration, one-third (17) reported that they were afraid, and seven women (12.5%) expressed the opinion that the injection in the thigh was more painful than the administration in the buttocks or arm. In conclusion, our study showed that women can be trained to successfully self-administer the monthly injectable contraceptive Cyclofem and generally respond positively to UniJect.


A Volatile and Dynamic Longitudinal Microbiome Is Associated With Less Reduction in Lung Function in Adolescents With Cystic Fibrosis.
Progressive impairment in lung function caused by chronic polymicrobial airway infection remains the major cause of death in patients with cystic fibrosis (CF). Cross-sectional studies suggest an association between lung function decline and specific lung microbiome ecotypes. However, longitudinal studies on the stability of the airway microbiome are missing for adolescents with CF constituting the age group showing the highest rate of decline in lung function. In this study, we analyzed longitudinal lung function data and sputum samples collected over a period of 3 to 5 years from 12 adolescents with CF. The sputum microbiome was analyzed using 16S rRNA gene sequencing. Our results indicate that the individual course of the lung microbiome is associated with longitudinal lung function. In our cohort, patients with a dynamic, diverse microbiome showed a slower decline of lung function measured by FEV1% predicted, whereas a more stable and less diverse lung microbiome was related to worse outcomes. Specifically, a higher abundance of the phyla Bacteroidetes and Firmicutes was linked to a better clinical outcome, while Proteobacteria were correlated with a decline in FEV1% predicted. Our study indicates that the stability and diversity of the lung microbiome and the abundance of Bacteroidetes and Firmicutes are associated with the lung function decline and are one of the contributing factors to the disease severity.