Analysis of the tissue-specific expression, developmental regulation, and linkage relationships of a rodent gene encoding heart fatty acid binding protein.
The rat contains at least three homologous cytosolic proteins that bind long chain fatty acids, termed liver (L-), intestinal (I-), and heart (H-) fatty acid binding protein (FABP). I-FABP mRNA is confined to the gastrointestinal tract while L-FABP mRNA is abundantly represented in hepatocytes as well as enterocytes. We have isolated a rat heart FABP cDNA clone and determined the pattern of H-FABP mRNA accumulation in a wide variety of tissues harvested from late fetal, suckling, weaning, and adult rats. RNA blot hybridizations and primer extension analysis disclosed that the distribution of H-FABP mRNA in adult rat tissues is different from that of I- or L-FABP mRNA. H-FABP mRNA is most abundant in adult heart. This mRNA was also present in an adult slow twitch (type I) skeletal muscle (soleus, 63% of the concentration in heart), testes (28%), a fast twitch skeletal muscle (psoas, 17%), brain (10%), kidney (5%), and adrenal gland (5%). H-FABP mRNA was not detected in adult small intestine, colon, spleen, lung, or liver RNA. Distinct patterns of developmental change in H-FABP mRNA accumulation were documented in heart, placenta, brain, kidney, and testes. Myocardial H-FABP mRNA levels rise rapidly during the 48 h prior to and after birth, reaching peak levels by the early weaning period. The postnatal increase in myocardial H-FABP mRNA concentration and its relative distribution in adult fast and slow twitch skeletal muscle are consistent with its previously proposed function in facilitating mitochondrial beta-oxidation of fatty acids. However, the presence of H-FABP mRNA in brain, a tissue which does not normally significantly oxidize fatty acids in late postnatal life, suggests that H-FABP may play a wider role in fatty acid metabolism than previously realized. Mouse-hamster somatic cell hybrids were utilized to map H-FABP. Using stringencies which did not produce cross-hybridization between L-, I-, and H-FABP DNA sequences, we found at least three loci in the mouse genome, each located on different chromosomes, which reacted with our cloned H-FABP cDNA. None of these H-FABP-related loci were linked to the gene which specifies a highly homologous adipocyte-specific protein termed aP2 or to genes encoding two other members of this protein family, cellular retinol binding protein and cellular retinol binding protein II.(ABSTRACT TRUNCATED AT 400 WORDS)


Radiographic, Bone Densitometry and Clinic Outcomes Assessments in Femoral Shaft Fractures Fixed by Plating or Locking Retrograde Nail.
Distal femoral shaft fractures are characterized by increasing incidence and complexity and are still considered a challenging problem (high morbidity and mortality). No consensus on best surgical option has been achieved. This study aims is to investigate radiographic, mineral bone densitometry and clinical outcomes of locking retrograde intramedullary (LRN) nailing and anatomical locking plate to surgically treat distal femoral shaft fractures in young adults based on the hypothesis that there is no statistical difference among the two surgical options in terms of results (radiographic, bone densitometry and outcomes assessment). Retrospective study: 30 patients divided into 2 groups (Group 1 LRN, Group 2 Nailing). Average age was 42.67±18.32 for Group 1 and 42.84 ±18.32 for Group 2 (range of age 18-65 for both groups). Gender Ratio (male: female) was 2.75 (11:4) for both Group 1 and 2. AO Classification, NUSS and RUSH score, VAS, Dexa scans, plain radiographs were used. Evaluation endpoint: 12 months after surgery (mean follow up 16.24). No statistical difference in terms of surgery time, transfusions, wound healing. Similar results with regard to average time of bone healing, RUSH scores, VAS, regression between RUSH and VAS, average correlation clinical-radiographic results and patients outcomes. Only one patient of each group had reduction of mineral bone densitometry values. No statistical difference about the use of LNR or Nailing for treatment of distal femur shaft fractures in terms of radiographic, bone densitometry and outcomes has been found accordingly to our results. Good subjective and objective results are provided by both techniques. The choice among the two techniques must be based on surgeons' experience, indications and subjective patients' aspects. The absence of relevant similar data in the published literature does not allow definitive validation (or rejection) of our hypothesis. A more powered study with bigger cohort is needed for definitive validation.


Inborn and experience-dependent models of categorical brain organization. A position paper.
The present review aims to summarize the debate in contemporary neuroscience between inborn and experience-dependent models of conceptual representations that goes back to the description of category-specific semantic disorders for biological and artifact categories. Experience-dependent models suggest that categorical disorders are the by-product of the differential weighting of different sources of knowledge in the representation of biological and artifact categories. These models maintain that semantic disorders are not really category-specific, because they do not respect the boundaries between different categories. They also argue that the brain structures which are disrupted in a given type of category-specific semantic disorder should correspond to the areas of convergence of the sensory-motor information which play a major role in the construction of that category. Furthermore, they provide a simple interpretation of gender-related categorical effects and are supported by studies assessing the importance of prior experience in the cortical representation of objects On the other hand, inborn models maintain that category-specific semantic disorders reflect the disruption of innate brain networks, which are shaped by natural selection to allow rapid identification of objects that are very relevant for survival. From the empirical point of view, these models are mainly supported by observations of blind subjects, which suggest that visual experience is not necessary for the emergence of category-specificity in the ventral stream of visual processing. The weight of the data supporting experience-dependent and inborn models is thoroughly discussed, stressing the fact observations made in blind subjects are still the subject of intense debate. It is concluded that at the present state of knowledge it is not possible to choose between experience-dependent and inborn models of conceptual representations.


A digital platform to support communication and organization in the general practice: Evaluation of healthcare usage and costs using claims data of a health insurer.
The pressure on general practitioners (GPs) is rising due to the increasing demand for care and a decreasing availability of GPs. eHealth is seen as one of the solutions to enhance accessibility and reduce workload. A platform supporting the organization and communication in general practice has been developed offering services, such as econsultations. This study aims to evaluate healthcare usage and costs of patients using this platform by comparing these outcomes (1) before and after implementation and (2) an intervention with a matched control group. This study is a retrospective observational cohort study. To evaluate the longitudinal impact of the implementation on healthcare usage, mixed model Poisson analyses were used with time as a factor term for the within-subject analysis and exposure to the platform as a factor term and an interaction term (i.e., exposure X 6-months) in the between-subject analysis. Cost analyses were done with mixed model analyses of variance over time. The total number of GP consultations significantly increased after compared to before implementation (i.e., Rate = 1.52; p < 0.001). The number of GP consultations was higher in the intervention compared to the control group (respectively, Rate = 1.23; p = 0.035). Healthcare costs increased for GP consultations after compared to before implementation (€13,57; p < 0.001). The costs for GP consults were significantly higher in the intervention compared to the control group (€7,06; p 0.018). This study showed a rise in GP consultations and costs when implementing a digital platform. This increase was presumably and partly caused by circumstances in one of the two included practices. Moreover, creating new options for contacting and communicating with the GP can enhance care accessibility and thereby driving an increase in consultations. This digital platform is a promising working method in general practice to facilitate patients and provide GPs with more flexibility.


Summation in autoshaping is affected by the similarity of the visual stimuli to the stimulation they replace.
Pigeons received autoshaping with 2 stimuli, A and B, presented in adjacent regions on a television screen. Conditioning with each stimulus was therefore accompanied by stimulation that was displaced from the screen whenever the other stimulus was presented. Test trials with AB revealed stronger responding if this displaced stimulation was similar to, rather than different from, A and B. For a further experiment the training just described included trials with A and B accompanied by an additional, similar, stimulus. Responding during test trials with AB was stronger if the additional trials signaled the presence rather than the absence of food. The results are explained with a configural theory of conditioning.


Racial variation in the control of diabetes among elderly medicare managed care beneficiaries.
To examine racial variation in the poor control of GHb, a GHb value >9.5%, or GHb not tested in 1999 among Medicare beneficiaries aged 65-75 years enrolled in managed care plans. The National Committee on Quality Assurance provides person-level data regarding diabetes care services and control for Medicare beneficiaries enrolled in managed care to the Centers for Medicare and Medicaid Services (CMS). We merged this information with information on each individual's race, as well as other person-level and plan-level characteristics obtained from CMS. Bivarate and multivariate analyses were performed. The overall rate of poor GHb control was 32.7%. The age- and sex-adjusted rate of poor control among whites was 32.0%. This rate was significantly higher than the rate among Asians (24.7%) but significantly lower than the rate among blacks (40.6%) and Hispanics (36.5%) (P < 0.001). An increase in the number of comprehensive diabetes care measures received by an individual was associated with a significantly lower percentage of individuals with poor GHb control in all race groups. After controlling for the individual-level, plan-level, and diabetes care measure variables, the difference in GHb control between Asians and whites disappeared. However, blacks and Hispanics continued to have significantly higher rates of poor control than whites. There is room for significant reduction in the number of patients with poor control of GHb among all races, particularly among blacks and Hispanics.


A model for space-time cluster detection using spatial clusters with flexible temporal risk patterns.
Maps of estimated disease rates over multiple time periods are useful tools for gaining etiologic insights regarding potential exposures associated with specific locations and times. In this paper, we describe an extension of the Gangnon-Clayton model for spatial clustering to spatio-temporal data. As in the purely spatial model, a large set of circular regions of varying radii centered at observed locations are considered as potential clusters, e.g. subregions with a different pattern of risk than the remainder of the study region. Within the spatio-temporal model, no specific parametric form is imposed on the temporal pattern of risk within each cluster. In addition to the clusters, the proposed model incorporates spatial and spatio-temporal heterogeneity effects and can readily accommodate regional covariates. Inference is performed in a Bayesian framework using MCMC. Although formal inferences about the number of clusters could be obtained using a reversible jump MCMC algorithm, we use local Bayes factors from models with a fixed, but overly large, number of clusters to draw inferences about both the number and the locations of the clusters. We illustrate the approach with two applications of the model to data on female breast cancer mortality in Japan and evaluate its operating characteristics in a simulation study.


Intact nitric oxide synthase II gene is required for interferon-beta-mediated suppression of growth and metastasis of pancreatic adenocarcinoma.
Previous studies have shown that enforced expression of IFN-beta suppressed tumor growth and metastasis. In this report, we determined whether the induction of nitric oxide synthase II (NOS II) gene is required for IFN-beta-mediated antitumor activity using syngeneic mice with intact (NOS II+/+) or genetically disrupted (NOS II-/-) NOS II gene. PANC02-H7 highly metastatic murine pancreatic adenocarcinoma cells were transfected with an IFN-beta expression vector or a control pcDNA3 vector. The parental PANC02-H7, control vector-transfected, and IFN-beta-transfected cells were orthotopically implanted into the pancreas of syngeneic NOS II+/+ and NOS II-/- C57BL/6J mice. In NOS II+/+ C57BL/ 6J, both parental and control vector-transfected cells grew progressively in pancreas and produced numerous liver metastases and a large amount of malignant ascites, whereas IFN-beta-secreting cells did not. In NOS II-/- C57BL/6J mice, however, IFN-beta-secreting cells grew much more aggressively. Higher NO induction was detected in NOS II+/+ mice that received injections with IFN-beta-secreting cells than with the control cells, but it was not detected in NOS II-/- mice. These data suggested that IFN-beta secreted from tumor cells stimulates NO production by host cells and suppresses tumor growth and metastasis.


Forensic peer specialists: Training, employment, and lived experience.
Despite calls for increased peer support services for individuals involved with the criminal justice system, little is known about the training, employment, and characteristics of forensic peer specialists (FPSs). Pennsylvania developed the nation's first FPS program and remains at the forefront of the field. This paper responds to three research questions regarding forensic peer support in Pennsylvania, assessing: (a) employment patterns and demographic characteristics; (b) work tasks and sites; and (c) challenges and benefits of working as an FPS. Data are drawn from a larger mixed-methods study of forensic peer support in Pennsylvania, including a detailed online survey of 117 FPS-trained workers, followed by 16 qualitative interviews with a subset of survey respondents. While all survey respondents completed forensic peer support training, only 36% (n = 44) have been employed as FPSs. 96% of survey respondents report having used mental health services, compared to 55% reporting past incarceration. FPSs spend the majority of their work time providing individual support to peer clients and work in multiple settings. Additionally, FPSs report both benefits and challenges related to their work. Study results suggest that FPS characteristics, practices, and work settings simultaneously align with and differ from nonspecialized peer support services. Program developers and advocates should focus attention on expanding employment opportunities, and FPSs with lived experience should be prioritized for these positions. Despite significant implementation barriers, FPSs are passionate about this work and promote the expansion of the field. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Examining the Association of Antidepressant Prescriptions With First Abortion and First Childbirth.
The repercussions of abortion for mental health have been used to justify state policies that limit access to abortion in the United States. Much earlier research has relied on self-report of abortion or mental health conditions or on convenience samples. This study uses data that rely on neither. To examine whether first-trimester first abortion or first childbirth is associated with an increase in women's initiation of a first-time prescription for an antidepressant. This study linked data and identified a cohort of women from Danish population registries who were born in Denmark between January 1, 1980, and December 30, 1994. Overall, 396 397 women were included in this study; of these women, 30 834 had a first-trimester first abortion and 85 592 had a first childbirth. First-time antidepressant prescription redemptions were determined and used as indication of an episode of depression or anxiety, and incident rate ratios (IRRs) were calculated comparing women who had an abortion vs women who did not have an abortion and women who had a childbirth vs women who did not have a childbirth. Of 396 397 women whose data were analyzed, 17 294 (4.4%) had a record of at least 1 first-trimester abortion and no children, 72 052 (18.2%) had at least 1 childbirth and no abortions, 13 540 (3.4%) had at least 1 abortion and 1 childbirth, and 293 511 (74.1%) had neither an abortion nor a childbirth. A total of 59 465 (15.0%) had a record of first antidepressant use. In the basic and fully adjusted models, relative to women who had not had an abortion, women who had a first abortion had a higher risk of first-time antidepressant use. However, the fully adjusted IRRs that compared women who had an abortion with women who did not have an abortion were not statistically different in the year before the abortion (IRR, 1.46; 95% CI, 1.38-1.54) and the year after the abortion (IRR, 1.54; 95% CI, 1.45-1.62) (P = .10) and decreased as time from the abortion increased (1-5 years: IRR, 1.24; 95% CI, 1.19-1.29; >5 years: IRR, 1.12; 95% CI, 1.05-1.18). The fully adjusted IRRs that compared women who gave birth with women who did not give birth were lower in the year before childbirth (IRR, 0.47; 95% CI, 0.43-0.50) compared with the year after childbirth (IRR, 0.93; 95% CI, 0.88-0.98) (P < .001) and increased as time from the childbirth increased (1-5 years: IRR, 1.52; 95% CI, 1.47-1.56; >5 years: IRR, 1.99; 95% CI, 1.91-2.09). Across all women in the sample, the strongest risk factors associated with antidepressant use in the fully adjusted model were having a previous psychiatric contact (IRR, 3.70; 95% CI, 3.62-3.78), having previously obtained an antianxiety medication (IRR, 3.03; 95% CI, 2.99-3.10), and having previously obtained antipsychotic medication (IRR, 1.88; 95% CI, 1.81-1.96). Women who have abortions are more likely to use antidepressants compared with women who do not have abortions. However, additional aforementioned findings from this study support the conclusion that increased use of antidepressants is not attributable to having had an abortion but to differences in risk factors for depression. Thus, policies based on the notion that abortion harms women's mental health may be misinformed.


Combinatorial library design for improving isobutanol production in Saccharomyces cerevisiae.
Saccharomyces cerevisiae is the dominant fermentative producer of ethanol in industry and a preferred host for production of other biofuels. That said, rewiring the metabolism of S. cerevisiae to produce other fermentation products, such as isobutanol, remains an academic challenge. Many studies report aerobic production of isobutanol, but ethanol remains a substantial by-product under these conditions due to the Crabtree effect. These studies indicate that the native isobutanol pathway is incapable of carrying sufficient flux to displace ethanol. In this report, we screened a combinatorial library of pathway enzymes to identify an isobutanol pathway cassette capable of supporting the growth of a non-ethanol producing S. cerevisiae. We began by identifying a diverse set of isobutanol pathway enzyme homologs and combined each open reading frame with varied-strength promoters in a combinatorial, pooled fashion. We applied a growth-coupled screen where a functional isobutanol pathway restored NAD+ regeneration during glucose catabolism that is otherwise repressed via the Crabtree effect. Using this screen, we isolated a cassette consisting of a mosaic of bacterial and cytosol-localized fungal enzymes that conferred under aerobic conditions the ability to produce 364 mg/L isobutanol (8.8% of the theoretical maximum yield). We next shifted the cofactor usage of the isolated ketol-acid reductoisomerase enzyme in the cassette from NADPH to NADH-preferring to improve redox balance. The approach used herein isolated isobutanol producing strains that approach the best in the literature without producing substantial ethanol titers. Still, the best isolated cassette was insufficient to support anaerobic growth in the absence of ethanol fermentation - indicating the presence of further fundamental gaps in our understanding of yeast fermentation.


Mental health training experiences among Haitian healthcare workers post-earthquake 2010.
After the 2010 earthquake in Haiti, the large number of persons with major limb damage, amputations, shock, trauma, anxiety and depression placed a severe strain on mental health (MH) services. This qualitative study describes the impact and acceptability of a Mental Health Training Program (MHTP) implemented in the north of Haiti after the earthquake. A total of 113 healthcare workers (HCWs) participated in a training program designed to build local MH care capacity. The training curriculum draws on literature related to MH and the impact of the Haiti earthquake. Two focus groups were conducted with 16 HCWs; discussions centred on the personal and professional impact and acceptability of the training program. Results demonstrated that the MHTP changed the HCWs' perceptions about MH issues and provided them with the knowledge and skills to respond to growing community MH needs. Acceptability of the MHTP was related to the content covered, to the delivery mode of the content and to the cultural appropriateness of the program. Disasters of different types will continue to occur and to impact MH in communities around the world. MH training will allow nurses to quickly and effectively respond to disasters. A coordinated emergency plan that is subject to frequent review, rehearsal and evaluation is also essential.


Sports-related injuries in New Zealand: National Insurance (Accident Compensation Corporation) claims for five sporting codes from 2012 to 2016.
To provide epidemiological data and related costs for sport-related injuries of five sporting codes (cricket, netball, rugby league, rugby union and football) in New Zealand for moderate-to-serious and serious injury claims. A retrospective analytical review using detailed descriptive epidemiological data obtained from the Accident Compensation Corporation (ACC) for 2012-2016. Over the 5 years of study data, rugby union recorded the most moderate-to-serious injury entitlement claims (25 226) and costs (New Zealand dollars (NZD$)267 359 440 (£139 084 749)) resulting in the highest mean cost (NZD$10 484 (£5454)) per moderate-to-serious injury entitlement claim. Rugby union recorded more serious injury entitlement claims (n=454) than cricket (t(4)=-66.6; P<0.0001); netball (t(4)=-45.1; P<0.0001); rugby league (t(4)=-61.4; P<0.0001) and football (t(4)=66.6; P<0.0001) for 2012-2016. There was a twofold increase in the number of female moderate-to-serious injury entitlement claims for football (RR 2.6 (95%CI 2.2 to 2.9); P<0.0001) compared with cricket, and a threefold increase when compared with rugby union (risk ratio (RR) 3.1 (95%CI 2.9 to 3.3); P<0.0001). Moderate-to-serious concussion claims increased between 2012 and 2016 for netball (RR 3.7 (95%CI 1.9 to 7.1); P<0.0001), rugby union (RR 2.0 (95% CI 1.6 to 2.4); P<0.0001) and football (RR 2.3 (95%CI 1.6 to 3.2); P<0.0001). Nearly a quarter of moderate-to-serious entitlement claims (23%) and costs (24%) were to participants aged 35 years or older. Rugby union and rugby league have the highest total number and costs associated with injury. Accurate sport exposure data are needed to enable injury risk calculations.


A Peptide-Coated Gold Nanocluster Exhibits Unique Behavior in Protein Activity Inhibition.
Gold nanoclusters (AuNCs) can be primed for biomedical applications through functionalization with peptide coatings. Often anchored by thiol groups, such peptide coronae not only serve as passivators but can also endow AuNCs with additional bioactive properties. In this work, we use molecular dynamics simulations to study the structure of a tridecapeptide-coated Au25 cluster and its subsequent interactions with the enzyme thioredoxin reductase 1, TrxR1. We find that, in isolation, both the distribution and conformation of the coating peptides fluctuate considerably. When the coated AuNC is placed around TrxR1, however, the motion of the highly charged peptide coating (+5e/peptide) is quickly biased by electrostatic attraction to the protein; the asymmetric coating acts to guide the nanocluster's diffusion toward the enzyme's negatively charged active site. After the AuNC comes into contact with TrxR1, its peptide corona spreads over the protein surface to facilitate stable binding with protein. Though individual salt bridge interactions between the tridecapeptides and TrxR1 are transient in nature, the cooperative binding of the peptide-coated AuNC is very stable, overall. Interestingly, the biased corona peptide motion, the spreading and the cooperation between peptide extensions observed in AuNC binding are reminiscent of bacterial stimulus-driven approaching and adhesion mechanisms mediated by cilia. The prevailing AuNC binding mode we characterize also satisfies a notable hydrophobic interaction seen in the association of thioredoxin to TrxR1, providing a possible explanation for the AuNC binding specificity observed in experiments. Our simulations thus suggest this peptide-coated AuNC serves as an adept thioredoxin mimic that extends an array of auxiliary structural components capable of enhancing interactions with the target protein in question.


Determining factors affecting job strain for nurse practitioners in acute care practice.
To investigate nurse practitioners' (NPs) experience of job strain in acute care settings, and to examine the factors that may affect job strain in that context. Descriptive design using a national survey was employed. A total of 1396 NPs completed online surveys that recorded demographic characteristics and included a Job Content Questionnaire (JCQ), a Condition for Work Effectiveness Questionnaire (CWEQ), and the Dempster Practice Behavior Scale (DPBS). Multiple logistic regressions were conducted to explore the factors associated with job strain types. NPs were classified into job strain categories of passive (24.4%), active (31.4%), low (19.2%), or high (24.9%). The passive job strain type was associated with overtime hours, organizational empowerment, and autonomy. The active job type was associated with higher organizational empowerment and autonomy. The high-strain and low-strain job types were both associated with overtime hours and autonomy. Nearly 25% of acute care NPs are in a high job strain type. Organizational empowerment and autonomy were two major factors associated with the passive and active job strain types. Overtime hours and autonomy were both associated with the high-strain and low-strain job types. Supportive hospital/nursing leadership should acknowledge the impact of NP practice as they can contribute to the operational efficacy of their organization. Hospital administrators should provide a supportive practice environment by empowering NPs, enhancing autonomy, and addressing working conditions for NPs to decrease the odds of having a passive or high-strain job type in practice.


Sexual outcome of patients undergoing thulium laser enucleation of the prostate for benign prostatic hyperplasia.
Treatment of patients with lower urinary tract symptoms (LUTS) secondary to benign prostatic hyperplasia (BPH) may affect the quality of sexual function and ejaculation. The effect of new surgical procedures, which are currently available to treat BPH, on erection and ejaculation, has been poorly studied. This study aimed to assess the effect of thulium laser enucleation of the prostate (ThuLEP) on sexual function and retrograde ejaculation in patients with LUTS secondary to BPH. We performed a prospective study in 110 consecutive patients who had undergone ThuLEP to analyze changes in sexual function and urinary symptoms. To evaluate changes in erection and ejaculation, and the effect of urinary symptoms on the quality of life (QoL), five validated questionnaires were used: the ICIQ-MLUTSsex, MSHQ-EjD, International Index of Erectile Function 5, International Prognostic Scoring System (IPSS) questionnaire, and QoL index of the intraclass correlation coefficients. Patients also underwent IPSS and flowmetry to assess the outcome of flow. Patients were evaluated before surgery and 3-6 months after ThuLEP, whereas those with previous abdominal surgery were excluded. The patients' mean age was 67.83 years. Postoperative urinary symptoms improved after surgery. No significant differences in erectile function before and after surgery were observed. As compared with other techniques described in the literature, the percentage of patients with conserved ejaculation increased by 52.7% after ThuLEP. ThuLEP positively affects urinary symptoms and their effect on the QoL of patients as assessed by questionnaire scores. While endoscopic management of BPH (e.g. transurethral resection of the prostate) causes retrograde ejaculation in most patients, those who undergo ThuLEP have conserved ejaculation and erectile function.


Layer I neurons of rat neocortex. I. Action potential and repetitive firing properties.
1. Whole cell patch-clamp techniques, combined with direct visualization of neurons, were used to study action potential (AP) and repetitive firing properties of layer I neurons in slices of rat neocortex. 2. Layer I neurons had resting membrane potentials (RMP) of -59.8 +/- 4.7 mV (mean +/- SD) and input resistances (RN) of 592 +/- 284 M Omega. Layer II/III pyramidal neurons had RMPs and RNs of -61.5 +/- 5.6 mV and 320 +/- 113 M omega, respectively. A double exponential function was needed to describe the charging curves of both neuron types. In layer I neurons, tau(0) was 45 +/- 22 ms and tau(1) was 5 +/- 3.3 ms whereas in layer II/III pyramidal neurons, tau(0) was 41 +/- 11 ms and tau(1) was 3 +/- 2.6 ms. Estimates of specific membrane resistance (Rm) for layer I and layer II/III cells were 45 +/- 22 and 41 +/- 11 k omega cm2, respectively (Cm was assumed to be 1 microF/cm2). 3. AP threshold was -41 +/- 2 mV in layer I neurons. Spike amplitudes, measured from threshold to peak, were 90.6 +/- 7.7 mV. AP durations, measured both at the base and half maximal amplitude, were 2.5 +/- 0.4 and 1.1 +/- 0.2 ms, respectively. AP 10-90% rise and repolarization times were 0.6 +/- 0.1 and 1.1 +/- 0.2 ms, respectively. In layer II/III pyramidal neurons, AP threshold was -41 +/- 2.5 mV and spike amplitude was 97 +/- 9.7 mV. AP duration at base and half maximal amplitude was 5.4 +/- 1.1 ms and 1.8 +/- 0.2 ms, respectively. AP 10-90% rise and decay times were 0.6 +/- 0.1 ms and 2.8 +/- 0.6 ms, respectively. 4. Layer I neurons were fast spiking cells that showed little frequency adaptation, a large fast afterhyperpolarization (fAHP), and no slow afterhyperpolarization (sAHP). Some cells had a medium afterhyperpolarization (mAHP) and a slow afterdepolarization (sADP). All pyramidal cells in layer II/III and "atypical" pyramidal neurons in upper layer II showed regular spiking behavior, prominent frequency adaptation, and marked sAHPs. 5. In both layer I neurons and layer II/III pyramidal neurons, changes in membrane potential did not greatly alter AP properties. The duration of APs evoked from -50 to -60 mV was only slightly longer, from -80 to -90 mV. The latency to first spike also was not solely dependent on membrane potential. 6. During repetitive firing, APs broadened in both layer I neurons and layer II/III pyramidal neurons. This was most prominent in pyramidal cells. Broadening was dependent on spike frequency and appeared to result from partial inactivation of both outward potassium and inward sodium currents. 7. In layer I neurons, removing Ca2+ from the bathing solution slightly prolonged spike duration and modestly increased AP firing frequency. These results indicate minimal involvement of Ca2+-dependent K+ currents in AP repolarization. fAHPs were reduced whereas sADPs were abolished. In layer II/III pyramidal neurons, removing Ca2+ reduced or blocked mAHPs and sAHPs and decreased or abolished frequency adaptation. 8. Low concentrations (50 microM) of 4-aminopyridine (4-AP) prolonged APs and induced burst-like firing in layer I neurons. In the presence of 4-AP, the spiking behavior of layer I neurons resembled that of regular spiking layer II/III pyramidal cells. At high concentrations (4 mM), 4-AP could induce a delayed depolarization (DD) after each spike in layer I neurons and in a minority of pyramidal neurons. 9. All layer I neurons had a prominent fAHP that was absent or very small in layer II/III pyramidal neurons. fAHP amplitude was inversely related to AP duration. The reduction of fAHPs by 4-AP or during repetitive firing was accompanied by AP prolongation, suggesting that the current underlying fAHP played an essential role in AP repolarization. The fAHP of layer I neurons could be effectively blocked by 4-AP but only slightly reduced by removing Ca2+ from bathing solution, indicating that the fAHP was mediated primarily by a voltage-dependent transient outward current.(ABSTRACT TRUNCATED)


Non-pharmacological interventions for chronic pain in multiple sclerosis.
Chronic pain is common and significantly impacts on the lives of persons with multiple sclerosis (pwMS). Various types of non-pharmacological interventions are widely used, both in hospital and ambulatory/mobility settings to improve pain control in pwMS, but the effectiveness and safety of many non-pharmacological modalities is still unknown. This review aimed to investigate the effectiveness and safety of non-pharmacological therapies for the management of chronic pain in pwMS. Specific questions to be addressed by this review include the following.Are non-pharmacological interventions (unidisciplinary and/or multidisciplinary rehabilitation) effective in reducing chronic pain in pwMS?What type of non-pharmacological interventions (unidisciplinary and/or multidisciplinary rehabilitation) are effective (least and most effective) and in what setting, in reducing chronic pain in pwMS? A literature search was performed using the specialised register of the Cochrane MS and Rare Diseases of the Central Nervous System Review Group, using the Cochrane MS Group Trials Register which contains CENTRAL, MEDLINE, Embase, CINAHL, LILACUS, Clinical trials.gov and the World Health Organization International Clinical Trials Registry Platform on 10 December 2017. Handsearching of relevant journals and screening of reference lists of relevant studies was carried out. All published randomised controlled trials (RCTs)and cross-over studies that compared non-pharmacological therapies with a control intervention for managing chronic pain in pwMS were included. Clinical controlled trials (CCTs) were eligible for inclusion. All three review authors independently selected studies, extracted data and assessed the methodological quality of the studies using the Grades of Recommendation, Assessment, Development and Evaluation (GRADE) tool for best-evidence synthesis. Pooling data for meta-analysis was not possible due to methodological, clinical and statistically heterogeneity of the included studies. Overall, 10 RCTs with 565 participants which investigated different non-pharmacological interventions for the management of chronic pain in MS fulfilled the review inclusion criteria. The non-pharmacological interventions evaluated included: transcutaneous electrical nerve stimulation (TENS), psychotherapy (telephone self-management, hypnosis and electroencephalogram (EEG) biofeedback), transcranial random noise stimulation (tRNS), transcranial direct stimulation (tDCS), hydrotherapy (Ai Chi) and reflexology.There is very low-level evidence for the use of non-pharmacological interventions for chronic pain such as TENS, Ai Chi, tDCS, tRNS, telephone-delivered self-management program, EEG biofeedback and reflexology in pain intensity in pwMS. Although there were improved changes in pain scores and secondary outcomes (such as fatigue, psychological symptoms, spasm in some interventions), these were limited by methodological biases within the studies. Despite the use of a wide range of non-pharmacological interventions for the treatment of chronic pain in pwMS, the evidence for these interventions is still limited or insufficient, or both. More studies with robust methodology and greater numbers of participants are needed to justify the effect of these interventions for the management of chronic pain in pwMS.


The impact of gestational diabetes on the development of fetal frontal lobe: A case-control study from a tertiary center.
To determine the effects of gestational diabetes mellitus (GDM) on fetal frontal lobe development. This study was conducted prospectively between May 2023 and August 2023 in Ankara City Hospital perinatology clinic. Maternal age, maternal body mass index (BMI), gestational week (GW), biparietal diameter (BPD), head circumference (HC), abdominal circumference (AC), femur length (FL), estimated fetal weight (EFW), frontal antero-posterior diameter (FAPD), occipito-frontal diameter (OFD), FAPD/OFD ratio, and FAPD/HC ratio, were compared between GDM (n = 40) and low risk controls (n = 56). The mean maternal age was found higher in the GDM group compared to control group (p = 0.002). Maternal BMI was significantly higher in the GDM group than the control group (p = 0.01). Abdominal circumference (AC) was significantly higher in the GDM group compared to control group (p = 0.04). EFW was significantly higher in the GDM group compared to control group (p = 0.04). FAPD/OFD ratio was found to be higher in the GDM group than in the control group (p = 0.001). Among GDM patients, no statistically significant difference was found in the ultrasound measurements between the groups receiving insulin treatment and those without treatment. According to the correlation analysis results a moderate, positive, and statistically significant correlation was present between FAPD/OFD and GDM. In perinatal outcomes, the rate of neonatal intensive care unit admission was significantly higher in the GDM group. Fetal frontal lobe development seems to be affected by GDM.


VTE risk assessment in cancer. Who needs prophylaxis and who does not?
Venous thromboembolism (VTE) in patients with cancer is associated with an increased morbidity and mortality, and its prevention is of major clinical importance. However, the VTE rates in the cancer population vary between 0.5% - 20%, depending on cancer-, treatment- and patient-related factors. The most important contributors to VTE risk are the tumor entity, stage and certain anti-cancer treatments. Cancer surgery represents a strong risk factor for VTE, and medical oncology patients are at increased risk of developing VTE, especially when receiving chemotherapy or immunomodulatory drugs. Also biomarkers have been investigated for their usefulness to predict risk of VTE (e.g. elevated leukocyte and platelet counts, soluble P-selectin, D-dimer, etc.). In order to identify cancer patients at high risk of VTE and to improve risk stratification, risk assessment models have been developed, which contain both clinical parameters and biomarkers. While primary thromboprophylaxis with low-molecular-weight-heparin (LMWH) is recommended postoperatively for a period of up to 4 weeks after major cancer surgery, the evidence is less clear for medical oncology patients. Thromboprophylaxis in hospitalized medical oncology patients is advocated, and is based on results of randomized controlled trials which evaluated the efficacy and safety of LMWH for prevention of VTE in hospitalized medically ill patients. In recent trials the benefit of primary thromboprophylaxis in cancer patients receiving chemotherapy in the ambulatory setting has been investigated. However, at the present stage primary thromboprophylaxis for prevention of VTE in these patients is still a matter of debate and cannot be recommended for all cancer outpatients.


The ketogenic and atkins diets effect on intractable epilepsy: a comparison.
Intractable epilepsy is a major difficulty in child neurology, because the numbers of drugs that are available for treatment are limited and new treatments such as diets must be tried. Now there are some diets available for treating patients with intractable epilepsy. The oldest diet is the classic ketogenic diet and one of the newest diets is the modified Atkins diet. Patients have a harder time accepting the classic ketogenic diet than the Atkins diet, which is easier to accept because the food tastes better. This study compares the efficacy of the ketogenic diet and the Atkins diet for intractable epilepsy in children. This study is a clinical trial survey with sample size of 40 children with refractory epilepsy who were patients at Mofid hospital in Tehran, Iran. Initially, from Jan 2005-Oct 2007, 20 children were treated with the Atkins diet, and then from Oct 2007-March 2010, the other group was treated with the classic ketogenic diet and the results were compared. In this study, response to treatment was greater than a 50% reduction in seizures and at the end of first, second, and third months for the ketogenic diet were 55%, 30%, and 70% and for the Atkins diet were 50%, 65%, and 70%, respectively. The results of this study show that there is no significant difference between the classic Ketogenic diet and the Atkins diet at the end of first, second, and third months and both had similar responses to the treatments.


Extended clinical phenotype, endocrine investigations and functional studies of a loss-of-function mutation A150V in the thyroid hormone specific transporter MCT8.
Thyroid hormones, besides having other functions, are known to be essential for the development of the human brain. Recently the monocarboxylate transporter 8 (MCT8) was identified as a thyroid hormone transporter which is expressed in different regions of the human brain. Here we describe in detail the clinical and biochemical features in response to thyroid hormone administration of a boy carrying an MCT8 mutation (A150V) in the second transmembrane domain. To study the functional impact of the mutation we performed triiodothyronine (T3) uptake, immunofluorescence and dimerization studies. Thyroid hormone (l-thyroxine (LT4) and LT3) administration did not result in any significant clinical changes; however, with high doses of LT4, alone or in combination with T3, TSH suppression was achieved. We could show a robust uptake of (125)I-T3 for wild type (WT) MCT8, whereas no specific uptake could be detected for the mutant A150V. Subcellular localization of WT and mutant MCT8 revealed a strong cell surface expression for the WT MCT8, in contrast to A150V, which is mostly retained intracellularly with only weak cell surface expression. We could also demonstrate for the first time that WT MCT8 as well as the mutant are able to form multimers. Our findings open a wide field of possible interaction within the central nervous system and will help to understand the crucial role of MCT8 in early fetal brain development.


Cardiac index measurements by transcutaneous Doppler ultrasound and transthoracic echocardiography in adult and pediatric emergency patients.
Non-invasive hemodynamic monitoring may facilitate resuscitation in critically ill patients. Validation studies examining a transcutaneous Doppler ultrasound technology, USCOM-1A, using pulmonary artery catheter as the reference standard showed varying results. In this study, we compared non-invasive cardiac index (CI) measurements by USCOM-1A with transthoracic echocardiography (TTE). This study was a prospective, observational cohort study at a university tertiary-care emergency department, enrolling a convenience sample of adult and pediatric patients. Paired measures of CI, stroke volume index (SVI), aortic outflow tract diameter (OTD), velocity time integral (VTI) were obtained using USCOM-1A and TTE. Pearson's correlation and Bland-Altman analyses were performed. One-hundred and sixteen subjects were enrolled, with obtainable USCOM-1A CI measurements for 99 subjects (55 adults age 50 +/- 20 years and 44 children age 11 +/- 4 years) in the final analysis. Cardiac, gastrointestinal and infectious illnesses were the most common presenting diagnostic categories. The reference standard TTE measurements of CI, SVI, OTD, and VTI in all subjects were 3.08 +/- 1.18 L/min/m(2), 37.10 +/- 10.91 mL/m(2), 1.92 +/- 0.36 cm, and 20.36 +/- 4.53 cm, respectively. Intra-operator reliability of USCOM-1A CI measurements showed a correlation coefficient of r = 0.79, with 11 +/- 22% difference between repeated measures. The bias and limits of agreement of USCOM-1A compared to TTE CI were 0.58 (-1.48 to 2.63) L/min/m(2). The percent difference in CI measurements with USCOM-1A was 31 +/- 28% relative to TTE measurements. The USCOM-1A hemodynamic monitoring technology showed poor correlation and agreement to standard transthoracic echocardiography measures of cardiac function. The utility of USCOM-1A in the management of critically ill patients remains to be determined.


Proteomic analysis revealed ROS-mediated growth inhibition of Aspergillus terreus by shikonin.
Aspergillus terreus is an emerging fungal pathogen in immunocompromised patients. Due to the intrinsic resistance of AmB against A. terreus and acquiring resistance to azoles, an alternative antifungal strategy needs investigation. Thus, we explored the activity of phytochemicals such as shikonin, gallic acid, coumaric acid and quercetin against A. terreus. Among these, shikonin showed significant inhibition at MIC50; 2 μg/ml. SEM analyses revealed delayed swelling and distorted cell wall organization in shikonin treated A. terreus conidia. Further, we performed differential proteome profiling using nLC-ESI-MS/MS, qRT-PCR and catalase assay with and without shikonin treated A. terreus. Protein data generated using Proteome Discoverer showed 882 differentially expressed proteins (680 up- and 202 down-regulated). GO analysis showed proteins from signaling pathways, oxidative stress, energy metabolism, and cytoskeleton organization. qRT-PCR of selected genes from ROS detoxification (catalase-peroxidase, superoxide dismutase), respiration (succinate-dehydrogenase, NADH-ubiquinone oxidoreductase), signaling (protein kinase C, Mitogen-activated protein kinase, cAMP-dependent protein kinase, ras-1), and 1, 3-β-glucanosyltransferase, rho-1, β-hexosaminidase showed correlation with expressed proteins. We also observed elevated reactive oxygen species using fluorescence ROS assay correlating with low catalase-peroxidase activity in shikonin treated A. terreus. Modulation of ROS homeostasis and the metabolic shift could be instrumental in shikonin- mediated growth inhibition of A. terreus. SIGNIFICANCE: Aspergillosis caused by A. terreus requires more attention due to the development of drug resistance. In this report, we employed nLC-ESI-MS/MS to identify the differential proteome in A. terreus in response to shikonin. The identified proteins involved in pathways influenced by shikonin could be helpful to understand the molecular mechanism on how shikonin/ phytochemicals or other antifungal agents inhibit fungal growth and may enable the discovery of novel or synergistic drug targets.


Novel 2-alkyl-1alpha,25-dihydroxy-19-norvitamin D3: efficient synthesis with Julia olefination, evaluation of biological activity and development of new analyzing system for co-activator recruitment.
New 2-alkyl-1alpha,25-dihydroxy-19-norvitamin D3 analogs were efficiently synthesized. The C2-alkyl-A-ring precursors were prepared as (3R,5R)-4-alkyl-3,5-dihydroxycyclohexanones from (-)-quinic acid based on radical allylation at the C4 position of methyl (-)-quinicate. The novel CD-ring coupling partner, with the elongated two carbon unit, was synthesized from 25-hydroxy Grundmann's ketone, and applied to modified Julia olefination to construct a diene unit between the A-ring and the CD-ring. The coupling yields, including a deprotection step, were moderate (47-62%). After the separation of the diastereomers based on C2 stereochemistry using HPLC, the structure (2alpha or 2beta) was determined by 1H NMR studies including NOE (nuclear Overhauser effect) experiments. It was also found that 1H NMR chemical shifts on the A-ring showed good correlation with DeLuca's 2-methyl- and 2-ethyl-1alpha,25-dihydroxy-19-norvitamin D3 analogs. Among the synthesized 19-norvitamin D3 analogs, 2alpha-(3-hydroxypropyl)-1alpha,25-dihydroxy-19-norvitamin D3 (8a) showed almost the same potency in binding to the bovine thymus vitamin D receptor (VDR) as the natural hormone (1), while its beta-isomer had only a 3% affinity. Both 2alpha-allyl- and 2alpha-propyl-1alpha,25-dihydroxy-19-norvitamin D3 and their 2beta-analogs possessed a weak affinity for the VDR. The strong VDR ligand 8a was ca. 36-fold more potent in induction of HL-60 cell differentiation than 1 and, interestingly, even the weaker ligand, 2beta-(3-hydroxypropyl)-1alpha,25-dihydroxy-19-norvitamin D3 (8b), showed a 6.7-fold higher potency in cell differentiation activity than that of 1. Next, a novel rapid analyzing system of ligand-induced co-factor recruitment on VDR was developed using fluorophore labelled CoA and HCHO fixing of the complex. The efficiency of recruitment of co-activators would explain the discrepancy between the biological activity and the VDR affinity of the ligand.


Genetics of resistance to the rust fungus Coleosporium ipomoeae in three species of morning glory (Ipomoea).
We examined the genetic basis of resistance to the rust pathogen Coleosporium ipomoea in three host species: Ipomoea purpurea, I. hederacea, and I. coccinea (Convolvulaceae). In crosses between resistant and susceptible individuals, second-generation selfed offspring segregated in ratios that did not differ statistically from the 3:1 ratio indicative of single-gene resistance with the resistant allele dominant. One out of three crosses between resistant individuals from two different populations revealed that resistance loci differed in the two populations, as evidenced by the production of susceptible individuals among the S(2) generation. These results suggest that gene-for-gene interactions contribute substantially to the dynamics of coevolution in this natural pathosystem. They also suggest that evolution of resistance to the same pathogen strain may involve different loci in different Ipomoea populations.


The versatile nature of coral-associated viruses.
A recent hypothesis considers that many coral pathologies are the result of a sudden structural alteration of the epibiotic bacterial communities in response to environmental disturbances. However, the ecological mechanisms that lead to shifts in their composition are still unclear. In the ocean, viruses represent a major bactericidal agent but little is known on their occurrence within the coral holobiont. Recent reports have revealed that viruses are abundant and diversified within the coral mucus and therefore could be decisive for coral health. However, their mode of action is still unknown, and there is now an urgent need to shed light on the nature of the relationships they might have with the other prokaryotic and eukaryotic members of the holobiont. In this opinion letter, we are putting forward the hypothesis that coral-associated viruses (mostly bacterial and algal viruses), depending on the environmental conditions might either reinforce coral stability or conversely fasten their decline. We propose that these processes are presumably based on an environmentally driven shift in infection strategies allowing viruses to regulate, circumstantially, both coral symbionts (bacteria or Symbiodinium) and surrounding pathogens.


[Clinical features and history of vasovagal syncope].
Aim of this investigation was to study special characteristics of natural course of vasovagal syncope (VVS). During 3 years we examined 212 patients (44% men) in accordance with recommendations of European Society of Cardiology using tilt tests according to Westminster or Italian protocols for confirmation of vasovagal genesis. Depending on results of initial investigation patients were divided into 2 groups: group 1 comprised 144 patients (68%) satisfying criteria of VVS; group 2 comprised other patients (n=41, 19%) with transitory loss of consciousness of unclear origin. Patients with VVS were significantly younger (mean age 35.1+/-13.6 and 44.4+/-13.,9 years, respectively; <0.01) with earlier appearance of first episode of fainting (16 vs. 39 years; <0,01). In most cases VVS appeared in the age younger than 35 years. Accuracy of anamnestic method for diagnosis of VVS was 99%. Forty one percent of patients with classical VVS had several potential causes of fainting (situational syncope, paroxysms of tachyarrhythmia, epilepcy). This could lead to mistakes at initial stage of diagnostics. Progressive clinical course was observed in 15% of patients and was associated either with syncopi of other, including nonvagal origin, or with increased frequency of previously stereotypical attacks. In every second patient with initially frequent recurrences of VVS we observed long spontaneous remissions. Nondrug methods of treatment were effective in 43% of these patients. Tilt test had high informative power for diagnostics of VVS. With this its informativity was low in patients with onset of fainting attacks in middle age and with atypical clinical picture.


Maternal stress during pregnancy is associated with moderate to severe depression in 11-year-old children.
Maternal stress during pregnancy has been associated with negative outcomes in children. We examined the risk factors for symptoms of depression in 11-year-old children, including the interaction between birthweight and other variables. We collected maternal, obstetric and demographic information from birth through to the age of 11. Approximately, half of the 609 children were born small-for-gestational-age (SGA). Information collected at 3.5 and 7 years of age included intelligence testing and parent-reported behavioural and emotional development. At 11 years of age, the children completed the Center for Epidemiological Studies Depression Scale for Children. Multivariable logistic regression analysis examined the relationship between self-reported symptoms of moderate to severe depression at the age of 11 and explanatory variables. Symptoms of moderate to severe depression were related to increasing maternal stress during pregnancy, young maternal age, lower intelligence test scores at 7-years-old and being bullied at school in the previous 6 months. There was also a significant interaction between maternal stress in pregnancy and symptoms of depression in 11-year-old children born SGA. Increasing maternal stress during pregnancy was associated with increased risk of symptoms of moderate to severe depression in 11-year-old children, especially those who were born SGA.


[Soluble expression of peptide containing MUC1/Y-specific epitope in Escherichia coli and preparation of the antibody].
MUC1 mucin is a high molecular weight, type I transmembrane glycoprotein. High and aberrant expression of MUC1 is observed in various types of tumors, which make it an ideal target for tumor biotherapy as well as a biomarker for tumor diagnosis and prognosis. MUC1/Y is an isoform of MUC1 generated by alternative splicing. Specific expression of MUC1/Y in breast cancer as well as its involvement in tumor cell signal transduction have been reported. In order to purify peptides containing MUC1/Y-specific epitope in E. coli and prepare MUC1/Y-specific antibody, DNA fragment encoding the MUC1/Y-specific peptide was amplified by PCR using MUC1/Y full length cDNA as the template and cloned into fusion expression vector pGEX-2T, resulting pGEX-Y30. DNA sequencing was performed to confirm the correct amplification and orientation of the target sequence. Competent E. coli DH5alpha was transformed with pGEX-Y30 and the expression was induced for 4-5 hours in 0.2 mmol/L IPTG at 30 degrees C and 37 degrees C. Expressed proteins were released from the cells by ultrasonication or B-PER II reagent treatments. The fusion protein GST-Y30 were purified by affinity and anion exchange columns and identified by SDS-PAGE and Western-blotting. Polyclonal antibody was prepared by immunizing rabbits with the GST-Y30 protein for 4 times with intervals of 3 weeks and purified by GST column. Western blotting, ELISA and immunohistochemistry analysis were carried out using the purified antibody to confirm its MUC1/Y-binding capacity and specificity. The expressed fusion protein GST-Y30 is about 31 kD in size and represented about 20% of total cellular proteins. The majority of the GST-Y30 protein existed as soluble form when the induction was carried out at both 30 degrees C and 37 degrees C. After the two-step purification, the purity of GST-Y30 was about 94%. The titer of polyserum generated by GST-Y30 immunization was 1:320,000 by ELISA. The antiserum showed MUC1/Y specificity and can recognize MUC1/Y on MCF7 cell. The MUC1/Y-specific polyclonal antibody can be used for studying the role of MUC1/Y in carcinogenesis.


LSD1/KDM1A mutations associated to a newly described form of intellectual disability impair demethylase activity and binding to transcription factors.
Genetic diseases often lead to rare and severe syndromes and the identification of the genetic and protein alterations responsible for the pathogenesis is essential to understand both the physiological and pathological role of the gene product. Recently, de novo variants have been mapped on the gene encoding for the lysine-specific histone demethylase 1 (LSD1)/lysine(K)-specific histone demethylase 1A in three patients characterized by a new genetic disorder. We have analyzed the effects of these pathological mutations on the structure, stability and activity of LSD1 using both in vitro and cellular approaches. The three mutations (Glu403Lys, Asp580Gly and Tyr785His) affect active-site residues and lead to a partial impairment of catalytic activity. They also differentially perturb the ability of LSD1 to engage transcription factors that orchestrate key developmental programs. Moreover, cellular data indicate a decrease in the protein cellular half-life. Taken together, these results demonstrate the relevance of LSD1 in gene regulation and how even moderate alterations in its stability, catalytic activity and binding properties can strongly affect organism development. This depicts a perturbed interplay of catalytic and non-catalytic processes at the origin of the pathology.


Interactions of Wnt/beta-catenin signaling and sonic hedgehog regulate the neurogenesis of ventral midbrain dopamine neurons.
Signaling mechanisms involving Wnt/beta-catenin and sonic hedgehog (Shh) are known to regulate the development of ventral midbrain (vMB) dopamine neurons. However, the interactions between these two mechanisms and how such interactions can be targeted to promote a maximal production of dopamine neurons are not fully understood. Here we show that conditional mouse mutants with region-specific activation of beta-catenin signaling in vMB using the Shh-Cre mice show a marked expansion of Sox2-, Ngn2-, and Otx2-positive progenitors but perturbs their cell cycle exit and reduces the generation of dopamine neurons. Furthermore, activation of beta-catenin in vMB also results in a progressive loss of Shh expression and Shh target genes. Such antagonistic effects between the activation of Wnt/beta-catenin and Shh can be recapitulated in vMB progenitors and in mouse embryonic stem cell cultures. Notwithstanding these antagonistic interactions, cell-type-specific activation of beta-catenin in the midline progenitors using the tyrosine hydroxylase-internal ribosomal entry site-Cre (Th-IRES-Cre) mice leads to increased dopaminergic neurogenesis. Together, these results indicate the presence of a delicate balance between Wnt/beta-catenin and Shh signaling mechanisms in the progression from progenitors to dopamine neurons. Persistent activation of beta-catenin in early progenitors perturbs their cell cycle progression and antagonizes Shh expression, whereas activation of beta-catenin in midline progenitors promotes the generation of dopamine neurons.


Expression of SERCA isoform with faster Ca2+ transport properties improves postischemic cardiac function and Ca2+ handling and decreases myocardial infarction.
Myocardial ischemia-reperfusion (I/R) injury is associated with contractile dysfunction, arrhythmias, and myocyte death. Intracellular Ca(2+) overload with reduced activity of sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA) is a critical mechanism of this injury. Although upregulation of SERCA function is well documented to improve postischemic cardiac function, there are conflicting reports where pharmacological inhibition of SERCA improved postischemic function. SERCA2a is the primary cardiac isoform regulating intracellular Ca(2+) homeostasis; however, SERCA1a has been shown to substitute SERCA2a with faster Ca(2+) transport kinetics. Therefore, to further address this issue and to evaluate whether SERCA1a expression could improve postischemic cardiac function and myocardial salvage, in vitro and in vivo myocardial I/R studies were performed on SERCA1a transgenic (SERCA1a(+/+)) and nontransgenic (NTG) mice. Langendorff-perfused hearts were subjected to 30 min of global ischemia followed by reperfusion. Baseline preischemic coronary flow and left ventricular developed pressure were significantly greater in SERCA1a(+/+) mice compared with NTG mice. Independent of reperfusion-induced oxidative stress, SERCA1a(+/+) hearts demonstrated greatly improved postischemic (45 min) contractile recovery with less persistent arrhythmias compared with NTG hearts. Morphometry showed better-preserved myocardial structure with less infarction, and electron microscopy demonstrated better-preserved myofibrillar and mitochondrial ultrastructure in SERCA1a(+/+) hearts. Importantly, intraischemic Ca(2+) levels were significantly lower in SERCA1a(+/+) hearts. The cardioprotective effect of SERCA1a was also observed during in vivo regional I/R with reduced myocardial infarct size after 24 h of reperfusion. Thus SERCA1a(+/+) hearts were markedly protected against I/R injury, suggesting that expression of SERCA 1a isoform reduces postischemic Ca(2+) overload and thus provides potent myocardial protection.


[Tailored therapy for rheumatic disease within reach].
Personalised medicine has the potential to increase therapeutic effectiveness, reduce side effects and lower cost. This approach has recently taken off in oncology where different malignancies may be treated with specific drugs based on genetic biomarkers or other tumour characteristics. This type of tailored therapy could also be developed for immune-mediated inflammatory disease, for which rheumatoid arthritis (RA) may serve as a prototype. While novel treatments are able to halt or even prevent disease progression, not all RA patients respond, and stratification of patient groups is needed. The identification of biomarkers predictive of the clinical response to specific treatments in subsets of patients may soon become reality in a variety of diseases.


Paracrine stimulation of preadipocyte-enriched cell cultures by mature adipocytes.
We developed a double-chamber system in which to examine the effects of mature adipocytes on the growth and differentiation of preadipocytes and other cells in the adipose tissue. In the present study we found that mature adipocytes from both lean and obese subjects release a factor that stimulates the growth of preadipocyte-enriched and dedifferentiated adipocyte-enriched cell cultures. This growth stimulation was dependent on both time of exposure to mature cells and the number of mature cells in the coculture. Proliferation of the preadipocyte-enriched (n = 4) and dedifferentiated adipocyte-enriched cultures (n = 5) in the presence of mature adipocytes from obese subjects [body mass index (BMI) > 35] was 4.1- and 2.9-fold more (P < 0.05) than that in the presence of adipocytes from lean subjects (BMI < or = 25). There was no difference in the growth of cultures enriched in preadipocytes or dedifferentiated adipocytes from lean or obese subjects in the absence of mature adipocytes. These observations demonstrate that mature adipocytes from obese patients stimulate the growth of preadipocyte-enriched cultures to a greater extent than those from lean individuals.


Double-network gels as polyelectrolyte gels with salt-insensitive swelling properties.
Polyelectrolyte gels exhibit intrinsic salt-sensitive swelling behaviour, which causes size instability in ionic environments. Thus, polyelectrolyte gels that show salt-insensitive swelling have been anticipated for applications in ionic environments, such as medical materials used in vivo. We found that double-network (DN) gels consisting of both a polyelectrolyte network and a non-ionic network are resistant to salt-sensitive swelling. This resistance is attributed to their lower osmotic pressure originating from mobile ions relative to the osmotic pressure of mixing at swelling equilibrium. Our investigation indicated that the two contrasting network structures within the DN gels are vital for achieving these properties, where the structures include a highly prestretched and sparse polyelectrolyte network and a coiled and dense non-ionic network. The salt-insensitivity of the DN gels will lead to their unique applications in ionic environments.


An siRNA library screen identifies CYLD and USP34 as deubiquitinases that regulate GPCR-p38 MAPK signaling and distinct inflammatory responses.
G protein-coupled receptors (GPCRs) are highly druggable and implicated in numerous diseases, including vascular inflammation. GPCR signals are transduced from the plasma membrane as well as from endosomes and controlled by posttranslational modifications. The thrombin-activated GPCR protease-activated receptor-1 is modified by ubiquitin. Ubiquitination of protease-activated receptor-1 drives recruitment of transforming growth factor-β-activated kinase-1-binding protein 2 (TAB2) and coassociation of TAB1 on endosomes, which triggers p38 mitogen-activated protein kinase-dependent inflammatory responses in endothelial cells. Other endothelial GPCRs also induce p38 activation via a noncanonical TAB1-TAB2-dependent pathway. However, the regulatory processes that control GPCR ubiquitin-driven p38 inflammatory signaling remains poorly understood. We discovered mechanisms that turn on GPCR ubiquitin-dependent p38 signaling, however, the mechanisms that turn off the pathway are not known. We hypothesize that deubiquitination is an important step in regulating ubiquitin-driven p38 signaling. To identify specific deubiquitinating enzymes (DUBs) that control GPCR-p38 mitogen-activated protein kinase signaling, we conducted a siRNA library screen targeting 96 DUBs in endothelial cells and HeLa cells. We identified nine DUBs and validated the function two DUBs including cylindromatosis and ubiquitin-specific protease-34 that specifically regulate thrombin-induced p38 phosphorylation. Depletion of cylindromatosis expression by siRNA enhanced thrombin-stimulated p38 signaling, endothelial barrier permeability, and increased interleukin-6 cytokine expression. Conversely, siRNA knockdown of ubiquitin-specific protease-34 expression decreased thrombin-promoted interleukin-6 expression and had no effect on thrombin-induced endothelial barrier permeability. These studies suggest that specific DUBs distinctly regulate GPCR-induced p38-mediated inflammatory responses.


Maternal vitamin A and β-carotene supplementation and risk of bacterial vaginosis: a randomized controlled trial in rural Bangladesh.
Bacterial vaginosis (BV) in pregnancy is linked to preterm birth, but its risk factors are not well understood. Micronutrient deficiencies may be associated with an increased risk of this condition. We assessed the effect of weekly vitamin A or β-carotene supplementation during pregnancy until 3 mo postpartum on BV risk in rural northeastern Bangladesh. In this cluster-randomized, placebo-controlled trial, 33 clusters (n = 33) were randomly assigned to 3 groups. Women (n = 1812) were examined for BV by using self-administered swabs and the Nugent scoring method in early pregnancy, at 32 wk of gestation, and at 3 mo postpartum. The prevalence of BV in early pregnancy, before supplementation, was 7.6% (95% CI: 6.3%, 9.1%) overall. Neither the prevalence nor the incidence of BV in the third trimester differed by supplement group. However, the prevalence (OR: 0.71; 95% CI: 0.52, 0.98) and incidence (RR: 0.58; 95% CI: 0.41, 0.81) of BV at 3 mo postpartum was lower among women in the vitamin A group (9.1% and 6.7%, respectively) than in the placebo group (12.4% and 11.8%, respectively), but not in the β-carotene group. Both vitamin A and β-carotene reduced the prevalence and incidence of BV at both time points (ie, third trimester and 3 mo postpartum) by 30-40% compared with placebo (all P < 0.05). Weekly vitamin A supplementation reduced the risk of maternal BV in this rural Bangladeshi population. Enhancement of vitamin A status before and during pregnancy may reduce the risk of BV in areas with vitamin A deficiency. This trial is registered at clinicaltrials.gov as NCT00198822.


Subtalar distraction arthrodesis using interpositional frozen structural allograft.
Subtalar bone-block distraction arthrodesis using structural autograft carries the risk of donor site morbidity. Recent reports suggest that structural allograft may be an attractive alternative to structural autograft in subtalar arthrodesis. This prospective study analyzes subtalar distraction arthrodesis using interpositional structural allograft. Between 2000 and 2006, 22 patients (24 feet; mean age, 45.6 years) underwent subtalar arthrodesis with interpositional fresh-frozen femoral head structural allograft. Indications included subtalar arthrosis, loss of heel height, and anterior ankle impingement. Clinical outcome was assessed using the AOFAS ankle-hindfoot scoring system. Time to union was determined by previously reported clinical findings and radiographic evidence for bridging trabeculation between host bone and structural allograft. Mean followup was 35.8 months for 20 patients (21 feet) available for followup evaluation. Union was achieved in 19 of 21 patients (90%) at a mean of 15.5 (range, 11 to 19) weeks. Mean AOFAS hindfoot score improved from 21 to 71 points (p < 0.05). Radiographic analysis suggested significant (p < 0.05) improvement in all measurements. Complications included nonunion (2), varus malalignment (1), persistent subfibular impingement (1), sural neuralgia (1), and prominent hardware (2). Both patients with nonunions had avascular bone at the arthrodesis site and used tobacco products. This study supports recent publications that subtalar arthrodesis using interpositional structural allograft can have a favorable outcome. Our clinical and radiographic results suggest that restoration of hindfoot function and dimensions with structural allograft are comparable to results reported for the same procedure using structural autograft. Level IV, prospective case series.


Regorafenib induces rapid and reversible changes in plasma nitric oxide and endothelin-1.
Hypertension is a toxicity of antiangiogenic therapies and a possible biomarker that identifies patients with superior cancer outcomes. Understanding its mechanism will aid in treatment and could lead to the development of other biomarkers for predicting toxicity and anticancer efficacy. Recent evidence implicates nitric oxide (NO) suppression and endothelin-1 (ET-1) stimulation as potential mechanisms leading to antiangiogenic therapy-induced hypertension. The aim of this study was to evaluate the effects of regorafenib, a novel broad-spectrum kinase inhibitor with activity against multiple targets, including vascular endothelial growth factor receptor 2 inhibition, on NO and ET-1 levels. Regorafenib was administered to 32 subjects with gastrointestinal stromal tumor on a 3-week-on, 1-week-off basis. Plasma levels of NO and ET-1 were measured at baseline, 2, 4, and 6 weeks of therapy. Data analysis was by Wilcoxon rank-sum and paired t-tests. Twenty subjects (63%) developed regorafenib-induced hypertension. Two weeks after starting regorafenib therapy, plasma ET-1 levels increased (25% increase, P < 0.05) and NO was suppressed (20% decrease, P < 0.05). These normalized after 1-week washout but ET-1 rose again by 30% (P < 0.05) and NO fell by 50% (P < 0.05) after restarting regorafenib. These findings indicate that regorafenib induces a coordinated and reversible suppression of NO and stimulation of ET-1. Whether NO and ET-1 might predict therapeutic efficacy in these patients requires further study.


Respiratory abnormalities in hypothyroidism.
While the respiratory and oxygen transport systems are complex, they are orderly, so that they can be studied logically and systematically. There are many interactions and potential abnormalities at various levels that may combine to give the ultimate defect of inadequate oxygen supply at the tissue level. An understanding of these many factors using hypothyroidism as a model should enable the clinician to approach the problem rationally and manage if effectively.


Liposome composition is important for retention of liposomal rhodamine in P-glycoprotein-overexpressing cancer cells.
Multidrug resistance (MDR) caused by high expression of P-glycoprotein (Pgp) in cancer patients remains one of the major obstacles to successful therapy of cancer. Earlier studies have shown that the incorporation of Pgp-substrate drugs in liposomes may provide a strategy to circumvent Pgp-mediated drug efflux. The present study investigated the impact of liposome composition on the efflux of Pgp-substrate incorporated in liposomes. Liposomes with varying compositions were loaded with rhodamine 123, a fluorescent probe frequently used as a Pgp-substrate, and the retention of rhodamine was compared in two breast cancer cell lines: wild-type cells with no detectable Pgp expression (MCF-7/WT) and Pgp-expressing cells resulting from stable transfection of the MDR1 gene (MCF-7/Pgp). Pgp-expression decreased the rhodamine retention in MCF-7 cells, suggesting that Pgp is functional. Liposome loading increased rhodamine retention in MCF-7/Pgp cells, but not in MCF-7/WT cells. Surface charge of liposomes did not affect the rhodamine retention, whereas the incorporation of cholesterol and polyethyleneglycol-attached lipids was effective in further increasing the rhodamine retention in MCF-7/Pgp cells. Since further study demonstrated that the rate of rhodamine release from liposomes tended to be inversely correlated with rhodamine retention by cells, it seems likely that more rigid liposomes are able to sequester rhodamine more efficiently, thereby inhibiting direct interactions of rhodamine with Pgp proteins. Taken together, these findings suggest that Pgp-mediated MDR in cancer cells may be more effectively modulated by optimizing the composition of liposomes for loading Pgp-substrate anti-cancer drugs.


Effect of Vitamin D Supplementation on Intracytoplasmic Sperm Injection Outcomes: A Randomized Double-Blind Placebo-Controlled Trial.
Despite numerous studies indicating an imperative role for reproduction, however, the role of Vitamin D supplementation on outcomes of assisted reproductive techniques remains controversial. This clinical trial was performed to evaluate the effect of Vitamin D supplementation 6 weeks prior to intracytoplasmic sperm injection (ICSI) on fertility indices. The present study was a double-blind clinical trial conducted on infertile women was randomly allocated into two groups: Vitamin D supplementation (42 participants) and placebo (43 participants). Serum Vitamin D was measured before and six to eight weeks after treatment, on the day of ovum pick up. Results were analyzed using SPSS16 and fertility indices were compared between the two groups. No significant difference was observed between the intervention and control groups regarding the mean number of oocytes retrieved, percentage mature oocyte, fertilization rate and the rate of good quality embryos (all P>0.05). But, percentages of the individual with suitable endometrium (7-14 mm thickness) were significantly higher in the Vitamin D compared to control group (P=0.011). The rate of chemical (47.6 vs. 25.5%, P=0.013) and clinical pregnancy rate (38.1 vs. 20.9%, P=0.019) were also significantly higher in the Vitamin D compared to control group. The present study reveals that consuming Vitamin D for 6 weeks prior to ICSI improves quality of endometrium, rate of chemical and clinical pregnancy (Registration Number: IRCT2015111124999N1).


Human Bone Marrow Stromal Cells Differentiate Into Corneal Tissue and Prevent Ocular Graft-Versus-Host Disease in Mice.
Clinical trials have assessed the use of human bone marrow stromal cells (hBMSCs) for the treatment of immune-related disorders such as graft-versus-host disease (GVHD). In the current study, we show that GFP(+)-transduced hBMSCs generated from bone marrow migrate and differentiate into corneal tissue after subconjunctival injection in mice. Interestingly, these hBMSCs display morphological features of epithelial, stromal, and endothelial cells and appear at different layers and with different morphologies depending on their position within the epithelium. Furthermore, these cells display ultrastructural properties, such as bundles of intermediate filaments, interdigitations, and desmosomes with GFP(-) cells, which confirms their differentiation into corneal tissues. GFP(+)-transduced hBMSCs were injected at different time points into the right eye of lethally irradiated mice undergoing bone marrow transplantation, which developed ocular GVHD (oGVHD). Remarkably, hBMSCs massively migrate to corneal tissues after subconjunctival injection. Both macroscopic and histopathological examination showed minimal or no evidence of GVHD in the right eye, while the left eye, where no hBMSCs were injected, displayed features of GVHD. Thus, in the current study, we confirm that hBMSCs may induce their therapeutic effect at least in part by differentiation and regeneration of damaged tissues in the host. Our results provide experimental evidence that hBMSCs represent a potential cellular therapy to attenuate oGVHD.


New developments in the comprehension of the biotransformation and transport of insulin-enhancing vanadium compounds in the blood serum.
The possible biotransformations in the blood serum of four representative insulin-enhancing vanadium compounds, [VO(6-mepic)(2)], cis-[VO(pic)(2)(H(2)O)], [VO(acac)(2)], and [VO(dhp)(2)], where 6-mepic, pic, acac, and dhp indicate the deprotonated forms of 6-methylpicolinic and picolinic acids, acetylacetone, and 1,2-dimethyl-3-hydroxy-4(1H)-pyridinone, were examined. In particular, the behavior of the quinary systems formed by the insulin-enhancing species, human serum apo-transferrin (hTf), human serum albumin (HSA), and lactate (lact) or citrate (citr) at physiological pH and conditions was studied. The results indicate that, besides the case in which the ligand is very weak like 6-mepic, the carrier can interact in some form with VO(2+) ion until its intake into the cell. In fact with stronger ligands like pic, acac, and dhp, VO(2+) is transported not only by transferrin but also as [VO(carrier)(2)] and as mixed species VO(2+)-hTF-carrier. There are two ways in which the undissociated form of a bis-chelated complex can interact with transferrin, one "specific" when the carrier possesses a carboxylate group and behaves like a synergistic anion, and another "non-specific" when an imidazole nitrogen of a histidine residue from hTf replaces an equatorially coordinated water molecule giving rise to a ternary species with cis-octahedral geometry and cis-VO(carrier)(2)(hTf) stoichiometry. It is found that also albumin can participate in the transport of an insulin-enhancing compound forming a mixed species cis-VO(carrier)(2)(HSA), when the carrier stabilizes in aqueous solution the cis-octahedral form, or the dinuclear compound (VO)(2)(d)HSA, when the carrier forms unstable complexes. These insights were confirmed through density functional theory (DFT) calculations.


The effect of age on ocular blood supply determined by pulsatile ocular blood flow and color Doppler ultrasonography.
Pulsatile ocular blood flow (POBF) assessment measures the choroidal circulation and provides diagnostic value to certain ocular diseases such as glaucoma. This technique assumes a constant ocular rigidity and is influenced by axial length, diurnal variation, and age. This study investigated the effect of age on POBF, with consideration of the above factors. Ocular blood supply in the ophthalmic artery was also determined using color Doppler ultrasonography. A total of 118 healthy subjects aged 19 to 75 years were recruited. They were divided into five groups (below age 30, 30 to 39, 40 to 49, 50 to 59, and 60 or above) of at least 20 subjects each. Only one eye of each subject, with axial length <24.5 mm, was considered. The subject's supine POBF was determined followed by a measurement of the blood flow velocity in the ophthalmic artery using color Doppler ultrasonography. All the measurements were at around the same time of day to eliminate any effect from diurnal variation. The scleral rigidity was measured using a Schiotz tonometer with 5.5- and 7.5-g weights. Linear regression analysis demonstrated a significant increase of scleral rigidity with age (Pearson correlation coefficient r = 0.26, p < 0.01) and a significant decrease of POBF with age (r = -0.35, p < 0.01). The reduction in peak systolic velocity in the ophthalmic artery with age was significant (r = -0.28, p < 0.01). Both the systolic and diastolic brachial pressure showed significant increase with age (r = 0.55, p < 0.01; r = 0.40, p < 0.01, respectively). Using multiple regression analysis, POBF showed a significant correlation with age (partial correlation r = -0.36, p < 0.01), but not with scleral rigidity or systolic or diastolic brachial pressure. The peak systolic velocity in the ophthalmic artery also showed significant correlation with age (partial correlation r = -0.29, p < 0.01). The reduction in POBF with age was significant. Although aging affects scleral rigidity and systemic blood pressure, multiple regression analysis indicates that the most influential factor affecting POBF is aging. The peak systolic velocity in the ophthalmic artery also decreased with age, indicating reduced ocular blood supply.


Macrophage interferon regulatory factor 4 deletion ameliorates aristolochic acid nephropathy via reduced migration and increased apoptosis.
Aristolochic acid (AA) is the causative nephrotoxic alkaloid in AA nephropathy, which results in a tubulointerstitial fibrosis. AA causes direct proximal tubule damage as well as an influx of macrophages, although the role of macrophages in pathogenesis is poorly understood. Here, we demonstrate that AA directly stimulates migration, inflammation, and ROS production in macrophages ex vivo. Cells lacking interferon regulatory factor 4 (IRF4), a known regulator of macrophage migration and phenotype, had a reduced migratory response, though effects on ROS production and inflammation were preserved or increased relative to WT cells. Macrophage-specific IRF4-knockout mice were protected from both acute and chronic kidney effects of AA administration based on functional and histological analysis. Renal macrophages from kidneys of AA-treated macrophage-specific IRF4-knockout mice demonstrated increased apoptosis and ROS production compared with WT controls, indicating that AA directly polarizes macrophages to a promigratory and proinflammatory phenotype. However, knockout mice had reduced renal macrophage abundance following AA administration. While macrophages lacking IRF4 can adopt a proinflammatory phenotype upon AA exposure, their inability to migrate to the kidney and increased rates of apoptosis upon infiltration provide protection from AA in vivo. These results provide evidence of direct AA effects on macrophages in AA nephropathy and add to the growing body of evidence that supports a key role of IRF4 in modulating macrophage function in kidney injury.


Prenatal Exposure to Environmental Tobacco Smoke and Hyperactivity Behavior in Chinese Young Children.
This study aimed to examine the association between prenatal environmental tobacco smoke (ETS) exposure and hyperactivity behaviors in young children. A cross-sectional study was undertaken among 21,243 participants from all of the kindergartens in Longhua District of Shenzhen, China. Multivariate logistic regression models and hierarchical linear models were employed to assess the associations. After adjusting for potential confounders of gender, preterm birth, birth asphyxiation, etc., prenatal ETS exposure was significantly associated with an increased risk of hyperactivity behaviors in young children (OR (95% CI) = 1.51 (1.28-1.77); β (95% CI) = 0.017 (0.013-0.020)). Along with increases in children's prenatal ETS exposure dose (measured by daily ETS exposure duration, daily cigarette consumption by household members, and overall score of prenatal ETS exposure), the children were also increasingly more likely to exhibit hyperactivity behaviors. Furthermore, children whose mothers had prenatal ETS exposure in any one or more of the pregnancy trimesters were more likely to exhibit hyperactivity behaviors as compared with those born to non-exposure mothers (all p < 0.05). Overall, prenatal ETS exposure could be associated with a detrimental impact on offspring's hyperactivity behaviors, and public health efforts are needed to reduce prenatal ETS exposure.


Demonstration of defective C3-receptor-mediated clearance by the reticuloendothelial system in patients with acquired immunodeficiency syndrome.
The function of macrophage C3 receptors was assessed in vivo by measuring the clearance of C3-sensitized autologous erythrocytes in seven acquired immunodeficiency syndrome (AIDS) patients, eight healthy homosexual men, eight healthy heterosexual men, and four infected controls. Healthy heterosexual men had an initial clearance of 50.1 +/- 2.0% of the inoculum, with a release of a small portion of these cells (10.9 +/- 1.3%) into the circulation. Healthy homosexual men had a greater initial clearance of 66.0 +/- 4.2% (P less than 0.01) followed by a similar release (14.0 +/- 3.3%). AIDS patients had an initial clearance of 60.6 +/- 7.5% but had a relatively large release of cells (25.6 +/- 3.2%) (P less than 0.005 vs. heterosexuals; P less than 0.05 vs. homosexuals), suggesting a failure of macrophage phagocytosis. Infected controls had an initial clearance of 59.4 +/- 4.9%, with a release of 19.6 +/- 3.8% (P = NS vs. AIDS). These data, in addition to Fc-receptor dysfunction, demonstrate a global reticuloendothelial system dysfunction in AIDS patients. This may contribute to their frequent infections with opportunistic pathogens and inappropriate immune responses against these microorganisms.


Quality control of the sheep bacterial artificial chromosome library, CHORI-243.
The sheep CHORI-243 bacterial artificial chromosome (BAC) library is being used in the construction of the virtual sheep genome, the sequencing and construction of the actual sheep genome assembly and as a source of DNA for regions of the genome of biological interest. The objective of our study is to assess the integrity of the clones and plates which make up the CHORI-243 library using the virtual sheep genome. A series of analyses were undertaken based on the mapping the sheep BAC-end sequences (BESs) to the virtual sheep genome. Overall, very few plate specific biases were identified, with only three of the 528 plates in the library significantly affected. The analysis of the number of tail-to-tail (concordant) BACs on the plates identified a number of plates with lower than average numbers of such BACs. For plates 198 and 213 a partial swap of the BESs determined with one of the two primers appear to have occurred. A third plate, 341, also with a significant deficit in tail-to-tail BACs, appeared to contain a substantial number of sequences determined from contaminating eubacterial 16 S rRNA DNA. Additionally a small number of eubacterial 16 S rRNA DNA sequences were present on two other plates, 111 and 338, in the library. The comparative genomic approach can be used to assess BAC library integrity in the absence of fingerprinting. The sequences of the sheep CHORI-243 library BACs have high integrity, especially with the corrections detailed above. The library represents a high quality resource for use by the sheep genomics community.


Utilization of Osseous Landmarks for Anatomic Anterior Cruciate Ligament Femoral Tunnel Placement.
Accurate placement of the femoral tunnel is critical for long-term clinical success following anterior cruciate ligament (ACL) reconstruction. The purpose of the present study is to evaluate the accuracy of femoral tunnel placement when referencing osseous landmarks during ACL reconstruction. We hypothesize that referencing osseous landmarks during ACL reconstruction consistently results in anatomic placement of the ACL femoral tunnel. This study was a retrospective case series. We reviewed 83 consecutive ACL reconstructions performed by a single surgeon. The lateral intercondylar ridge and lateral bifurcate ridge were referenced intraoperatively for anatomic placement of the ACL femoral tunnel during single-bundle reconstruction. Using these landmarks, the femoral tunnel was placed in the center of the anteromedial bundle footprint on the lateral wall of the intercondylar notch. We reviewed all operative notes and intraoperative arthroscopic images to assess tunnel placement. Postoperative anteroposterior and lateral radiographs were obtained in all patients. Anatomic placement was confirmed by review of lateral radiographs utilizing both the quadrant method (QM) and Blumensaat-ridge ratio (BRR). We used a total of 80 patients for our study. Review of arthroscopic images confirmed anatomic placement of the ACL femoral tunnel in all patients. All patients demonstrated that the femoral tunnel was placed anatomically according to the BRR method. Using the QM, all femoral tunnels were placed anatomically except for one tunnel that was placed slightly anteriorly. There was excellent agreement between the two radiographic measurement techniques. The principal finding of this study indicates that the lateral intercondylar ridge and the lateral bifurcate ridge are reliable landmarks for anatomic placement of the ACL femoral tunnel. Referencing osseous landmarks during surgery can help surgeons avoid nonanatomic placement of the ACL femoral tunnel, especially in cases where the soft-tissue footprint is no longer present. Furthermore, both the radiographic QM and the BRR are valid techniques to assess for anatomic ACL femoral tunnel placement both intraoperatively and postoperatively.


Understanding the fine-scale heterogeneity and spatial drivers of malaria transmission in Kenya using model-based geostatistical methods.
Malaria remains a public health concern. Monitoring the fine-scale heterogeneity of the malaria burden enables more targeted control efforts. Although malaria indicator surveys (MIS) have been crucial in evaluating the progress of malaria control interventions, they are only designed to provide a cross-sectional national and regional malaria disease burden. Recent advances in geostatistical methods allow us to interpolate national survey data to describe subnational disease burden that is crucial in informing targeted control. A binomial geostatistical model employing Markov chain Monte Carlo (MCMC) parameter estimation methods is used to understand the spatial drivers of malaria risk in Kenya and to predict malaria risk at a fine-scale resolution, including identifying hotspots. A total of 11,549 children aged six months to 14 years from 207 clusters were sampled in this survey and used in the present analysis. The national malaria prevalence based on the data was 8.4%, with the highest in the lake endemic zone (18.1%) and the lowest in the low-risk zone (<1%). The analysis shows that elevation, proportion of insectcide treated net (ITN) distributed, rainfall, temperature and urbanization covariates are all significant predictors of malaria transmission. The 5x5 Km resolution maps show that malaria is heterogeneous in Kenya, with hotspot areas in the lake endemic area, the coastal areas, and some parts of the shores of Lake Turkana and Kajiado. The high-resolution malaria prevalence maps produced as part of the analysis have shown that Kenya has additional malaria hotspots, especially in areas least expected. These findings call for a rethinking of malaria burden classification in some regions for effective planning, implementation, resource mobilization, monitoring, and evaluation of malaria interventions in the country.


Systematic review and meta-analysis on management of hemodialysis catheter-related bacteremia.
Hemodialysis catheter-related bacteremia is a common clinical problem with several management options. We performed a systematic review and meta-analysis to determine cure proportions with systemic antibiotics, antibiotic lock solution, and guidewire exchange. We searched databases and registries; conference proceedings from relevant medical societies; and article reference lists. Data regarding management approach, cure, follow-up, recurrence, complications, and microbiology were abstracted and pooled from 28 selected publications. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated from a mixed effects logistic regression model. In total, 1596 patients with tunneled hemodialysis catheter-related bacteremia were divided into groups on the basis of treatment with systemic antibiotics (n=697), antibiotic lock solution (n=546), or guidewire exchange (n=353). Antibiotic lock solution and guidewire exchange had similar cure proportions that were superior to systemic antibiotics alone (OR, 2.08; 95% CI, 1.25 to 3.45; P<0.01 for antibiotic lock solution; OR, 2.88; 95% CI, 1.82 to 4.55; P<0.001 for guidewire exchange versus systemic antibiotics). Cure proportions were highest for coagulase-negative staphylococci followed by gram-negative rods and Staphylococcus aureus. Among S. aureus infections, guidewire exchange led to a higher cure proportion than systemic antibiotics or antibiotic lock solution (OR, 3.33; 95% CI, 1.17 to 9.46; P=0.02; OR, 4.72; 95% CI, 1.79 to 12.46; P=0.002, respectively). Thus, results of this study suggest that tunneled hemodialysis catheter-related bacteremia should be treated with either guidewire exchange or antibiotic lock solution. Future studies should address prospectively whether one strategy is better than the other overall and for specific pathogens.


Types and clinical outcomes of chemical ingestion in emergency departments in South Korea (2011-2016).
This study aims to provide basic data on the types and frequency of chemical ingestions and the clinical outcomes of chemical ingestion injury. This study retrospectively analyzed the data obtained from the Emergency Department-Based Injury In-depth Surveillance of the Korea Centers for Disease Control and Prevention (South Korea) from 2011 to 2016. Patients ingesting chemicals aged ≥ 18 years were included, but those ingesting unknown chemical substances or with unknown clinical outcomes were excluded. This study included 2,712 (47.2% were men and 52.8% were women, mean age, 47.05 years) patients ingesting chemicals. Unintentional and intentional ingestions were reported in 1,673 (61.7%) and 1,039 (38.3%), respectively. The most commonly ingested chemical substances were hypochlorites, detergents, ethanol, and acetic acid. In the unintentional ingestion group, the most common chemicals upon admission were hypochlorites (74), glacial acetic acid (60), and detergent (33). The admission rates were 60% for glacial acetic acid, 58.3% ethylene glycol, and 30.4% other alkali agents. In the intentional ingestion group, the most common chemicals upon admission were hypochlorites (242), glacial acetic acid (79), ethylene glycol (42), and detergent (41). The admission rates were 91.9% for glacial acetic acid, 87.5% ethylene glycol, 85.7% potassium cyanide, and 81.4% hydrochloric acid. In total, 79 deaths (10 unintentional ingestions, 69 intentional ingestion) were reported, and glacial acetic acid had an odds ratio of 9.299 for mortality. We compared the intentional and unintentional ingestion groups, and analyzed the factors affecting hospital admission and mortality in each group. The types and clinical outcomes of chemical ingestion varied depending on the purpose of chemical ingestion. The findings are considered beneficial in establishing treatment policies for patients ingesting chemicals.


Visible-light-induced electron transport from small to large nanoparticles in bimodal gold nanoparticle-loaded titanium(IV) oxide.
A key to realizing the sustainable society is to develop highly active photocatalysts for selective organic synthesis effectively using sunlight as the energy source. Recently, metal-oxide-supported gold nanoparticles (NPs) have emerged as a new type of visible-light photocatalysts driven by the excitation of localized surface plasmon resonance of Au NPs. Here we show that visible-light irradiation (λ>430 nm) of TiO2 -supported Au NPs with a bimodal size distribution (BM-Au/TiO2 ) gives rise to the long-range (>40 nm) electron transport from about 14 small (ca. 2 nm) Au NPs to one large (ca. 9 nm) Au NP through the conduction band of TiO2 . As a result of the enhancement of charge separation, BM-Au/TiO2 exhibits a high level of visible-light activity for the one-step synthesis of azobenzenes from nitrobenzenes at 25 °C with a yield greater than 95 % and a selectivity greater than 99 %, whereas unimodal Au/TiO2 (UM-Au/TiO2 ) is photocatalytically inactive.