Surgical prevention and management of vascular complications of kidney transplantation.
The main surgical changes in kidney procurement, preparation, and transplantation procedures occurred 20 years ago and were undertaken despite the inability to design randomized studies. The objective was to assess the evolution of vascular complications after kidney transplantation in a setting of surgical preventive measures in a historical series. A monocentric series of 3129 consecutive kidney transplantations performed over 3 decades was reviewed. The occurrence of arterial or venous thromboses, stenoses, and aneurysms was analyzed in relation with kidney procurement, preparation, and transplantation techniques. Vascular complications occurred in 13.5% of the recipients with a mean 3-year decrease in kidney graft function. The transplantation of a right kidney without renal vein extension, multiple renal arteries, ex vivo vascular repairs, and end-to-end arterial anastomoses were the unfavorable surgical vascular factors. It was possible to manage Transplant Renal Artery Stenosis (TRAS) nonsurgically in 80% of the cases. The prevention of vascular complications begins from the time of organ procurement by skilled surgeons. The aims of organ preparation are to evaluate the vascular risk, select the organs, and to simplify the anatomical constraints of vascular implantations. The three surgical steps of kidney transplantation are determinant in postoperative vascular complications and the duration of graft function.


LncRNA HULC promotes non-small cell lung cancer cell proliferation and inhibits the apoptosis by up-regulating sphingosine kinase 1 (SPHK1) and its downstream PI3K/Akt pathway.
LncRNA HULC has been proved to have important functions in the pathogenesis of several types of cancers. While its involvement in non-small cell lung cancer (NSCLC), which is one of the most common malignancies, still hasn't been reported to date. Therefore, we aimed to investigate the role of HULC in NSCLC and to explore the possible mechanisms. Tumor tissues and adjacent healthy tissues were collected from NSCLC patients, and blood samples were collected from both NSCLC patients and healthy controls. Expression of HULU in those tissues was detected by qRT-PCR. All patients were followed up for 5 years. Diagnostic and prognostic values of serum HULU for NSCLC were investigated by ROC curve analysis and survival curve analysis, respectively. HULC overexpression NSCLC cell lines were established and its effects on cell proliferation as well as apoptosis were investigated by CCK-8 assay and MTT assay, respectively. Effects of HULC overexpression on sphingosine kinase 1 (SPHK1) and its downstream PI3K/Akt pathway were investigated by Western blot. HULC expression level was increased in tumor tissues compared with adjacent healthy tissues in most patients. Serum level of HULC was higher in cancer patients than that in healthy control. Serum level of HULC was increased with the increased stage of primary tumor (T stage). Serum HULC can be used to accurately predict NSCLC and its prognosis. HULC overexpression promoted tumor cell proliferation, but inhibited cell apoptosis. HULC overexpression also increased expression level of SPHK1 and phosphorylation level of Akt in NSCLC cell, but showed on significant effects on Akt expression. Treatment with SPHK1 inhibitor and Akt reduced the effects of HULC overexpression on proliferation and apoptosis of NSCLC cells. But the treatment showed no significant effects on HULC expression. SPHK1 inhibitor treatment inhibited phosphorylation of Akt, while Akt inhibitor treatment showed no significant effects on SPHK1 expression. LncRNA HULC overexpression can promote NSCLC cell proliferation and inhibit cell apoptosis by up-regulating sphingosine kinase 1 (SPHK1) and further induce the activation of its downstream PI3K/Akt pathway.


Sample seal-and-drop device and methodology for high temperature oxide melt solution calorimetric measurements of PuO2.
Thermodynamic properties of refractory materials, such as standard enthalpy of formation, heat content, and enthalpy of reaction, can be measured by high temperature calorimetry. In such experiments, a small sample pellet is dropped from room temperature into a calorimeter operating at high temperature (often 700 °C) with or without a molten salt solvent present in an inert crucible in the calorimeter chamber. However, for hazardous (radioactive, toxic, etc.) and/or air-sensitive (hygroscopic, sensitive to oxygen, pyrophoric, etc.) samples, it is necessary to utilize a sealed device to encapsulate and isolate the samples, crucibles, and solvent under a controlled atmosphere in order to prevent the materials from reactions and/or protect the personnel from hazardous exposure during the calorimetric experiments. We have developed a sample seal-and-drop device (calorimetric dropper) that can be readily installed onto the dropping tube of a calorimeter such as the Setaram AlexSYS Calvet-type high temperature calorimeter to fulfill two functions: (i) load hazardous or air-sensitive samples in an air-tight, sealed container and (ii) drop the samples into the calorimeter chamber using an "off-then-on" mechanism. As a case study, we used the calorimetric dropper for measurements of the enthalpy of drop solution of PuO2 in molten sodium molybdate (3Na2O·4MoO3) solvent at 700 °C. The obtained enthalpy of -52.21 ± 3.68 kJ/mol is consistent with the energetic systematics of other actinide oxides (UO2, ThO2, and NpO2). This capability has thus laid the foundation for thermodynamic studies of other Pu-bearing phases in the future.


Generation and Characterization of a Nanobody Against SARS-CoV.
The sudden emergence of severe acute respiratory syndrome coronavirus (SARS-CoV) has caused global panic in 2003, and the risk of SARS-CoV outbreak still exists. However, no specific antiviral drug or vaccine is available; thus, the development of therapeutic antibodies against SARS-CoV is needed. In this study, a nanobody phage-displayed library was constructed from peripheral blood mononuclear cells of alpacas immunized with the recombinant receptor-binding domain (RBD) of SARS-CoV. Four positive clones were selected after four rounds of bio-panning and subjected to recombinant expression in E. coli. Further biological identification demonstrated that one of the nanobodies, S14, showed high affinity to SARS-CoV RBD and potent neutralization activity at the picomole level against SARS-CoV pseudovirus. A competitive inhibition assay showed that S14 blocked the binding of SARS-CoV RBD to either soluble or cell-expressed angiotensin-converting enzyme 2 (ACE2). In summary, we developed a novel nanobody targeting SARS-CoV RBD, which might be useful for the development of therapeutics against SARS.


Providing Care for Children in Immigrant Families.
Children in immigrant families (CIF), who represent 1 in 4 children in the United States, represent a growing and ever more diverse US demographic that pediatric medical providers nationwide will increasingly encounter in clinical care. Immigrant children are those born outside the United States to non-US citizen parents, and CIF are defined as those who are either foreign born or have at least 1 parent who is foreign born. Some families immigrate for economic or educational reasons, and others come fleeing persecution and seeking safe haven. Some US-born children with a foreign-born parent may share vulnerabilities with children who themselves are foreign born, particularly regarding access to care and other social determinants of health. Therefore, the larger umbrella term of CIF is used in this statement. CIF, like all children, have diverse experiences that interact with their biopsychosocial development. CIF may face inequities that can threaten their health and well-being, and CIF also offer strengths and embody resilience that can surpass challenges experienced before and during integration. This policy statement describes the evolving population of CIF in the United States, briefly introduces core competencies to enhance care within a framework of cultural humility and safety, and discusses barriers and opportunities at the practice and systems levels. Practice-level recommendations describe how pediatricians can promote health equity for CIF through careful attention to core competencies in clinical care, thoughtful community engagement, and system-level support. Advocacy and policy recommendations offer ways pediatricians can advocate for policies that promote health equity for CIF.


mTORC1 signaling controls mammalian skeletal growth through stimulation of protein synthesis.
Much of the mammalian skeleton is derived from a cartilage template that undergoes rapid growth during embryogenesis, but the molecular mechanism of growth regulation is not well understood. Signaling by mammalian target of rapamycin complex 1 (mTORC1) is an evolutionarily conserved mechanism that controls cellular growth. Here we report that mTORC1 signaling is activated during limb cartilage development in the mouse embryo. Disruption of mTORC1 signaling through deletion of either mTOR or the associated protein Raptor greatly diminishes embryonic skeletal growth associated with severe delays in chondrocyte hypertrophy and bone formation. The growth reduction of cartilage is not due to changes in chondrocyte proliferation or survival, but is caused by a reduction in cell size and in the amount of cartilage matrix. Metabolic labeling reveals a notable deficit in the rate of protein synthesis in Raptor-deficient chondrocytes. Thus, mTORC1 signaling controls limb skeletal growth through stimulation of protein synthesis in chondrocytes.


Adult learning and video training in health care.
Research has shown that the use of video in health care training can be used effectively, but only in selected situations. The author has given some guidelines on the use of video as a teaching media by broadly grouping its effective use in three main areas. Classification of the groups is given.


[Depressive symptoms diagnosis and treatment of schizophrenia and schizophrenia spectrum disorders in Russian clinical practice].
A cohort study of depression in schizophrenia and schizophrenia spectrum disorders was conducted in four Russian regional mental health services with the use of simultaneous one-day census on October 20, 2011. The total number of patients in this study was 1269: 753 patients were receiving outpatient treatment, 143 patients were served in day clinics and 373 patients were in-patients in psychiatric hospitals. Depressive symptoms were present in 6.0% of outpatients, 50.3% of day clinic patients, and 36.5% of inpatients. The average duration of current depressive symptoms in weeks was 10.0, 8.1, 9.0, respectively. The periods of depressive symptoms during the last 2 years were revealed in 30.8% of outpatients, 67.8% of day clinic patients, and 42.1% of inpatients with the average duration of each such period in weeks 10.9, 12.2, 8.3, respectively. In outpatient settings, 28.8% of patients received antidepressants in addition to antipsychotic therapy. The proportion of inpatients receiving this treatment was 42.1%, and two thirds of patients received antidepressants in day clinics.


Risk Factors and Time to Recurrent Ipsilateral and Contralateral Patellar Dislocations.
Previous studies have reported variable rates of recurrent lateral patellar instability mainly because of limited cohort sizes. In addition, there is currently a lack of information on contralateral patellar instability. To evaluate the rate of recurrent ipsilateral patellar dislocations and contralateral patellar dislocations after a first-time lateral patellar dislocation. Additionally, risk factors associated with recurrent dislocations (ipsilateral or contralateral) and time to recurrence were investigated. Cohort study; Level of evidence, 3. This population-based study included 584 patients with a first-time lateral patellar dislocation occurring between 1990 and 2010. A retrospective review was conducted to gather information about the injury, subsequent dislocations (ipsilateral or contralateral), and structural characteristics including trochlear dysplasia, patella alta, and tibial tubercle to trochlear groove (TT-TG) distance. Risk factors were assessed to delineate associations with subsequent dislocations and time to recurrence. At a mean follow-up of 12.4 years, 173 patients had ipsilateral recurrence, and 25 patients had a subsequent contralateral dislocation. At 20 years, the cumulative incidence of ipsilateral recurrence was 36.0%, while the cumulative incidence of contralateral dislocations was 5.4%. Trochlear dysplasia (odds ratio [OR], 18.1), patella alta (OR, 10.4), age <18 years at the time of the first dislocation (OR, 2.4), elevated TT-TG distance (OR, 2.1), and female sex (OR, 1.5) were associated with recurrent ipsilateral dislocations. Time to recurrence was significantly decreased in patients with trochlear dysplasia (23.0 months earlier time to recurrence; P < .001), elevated TT-TG distance (18.5 months; P < .001), patella alta (16.4 months; P = .001), and age <18 years at the time of the first dislocation (15.4 months; P < .001). Risk factors for subsequent contralateral dislocations included patella alta and trochlear dysplasia. At 20 years after a first-time lateral patellar dislocation, the cumulative incidence of recurrent ipsilateral patellar dislocations was 36.0%, compared with 5.4% for contralateral dislocations. Trochlear dysplasia, elevated TT-TG distance, patella alta, age <18 years at the time of the first dislocation, and female sex were associated with ipsilateral recurrence. Trochlear dysplasia, elevated TT-TG distance, patella alta, and age <18 years at the time of the first dislocation were predictive of a statistically significant decrease in time to recurrence.


Effects of the extract of Spirostachys venenifera Pax on the immune response in mice.
The effects of the methanol extract of Spirostachys venenifera Pax (SVP) on the immune response in mice was investigated. Keyhole limpet haemocyanin (KLH) was the antigen. An enzyme linked immunosorbent assay (ELISA) was used to monitor serum antibody levels in mice exposed to various doses of SVP at a dosage of 0.1 mg/Kg. In mice in vivo, SVP caused a significant suppression of the immune response compared to control mice that received no SVP. A haemolytic plaque assay was used to monitor the effect of SVP on antibody producing B lymphocytes from the peritoneum and from the spleen in vitro. SVP caused a reduction in the number of plaque forming cells (PFC) at 0.1 ug/ml with peritoneal B cells. There was further reduction of PFC with higher doses of SVP. With the spleen B cells, clear reduction in PFC was seen only with the higher doses of SVP, from 1 ug/ml upwards to 100 ug/ml. The prospect of developing a new immunosuppressive agent from SVP is a welcome possibility that can add to the newer immunosuppressive agents with greater efficacy and less side effects in comparison to the traditional ones.


Risk Factors for Opioid-Use Disorder and Overdose.
Opioid analgesics are recognized as a legitimate medical therapy for selected patients with severe chronic pain that does not respond to other therapies. However, opioids are associated with risks for patients and society that include misuse, abuse, diversion, addiction, and overdose deaths. Therapeutic success depends on proper candidate selection, assessment before administering opioid therapy, and close monitoring throughout the course of treatment. Risk assessment and prevention include knowledge of patient factors that may contribute to misuse, abuse, addiction, suicide, and respiratory depression. Risk factors for opioid misuse or addiction include past or current substance abuse, untreated psychiatric disorders, younger age, and social or family environments that encourage misuse. Opioid mortality prevalence is higher in people who are middle aged and have substance abuse and psychiatric comorbidities. Suicides are probably undercounted or frequently misclassified in reports of opioid-related poisoning deaths. Greater understanding and better assessment are needed of the risk associated with suicide risk in patients with pain. Clinical tools and an evolving evidence base are available to assist clinicians with identifying patients whose risk factors put them at risk for adverse outcomes with opioids.


Role of Mg2+ Ions in DNA Hydrolysis by EcoRV, Studied by the 3D-Reference Interaction Site Model and Molecular Dynamics.
The role of Mg2+ ions during precursor formation in DNA hydrolysis by the homodimeric restriction enzyme EcoRV was elucidated based on the 3D-reference interaction site model (RISM) theory and the molecular dynamics (MD) simulation. From an analysis of the spatial distribution of Mg2+ in an active site using 3D-RISM, we identified a new position for Mg2+ in the X-ray EcoRV-DNA complex structure ( 1rvb ). We refer to the position as site IV†. Site IV† is almost at the same position as that of a Ca2+ ion in the superimposed X-ray crystallographic active-site structure of the PvuII-DNA complex ( 1f0o ). 3D-RISM was also used to locate the position of water molecules, including the water nucleophile at the active site. MD simulations were carried out with the initial structure having two Mg2+ ions at site IV† and at site I*, experimentally identified by Horton et al., to find a stable complex structure in which the DNA fragment was rearranged to orient the scissile bond direction toward the water nucleophile. The equilibrium active-site structure of the EcoRV-DNA complex obtained from the MD simulation was similar to the superimposed X-ray crystallographic structure of the BamHI-DNA complex ( 2bam ). In the active-site structure, two metal ions have almost the same position (≤1.0 Å) as that of 2bam , and the scissile phosphate is twisted to orient the scissile bond toward the water nucleophile, as is the case in 2bam . We propose the equilibrium active-site structure obtained in this study as a precursor for the hydrolysis reaction of EcoRV.


Leishmania lipophosphoglycan (LPG) activates NK cells through toll-like receptor-2.
Toll-like receptors (TLRs) mediate the cellular response to conserved molecular patterns shared by microorganisms. We report that TLR-2 on human NK cells is upregulated and stimulated by Leishmania major lipophosphoglycan (LPG), a phosphoglycan belonging to a family of unique Leishmania glycoconjugates. We found that purified L. major LPG upregulates both mRNA and the membrane expression of TLR-2 in NK cells. Additionally, IFN-gamma and TNF-alpha production and nuclear translocation of NF-kappaB was enhanced. The activation effect was more intense with LPG purified from infectious metacyclic parasites than from noninfectious procyclic Leishmania. Since the difference between the molecules derived from these two stages of the parasite growth cycle lies exclusively in the number of phosphosaccharide repeat domains and in the composition of glycan side chains that branch off these domains, we propose that TLR-2 possibly distinguishes between phosphorylated glycan repeats on LPG molecules. The effect of LPG on cytokine production and on membrane expression of TLR-2 could be blocked with F(ab')2 fragments of the mAb against LPG (WIC 79.3). Confocal microscopy demonstrated the co-localization of LPG and TLR-2 on the NK cell membrane. Binding of LPG to TLR-2 in NK cells was demonstrated by immunoprecipitations done with anti-TLR-2 and anti-LPG mAb followed by immunoblotting with anti-LPG and anti-TLR-2, respectively. Both antibodies recognized the immune complexes. These results suggest that NK cells are capable of recognition of, and activation by, Leishmania LPG through TLR-2, enabling them to participate autonomously in the innate immune system and thereby increasing the effective destruction of the parasite.


[Mortality and biologic aging in the human].
Summarizing it can be said that the statistical approach proposed in the present paper constitutes a starting point for one of the possible alternatives regarding quantitative assessments of the relationship between features and the vitality level and the mortality of man. The other alternatives and modifications that permit to increase the information content of the calculations should also be used to obtain an idea of the overall system.


Alteration of the selectivity of DNA cleavage by a deglycobleomycin analogue containing a trithiazole moiety.
The bleomycin (BLM) group of antitumor antibiotics effects DNA cleavage in a sequence-selective manner. Previous studies have indicated that the metal-binding and bithiazole moieties of BLM are both involved in the binding of BLM to DNA. The metal-binding domain is normally the predominant structural element in determining the sequence selectivity of DNA binding, but it has been shown that replacement of the bithiazole moiety with a strong DNA binder can alter the sequence selectivity of DNA binding and cleavage. To further explore the mechanism by which BLM and DNA interact, a trithiazole-containing deglycoBLM analogue was synthesized and tested for its ability to relax supercoiled DNA and cleave linear duplex DNA in a sequence-selective fashion. Also studied was cleavage of a novel RNA substrate. Solid-phase synthesis of the trithiazole deglycoBLM A(5) analogue was achieved using a TentaGel resin containing a Dde linker and elaborated from five key intermediates. The ability of the resulting BLM analogue to relax supercoiled DNA was largely unaffected by introduction of the additional thiazole moiety. Remarkably, while no new sites of DNA cleavage were observed for this analogue, there was a strong preference for cleavage at two 5'-GT-3' sites when a 5'-(32)P end-labeled DNA duplex was used as a substrate. The alteration of sequence selectivity of cleavage was accompanied by some decrease in the potency of DNA cleavage, albeit without a dramatic diminution. In common with BLM, the trithiazole analogue of deglycoBLM A(5) effected both hydrolytic cleavage of RNA in the absence of added metal ion and oxidative cleavage in the presence of Fe(2+) and O(2). In comparison with BLM A(5), the relative efficiencies of hydrolytic cleavage at individual sites were altered.


Diurnal variation in stature: is stretching the answer?
To investigate the extent and timing of diurnal variation in stature and to examine the effectiveness of the stretched technique in reducing the loss in height. A Southampton school. Fifty three children, divided into two groups, were measured by two independent auxologists using a Leicester height measure. Each child was measured four times, at 0900, 1100, 1300, and 1500, using both an unstretched and a stretched technique. Height loss after each of the three time intervals for both unstretched and stretched modes. There was a clear decrease in stature during the morning, but no further loss occurred after the subjects had been up for around six hours. The mean height losses for the unstretched (stretched) modes were 0.31 cm (0.34 cm) and 0.20 cm (0.23 cm) for the periods 0900 to 1100 and 1100 to 1300, respectively, but only 0.045 cm (-0.019 cm) from 1300 to 1500. Stretching did not reduce the effects of diurnal variation, but significantly affected the recorded height by an average of 0.28 cm. There was no significant difference in reproducibility using either technique (SD 0.30 cm stretched v 0.31 cm unstretched). Diurnal variation in stature may substantially affect the reliability of height data and careful consideration should be given to the timing of repeat measurements. As most height loss occurs in the morning, afternoon clinic appointments would be preferable. The standard stretched technique does not appear to reduce diurnal variation, nor does it affect precision. Measurements made using an unstretched method are recommended to avoid interobserver differences, known to occur where different observers are used.


Prevalence and correlates of hunger among primary and secondary school children in Malawi: results from the 2009 Global School-based Health Survey.
Education is important in improving economies and creating literate, self-reliant and healthy societies. However, hunger is a barrier to basic education in Malawi. Hunger is also associated with a number of health risk behaviours, such as bullying, suicide ideation and unhygienic behaviours that may jeopardize the future of children. There are, however, limited data on the prevalence and associated factors of hunger among school children in Malawi. The study used data from the Malawi Global School-Based Health Survey conducted in 2009 to estimate the prevalence of self-reported hunger within the last 30 days among primary and secondary school age group. It also assessed the association between self-reported hunger and some selected list of independent variables using frequency distribution, chi-squared test and logistic regression. A total of 2359 students were available for analysis. The overall self-reported prevalence of hunger within the last 30 days was 12.5% (18.9% (172) in the rural and 8.3% (115) in urban areas; and 11.9%(123) for male and 12.5(148) for female children). In the final analysis, geographical location, eating fruits, having been bullied, suicide ideation, and washing hands with soap were significantly associated with hunger. Hunger in both primary and secondary school children in Malawi is a major social problem. The design of school feeding programmes aimed to reduce hunger should incorporate the factors identified as associated with hunger.


Management of hepatitis C virus infection in the Asia-Pacific region: an update.
The Asia-Pacific region has disparate hepatitis C virus (HCV) epidemiology, with prevalence ranging from 0·1% to 4·7%, and a unique genotype distribution. Genotype 1b dominates in east Asia, whereas in south Asia and southeast Asia genotype 3 dominates, and in Indochina (Vietnam, Cambodia, and Laos), genotype 6 is most common. Often, availability of all-oral direct-acting antivirals (DAAs) is delayed because of differing regulatory requirements. Ideally, for genotype 1 infections, sofosbuvir plus ledipasvir, sofosbuvir plus daclatasvir, or ombitasvir, paritaprevir, and ritonavir plus dasabuvir are suitable. Asunaprevir plus daclatasvir is appropriate for compensated genotype 1b HCV if baseline NS5A mutations are absent. For genotype 3 infections, sofosbuvir plus daclatasvir for 24 weeks or sofosbuvir, daclatasvir, and ribavirin for 12 weeks are the optimal oral therapies, particularly for patients with cirrhosis and those who are treatment experienced, whereas sofosbuvir, pegylated interferon, and ribavirin for 12 weeks is an alternative regimen. For genotype 6, sofosbuvir plus pegylated interferon and ribavirin, sofosbuvir plus ledipasvir, or sofosbuvir plus ribavirin for 12 weeks are all suitable. Pegylated interferon plus ribavirin has been replaced by sofosbuvir plus pegylated interferon and ribavirin, and all-oral therapies where available, but cost and affordability remain a major issue because of the absence of universal health coverage. Few patients have been treated because of multiple barriers to accessing care. HCV in the Asia-Pacific region is challenging because of the disparate epidemiology, poor access to all-oral therapy because of availability, cost, or regulatory licensing. Until these problems are addressed, the burden of disease is likely to remain high.


Breathalyser-Based eHealth Data Suggest That Self-Reporting of Abstinence Is a Poor Outcome Measure for Alcohol Use Disorder Clinical Trials.
To evaluate the efficacy and monitoring capabilities of a breathalyser-based eHealth system for patients with alcohol use disorder (AUD) and to investigate the quality and validity of timeline follow-back (TLFB) as outcome measure in clinical trials and treatment. Patients (n = 115) were recruited to clinical trials from a 12-step aftercare programme (12S-ABS) and from hospital care with abstinence (HC-ABS) or controlled drinking (HC-CDR) as goal and randomly divided into an eHealth and a control group. The effect of the eHealth system was analysed with TLFB-derived primary outcomes-change in number of abstinent days (AbsDay) and heavy drinking days (HDDs) compared to baseline-and phosphatidyl ethanol (PEth) measurements. Validity and quality of TLFB were evaluated by comparison with breath alcohol content (BrAC) and eHealth digital biomarkers (DBs): Addiction Monitoring Index (AMI) and Maximum Time Between Tests (MTBT). TLFB reports were compared to eHealth data regarding reported abstinence. The primary outcome (TLFB) showed no significant difference between eHealth and control groups, but PEth did show a significant difference especially at months 2 and 3. Self-reported daily abstinence suffered from severe quality issues: of the 28-day TLFB reports showing full abstinence eHealth data falsified 34% (BrAC measurements), 39% (MTBT), 54% (AMI) and 68% (BrAC/MTBT/AMI). 12S-ABS and HC-ABS patients showed severe under-reporting. No effect of the eHealth system was measured with TLFB, but a small positive effect was measured with PEth. The eHealth system revealed severe quality problems with TLFB, especially regarding abstinence-should measurement-based eHealth data replace TLFB as outcome measure for AUD?


Cerebral perfusion during major cardiac surgery in children.
Six children undergoing major cardiac surgery had extensive cerebral monitoring during cardiopulmonary bypass (CPB). The monitoring included continuous recording of arterial blood pressure (BP), central venous pressure (CVP), cerebral electrical activity by a cerebral function monitor (CFM), and middle cerebral artery (MCA) flow velocity by the transcranial pulsed Doppler (TCD) technique. Introduction of the precooled blood containing priming solution resulted in rapid fall in BP as well as MCA velocities in these children at the start of CPB. During steady-state CPB at 20 degrees C, MCA flow velocities were reduced in five of six children, range 45%-105% of pre-bypass value. These flow velocity values were recorded at cerebral perfusion pressures (CPP = BP - CVP) in the range of 14-26 mmHg. This reduced cerebral perfusion during steady-state CPB appears to be more than sufficient to meet the cerebral metabolic demands at the particular temperature. The reduced cerebral perfusion is in contrast to the enhanced perfusion found in adults during moderately hypothermic (28 degrees-30 degrees C) low-flow, low-pressure CPB previously reported. It was presumably due to the reduced temperature, reduced perfusion pressure, and less hemodilution. During periods of constant temperature, hematocrit, and partial pressure of carbon dioxide (PaCO2), MCA flow velocities varied passively with changes in CPP, demonstrating that cerebral autoregulation was not operative. Transcranial Doppler appears to be a suitable tool for investigating CPB techniques optimal with respect to cerebral circulation.


Biased guessing in a complete-identification visual-working-memory task: Further evidence for mixed-state models.
Research is reported that provides evidence for a significant role of mixed states and guessing processes in tasks of visual working memory (VWM). Subjects engaged in a complete-identification VWM task. The stimulus set consisted of 16 colors roughly equally spaced around a color circle. On each trial, a memory-set drawn from the colors was briefly presented, followed by a location probe. Subjects attempted to reproduce the color of the probed item by clicking on the appropriate response button of a discrete color wheel. The key manipulation was to vary payoffs for alternative correct responses across trials. Analysis of the resulting matrices of individual-subject identification-confusion data provided evidence for a systematic guessing process: On trials in which subjects had no memory for the probed stimulus, they guessed with high probability using the high-payoff response. Formal modeling corroborated this interpretation. Mixed-state models that assumed that performance involved a combination of memory-based responding and biased guessing yielded accurate and easy-to-interpret accounts of the identification data; by comparison, variable-resources (VR) models without a guessing state struggled to account for the data, including versions with bias parameters for the high-payoff response. The authors argue that the work adds to recent converging sources of evidence that point to a significant role of discrete, mixed states in VWM. The authors also suggest directions for development of extended VR models with sophisticated knowledge-rich decision rules for the complete-identification task. (PsycINFO Database Record


Association or Causation? Exploring the Oral Microbiome and Cancer Links.
Several epidemiological investigations have found associations between poor oral health and different types of cancer, including colorectal, lung, pancreatic, and oral malignancies. The oral health parameters underlying these relationships include deficient oral hygiene, gingival bleeding, and bone and tooth loss. These parameters are related to periodontal diseases, which are directly and indirectly mediated by oral bacteria. Given the increased accessibility of microbial sequencing platforms, many recent studies have investigated the link between the oral microbiome and these cancers. Overall, it seems that oral dysbiotic states can contribute to tumorigenesis in the oral cavity as well as in distant body sites. Further, it appears that certain oral bacterial species can contribute to carcinogenesis, in particular, Fusobacterium nucleatum and Porphyromonas gingivalis, based on results from epidemiological as well as mechanistic studies. Yet, the strength of the findings from these investigations is hampered by the heterogeneity of the methods used to measure oral diseases, the treatment of confounding factors, the study design, the platforms employed for microbial analysis, and types of samples analyzed. Despite these limitations, there is an overall indication that the presence of oral dysbiosis that leads to oral diseases may directly and/or indirectly contribute to carcinogenesis. Proper methodological standardized approaches should be implemented in future epidemiological studies as well as in the mechanistic investigations carried out to explore these results.


Premise Plumbing Pipe Materials and In-Building Disinfectants Shape the Potential for Proliferation of Pathogens and Antibiotic Resistance Genes.
In-building disinfectants are commonly applied to control the growth of pathogens in plumbing, particularly in facilities such as hospitals that house vulnerable populations. However, their application has not been well optimized, especially with respect to interactive effects with pipe materials and potential unintended effects, such as enrichment of antibiotic resistance genes (ARGs) across the microbial community. Here, we used triplicate convectively mixed pipe reactors consisting of three pipe materials (PVC, copper, and iron) for replicated simulation of the distal reaches of premise plumbing and evaluated the effects of incrementally increased doses of chlorine, chloramine, chlorine dioxide, and copper-silver disinfectants. We used shotgun metagenomic sequencing to characterize the resulting succession of the corresponding microbiomes over the course of 37 weeks. We found that both disinfectants and pipe material affected ARG and microbial community taxonomic composition both independently and interactively. Water quality and total bacterial numbers were not found to be predictive of pathogenic species markers. One result of particular concern was the tendency of disinfectants, especially monochloramine, to enrich ARGs. Metagenome assembly indicated that many ARGs were enriched specifically among the pathogenic species. Functional gene analysis was indicative of a response of the microbes to oxidative stress, which is known to co/cross-select for antibiotic resistance. These findings emphasize the need for a holistic evaluation of pathogen control strategies for plumbing.


A comparative analysis of restriction enzyme digests of the DNA of herpes simplex virus isolated from genital and facial lesions.
The restriction endonuclease (RE) cleavage patterns of the DNA of herpes simplex virus (HSV) genital isolates from two geographically distinct groups and from one group of facial isolates were examined. Eleven of 21 genital isolates from females, 1 of 27 genital isolates from males and all 17 of the facial isolates were HSV type 1 (HSV-1). The groups of isolates of the same serotype could not be distinguished by significant differences in the frequency of variable restriction endonuclease sites or molecular weight of variable length fragments. Simultaneous consideration of two or more variable sites has disclosed some which are apparently correlated in the HSV-1 isolates, and which could provide a useful marker for phylogenetic relationships. However, most pairs showed no correlation, while certain sites appeared more closely correlated in the genital than in the facial isolates. All 39 HSV-2 isolates could be distinguished from each other on the basis of a combination of variable RE sites and variable length fragments. Bg/II RE sites appeared less variable than other RE sites.


Germline mutations of the DNA repair pathways in uterine serous carcinoma.
Treatment options are limited for patients with uterine serous carcinoma (USC). Knowledge of USC's somatic mutation landscape is rapidly increasing, but its role in hereditary cancers remains unclear. We aim to evaluate the frequency and characteristics of germline mutations in genes commonly implicated in carcinogenesis, including those within homologous recombination (HR) and mismatch repair (MMR) pathways in patients with pure USC. By using targeted capture exome sequencing, 43 genes were analyzed in a cohort of 7 consecutive patients with paired tumor and non-tumor USC samples in our institutional tumor repository. Mutations predicted to have damaging effects on protein function are validated by Sanger Sequencing. We found 21 germline mutations in 11 genes in our USC cohort. Five patients harbored 7 germline mutations (33.3%) within genes involved in the HR pathway, RAD51D being the most common. Four patients had 9 (42.8%) germline mutations in hereditary colon cancer genes, most commonly MLH. All patients (42.7%) who are platinum-sensitive had HR germline mutations (RAD50, NBN, ATM). Patients with HER2 overexpression (2/7, 28.6%) had germline HR mutations and were platinum-sensitive. Three patients in our cohort reported a personal history of breast cancer, one with HR germline mutation, and 2 in patients with germline mutations in HCC genes. In addition, 5 out of 7 patients had germline mutations in genes associated with growth factor signaling pathway. A significant proportion of our cohort harbor germline mutations in DNA repair genes. This may be associated with the high rate of breast cancer in our patients and their family, and suggests a targeted cohort for genetic counseling. If validated in a larger cohort, our findings may allow clinicians to expand therapeutic options to include targeted therapies and inclusion of USC patient in preventative and genetic counseling.


GRIPT: a novel case-control analysis method for Mendelian disease gene discovery.
Despite rapid progress of next-generation sequencing (NGS) technologies, the disease-causing genes underpinning about half of all Mendelian diseases remain elusive. One main challenge is the high genetic heterogeneity of Mendelian diseases in which similar phenotypes are caused by different genes and each gene only accounts for a small proportion of the patients. To overcome this gap, we developed a novel method, the Gene Ranking, Identification and Prediction Tool (GRIPT), for performing case-control analysis of NGS data. Analyses of simulated and real datasets show that GRIPT is well-powered for disease gene discovery, especially for diseases with high locus heterogeneity.


OrthoSNAP: A tree splitting and pruning algorithm for retrieving single-copy orthologs from gene family trees.
Molecular evolution studies, such as phylogenomic studies and genome-wide surveys of selection, often rely on gene families of single-copy orthologs (SC-OGs). Large gene families with multiple homologs in 1 or more species-a phenomenon observed among several important families of genes such as transporters and transcription factors-are often ignored because identifying and retrieving SC-OGs nested within them is challenging. To address this issue and increase the number of markers used in molecular evolution studies, we developed OrthoSNAP, a software that uses a phylogenetic framework to simultaneously split gene families into SC-OGs and prune species-specific inparalogs. We term SC-OGs identified by OrthoSNAP as SNAP-OGs because they are identified using a splitting and pruning procedure analogous to snapping branches on a tree. From 415,129 orthologous groups of genes inferred across 7 eukaryotic phylogenomic datasets, we identified 9,821 SC-OGs; using OrthoSNAP on the remaining 405,308 orthologous groups of genes, we identified an additional 10,704 SNAP-OGs. Comparison of SNAP-OGs and SC-OGs revealed that their phylogenetic information content was similar, even in complex datasets that contain a whole-genome duplication, complex patterns of duplication and loss, transcriptome data where each gene typically has multiple transcripts, and contentious branches in the tree of life. OrthoSNAP is useful for increasing the number of markers used in molecular evolution data matrices, a critical step for robustly inferring and exploring the tree of life.


Expression of urea transporters is affected by dietary nitrogen restriction in goat kidney.
Ruminants are known to be able to very effectively recycle urinary urea and reuse it as a source of N for ruminal microbes. It is presumed that urea recycling is accomplished by specialized urea transporters (UT) which are localized in the kidney. This could be especially important in times of increased N requirement, such as during growth or during reduced dietary N intake. The aim of our study was to characterize and to localize UT in the goat (capra hircus) kidney and to investigate its response to reduced dietary N intake in growing goats. Therefore, 12 growing, male goats were fed either a diet containing high (17% CP in complete diet) or low (9% CP in complete diet) N content for 6 wk. After harvesting, blood and kidney samples were taken and analyzed. The mRNA of the different UT isoforms, UT-A1, UT-A2 and UT-B, were detected semiquantitatively in renal tissue by Northern blot analysis. For UT-A2 and UT-B, no statistically significant effect of dietary N restriction on renal mRNA expression could be detected (UT-A2: P = 0.26, UT-B: P = 0.07). However, renal mRNA abundance of UT-A1 significantly increased in the kidney of low-N-fed goats (P = 0.01). Furthermore, protein amounts of UT-B were verified by western blotting; and the localization of UT-A2 and UT-B protein was demonstrated by immunohistochemistry. No significant differences in protein amounts of UT-B could be observed comparing the 2 feeding groups (P = 0.78). The UT-B was localized in renal medulla and papilla, whereas UT-A2 was only found in renal medulla. In addition, comparison of UT-A and UT-BAA sequences of monogastric animals and ruminants showed a high degree of homology, indicating a similar function of the transporters among these species. In summary, we conclude that in ruminants, urea reabsorption in the kidney is most likely increased in response to a low-N diet via an upregulation of UT-A1 mRNA expression. Hypothetically, the reabsorbed urea can then be returned to the rumen via the bloodstream and thus be reused as a source of N for protein synthesis of ruminal microbial community.


Fasting enhances glycogen synthase activation in hepatocytes from insulin-resistant genetically obese (fa/fa) rats.
Glycogen synthase activation and phosphorylase inactivation by glucose were studied in hepatocytes isolated from fed or overnight-fasted lean or genetically obese (fa/fa) rats. In cells from fed animals, both the time course and dose-response to glucose of synthase activation were the same in both groups, despite higher levels of phosphorylase a in hepatocytes from obese animals. In contrast, in cells from fasted obese animals synthase activation with or without glucose was enhanced severalfold over that of lean controls, despite similar levels of phosphorylase a and of total (a + b) synthase activities. In both nutritional conditions glucose 6-phosphate concentrations were 2-3-fold higher in obese-rat hepatocytes than in lean-rat cells. In addition, synthase activation was transient in the fasted lean group, but was sustained in obese-rat hepatocytes. The rate of synthase activation was, however, comparable in lean- and obese-rat liver Sephadex G-25 filtrates, irrespective of the nutritional state of the donor rats. It is concluded that enhanced synthase activation in hepatocytes from starved obese rats might be due to an unbalanced synthase interconversion brought about by elevated glucose 6-phosphate concentrations and impaired kinase [van de Werve & Massillon (1990) Biochem. J. 269, 795-799], rather than to an intrinsic change in synthase phosphatase.


Cannabis poisoning under the age of 3 years.
The increased use of cannabis among adults and the general perception that cannabis is harmless has been accompanied by a rise in the number of acute poisoning cases among children. We present a retrospective, descriptive study of patients aged 3 years or younger who were treated for cannabis poisoning in a pediatric emergency department between 2012 and 2017. Epidemiologic and clinical data as well as toxicologic findings are reported. We found 10 cases of cannabis poisoning in our records. Nine of the patients had diminished consciousness. Only 2 families initially reported exposure to cannabis when interviewed. Repeated exposure was demonstrated by analysis of hair from 2 patients. We conclude that cannabis poisoning should be included in the differential diagnosis of young children with acute neurologic symptoms of unknown origin. When cannabis is detected in urine samples collected in the pediatric emergency department, more specific diagnostic techniques should be used to assess the possibility of repeated exposure.


10.5 T MRI static field effects on human cognitive, vestibular, and physiological function.
To perform a pilot study to quantitatively assess cognitive, vestibular, and physiological function during and after exposure to a magnetic resonance imaging (MRI) system with a static field strength of 10.5 Tesla at multiple time scales. A total of 29 subjects were exposed to a 10.5 T MRI field and underwent vestibular, cognitive, and physiological testing before, during, and after exposure; for 26 subjects, testing and exposure were repeated within 2-4 weeks of the first visit. Subjects also reported sensory perceptions after each exposure. Comparisons were made between short and long term time points in the study with respect to the parameters measured in the study; short term comparison included pre-vs-isocenter and pre-vs-post (1-24 h), while long term compared pre-exposures 2-4 weeks apart. Of the 79 comparisons, 73 parameters were unchanged or had small improvements after magnet exposure. The exceptions to this included lower scores on short term (i.e. same day) executive function testing, greater isocenter spontaneous eye movement during visit 1 (relative to pre-exposure), increased number of abnormalities on videonystagmography visit 2 versus visit 1 and a mix of small increases (short term visit 2) and decreases (short term visit 1) in blood pressure. In addition, more subjects reported metallic taste at 10.5 T in comparison to similar data obtained in previous studies at 7 T and 9.4 T. Initial results of 10.5 T static field exposure indicate that 1) cognitive performance is not compromised at isocenter, 2) subjects experience increased eye movement at isocenter, and 3) subjects experience small changes in vital signs but no field-induced increase in blood pressure. While small but significant differences were found in some comparisons, none were identified as compromising subject safety. A modified testing protocol informed by these results was devised with the goal of permitting increased enrollment while providing continued monitoring to evaluate field effects.


US County-Level Variation in Preterm Birth Rates, 2007-2019.
Preterm birth is a leading cause of preventable neonatal morbidity and mortality. Preterm birth rates at the national level may mask important geographic variation in rates and trends at the county level. To estimate age-standardized preterm birth rates by US county from 2007 to 2019. This serial cross-sectional study used data from the National Center for Health Statistics composed of all live births in the US between 2007 and 2019. Data analyses were performed between March 22, 2022, and September 29, 2022. Age-standardized preterm birth (<37 weeks' gestation) and secondarily early preterm birth (<34 weeks' gestation) rates by county and year calculated with a validated small area estimation model (hierarchical bayesian spatiotemporal model) and percent change in preterm birth rates using log-linear regression models. Between 2007 and 2019, there were 51 044 482 live births in 2383 counties. In 2007, the national age-standardized preterm birth rate was 12.6 (95% CI, 12.6-12.7) per 100 live births. Preterm birth rates varied significantly among counties, with an absolute difference between the 90th and 10th percentile counties of 6.4 (95% CI, 6.2-6.7). The gap between the highest and lowest counties for preterm births was 20.7 per 100 live births in 2007. Several counties in the Southeast consistently had the highest preterm birth rates compared with counties in California and New England, which had the lowest preterm birth rates. Although there was no statistically significant change in preterm birth rates between 2007 and 2019 at the national level (percent change, -5.0%; 95% CI, -10.7% to 0.9%), increases occurred in 15.4% (95% CI, 14.1%-16.9%) of counties. The absolute and relative geographic inequalities were similar across all maternal age groups. Higher quartile of the Social Vulnerability Index was associated with higher preterm birth rates (quartile 4 vs quartile 1 risk ratio, 1.34; 95% CI, 1.31-1.36), which persisted across the study period. Similar patterns were observed for early preterm birth rates. In this serial cross-sectional study of county-level preterm and early preterm birth rates, substantial geographic disparities were observed, which were associated with place-based social disadvantage. Stability in aggregated rates of preterm birth at the national level masked increases in nearly 1 in 6 counties between 2007 and 2019.


The role of adipose tissue in mediating the beneficial effects of dietary fish oil.
Fish oil improves several features of metabolic syndrome (MetS), such as dyslipidemia, insulin resistance and hepatic steatosis. Fish oil may mediate some of its beneficial effects by modulating the storage and/or secretory functions of adipose tissue (AT). The storage of triglycerides in AT is regulated by the availability of free fatty acids and the degree of lipolysis in AT. Fish oil has been shown to reduce lipolysis in several studies, indicating improved triglyceride storage. Importantly, AT secretes a variety of adipokines and fish oil feeding is associated with remarkable changes in the plasma levels of two key adipokines, adiponectin and leptin. Much attention has been focused on the contribution of adiponectin in fish oil-mediated improvements in MetS. However, emerging evidence also indicates a role of leptin in modulating the components of the MetS upon fish oil feeding. In addition to improving the storage and secretory functions of AT, fish oil, and the n-3 fatty acids found in fish oil, has been shown to reduce inflammation in AT. These effects may be in part a result of activation of peroxisome proliferator-activated receptor γ or inhibition of Toll-like receptor 4. Thus, there is compelling evidence that fish oil mediates its beneficial effects on MetS by improving AT storage and secretory functions and by reducing inflammation.


Asthma control and concordance of opinions between patients and pulmonologists.
Patient-physician opinion concordance could play a key role in asthma control. There have been no studies evaluating this association in large samples of patients. To determine opinion concordance between asthma patients and their pulmonologists on the impact of the disease and to correlate concordance to asthma control. This was a cross-sectional multicentre study including 1160 patients and 300 pulmonologists. Patient-physician concordance rates were assessed by two semi-structured qualitative questionnaires: (1) impact of the disease and (2) treatment satisfaction. Subsequently, participating pulmonologists determined the concordance between their perceptions and their patient's. Sociodemographic and clinical data were recorded for all patients. In 53.6% of cases, asthma was controlled. The rate of patient-pulmonologist concordance on disease impact on patient daily life was 57%, with physicians underestimating the impact (compared to patients) in 26% of cases. Concordance on satisfaction with treatment was 56%, with physicians underestimating satisfaction in 26% of cases. Patient-physician discordance rates were significantly lower among patients with controlled asthma (29 and 32.1%) than those with poor control (73.7 and 73.1%). Patient-pulmonologist concordance on perceptions of disease impact is low, particularly in uncontrolled asthma. This poor concordance should be addressed in education programmes, particularly for patients with uncontrolled symptoms.


Eye movement desensitization and reprocessing therapy versus supportive therapy in affective relapse prevention in bipolar patients with a history of trauma: study protocol for a randomized controlled trial.
Up to 60% of patients with bipolar disorder (BD) have a history of traumatic events, which is associated with greater episode severity, higher risk of comorbidity and higher relapse rates. Trauma-focused treatment strategies for BD are thus necessary but studies are currently scarce. The aim of this study is to examine whether Eye Movement Desensitization and Reprocessing (EMDR) therapy focusing on adherence, insight, de-idealisation of manic symptoms, prodromal symptoms and mood stabilization can reduce episode severity and relapse rates and increase cognitive performance and functioning in patients with BD. This is a single-blind, randomized controlled, multicentre trial in which 82 patients with BD and a history of traumatic events will be recruited and randomly allocated to one of two treatment arms: EMDR therapy or supportive therapy. Patients in both groups will receive 20 psychotherapeutic sessions, 60 min each, during 6 months. The primary outcome is a reduction of affective episodes after 12 and 24 months in favour of the EMDR group. As secondary outcome we postulate a greater reduction in affective symptoms in the EMDR group (as measured by the Bipolar Depression Rating Scale, the Young Mania Rating Scale and the Clinical Global Impression Scale modified for BD), and a better performance in cognitive state, social cognition and functioning (as measured by the Screen for Cognitive Impairment in Psychiatry, The Mayer-Salovey-Caruso Emotional Intelligence Test and the Functioning Assessment Short Test, respectively). Traumatic events will be evaluated by The Holmes-Rahe Life Stress Inventory, the Clinician-administered PTSD Scale and the Impact of Event Scale. The results of this study will provide evidence whether a specific EMDR protocol for patients with BD is effective in reducing affective episodes, affective symptoms and functional, cognitive and trauma symptoms. The trial is registered at ClinicalTrials.gov, identifier: NCT02634372 . Registered on 3 December 2015.


Vasoconstriction of the basilar artery in rabbits: the vasoconstrictor activity of blood, CSF and blood-CSF mixtures incubated for varying periods of time.
After subarachnoid haemorrhage, the amount of blood in the subarachnoid space and the time elapsed since the haemorrhage seem important factors in predicting the occurrence of vasospasm. In this experiment we studied the responses of the rabbit basilar artery to acute topical application of samples of human cerebrospinal fluid (CSF), mixtures of blood and CSF incubated at 37 degrees C for 0 to 12 days. The degree and the duration of the vasoconstriction depended on the time of incubation and on the concentration of blood in the samples. Samples incubated for five and seven days caused severe and long-lasting vasoconstrictions. Samples incubated for 0, 1, 3, 8, 10 and 12 days caused slight or moderate and short-lasting vasoconstriction. Samples incubated for 5, 7, 8, and 10 days showed a linear relationship between the concentration of blood and the degree of constriction up to the blood-CSF ratio of 0.6 to 1 (blood content of 37.5%); further increases of blood concentration did not cause further vasoconstriction. This phenomenon may be related to the saturation of arterial receptors. The course of the vasoconstriction using samples with different blood concentrations suggests the existence of more vasoactive substances. This work seems to confirm the importance of blood concentration in the CSF and of time as factors governing the development of vasospasm.


Time-Restricted Fasting Improves Liver Steatosis in Non-Alcoholic Fatty Liver Disease-A Single Blinded Crossover Trial.
Non-alcoholic fatty liver disease (NAFLD) is associated with visceral adiposity. We assessed the effectiveness of time-restricted fasting (TRF) for 16 h daily without calorie restrictions compared to standard care (SC; diet and lifestyle advice) in improving visceral adiposity and steatosis via controlled attenuation parameter (CAP). In a prospective single-blind randomized controlled trial, 32 participants with NAFLD were randomly assigned to TRF or SC for 12 weeks. The secondary endpoints were changes in liver stiffness, anthropometry, blood pressure, and other metabolic factors. Twenty-eight participants completed the first arm of the study (TRF = 14, SC = 14), with 23 completing the crossover arm (TRF = 10, SC = 13). The baseline demographics were similar between the groups. Intermittent fasting caused a significant decrease in hepatic steatosis (p = 0.038), weight (p = 0.005), waist circumference (p = 0.001), and BMI (p = 0.005) compared to standard care. Intermittent fasting also resulted in additional within-group changes that were not seen in the standard care intervention. TRF offers superior improvements in patients with NAFLD, improving steatosis, weight, and waist circumference despite a lack of change in overall caloric intake. Time-restricted fasting should be considered as a primary weight loss intervention in the context of NAFLD. ACTRN12613000935730.


A molecular model and Monte Carlo simulation of flavivirus envelope building block.
A molecular model of mature dengue virus envelope building block consisting of two E and two M protein subunits is constructed. Monte Carlo optimisation is performed for the model in the implicit membrane system. Interactions between the ectodomains and membrane parts of E and M proteins are studied, and possible role of conserved residues is suggested. The opening of E protein detergent binding site and its fixation in the open state by the M non-structured loop is observed.


[Comparison of two transportation media for study of normal individual conjunctival microbiota].
To compare the microbiological profile of normal microbiota of healthy people obtained from conjunctival smear using dry swab in Stuart's transport medium and wet swab transported in test tube sealed with cotton. A prospective study with random samples, performed at the Departments of Ophthalmology and Pathology of Santa Casa Misericórdia de São Paulo, in August of 2006. Eighty normal eyes of 40 healthy individuals were analyzed. Samples were collected in the right eye with a dry swab and stored in Stuart's medium, where all microbiological material is kept immersed in the medium and the tube is hermetically sealed. In the left eye, the conjunctival material was collected using a swab embedded in saline solution 0.9%, and stored in dry and sterile test tubes sealed with cotton. The samples were analyzed within 2 hours at most after collection. Out of 40 samples collected with wet swab and transported in dry tube, bacteria were observed in 10 (25%), whereas of 40 samples collected with dry swab and transported in Stuart's medium, 12 (30%) had bacteria. The results of the microbiological profile of normal conjunctival microbiota using dry swab in Stuart's medium were statistically similar (p=0.85) to those obtained in wet swab in dry tube for spreading performed within 2 hours after collection of conjunctival specimen.


Vitamin K status among children with cystic fibrosis and its relationship to bone mineral density and bone turnover.
The aim of this study was to assess vitamin K status in an unselected population of children with cystic fibrosis (CF) and to investigate any vitamin K effect on bone turnover and bone mineral status. Children > or =5 years of age who were attending the CF unit were invited to enter the study. Fasting blood samples were analyzed for levels of vitamin K1 and prothrombin produced in vitamin K absence; total, undercarboxylated, and carboxylated osteocalcin (OC); and bone-specific alkaline phosphatase and procollagen I carboxy-terminal propeptide (bone formation markers). Levels of N-telopeptide and free pyridinoline and deoxypyridinoline (bone breakdown products) were measured in urine samples. Bone mineral density and bone mineral content were measured at the lumbar spine and for the total body with a GE Lunar Prodigy densitometer. Statistical analyses were performed with Minitab version 9.1. One hundred six children entered the study. Sixty-five of 93 children (70%) from whom blood samples were obtained showed suboptimal vitamin K status, on the basis of low serum vitamin K1 levels, increased prothrombin produced in vitamin K absence levels, or both abnormalities. Vitamin K1 levels showed a significant negative correlation with undercarboxylated OC levels but showed no significant correlation with any marker of bone turnover or measurement of bone mineral status. Undercarboxylated OC levels were correlated significantly with bone turnover markers, which themselves showed a significant negative correlation with measurements of bone mineral density and content. There were no significant correlations between carboxylated or undercarboxylated OC levels and bone density measurements. Vitamin K1 deficiency is common among children with CF, and routine supplements should be considered. Through its role in the carboxylation of OC, vitamin K deficiency may be associated with an uncoupling of the balance between bone resorption and bone formation. A cause-effect relationship between vitamin K deficiency and low bone mass has not been proved.


[Valproate (depakine chrono) in adult patients with partial epilepsy: results of a multicentral prospective non-comparative study].
We present results of an open multicentral prospective non-comparative clinical study on the efficacy and tolerability of depakine-chrono in patients with newly or recently diagnosed partial epilepsy (VIPe-1) that has been carried out in the Russian Federation. The study included 299 patients. It's duration was 6 months. Efficacy and tolerability of the drug used in mean dose 1479.0±448.7 mg/day were evaluated. After 6 month of treatment, 65.9% (95% CI 60.2 -71.2%) of patients were seizure-free. There was no significant difference in seizure-free rate in patients with different types of focal seizures. The improvement assessed with the Clinical Global Impression Scale (CGI) was seen in 73.3% of patients. Adverse events (mild or moderate) were recorded in 8% of patients. The most common adverse events were weight gain in (1.7%), hair loss (1.7%) and gastrointestinal disorders (1.3%). A serious adverse event (thrombocytopenia) occurred only in one case (0.3%). The results confirmed the high efficacy and good tolerability of depakine-chrono as first line monotherapy in adult patients with partial epilepsy.


Estimating subject-specific dependent competing risk profile with censored event time observations.
In a longitudinal study, suppose that the primary endpoint is the time to a specific event. This response variable, however, may be censored by an independent censoring variable or by the occurrence of one of several dependent competing events. For each study subject, a set of baseline covariates is collected. The question is how to construct a reliable prediction rule for the future subject's profile of all competing risks of interest at a specific time point for risk-benefit decision making. In this article, we propose a two-stage procedure to make inferences about such subject-specific profiles. For the first step, we use a parametric model to obtain a univariate risk index score system. We then estimate consistently the average competing risks for subjects who have the same parametric index score via a nonparametric function estimation procedure. We illustrate this new proposal with the data from a randomized clinical trial for evaluating the efficacy of a treatment for prostate cancer. The primary endpoint for this study was the time to prostate cancer death, but had two types of dependent competing events, one from cardiovascular death and the other from death of other causes.


Intensity-modulated radiation therapy dose verification using fluence and portal imaging device.
Patient-specific quality assurance for intensity-modulated radiation therapy (IMRT) dose verification is essential. The aim of this study is to provide a new method based on the relative error distribution by comparing the fluence map from the treatment planning system (TPS) and the incident fluence deconvolved from the electronic portal imaging device (EPID) images. This method is validated for 10 head and neck IMRT cases. The fluence map of each beam was exported from the TPS and EPID images of the treatment beams were acquired. Measured EPID images were deconvolved to the incident fluence with proper corrections. The relative error distribution between the TPS fluence map and the incident fluence from the EPID was created. This was also created for a 2D diode array detector. The absolute point dose was measured with an ionization chamber, and the dose distribution was measured by a radiochromic film. In three cases, MLC leaf positions were intentionally changed to create the dose error as much as 5% against the planned dose and our fluence-based method was tested using gamma index. Absolute errors between the predicted dose of 2D diode detector and of our method and measurements were 1.26% ± 0.65% and 0.78% ± 0.81% respectively. The gamma passing rate (3% global / 3 mm) of the TPS was higher than that of the 2D diode detector (p< 0.02), and lower than that of the EPID (p < 0.04). The gamma passing rate (2% global / 2 mm) of the TPS was higher than that of the 2D diode detector, while the gamma passing rate of the TPS was lower than that of EPID (p < 0.02). For three modified plans, the predicted dose errors against the measured dose were 1.10%, 2.14%, and -0.87%. The predicted dose distributions from the EPID were well matched to the measurements. Our fluence-based method provides very accurate dosimetry for IMRT patients. The method is simple and can be adapted to any clinic for complex cases.


Accuracy of de novo assembly of DNA sequences from double-digest libraries varies substantially among software.
Advances in DNA sequencing have made it feasible to gather genomic data for non-model organisms and large sets of individuals, often using methods for sequencing subsets of the genome. Several of these methods sequence DNA associated with endonuclease restriction sites (various RAD and GBS methods). For use in taxa without a reference genome, these methods rely on de novo assembly of fragments in the sequencing library. Many of the software options available for this application were originally developed for other assembly types and we do not know their accuracy for reduced representation libraries. To address this important knowledge gap, we simulated data from the Arabidopsis thaliana and Homo sapiens genomes and compared de novo assemblies by six software programs that are commonly used or promising for this purpose (ABySS, CD-HIT, Stacks, Stacks2, Velvet and VSEARCH). We simulated different mutation rates and types of mutations, and then applied the six assemblers to the simulated data sets, varying assembly parameters. We found substantial variation in software performance across simulations and parameter settings. ABySS failed to recover any true genome fragments, and Velvet and VSEARCH performed poorly for most simulations. Stacks and Stacks2 produced accurate assemblies of simulations containing SNPs, but the addition of insertion and deletion mutations decreased their performance. CD-HIT was the only assembler that consistently recovered a high proportion of true genome fragments. Here, we demonstrate the substantial difference in the accuracy of assemblies from different software programs and the importance of comparing assemblies that result from different parameter settings.


Exogenous Sucrose Confers Low Light Tolerance in Tomato Plants by Increasing Carbon Partitioning from Stems to Leaves.
Low light (LL) stress adversely affects plant growth and productivity. The role of exogenous sucrose in enhancing plant LL tolerance was investigated by spraying sucrose on tomato (Solanum lycopersicum L.) leaves. This study employed physiological and molecular approaches to identify the underlying mechanisms. Exogenous sucrose activated sucrose hydrolysis-related enzyme activity and upregulated genes encoding sucrose and hexose transporters in mature leaves, decreasing endogenous sucrose levels and promoting sucrose unloading during LL. Stem-related genes associated with sucrose synthesis and transport were also upregulated, enhancing sucrose phloem loading. Furthermore, sucrose from stems activated sucrose unloading in sink leaves, forming a feed-forward loop to sustain sucrose flow during LL. This led to increased nonstructural carbohydrates (NSCs), improved energy metabolism, and enhanced protein synthesis in leaves, ultimately boosting photosynthesis and fruit yield after light recovery. These findings highlight how exogenous sucrose enhances LL tolerance in tomatoes by increasing the transport of NSCs from stems to leaves.


Association of tumor necrosis factor-alpha polymorphisms and ozone-induced change in lung function.
Ozone is a major air pollutant with adverse health effects which exhibit marked inter-individual variability. In mice, regions of genetic linkage with ozone-induced lung injury include the tumor necrosis factor-alpha (TNF), lymphotoxin-alpha (LTA), Toll-like receptor 4 (TLR4), superoxide dismutase (SOD2), and glutathione peroxidase (GPX1) genes. We genotyped polymorphisms in these genes in 51 individuals who had undergone ozone challenge. Mean change in FEV1 with ozone challenge, as a percentage of baseline, was -3% in TNF -308G/A or A/A individuals, compared with -9% in G/G individuals (p = 0.024). When considering TNF haplotypes, the smallest change in FEV1 with ozone exposure was associated with the TNF haplotype comprising LTA +252G/TNF -1031T/TNF -308A/TNF -238G. This association remained statistically significant after correction for age, sex, disease, and ozone concentration (p = 0.047). SOD2 or GPX1 genotypes were not associated with lung function, and the TLR4 polymorphism was too infrequent to analyze. The results of this study support TNF as a genetic factor for susceptibility to ozone-induced changes in lung function in humans, and has potential implications for stratifying health risks of air pollution.


Typhoid fever in travelers: who should be targeted for prevention?
To clarify indications for typhoid vaccination, we reviewed laboratory-confirmed cases of typhoid fever reported to the United States Centers for Disease Control and Prevention between 1994 and 1999. To estimate the risk of adverse events associated with typhoid vaccination, we reviewed reports to the Vaccine Adverse Event Reporting System for the same period. Acute Salmonella enterica serotype Typhi infection was reported for 1393 patients. Of these patients, recent travel was reported by 1027 (74%), only 36 (4%) of whom reported having received a vaccination. Six countries accounted for 76% of travel-associated cases (India [30%], Pakistan [13%], Mexico [12%], Bangladesh [8%], The Philippines [8%], and Haiti [5%]). For 626 travelers who traveled to a single country, the length of stay was <or=1 week for 31 (5%), <or=2 weeks for 100 (16%), <or=3 weeks for 169 (27%), <or=4 weeks for 232 (37%), <or=5 weeks for 338 (54%), and <or=6 weeks for 376 (60%). Reports of serious adverse events due to typhoid vaccination were very rare. Vaccination should be considered even for persons planning short-term travel to high-risk areas.


TGF-beta-induced RhoA and p160ROCK activation is involved in the inhibition of Cdc25A with resultant cell-cycle arrest.
The ability of the transforming growth factor beta (TGF-beta) signaling pathways to inhibit proliferation of most cells while stimulating proliferation of others remains a conundrum. In this article, we report that the absence of RhoA and p160ROCK activity in fibroblastic NIH 3T3 cells and its presence in epithelial NMuMG cells can at least partially explain the difference in the TGF-beta growth response. Further, evidence is presented for TGF-beta-stimulated p160ROCK translocation to the nucleus and inhibitory phosphorylation of the cyclin-dependent kinase-activating phosphatase, Cdc25A. The resultant suppression of Cdk2 activity contributes to G1/S inhibition in NMuMG cells. These data provide evidence that signaling through RhoA and p160ROCK is important in TGF-beta inhibition of cell proliferation and links signaling components for epithelial transdifferentiation with regulation of cell-cycle progression.


Distribution of lipid variables in subjects in Belfast, Northern Ireland and Taiyuan, P R China.
To compare the plasma lipid, lipoprotein and apolipoprotein profiles of people of Chinese and European origin, a sample of 151 male Chinese subjects was selected from Taiyuan, Shanxi Province, P R China to 202 broadly age-matched subjects in Belfast who were selected as controls in a case-control study of myocardial infarction. Mean total cholesterol (TC) was 6.15 mmol/l in Belfast and 4.28 mmol/l in Taiyuan. Low density lipoprotein cholesterol (LDL) was much higher in Belfast. High density lipoprotein cholesterol (LDL) was also higher in Belfast than in Taiyuan but the ratio of HDL to TC was lower in Belfast than Taiyuan. Triglyceride (TG) levels were similar and this was reflected in the relatively high level of apolipoprotein E (apo E) in the Chinese sample.


Taibaiella koreensis sp. nov., isolated from soil of a ginseng field.
A Gram-staining-negative, strictly aerobic, motile (by gliding), non-spore-forming and rod-shaped bacterial strain, designated THG-DT86(T), was isolated from soil of a ginseng field of Pocheon province in the Republic of Korea and its taxonomic position was investigated by a polyphasic approach. Growth occurred at 10-35 °C, at pH 6.5-8.5 and with 0-1.5 % (w/v) NaCl on trypticase soy agar. Flexirubin-type pigments were found to be present. On the basis of 16S rRNA gene sequence similarity, strain THG-DT86(T) was shown to belong to the genus Taibaiella and was related to Taibaiella smilacinae PTJT-5(T) (95.3 %). The G+C content of the genomic DNA was 50.1 mol%. The only isoprenoid quinone detected in strain THG-DT86(T) was menaquinone-7 (MK-7) and the only polyamine was homospermidine. The predominant fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH, C16 : 0, iso-C15 : 1 G and iso-C17 : 0, and the major polar lipids were phosphatidylethanolamine, an unidentified aminophosphoglycolipid and an unidentified aminophospholipid. Phenotypic data and phylogenetic inference supported the affiliation of strain THG-DT86(T) to the genus Taibaiella, and a number of biochemical tests differentiated strain THG-DT86(T) from the recognized species of the genus Taibaiella. Therefore, the novel isolate represents a novel species, for which the name Taibaiella koreensis sp. nov. is proposed, with THG-DT86(T) as the type strain ( = KACC 17171(T) = JCM 18823(T)).


Adverse behavioral reactions attributed to triazolam in the Food and Drug Administration's Spontaneous Reporting System.
Reports of adverse behavioral reactions to triazolam, a triazolobenzodiazepine ultra-short-acting hypnotic, were examined in the postmarketing surveillance Spontaneous Reporting System of the Food and Drug Administration. Reports for triazolam of confusion, amnesia, bizarre behavior, agitation, and hallucinations were compared with reports of these reactions for temazepam, another short-acting hypnotic. Analysis of individual case reports from marketing through 1985 for triazolam vs temazepam showed 133 vs two for confusion, 109 vs three for amnesia, 59 vs two for bizarre behavior, 58 vs four for agitation, and 40 vs one for hallucinations. Considering extent of use, reporting rates for triazolam were 22 to 99 times those for temazepam, depending on the reaction. Reactions to triazolam tended to occur at higher doses and in older patients. This and an updated analysis of aggregate reports for the first 7 years of marketing of each drug with reporting rates and adjustment for various factors suggest a higher occurrence of these reactions with triazolam, but selection factors cannot be completely ruled out. When treating insomnia, physicians should emphasize sleep hygiene practices as alternatives to drug therapy; if drug therapy is required, they should prescribe hypnotics at the lowest recommended doses for the shortest clinically necessary durations and discontinue medication use should any adverse reactions occur.


The Importance of School-based Healthy Living Initiatives: Introducing the Health and Wellness Academy Concept.
Over the last 15 years, the number of school and community based health-intervention programs in the United States has grown. Many of these programs aim to prevent non-communicable chronic disease diagnoses (e.g., obesity, cardiovascular disease and type-2 diabetes). The Department of Physical Therapy in the College of Applied Health Sciences (CAHS) at the University of Illinois at Chicago (UIC) created a school-based wellness program (SBWP) that focuses on nutrition and physical activity, providing tailored experiences that motivate adolescents to make healthier lifestyle choices. The SBWP began as a camp for children in the surrounding neighborhoods and implemented healthy living practices utilizing students from Departments in the CAHS. From this camp, the Health and Wellness Academy (HWA) evolved. This paper provides a review of school-based initiatives and introduces the UIC HWA, an innovative and reproducible approach that can bring positive environmental change by improving health outcomes for children and their families.


Circulating Levels of Brain-Derived Neurotrophic Factor and History of Suicide Attempts in Women.
A growing body of research examining biological factors associated with suicidal behaviors highlights the role of brain-derived neurotropic factor (BDNF), involved in neurogenesis and synaptic plasticity. There is evidence suggesting that suicide attempters have lower BDNF levels than those with no history of suicide attempts. The key question addressed in the current investigation is whether differences in circulating BDNF levels persist beyond the current suicidal episode and would be observed in those with a past history of suicide attempts (SA). Plasma levels of BDNF were assessed in 73 women from the community. We found that women with a history of SA exhibited lower levels of BDNF than women with no SA history and this difference was maintained after statistically controlling for the influence of other potential psychiatric or demographic factors. These findings support and extend existing research by suggesting that circulating BDNF levels are decreased among individuals with a history of SA compared to individuals with no history of SA. This relation appeared to be specific to women's history of SA and was not explained by other potential psychiatric or demographic factors, which further highlights the role of BDNF as a promising biomarker for suicidal behavior.


[Levels of physical activity and excess body weight in university students].
University students are considered a vulnerable group due to their adoption of unhealthy diets and lifestyles. To determine the main risk factors associated with low level of physical activity and excess body weight in university students. This cross-sectional study included 358 university students from Talca, (53% female). Sociodemographic, academic, health, lifestyle, nutritional status, physical activity and sedentary behavior variables were analyzed by logistic regression. Men were more likely to present excess body weight than women (Odds ratio (OR): 2.16 [95% CI: 1.17-4.01], p = 0.01). Similarly, those with low level of physical activity (OR: 2.13 [95% CI: 1.10-414], p = 0.03), musculoskeletal injuries (OR: 2.43 [95% CI: 1.09-6.34], p = 0.05) and smokers (OR: 2.09 [95% CI: 1.05-4.14], p = 0.04) were more likely to have excess body weight. Individuals with excess body weight (OR: 2.94 [95% CI: 1.31-6.61], p < 0.01), high sedentary behaviors (OR: 2.01 [95% CI: 1.04-4.55], p = 0.04), those who spent more than 7 hours per week studying (OR: 2.11 [95% CI: 1.06-4.66], p = 0.04), and those with presence of musculoskeletal injuries (OR: 1.49 [95% CI: 1.06- 3.66], p = 0.04) were more likely to be physically inactive. In addition, men were 73% less likely to have low physical activity levels compared to women (OR: 0.27 [95% CI: 0.11-0.56], p < 0.01). University students have several modifiable risk factors associated with low levels of physical activity and excess weight. Healthy lifestyles promotion should modify these risk behaviors.


Regulation of cancer-immunity cycle and tumor microenvironment by nanobiomaterials to enhance tumor immunotherapy.
In the past decade, we have witnessed the revolution in cancer therapy, especially in the rapid development of cancer immunotherapy. In particular, the introduction of nanomedicine has achieved great improvement in breaking the limitations of and immunological tolerance caused by clinic-approved immunotherapies (cancer vaccine, CAR-T, and immune checkpoint blockade) to enhance immunogenicity, antigen presentation and T lymphocyte infiltration for eradicating the primary tumors and distant metastases simultaneously. However, some fundamental but significant issues still need to be thoroughly clarified before the combination of nanomedicine and immunotherapy moves toward clinical translation such as biological safety and synergistic mechanisms of nanomaterials in the systematic immune responses. Therefore, in this review, the role of nanomaterials in cancer immunotherapy is summarized, mainly focusing on the effective activation and long-term stimulation of both the innate and the adaptive immune responses and regulation of or remodeling the tumor microenvironment, especially the tumor immunosuppressive microenvironment. Also, we elaborate on the targets and challenges of nanomaterials in the cancer-immunity cycle, summarize several main strategies to convert the cold tumor immune microenvironment to the hot one, and illustrate the progress in regulation of tumor immune microenvironment by targeting specific immunosuppressive cells. Finally, we prospect the nano-combined immunotherapy strategies in tumor-targeting, normalization of tumor immune environment and modification of macrophages. This article is characterized under: Therapeutic Approaches and Drug Discovery > Emerging Technologies Diagnostic Tools > In Vivo Nanodiagnostics and Imaging Nanotechnology Approaches to Biology > Nanoscale Systems in Biology Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease.


Is the glucocorticoid-induced hypertriglyceridemia in uremic rats caused by decreased tissue lipoprotein lipase activity?
The glucocorticoid-induced hypertriglyceridemia in uremic rats is partly due to defective removal of triglyceride (Tg)-rich lipoproteins. To determine whether this was due to decreased lipoprotein lipase (LPL), the key enzyme involved in the hydrolysis of Tg-rich lipoproteins, the LPL activity in soleus, heart and adipose tissue and the triglyceride secretion rates (TgSR) of uremic rats treated with Triamcinolone or placebo were studied. Sprague-Dawley rats were made uremic by a 2-stage 5/6 nephrectomy and studied three weeks later. During the 3rd week, one group was treated with a five day course of Triamcinolone (0.5 mg/kg body weight i.m.) and the other given an equal volume of saline. Triamcinolone aggravated the hypertriglyceridemia in uremic rats. This was associated with an increase in soleus and heart LPL with no change in adipose tissue LPL. The TgSR was also increased in the Triamcinolone-treated rats. These data suggest that decreased LPL activity is not the cause of hypertriglyceridemia in these rats.


Millimeter-Wave Heating in In Vitro Studies: Effect of Convection in Continuous and Pulse-Modulated Regimes.
Shallow penetration of millimeter waves (MMW) and non-uniform illumination in in vitro experiments result in a non-uniform distribution of the specific absorption rate (SAR). These SAR gradients trigger convective currents in liquids affecting transient and steady-state temperature distributions. We analyzed the effect of convection on temperature dynamics during MMW exposure in continuous-wave (CW) and pulsed-wave (PW) amplitude-modulated regimes using micro-thermocouples. Temperature rise kinetics are characterized by the occurrence of a temperature peak that shifts to shorter times as the SAR of the MMW exposure increases and precedes initiation of convection in bulk. Furthermore, we demonstrate that the liquid volume impacts convection. Increasing the volume results in earlier triggering of convection and in a greater cooling rate after the end of the exposure. In PW regimes, convection strongly depends on the pulse duration that affects the heat pulse amplitude and cooling rate. The latter results in a change of the average temperature in PW regime. Bioelectromagnetics. 2019;40:553-568. © 2019 Bioelectromagnetics Society.


Optimization of Fe2 +/solids content ratio for a novel sludge heavy metal bioleaching process.
A novel bioleaching process, preincubation bioleaching (PB) has been recently developed in our laboratory for decontamination of heavy-metal-laden sewage sludge by removing the toxic low molecular weight organic acids using an isolated degradative yeast species. The objective of the present study was to optimize Fe2+ /solids content ratio required for removing heavy metals from sewage sludge by using this novel process at high solids content. Results from the present study showed that regardless of the Fe2+ /solids content ratio iron oxidation and sludge acidification could be achieved within 2 days confirming the effectiveness of PB in reducing the inhibitory organic acids. When PB was conducted at an elevated solids content of 3%, Fe2 +/solids content ratio could be reduced to 0.75:1, 1:1 and 1.25:1 for solubilization of Zn, Cu and Cr respectively. Under this optimized bioleaching conditions, the following solubilization efficiencies were obtained after 8 days of bioleaching: 100% Zn, 96% Cu, 60% Cr with no reduction in nutrient contents. Hence, bioleaching at elevated solids contents represents a more economical way for decontamination of heavy-metal-laden sewage sludge while preserving its fertility values.


Is the Universe homogeneous?
The standard model of cosmology is based on the existence of homogeneous surfaces as the background arena for structure formation. Homogeneity underpins both general relativistic and modified gravity models and is central to the way in which we interpret observations of the cosmic microwave background (CMB) and the galaxy distribution. However, homogeneity cannot be directly observed in the galaxy distribution or CMB, even with perfect observations, since we observe on the past light cone and not on spatial surfaces. We can directly observe and test for isotropy, but to link this to homogeneity we need to assume the Copernican principle (CP). First, we discuss the link between isotropic observations on the past light cone and isotropic space-time geometry: what observations do we need to be isotropic in order to deduce space-time isotropy? Second, we discuss what we can say with the Copernican assumption. The most powerful result is based on the CMB: the vanishing of the dipole, quadrupole and octupole of the CMB is sufficient to impose homogeneity. Real observations lead to near-isotropy on large scales--does this lead to near-homogeneity? There are important partial results, and we discuss why this remains a difficult open question. Thus, we are currently unable to prove homogeneity of the Universe on large scales, even with the CP. However, we can use observations of the cosmic microwave background, galaxies and clusters to test homogeneity itself.


A simplified method for immunological typing of trachoma-inclusion conjunctivitis-lymphogranuloma venereum organisms.
Responding to the pressure of large numbers of trachoma-inclusion conjunctivitis (TRIC)-lymphogranuloma venereum (LGV) isolates from field studies requiring serotyping, we have developed a simplified, less-precise method that utilizes cell culture-grown organisms to produce mouse antisera which is tested agaist prototype TRIC-LGV antigens in the micro-immunofluorescence test. Cell cultures with as few as 5 to 15% of cells showing inclusions produced adequate antibody in mice 4 days after single injection. Knowledge of the reaction of prototype antisera with the antigens has allowed typing of most isolates tested from the pattern of cross-reaction of their antiserum.


[Botulinum toxin A: experience in the treatment of 115 patients].
Botulinum toxin A is the more efficient therapy of focal dystonias and hemifacial spasm. Our experience with botulinum toxin A injections in 115 patients is reported. Marked or total improvement was achieved in all 45 patients with hemifacial spasm, in 70% of 20 patients with essential blepharospasm and in 71.4% of 14 patients with Meige's syndrome. In 65.2% of 23 patients with cervical dystonia marked but no total improvement was obtained. The worse results were seen in the 6 patients with hand dystonia (writers cramp), in whom marked improvement was obtained in just two. Mild and transient complications occurred in up to 24.4%, eyelid ptosis and eyelid weakness being the most frequent. One patient with Meige's syndrome had an aspiration pneumonia following dysphagia. Our results are in agreement with others, showing that botulinun toxin A is a useful and safe treatment for these conditions.


Immune Profiles Identification by Vaccinomics After MVA Immunization in Randomized Clinical Study.
Our previous work has demonstrated the benefits of transcutaneous immunization in targeting Langerhans cells and preferentially inducing CD8 T-cell responses. In this randomized phase Ib clinical trial including 20 HIV uninfected volunteers, we compared the safety and immunogenicity of the MVA recombinant vaccine expressing HIV-B antigen (MVA-B) by transcutaneous and intramuscular routes. We hypothesized that the quality of innate and adaptive immunity differs according to the route of immunization and explored the quality of the vector vaccine-induced immune responses. We also investigated the early blood transcriptome and serum cytokine levels to identify innate events correlated with the strength and quality of adaptive immunity. We demonstrate that MVA-B vaccine is safe by both routes, but that the quality and intensity of both innate and adaptive immunity differ significantly. Transcutaneous vaccination promoted CD8 responses in the absence of antibodies and slightly affected gene expression, involving mainly genes associated with metabolic pathways. Intramuscular vaccination, on the other hand, drove robust changes in the expression of genes involved in IL-6 and interferon signalling pathways, mainly those associated with humoral responses, and also some levels of CD8 response. Thus, vaccine delivery route perturbs early innate responses that shape the quality of adaptive immunity. http://ClinicalTrials.gov, identifier PER-073-13.


Development of a novel (poly)phenol-rich diet score and its association with urinary (poly)phenol metabolites.
Background: Estimating (poly)phenol intake is challenging due to inadequate dietary assessment tools and limited food content data. Currently, a priori diet scores to characterise (poly)phenol-rich diets are lacking. This study aimed to develop a novel (poly)phenol-rich diet score (PPS) and explore its relationship with circulating (poly)phenol metabolites. Methods: A total of 543 healthy free-living participants aged 18-80 years completed a food frequency questionnaire (FFQ) (EPIC-Norfolk) and provided 24 h urine samples. The PPS was developed based on the relative intake (quintiles) of 20 selected (poly)phenol-rich food items abundant in the UK diet, including tea, coffee, red wine, whole grains, chocolate and cocoa products, berries, apples and juice, pears, grapes, plums, citrus fruits and juice, potatoes and carrots, onions, peppers, garlic, green vegetables, pulses, soy and soy products, nuts, and olive oil. Foods included in the PPS were chosen based on their (poly)phenol content, main sources of (poly)phenols, and consumption frequencies in the UK population. Associations between the PPS and urinary phenolic metabolites were investigated using linear models adjusting energy intake and multiple testing (FDR adjusted p < 0.05). Result: The total PPS ranged from 25 to 88, with a mean score of 54. A total of 51 individual urinary metabolites were significantly associated with the PPS, including 39 phenolic acids, 5 flavonoids, 3 lignans, 2 resveratrol and 2 other (poly)phenol metabolites. The total (poly)phenol intake derived from FFQs also showed a positive association with PPS (stdBeta 0.32, 95% CI (0.24, 0.40), p < 0.01). Significant positive associations were observed in 24 of 27 classes and subclasses of estimated (poly)phenol intake and PPS, with stdBeta values ranging from 0.12 (0.04, 0.20) for theaflavins/thearubigins to 0.43 (0.34, 0.51) for flavonols (p < 0.01). Conclusion: High adherence to the PPS diet is associated with (poly)phenol intake and urinary biomarkers, indicating the utility of the PPS to characterise diets rich in (poly)phenols at a population level.


Axonal filopodial asymmetry induced by synaptic target.
During vertebrate neuromuscular junction (NMJ) assembly, motor axons and their muscle targets exchange short-range signals that regulate the subsequent steps of presynaptic and postsynaptic specialization. We report here that this interaction is in part mediated by axonal filopodia extended preferentially by cultured Xenopus spinal neurons toward their muscle targets. Immunoblotting and labeling experiments showed that basic fibroblast growth factor (bFGF) was expressed by muscle and associated with the cell surface, and treatment of cultured spinal neurons with recombinant bFGF nearly doubled the normal density of filopodia in neurites. This effect of bFGF was abolished by SU5402, a selective inhibitor of FGF-receptor 1 (FGFR1), and forced expression of wild-type or dominant-negative FGFR1 in neurons enhanced or suppressed the assembly of filopodia, respectively. Significantly, in nerve-muscle cocultures, knocking down bFGF in muscle decreased both the asymmetric extension of filopodia by axons toward muscle and the assembly of NMJs. In addition, neurons expressing dominant-negative FGFR1 less effectively triggered the aggregation of muscle acetylcholine receptors at innervation sites than did control neurons. These results suggest that bFGF activation of neuronal FGFR1 generates filopodial processes in neurons that promote nerve-muscle interaction and facilitate NMJ establishment.


BRCA1/2 pathogenic variants in triple-negative versus luminal-like breast cancers: genotype-phenotype correlation in a cohort of 531 patients.
Several available data suggest the association between specific molecular subtypes and BRCA1/2 mutational status. Previous investigations showed the association between BRCA1/2 pathogenic variants (PVs) in specific genomic regions and phenotypic variations of cancer relative risk, while the role of PV type and location in determining the breast cancer (BC) phenotypic features remains still unclear. The aim of this research was to describe the germline BRCA1/2 PVs in triple-negative breast cancer (TNBC) versus luminal-like BC and their potential leverage on BC phenotype. We retrospectively collected and analyzed all clinical information of 531 patients with BC genetically tested for germline BRCA1/2 PVs by Next-Generation Sequencing analysis at University Hospital Policlinico "P. Giaccone" of Palermo (Sicily) from January 2016 to February 2020. Our results corroborate the evidence that BRCA1-related tumors often have a profile which resembles the TNBC subtype, whereas BRCA2-associated tumors have a profile that resembles luminal-like BC, especially the Luminal B subtype. Interestingly, our findings suggest that the PVs identified in TNBC were not largely overlapping with those in luminal-like tumors. Differences in the frequency of two PVs potentially associated with different molecular tumor subtypes were observed. BRCA1-633delC was detected with relatively higher prevalence in patients with TNBC, whereas BRCA2-1466delT was found mainly in Luminal B tumors, but in no TNBC patient. Future studies examining the type and location of BRCA1/2 PVs within different molecular subtypes are required to verify our hypothesis and could provide an interesting insight into the complex topic of genotype-phenotype correlations. Additionally, a more in-depth understanding of the potential correlations between BRCA PVs and clinical and phenotypic features of hereditary BC syndrome patients could be the key to develop better strategies of prevention and surveillance in BRCA-positive carriers without disease.