The beta-beta-alpha fold: explorations in sequence space.
The computational redesign of the second zinc finger of Zif268 to produce a 28 residue peptide (FSD-1) that assumes a betabetaalpha fold without metal binding was recently reported. In order to explore the tolerance of this metal-free fold towards sequence variability, six additional peptides resulting from the ORBIT computational protein design process were synthesized and characterized. The experimental stabilities of five of these peptides are strongly correlated with the energies calculated by ORBIT. However, when a peptide with a mutation in the beta-turn is examined, the calculated stability does not accurately predict the experimentally determined stability. The NMR solution structure of a peptide incorporating this mutation (FSD-EY) reveals that the register between the beta-strands is different from the model structure used to select and score the sequences. FSD-EY has a type I' turn instead of the target EbaaagbE turn (rubredoxin knuckle). Two additional peptides that have improved side-chain to backbone hydrogen bonding and turn propensity for the target turn were characterized. Both are of stability comparable to that of FSD-1. These results demonstrate the robustness of the ORBIT protein design methods and underscore the need for continued improvements in negative design.


Exposure apportionment: ranking food items by their contribution to dietary exposure.
This paper identifies and ranks food items by estimating their contribution to the dietary exposure of the US population and 19 subpopulation groups. Contributions to dietary exposures to arsenic, cadmium, chromium, lead, nickel, benzene, chlorpyrifos, and diazinon are estimated using either the Dietary Exposure Potential Model (DEPM) approach, the National Human Exposure Assessment Survey Arizona (NHEXAS-AZ) approach or the combination of the two. The DEPM is a computer model that uses several national databases of food consumption and residue concentrations for estimating dietary. The DEPM approach ranks the contribution of food items to the total dietary exposure using two methods, the direct method that ranks contributions by population exposure magnitude and the weighted method that ranks by subpopulation exposure magnitude. The DEPM approach identifies highly exposed subpopulations and a relatively small number of food items contributing the most to dietary exposure. The NHEXAS-AZ approach uses the NHEXAS-AZ database containing food consumption data for each subject and chemical residues of a composite of food items consumed by each subject in 1 day during the sampling week. These data are then modeled to obtain estimates of dietary exposure to chemical residues. The third approach uses the NHEXAS-AZ consumption data with residue values from the national residue database. This approach also estimates percent contributions to exposure of each ranked food item for the Arizona population. Dietary exposures estimated using the three approaches are compared. The DEPM results indicate groups with highest dietary exposures include Nonnursing Infants, Children 1-6, Hispanic, Non-Hispanic White, Western, Northeast and Poverty 0-130%. The use of the Combined National Residue Database (CNRD) identifies 43 food items as primary contributors to total dietary exposure; they contribute a minimum of 68% of the total dietary exposure to each of the eight chemical residues. The percent contribution of ranked food items estimated using the NHEXAS samples is smaller than those obtained from the western US population via the DEPM. This indicates differences in consumption characteristics of the two groups with respect to the ranked food items. Six of 15 food items consumed by the NHEXAS-AZ subjects per day are ranked food items contributing between 56% and 70% of the estimated NHEXAS-AZ dietary exposure to each of the eight chemical residues. The difference between total dietary exposure estimates from the DEPM and NHEXAS-AZ approaches varies by chemical residue and is attributable to differences in sampling and analytical methods, and geographic areas represented by the data. Most metal exposures estimated using the NHEXAS consumption data with the CNRD have lower values than those estimated via the other approaches, possibly because the NHEXAS-AZ residue values are higher than the CNRD values. In addition, exposure estimates are seemingly affected by the difference in demographic characteristics and factors that affect types and amounts of food consumed. Efficient control strategies for reducing dietary exposure to chemical residues may be designed by focusing on the relatively small number of food items having similar ingredients that contribute substantively to the total ingestion exposure.


Infection dynamics and persistence of hepatitis E virus on pig farms - a review.
Hepatitis E virus (HEV) genotype 3 and 4 is a zoonosis that causes hepatitis in humans. Humans can become infected by consumption of pork or contact with pigs. Pigs are the main reservoir of the virus worldwide and the virus is present on most pig farms. Though HEV is present on most farms, the proportion of infected pigs at slaughter and thus the level of exposure to consumers differs between farms and countries. Understanding the cause of that difference is necessary to install effective measures to lower HEV in pigs at slaughter. Here, HEV studies are reviewed that include infection dynamics of HEV in pigs and on farms, risk factors for HEV farm prevalence, and that describe mechanisms and sources that could generate persistence on farms. Most pigs become infected after maternal immunity has waned, at the end of the nursing or beginning of the fattening phase. Risk factors increasing the likelihood of a high farm prevalence or proportion of actively infected slaughter pigs comprise of factors such as farm demographics, internal and external biosecurity and immunomodulating coinfections. On-farm persistence of HEV is plausible, because of a high transmission rate and a constant influx of susceptible pigs. Environmental sources of HEV that enhance persistence are contaminated manure storages, water and fomites. As HEV is persistently present on most pig farms, current risk mitigation should focus on lowering transmission within farms, especially between farm compartments. Yet, one should be aware of the paradox of increasing the proportion of actively infected pigs at slaughter by reducing transmission insufficiently. Vaccination of pigs may aid HEV control in the future.


Sphingoid bases inhibit acid-induced demineralization of hydroxyapatite.
Calcium hydroxyapatite (HAp), the main constituent of dental enamel, is inherently susceptible to the etching and dissolving action of acids, resulting in tooth decay such as dental caries and dental erosion. Since the prevalence of erosive wear is gradually increasing, there is urgent need for agents that protect the enamel against erosive attacks. In the present study we studied in vitro the anti-erosive effects of a number of sphingolipids and sphingoid bases, which form the backbone of sphingolipids. Pretreatment of HAp discs with sphingosine, phytosphingosine (PHS), PHS phosphate and sphinganine significantly protected these against acid-induced demineralization by 80 ± 17%, 78 ± 17%, 78 ± 7% and 81 ± 8%, respectively (p < 0.001). On the other hand, sphingomyelin, acetyl PHS, octanoyl PHS and stearoyl PHS had no anti-erosive effects. Atomic force measurement revealed that HAp discs treated with PHS were almost completely and homogeneously covered by patches of PHS. This suggests that PHS and other sphingoid bases form layers on the surface of HAp, which act as diffusion barriers against H(+) ions. In principle, these anti-erosive properties make PHS and related sphingosines promising and attractive candidates as ingredients in oral care products.


Faculty and resident perspectives on ambulatory care education: A collective case study of family medicine, psychiatry, and surgery.
Ambulatory care (AC) experiences within medical education are garnering increasing attention. We sought to understand how faculty and residents' describe their experiences of AC and ambulatory care education (ACEduc) within, between, and across disciplinary contexts. We designed a Stakian collective case study, applying constructivist grounded theory analytic methods. Using purposive and snowball sampling, we interviewed 17 faculty and residents across three instrumental cases: family medicine, psychiatry, surgery. Through constant comparative analysis, we identified patterns within, between, and across cases. Family medicine and psychiatry saw AC as an inherent part of continuous, longitudinal care; surgery equated AC with episodic experiences in clinic, differentiating it from operating. Across cases, faculty and residents cautiously valued ACEduc, and in particular, considered it important to develop non-medical expert competencies (e.g., communication). However, surgery residents described AC and ACEduc as less interesting and a lower priority than operating. Educational structures mediated these views. Differences between cases highlight a need for further study, as universal assumptions about ACEduc's purposes and approaches may need to be tempered by situated, contextually-rich perspectives. How disciplinary culture, program structure, and systemic structure influence ACEduc warrant further consideration as does the educational potential for explicitly framing learners' perspectives.


IgG antibody levels to meningococcal porins in patient sera: comparison of immunoblotting and ELISA measurements.
IgG antibody levels to the meningococcal PorA and PorB proteins in 56 acute and convalescent phase sera from 25 patients with meningococcal disease were compared by immunoblotting and ELISA. Heat-treated outer membrane vesicles from strain 44/76 (B:15:P1.7, 16) served as antigens for immunoblotting, whereas purified P1.7,16 PorA and P15 PorB from the same strain were used as antigens in the ELISA. In the blotting method, IgG binding to the porins was determined by digital scanning of the immunoreactive bands and calculated relative to the PorA binding of a reference serum on each blot. The coefficient of variation for the reference serum was 21.6% (a total of 144 strips) with smaller variations for each day's experiments. Blotting of all 56 sera at the standard 1/200 dilution measured anti-PorA and anti-PorB levels that correlated with those obtained by ELISA (Spearman rank-order correlation coefficient r(s)=0.48; P<0.001). At this dilution, the anti-PorA (r(s)=0.52; P<0. 004) and anti-PorB (r(s)=0.60; P<0.001) levels of the convalescent phase sera (n=29) corresponded with the ELISA measurements, whereas no correlation was found with the results for the acute phase sera, which mostly had low ELISA antibody levels (<2 microg/ml IgG). A corresponding blot analysis of convalescent sera from the seven patients, who had received the 44/76 outer membrane vesicle vaccine, demonstrated a high correlation coefficient for the anti-PorA levels (r(s)=0.95; P<0.001) vs. the ELISA results. No such correlation was observed for the PorB response in these sera, being nine-fold higher than the PorA response, because of a prozone effect on the blots at the standard dilution. However, blotting at a higher serum dilution (1/2000) resulted in anti-PorB levels that also correlated strongly (r(s)=0.93 P<0.001) with the ELISA measurements.


Comparisons of the P greater than V sign on the WISC-R and WAIS-R in delinquent males.
Comparisons of the WISC-R and WAIS-R were made in a sample of male delinquents (N = 80) to determine whether they would show differences in the degree to which the P greater than V sign occurred on them. A larger split between mean Verbal and mean Performance IQ was found on the WISC-R than on the WAIS-R. Frequency analyses revealed no differences in the number of persons who showed the P greater than V sign on each test. Age and the P greater than V sign were not related significantly. Results suggest that the WISC-R and WAIS-R may not be equivalent measures of this sign, and caution should be exercised in applying results of research on the WISC and WAIS to the revisions of those tests.


Crystal structures of the human p56lck SH2 domain in complex with two short phosphotyrosyl peptides at 1.0 A and 1.8 A resolution.
src homology 2 (SH2) domains are modules of about 100 amino acid residues and bind to phosphotyrosine-containing motifs in a sequence-specific manner. They play important roles in intracellular signal transduction and represent potential targets for pharmacological intervention. The protein tyrosine kinase p56lck is a member of the src family and is involved in T-cell activation. The crystal structure of its SH2 domain with an 11-residue peptide showed that the phosphotyrosine and the Ile residue at the pY + 3 position are recognized by the SH2 domain. We present here the crystal structure of the SH2 domain of human p56lck in complex with the short phosphotyrosyl peptide Ac-pTyr-Glu-Glu-Ile (pYEEI peptide) at 1.0 A resolution. The structural analysis at atomic resolution reveals that residue Arg134 (alphaA2), which interacts with the phosphotyrosine side-chain, is present in two conformations in the complex. The structure at 1.8 A resolution of the complex with the phosphotyrosyl peptide Ac-pTyr-Glu-Glu-Gly (pYEEG peptide), which is 11 fold less potent, shows another binding mode for the pY + 3 residue as well as rearrangements of the side-chain of Arg196 (EF3) and one of the water molecules at the base of the pY + 3 pocket. The structure of the complex with the short pYEEI peptide at atomic resolution represents a good starting point for the design and optimization of new inhibitors. Comparative structural analysis of many different inhibitor complexes will be an important component of this drug discovery process.


Introduction to CAUSES: Description of Weather and Climate Models and Their Near-Surface Temperature Errors in 5 day Hindcasts Near the Southern Great Plains.
We introduce the Clouds Above the United States and Errors at the Surface (CAUSES) project with its aim of better understanding the physical processes leading to warm screen temperature biases over the American Midwest in many numerical models. In this first of four companion papers, 11 different models, from nine institutes, perform a series of 5 day hindcasts, each initialized from reanalyses. After describing the common experimental protocol and detailing each model configuration, a gridded temperature data set is derived from observations and used to show that all the models have a warm bias over parts of the Midwest. Additionally, a strong diurnal cycle in the screen temperature bias is found in most models. In some models the bias is largest around midday, while in others it is largest during the night. At the Department of Energy Atmospheric Radiation Measurement Southern Great Plains (SGP) site, the model biases are shown to extend several kilometers into the atmosphere. Finally, to provide context for the companion papers, in which observations from the SGP site are used to evaluate the different processes contributing to errors there, it is shown that there are numerous locations across the Midwest where the diurnal cycle of the error is highly correlated with the diurnal cycle of the error at SGP. This suggests that conclusions drawn from detailed evaluation of models using instruments located at SGP will be representative of errors that are prevalent over a larger spatial scale.


Increasing the Vase Life of Chrysanthemum Cut Flowers by Using Silver and Zinc Nanoparticles.
Cut flowers are horticultural products that have great potential to be developed. Efforts to maintain quality and extend the shelf life of cut flowers are very important to obtain a product that is accepted in the market. The main problems of chrysanthemum cut flowers are the leaves easily turning yellow, wilting, and failure to fully open flowers. This study aimed to obtain the best pulsing solution formulation that increases vase life and maintains the freshness of chrysanthemum cut flowers. Pulsing solution treatment was carried out on chrysanthemum cut flowers during the evaluation period. Pulsing solution treatment consisted of control, AgNO3, nano-Ag (NAg), ZnO, and nano-Zn (NZn). The results showed that NAg20 treatment increased the vase life of chrysanthemum cut flowers up to 23 days, which was 19 days longer than the control. Nano-Ag inhibits bacterial growth, flower wilting, color degradation, and carotenoids. In addition, nano-Ag increased the size of the bloom-flower diameter. Considering the results of all postharvest quality parameters mentioned above, NAg20 extends the vase life of chrysanthemum cut flowers.


Automatic monitoring of obstructive sleep apnea based on multi-modal signals by phone and smartwatch.
Obstructive Sleep Apnea (OSA) is the most common sleep-related breathing disorder, with an overall population prevalence ranging from 9% to 38%, and it is associated with many cardiovascular diseases. The diagnosis of OSA requires polysomnography (PSG) testing, which is unsuitable for large-scale preliminary screening due to its high cost and discomfort to wear. Therefore, a simple and inexpensive screening method would be of great value. This study presents a novel at-home OSA screening method using a smartwatch and a smartphone to obtain several physiological signals, snoring segments, and questionnaire information during a whole night's sleep. The proposed method can distinguish four OSA risk levels based on machine learning (ML) classifications; the system was validated by conducting an in-hospital study on 350 subjects with sleep disorders. The estimated OSA risk levels are in good agreement with the OSA severity diagnosed by PSG (correlation with apnea-hypopnea index (AHI) = 0.92), and an encouraging classification performance is achieved (accuracy = 88.1%, 84.5%, 85.1%, sensitivity = 89.1%, 84.2%, 85.6% for mild, moderate and severe OSA). These findings reveal that wearable devices have the potential for large-scale OSA screening.


[Radiation doses and allozyme variability in the population of the northern red-backed vole (Clethrionomys rutilus) from the east urals radioactive trace zone].
The paper estimates the external (due to radionuclides accumulated in the soil) and internal (due to incorporated radionuclides) exposure of rodents that live in the head of the East Urals Radioactive Trace (EURT). It is shown that in the last decades the dose rates were lower than the values that lead to doubling the number of mutations in the allozyme loci in mammals. The variability ofthe eight allozyme loci in populations of northern red-backed voles from the EURT zone and their neighboring plots, as well as the territories of the Urals and Trans-Urals with background levels of radioactive contamination, are analyzed. No differences in the pattern and frequency of allozymes that would distinguish the EURT samples from a number of other populations of the Urals, were found. In the control sample "Sysert", "unique" for the Ural populations of northern red-backed voles, alleles of the loci Got and Sod were marked, conspecific to a closely related species--the bank vole. This fact can be regarded as evidence of recent cross-species hybridization.


SIRT3 interacts with the daf-16 homolog FOXO3a in the mitochondria, as well as increases FOXO3a dependent gene expression.
Cellular longevity is a complex process relevant to age-related diseases including but not limited to chronic illness such as diabetes and metabolic syndromes. Two gene families have been shown to play a role in the genetic regulation of longevity; the Sirtuin and FOXO families. It is also established that nuclear Sirtuins interact with and under specific cellular conditions regulate the activity of FOXO gene family proteins. Thus, we hypothesize that a mitochondrial Sirtuin (SIRT3) might also interact with and regulate the activity of the FOXO proteins. To address this we used HCT116 cells overexpressing either wild-type or a catalytically inactive dominant negative SIRT3. For the first time we establish that FOXO3a is also a mitochondrial protein and forms a physical interaction with SIRT3 in mitochondria. Overexpression of a wild-type SIRT3 gene increase FOXO3a DNA-binding activity as well as FOXO3a dependent gene expression. Biochemical analysis of HCT116 cells over expressing the deacetylation mutant, as compared to wild-type SIRT3 gene, demonstrated an overall oxidized intracellular environment, as monitored by increase in intracellular superoxide and oxidized glutathione levels. As such, we propose that SIRT3 and FOXO3a comprise a potential mitochondrial signaling cascade response pathway.


A patch test study to evaluate the allergenicity of a metallic jewelry alloy in patients allergic to cobalt.
Cobalt is a common inducer of allergic contact dermatitis. A new jewelry alloy containing cobalt was developed and tested on 30 individuals with documented allergy to cobalt. Of these subjects, 5 (18%) were found to be allergic to the new alloy after 7 or 8 days of exposure. In all cases, the reaction was much less severe than allergy to the 1% cobalt chloride patch test material. This new alloy should be better tolerated by individuals allergic to cobalt, perhaps because the cobalt is tightly bound in the alloy by the other main component, platinum.


Alternating optimization for G × E modelling with weighted genetic and environmental scores: Examples from the MAVAN study.
Motivated by the goal of expanding currently existing Genotype × Environment interaction (G × E) models to simultaneously include multiple genetic variants and environmental exposures in a parsimonious way, we developed a novel method to estimate the parameters in a G × E model, where G is a weighted sum of genetic variants (genetic score) and E is a weighted sum of environments (environmental score). The approach uses alternating optimization, an iterative process where the genetic score weights, the environmental score weights, and the main model parameters are estimated in turn, assuming the other parameters are constant. This technique can be used to construct relatively complex interaction models that are constrained to a particular structure, and hence contain fewer parameters. We present the model as a 2-way interaction longitudinal mixed model, for which ordinary linear regression is a special case, but it can easily be extended to be compatible with k-way interaction models and generalized linear mixed models. The model is implemented in R (LEGIT package) and using SAS macros (LEGIT_SAS). Through simulations, we demonstrate the power and validity of this approach even with small sample sizes. Furthermore, we present examples from the Maternal Adversity, Vulnerability, and Neurodevelopment (MAVAN) study where we improve significantly upon already existing models using alternating optimization. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


[Ultrasound transit-time method for evaluation of regional myocardial function (author's transl)].
The validity of an ultrasound transit-time method for defining local function was evaluated under experimental conditions in the anaesthetized open chest dog. Miniaturized piezoelectric crystals (1,5-2 mm phi) functioning as emitters and receivers were inserted into different depths of the myocardium to study the contraction patterns of defined muscle segments in different muscle layers and the dynamic changes of left ventricular wall thickness during the heart cycle. Changes in myocardial function were induced by coronary stenosis or occlusion and by the application of drugs. This method was shown to be a sensible and reliable tool for the evaluation of local myocardial function.


E-Cadherin Protein as a Potential Marker for Gastric Cancer and Its Association with Helicobacter Pylori- Induced Gastritis and Gastric Ulcer.
Gastric cancer is still the main health threat being the third leading cause of deaths from cancers in the world. The major risk behind the gastric cancer is that it remains asymptomatic in the early stages and in (97%) cases it metastasizes to other organs. Gastric cancer is a multifactorial disease in which Helicobacter pylori (H. pylori) has been known as a risk factor. However, patients with gastritis, especially atrophic gastritis and gastric ulcer have been shown to be at an increased risk for developing gastric cancer. This study included measuring the serum levels of E-Cadherin protein, carbohydrate antigen 19-9 (CA19-9) and carcinoembryonic antigen (CEA) in 30 patients diagnosed with gastritis, 20 gastric ulcer patients, 20 gastric cancer patients and in 20 healthy volunteers serving as the control group. Infection with H. pylori was diagnosed by serology (IgA and IgG antibodies) as well as by rapid urease test (RUT) and histology. The results showed that 50 (71.4%) of the patients were positive for H. pylori. Levels of E-Cadherin were increased significantly in all patients in comparison to the control group with a large significant increase in the gastric cancer group. The levels of E-Cadherin were also significantly increased in H. pylori infected patients compared to H. pylori negative patients. A non-significant difference in the levels of CA19-9 and CEA was observed in all patients in comparison to healthy controls. This study concluded that serum E-Cadherin could be considered as a potential marker in diagnosis of gastric cancer.


Spontaneous lung metastasis formation of human Merkel cell carcinoma cell lines transplanted into scid mice.
Merkel cell carcinoma (MCC) is an aggressive skin cancer entity that frequently leads to rapid death due to its high propensity to metastasize. The etiology of most MCC cases is linked to Merkel cell polyomavirus (MCPyV), a virus which is monoclonally integrated in up to 95% of tumors. While there are presently no animal models to study the role of authentic MCPyV infection on transformation, tumorigenesis or metastasis formation, xenograft mouse models employing engrafted MCC-derived cell lines (MCCL) represent a promising approach to study certain aspects of MCC pathogenesis. Here, the two MCPyV-positive MCC cell lines WaGa and MKL-1 were subcutaneously engrafted in scid mice. Engraftment of both MCC cell lines resulted in the appearance of circulating tumor cells and metastasis formation, with WaGa-engrafted mice showing a significantly shorter survival time as well as increased numbers of spontaneous lung metastases compared to MKL-1 mice. Interestingly, explanted tumors compared to parental cell lines exhibit an upregulation of MCPyV sT-Antigen expression in all tumors, with WaGa tumors showing significantly higher sT-Antigen expression than MKL-1 tumors. RNA-Seq analysis of explanted tumors and parental cell lines furthermore revealed that in the more aggressive WaGa tumors, genes involved in inflammatory response, growth factor activity and Wnt signalling pathway are significantly upregulated, suggesting that sT-Antigen is the driver of the observed differences in metastasis formation.


Biomechanics of large femoral heads: what they do and don't do.
The stability and durability of total hip reconstruction is dependent on many factors that include the design and anatomic orientation of prosthetic components. An analysis of femoral component head size and acetabular component orientation shows an interdependency of these variables and joint stability. Increased femoral component head size can increase hip stability by increasing the prosthetic impingement-free range of hip motion and by increasing the inferior head displacement required before hip dislocation. Increasing the femoral head size from 22 mm to 40 mm increases the required displacement for dislocation by about 5 mm with the acetabular component at 45 degrees of abduction; however, increasing acetabular component abduction greatly diminishes this stability advantage of larger femoral heads. Vertical acetabular component orientation and femoral component head subluxation are each predicted to more than double the tensile stress with acetabular component polyethylene compared with components at 45 degrees of abduction. With a desirable acetabular component orientation, the use of larger femoral heads may result in improved joint stability and durable use of polyethylene. With high abduction acetabular component orientation, the use of larger femoral heads contributes little to joint stability and contributes to elevated stress within the polyethylene that may result in implant failure.


Aarskog-Scott syndrome: An unusual cause of scoliosis.
A 16-year-old boy who had been diagnosed previously as Aarskog-Scott syndrome (AAS), referred to our clinic with shoulder asymmetry for 1 year. Results of spine examination showed a 52° right thoracic curve at T3-T11. Surgery was planned, and T1-L1 posterior instrumentation and fusion were performed. After surgery, satisfactory correction was achieved, and during 10 years follow-up, the patient had no complaints. AAS is a X-linked genetic disorder with facial, genital, and skeletal manifestations. Scoliosis is not reported as a typical finding of AAS, and there is no reported case in the English literature. Due to mutation affecting the developing skeleton tissue, spinal deformities may develop. In our case, concave side fusion was seen at the deformity. Although we do not know any specific pattern of the scoliotic deformity of this syndrome, surgical correction of the deformity can be difficult because of the premature fusion at these levels.


Roxadustat protects rat renal tubular epithelial cells from hypoxia-induced injury through the TGF-β1/Smad3 signaling pathway.
Roxadustat is used to treat renal anemia. The renoprotective effect of roxadustat needs to be further confirmed, and the mechanism of action is unknown. This study aims to evaluate the effect and mechanism of roxadustat in hypoxia-related nephropathy with the renal tubular epithelial cell line NRK-52E. The cell Counting Kit-8 (CCK-8) assay was employed to assess cellular proliferation in the current investigation. Flow cytometry was used to conduct cell apoptosis analysis. The utilization of electron microscopy facilitated the identification of changes in cellular ultrastructure. Immunofluorescence was used to detect the expression trend of hypoxia-inducible factor-1α (HIF-1α). The connective tissue growth factor (CTGF), transforming growth factor-β1 (TGF-β1), Smad family member 3 (Smad3), p-Smad3, α-smooth muscle actin (α-SMA), collagen I, and HIF-1α were assessed by western blotting. Real-time fluorescent quantitative PCR (RT-qPCR) was used to measure TGF-β1 and Smad3 mRNA. Significant growth inhibition and increased apoptosis were observed in NRK-52E cells cultured under hypoxic conditions (1% and 5% O2), which can be rescued by roxadustat. From a morphological perspective, it has been observed that roxadustat can counteract cellular damage features produced by hypoxia. These features include the contraction of the nuclear envelope and an increase in the formation of apoptotic bodies. Roxadustat increases HIF-1α expression acutely at 24 h, followed by a gradual reduction of HIF-1α expression to levels significantly below that of the hypoxia group by 72 h. Roxadustat can also inhibit hypoxia-induced increased expression of CTGF, TGF-β1, p-Smad3, α-SMA, collagen I, and HIF-1α. Combined treatment with roxadustat and siRNA against TGF-β1 synergistically reduced the expression of CTGF and HIF-1α, while the effect on TGF-β1 and p-Smad3 were comparable to that of the individual treatment alone. Comparably, the combined administration of roxadustat and siRNA targeting Smad3 had a synergistic impact on diminishing the expression of CTGF. These findings indicate that roxadustat attenuates experimental renal fibrosis likely by inhibiting the TGF-β1/Smad3 pathways, while its effect on CTGF and HIF-1α may involve other signaling pathways.


Caregiver acceptability of the guidelines for managing young infants with possible serious bacterial infections (PSBI) in primary care facilities in rural Bangladesh.
Many infants with possible serious bacterial infections (PSBI) do not receive inpatient treatment because hospital care may not be affordable, accessible, or acceptable for families. In 2015, WHO issued guidelines for managing PSBI in young infants (0-59 days) with simpler antibiotic regimens when hospital care is not feasible. Bangladesh adopted WHO's guidelines for implementation in outpatient primary health centers. We report results of an implementation research study that assessed caregiver acceptability of the guidelines in three rural sub-districts of Bangladesh during early implementation (October 2015-August 2016). We included 19 outpatient primary health centers involved in the initial rollout of the infection management guidelines. We extracted data for all PSBI cases (N = 192) from facility registers to identify gaps in referral feasibility, simplified antibiotic treatment, and follow-up. Focus group discussions (FGD) and in-depth interviews (IDI) were conducted with both caregivers (6 FGDs; 23 IDIs) and providers (2 FGDs; 28 IDIs) to assess caregiver acceptability of the guidelines. Referral to the hospital was not feasible for many families (83.3%; N = 160/192) and acceptance varied by infection severity. Barriers to referral feasibility included economic and household factors, and previous experiences with poor quality of care at the sub-district hospital. Conversely, providers and caregivers indicated high acceptability of simplified antibiotic treatment. 80% (N = 96/120) of infants with clinical severe infection for whom referral was not feasible returned to the facility for the second antibiotic injection. Some providers reported developing local solutions-including engaging informal providers in treatment of the infant-to address organizational barriers and promote treatment compliance. Follow-up of young infants receiving simplified treatment is critical, but only 67.4% (N = 87/129) of infants received fourth day follow-up. Some providers' reported deviations from the guidelines that shifted responsibility of follow-up to the caregiver, which may have contributed to lapses. Caregivers' perception of trust and communication with providers were influential in caregiver acceptability of care. Few caregivers accepted referral to the sub-district hospital, suggesting low acceptability of this option. When referral was not feasible, many caregivers reported satisfaction with simplified antibiotic treatment. Local solutions described by providers require further examination in this context to assess the safety and potential value of these strategies in outpatient treatment. Our findings suggest strengthening providers' interpersonal skills could improve caregiver acceptability of the guidelines.


[Three-dimensional finite element analysis of the stress distribution of bone tissue around porous titanium implant].
Objective: To analyze the stress distribution of different types of bone tissue around porous titanium implant in different mechanical loads and to further evaluate the biomechanical properties of porous titanium implant. Methods: Finite element (FE) models of implant restorations for the maxillary first premolar was established, and the diameter of implants in the models was 4.1 mm. Five models was constructed according to diameter of implant central pillar and the thickness of outer porosity: solid group (group A), central pillar 1.5 and 3.1 mm and outer porosity 30% (group B and C), central pillar 1.5 and 3.1 mm and outer porosity 40% (group D and E). Different loads (150 N vertical force, 50 N lateral force) were applied to the occlusal surface of implant restorations in type Ⅲ bone and maximal von Mises stress was evaluated. Meanwhile, a couple of simplified maxillary part models varied in four types of bone were constructed with the implants bearing load of simulation ultimate force to evaluate the stress distribution of different types of bone. Results: With different mechanical loading, the stress value of bone tissue around porous implant (group B-E) was greater than that in the solid structure (group A). Under the load of simulation ultimate force, the maximum stress of the bone rised with the increase of porosity and thickness of the porous implant. And the maximum stress value of the surrounding bone tissue changed with the change of bone. Under vertical loading, the maximal von Mises stress of the bone around solid implants of group A was 17.56 MPa, which was a little lower than that of group B and C. And the maximal equivalent von Mises stress of group D and E was 69.24 MPa. The results of lateral force and simulation ultimate force loading were similar. The stress of the bone tissue around implant increased with the decrease of bone quality. The maximum stress value of group D implant was 134.95 MPa. Conclusions: Porous structure of the implant is conducive to transmit stress to surrounding bone tissue and increases the mechanical stimulation of bone. However, if the value and direction of load are inappropriate or quality of bone is poor, pathological stress may be produced.


Diagnostic issues affecting the epidemiology of fetal alcohol spectrum disorders.
Epidemiological measures of the prevalence of fetal alcohol spectrum disorders (FASD) vary greatly in the literature. Irrespective of the methodology, the criteria to define a 'case' are set by the researchers. Hence, estimates of the prevalence of FASD primarily depend on the diagnostic criteria currently available. The problem lies therein - the aforementioned criteria are ill-defined. A critical analysis of the diagnostic criteria from the Institute of Medicine, Hoyme, 4-Digit Diagnostic Code and Canadian guidelines was performed, with particular attention focused on the inconsistencies in specificities of the fetal alcohol syndrome (FAS) facial phenotype. To date, the Canadian guidelines represent the only guidelines that have pushed for a uniform diagnostic capacity through harmonizing the IoM and 4-Digit Diagnostic Code criteria. In the absence of a reliable biochemical marker of effect to confirm maternal drinking during pregnancy, the importance and dependence on diagnostic guidelines for FASD is understated. With the availability of four published guidelines for diagnoses across the spectrum of FASD, there is a need to reach a set standard globally. There are profound implications of relaxed and strict diagnostic approaches on FAS prevalence reporting in the literature. This review exposes the clinical burden of diagnosing the range of FASD with disputing diagnostic criteria. Discrepancies in the criteria pose a danger to the validity of FASD diagnoses with respect to inaccurate estimates of incidence and prevalence. In turn, these discrepancies risk compromising the future healthcare of affected individuals with regards to intervention, counselling and treatment.


The influence of ageing on the development and management of rheumatoid arthritis.
The population of elderly individuals with rheumatoid arthritis (RA) is expanding, due mainly to increasing life expectancy. A variety of theories have been proposed to explain the ageing process, including accumulation of DNA damage and resultant changes in biological processes. Such changes can influence the development and/or course of disease. Furthermore, alterations in biological function determine the biological age-as opposed to chronological age-of an individual, which strongly influences their ability to cope with disease. Moreover, comorbidities are more frequent in elderly individuals. Together, these factors complicate treatment of disease and necessitate careful patient management. Indeed, although evidence from clinical trials suggests that DMARDs and biologic agents have good efficacy and are well tolerated in elderly patients with RA, such individuals are often undertreated and inadequately managed. Unfortunately, insufficient data are available for the development of evidence-based guidelines for this population, as elderly patients are often excluded from clinical trials owing to age restrictions or comorbidities. Thus, additional clinical studies in elderly patients are warranted, with treatment regimens tailored according to vitality or frailty parameters. This Review focuses on the pathophysiological aspects of ageing and their implications for the management of RA in elderly patients.


RFS+: A Clinically Adaptable and Computationally Efficient Strategy for Enhanced Brain Tumor Segmentation.
Automated brain tumor segmentation has significant importance, especially for disease diagnosis and treatment planning. The study utilizes a range of MRI modalities, namely T1-weighted (T1), T1-contrast-enhanced (T1ce), T2-weighted (T2), and fluid-attenuated inversion recovery (FLAIR), with each providing unique and vital information for accurate tumor localization. While state-of-the-art models perform well on standardized datasets like the BraTS dataset, their suitability in diverse clinical settings (matrix size, slice thickness, manufacturer-related differences such as repetition time, and echo time) remains a subject of debate. This research aims to address this gap by introducing a novel 'Region-Focused Selection Plus (RFS+)' strategy designed to efficiently improve the generalization and quantification capabilities of deep learning (DL) models for automatic brain tumor segmentation. RFS+ advocates a targeted approach, focusing on one region at a time. It presents a holistic strategy that maximizes the benefits of various segmentation methods by customizing input masks, activation functions, loss functions, and normalization techniques. Upon identifying the top three models for each specific region in the training dataset, RFS+ employs a weighted ensemble learning technique to mitigate the limitations inherent in each segmentation approach. In this study, we explore three distinct approaches, namely, multi-class, multi-label, and binary class for brain tumor segmentation, coupled with various normalization techniques applied to individual sub-regions. The combination of different approaches with diverse normalization techniques is also investigated. A comparative analysis is conducted among three U-net model variants, including the state-of-the-art models that emerged victorious in the BraTS 2020 and 2021 challenges. These models are evaluated using the dice similarity coefficient (DSC) score on the 2021 BraTS validation dataset. The 2D U-net model yielded DSC scores of 77.45%, 82.14%, and 90.82% for enhancing tumor (ET), tumor core (TC), and the whole tumor (WT), respectively. Furthermore, on our local dataset, the 2D U-net model augmented with the RFS+ strategy demonstrates superior performance compared to the state-of-the-art model, achieving the highest DSC score of 79.22% for gross tumor volume (GTV). The model utilizing RFS+ requires 10% less training dataset, 67% less memory and completes training in 92% less time compared to the state-of-the-art model. These results confirm the effectiveness of the RFS+ strategy for enhancing the generalizability of DL models in brain tumor segmentation.


[Transcutaneous transcatheter use of laser recanalization of coronary arteries in patients with ischemic heart disease].
The paper discusses the potential possibility and effectiveness of X-ray endovascular laser recanalization (ELR) of the coronary arteries in order to treat coronary atherosclerosis in patients with coronary heart disease. The intervention was performed in 4 patients (into the anterior interventricular artery in 3 and into the right coronary artery in 1). In 3 of 4 cases, X-ray ELR proved to be successful, in one case the intervention failed due to technological reasons. Recanalization of a completely occluded segment of the coronary artery with a residual stenosis of no more than 40% was observed in two cases. Laser recanalization of profound local coronary stenosis was made in the mid-third of the vessel in one case. It can be stated that X-ray ELR of the coronary artery may extend the scope of X-ray surgical therapeutical tools of the treatment of coronary atherosclerosis. At the same time, accumulation of clinical experience and further improvement of laser and laser catheter engineering are essential in defining the value and possible scope for the application of this method.


The impact of non-pharmacologic interventions on respiratory syncytial virus hospitalization during the COVID-19 pandemic: A retrospective study from Saudi Arabia.
Acute respiratory tract infections (ARTIs) are frequently observed in infants and young children. The dynamics and transmission of ARTIs have been significantly impacted by the global COVID-19 pandemic. This study investigates the change in admission rates of Respiratory Syncytial virus (RSV) in a hospital in Saudi Arabia. The study included hospitalized pediatric patients who underwent RSV testing in three periods: pre-pandemic (2019), during the pandemic (2020-2021), and the immediate post-pandemic (2022). RSV testing was conducted using either PCR or antigen tests. Between January 2019 to December 2022, out of 927 tested patients, 173 (18.7%) were positive for RSV. The yearly positivity rates were as follows: 42 (35.6%) of 118, 24 (33.3%) of 72, 15 (18.5%) of 81, and 92 (14%) of 656, yearly from 2019 to 2022, respectively (P < 0.00001). Among all cases, 150 (16.2%) required hospitalization, including 94 (62.7%) males and 56 (37.3%) females. The admission rate to the intensive care unit (ICU) was 25 (16.7%), and mechanical ventilation was required for 10 (6.6%) patients. The overall case fatality rate was 0.7%. A Binary logistic regression analysis showed upper respiratory tract symptoms were more common in patients admitted in 2019 compared to 2022 (odd ratio:20.9, 95% CI: 4.2-104.1, P value < 0.0001). The study showed that there were differences in RSV infection before and after COVID-19. Understanding how COVID-19 mitigation measures affect RSV transmission can aid in the development of effective prevention and control strategies.


Can we help just by good intentions? A meta-analysis of experiments on distant intention effects.
In recent years, several clinical trials have assessed effects of distant healing. The basic question raised by these studies is whether a positive distant intention can be related to some outcome in a target person. There is a specific simple experimental setup that tests such a basic assumption. The task is to focus attention and to indicate unwanted mind wandering by a button press while at the same time a second remote person is either supporting this performance or not according to a randomized schedule. A meta-analysis was conducted to assess the overall effect of this experimental approach. A systematic literature search yielded 11 eligible studies, with 576 single sessions and almost identical design, that were conducted on three different continents. Study parameters were extracted and combined with a random-effects model. The model yielded an overall effect size of d=0.11 (p=0.03). Furthermore, there was a significant difference of the frequency of button presses between studies conducted in Indonesia and the Western hemisphere (p<0.001). Two (2) similar experimental setups applying electrodermal activity as dependent variable meta-analyzed earlier showed almost identical effect sizes. This can be considered as mutual validation of the three data sets. The hypothesis of the positive effect of benevolent intentions is supported by the data presented. It is concluded that especially the intentional aspect common to all three different tasks may be responsible for these unorthodox findings. These finding may have implications for distant healing research and health care as well as for meditation performance.


Can acupuncture treatment be double-blinded? An evaluation of double-blind acupuncture treatment of postoperative pain.
Blinding protects against bias but the success of blinding is seldom assessed and reported in clinical trials including studies of acupuncture where blinding represents a major challenge. Recently, needles with the potential for double-blinding were developed, so we tested if acupuncture can be double-blinded in a randomized study of sixty-seven patients with acute pain ≥ 3 (0-10 scale following third molar removal) who received active acupuncture with a penetrating needle or placebo acupuncture with a non-penetrating needle. To test if acupuncture was administered double-blind, patients and acupuncturists were asked about perceived treatment allocation at the end of the study. To test if there were clues which led to identification of the treatment, deep dull pain associated with needle application and rotation (termed "de qi" in East Asian medicine), and patients' pain levels were assessed. Perceived treatment allocation depended on actual group allocation (p < 0.015) for both patients and acupuncturists, indicating that the needles were not successful in double-blinding. Up to 68% of patients and 83% of acupuncturists correctly identified the treatment, but for patients the distribution was not far from 50/50. Also, there was a significant interaction between actual or perceived treatment and the experience of de qi (p = 0.027), suggesting that the experience of de qi and possible non-verbal clues contributed to correct identification of the treatment. Yet, of the patients who perceived the treatment as active or placebo, 50% and 23%, respectively, reported de qi. Patients' acute pain levels did not influence the perceived treatment. In conclusion, acupuncture treatment was not fully double-blinded which is similar to observations in pharmacological studies. Still, the non-penetrating needle is the only needle that allows some degree of practitioner blinding. The study raises questions about alternatives to double-blind randomized clinical trials in the assessment of acupuncture treatment.


Acute effects of an oral supplement of (-)-epicatechin on postprandial fat and carbohydrate metabolism in normal and overweight subjects.
Postprandial hyperglycemia, in particular when accompanied by excessive hypertriglyceridemia, is associated with increased cardiovascular risk, mainly in overweight or obese subjects, as it favors oxidative stress, systemic inflammation and endothelial dysfunction. Thus, treatments that favorably modulate metabolism by reducing steep increases in postprandial serum glucose and triglycerides, are of considerable interest. Evidence suggests that (-)-epicatechin (EPI) is responsible for reductions in cardiometabolic risk associated with chocolate consumption; these effects may be associated with favorable effects of EPI on postprandial metabolism. The aims of this study were to assess the effects of EPI on postprandial metabolism in normal-weight and overweight/obese subjects. Twenty adult volunteers (normal and overweight) underwent oral metabolic tolerance tests in the absence and presence of oral EPI (1 mg kg(-1)). Metabolic responses were examined using indirect calorimetry and determining blood glucose and triglycerides at 0, 2 and 4 hours after metabolic load ingestion. Results show that EPI increased postprandial lipid catabolism, as evidenced by a significant decrease in the respiratory quotient, which implies an increase in fat oxidation. The effect was associated with significantly lower postprandial plasma glucose and triglycerides concentrations. The effects were more prominent in overweight subjects. In conclusion, EPI modulates postprandial metabolism by enhancing lipid oxidation accompanied by reductions in glycemia and triglyceridemia.


ECOLOGY: Forest Fire Plan Kindles Debate.
Forest fires burning in the western United States have already scorched over 2.5 million hectares this summer. Now a federal proposal to prevent them by paying loggers to cut smaller trees is generating heat among ecologists, who say the approach may not be right for all forests--or all fires.


The ARTS of p53-dependent mitochondrial apoptosis.
The tumor-suppressive activity of p53 is largely attributed to its ability to induce cell death, including apoptosis, through transcription-dependent and transcription-independent mechanisms. On the one hand, nuclear p53 transcriptionally activates the expression of a myriad of pro-apoptotic BCL-2 family genes, such as NOXA, PUMA, BID, BAD, BIK, BAX, etc., whereas it inactivates the expression of anti-apoptotic BCL-2, BCL-XL, and MCL1, leading to mitochondrial apoptosis. On the other hand, cytoplasmic p53 also promotes mitochondrial apoptosis by directly associating with multiple BCL-2 family proteins in the mitochondria. Apoptosis-related protein in TGF-β signaling pathway (ARTS), a mitochondria-localized pro-apoptotic protein encoded by an alternative spliced variant of the SEPT4 gene, triggers apoptosis by facilitating proteasomal degradation of BCL-2 and XIAP upon pro-apoptotic stimuli. We recently identified SEPT4/ARTS as a new p53 target gene in response to genotoxic stress. ARTS in turn binds to p53, drives its mitochondrial localization, and enhances the interaction between p53 and BCL-XL, thereby promoting mitochondrial apoptosis. This review will illustrate the mechanisms of p53-induced mitochondrial apoptosis, offer some recently discovered new insights into the functions of ARTS in regulating mitochondrial cell death, and discuss the clinical significance of ARTS in cancer and non-cancer diseases.


Ubiquitin ligase SYVN1/HRD1 facilitates degradation of the SERPINA1 Z variant/α-1-antitrypsin Z variant via SQSTM1/p62-dependent selective autophagy.
SERPINA1/AAT/α-1-antitrypsin (serpin family A member 1) deficiency (SERPINA1/ AAT-D) is an autosomal recessive disorder characterized by the retention of misfolded SERPINA1/AAT in the endoplasmic reticulum (ER) of hepatocytes and a significant reduction of serum SERPINA1/AAT level. The Z variant of SERPINA1/AAT, containing a Glu342Lys (E342K) mutation (SERPINA1E342K/ATZ), the most common form of SERPINA1/AAT-D, is prone to misfolding and polymerization, which retains it in the ER of hepatocytes and leads to liver injury. Both proteasome and macroautophagy/autophagy pathways are responsible for disposal of SERPINA1E342K/ATZ after it accumulates in the ER. However, the mechanisms by which SERPINA1E342K/ATZ is selectively degraded by autophagy remain unknown. Here, we showed that ER membrane-spanning ubiquitin ligase (E3) SYVN1/HRD1 enhances the degradation of SERPINA1E342K/ATZ through the autophagy-lysosome pathway. We found that SYVN1 promoted SERPINA1E342K/ATZ, especially Triton X 100-insoluble SERPINA1E342K/ATZ clearance. However, the effect of SYVN1 in SERPINA1E342K/ATZ clearance was impaired after autophagy inhibition, as well as in autophagy-related 5 (atg5) knockout cells. On the contrary, autophagy induction enhanced SYVN1-mediated SERPINA1E342K/ATZ degradation. Further study showed that SYVN1 mediated SERPINA1E342K/ATZ ubiquitination, which is required for autophagic degradation of SERPINA1E342K/ATZ by promoting the interaction between SERPINA1E342K/ATZ and SQSTM1/p62 for formation of the autophagy complex. Interestingly, SYVN1-mediated lysine 48 (K48)-linked polyubiquitin chains that conjugated onto SERPINA1E342K/ATZ might predominantly bind to the ubiquitin-associated (UBA) domain of SQSTM1 and couple the ubiquitinated SERPINA1E342K/ATZ to the lysosome for degradation. In addition, autophagy inhibition attenuated the suppressive effect of SYVN1 on SERPINA1E342K/ATZ cytotoxicity, and the autophagy inducer rapamycin enhanced the suppressive effect of SYVN1 on SERPINA1E342K/ATZ-induced cell apoptosis. Therefore, this study proved that SYVN1 enhances SERPINA1E342K/ATZ degradation through SQSTM1-dependent autophagy and attenuates SERPINA1E342K/ATZ cytotoxicity.


Cross-sectional study of malnutrition and associated factors among school aged children in rural and urban settings of Fogera and Libo Kemkem districts, Ethiopia.
Little information is available on malnutrition-related factors among school-aged children ≥5 years in Ethiopia. This study describes the prevalence of stunting and thinness and their related factors in Libo Kemkem and Fogera, Amhara Regional State and assesses differences between urban and rural areas. In this cross-sectional study, anthropometrics and individual and household characteristics data were collected from 886 children. Height-for-age z-score for stunting and body-mass-index-for-age z-score for thinness were computed. Dietary data were collected through a 24-hour recall. Bivariate and backward stepwise multivariable statistical methods were employed to assess malnutrition-associated factors in rural and urban communities. The prevalence of stunting among school-aged children was 42.7% in rural areas and 29.2% in urban areas, while the corresponding figures for thinness were 21.6% and 20.8%. Age differences were significant in both strata. In the rural setting, fever in the previous 2 weeks (OR: 1.62; 95% CI: 1.23-2.32), consumption of food from animal sources (OR: 0.51; 95% CI: 0.29-0.91) and consumption of the family's own cattle products (OR: 0.50; 95% CI: 0.27-0.93), among others factors were significantly associated with stunting, while in the urban setting, only age (OR: 4.62; 95% CI: 2.09-10.21) and years of schooling of the person in charge of food preparation were significant (OR: 0.88; 95% CI: 0.79-0.97). Thinness was statistically associated with number of children living in the house (OR: 1.28; 95% CI: 1.03-1.60) and family rice cultivation (OR: 0.64; 95% CI: 0.41-0.99) in the rural setting, and with consumption of food from animal sources (OR: 0.26; 95% CI: 0.10-0.67) and literacy of head of household (OR: 0.24; 95% CI: 0.09-0.65) in the urban setting. The prevalence of stunting was significantly higher in rural areas, whereas no significant differences were observed for thinness. Various factors were associated with one or both types of malnutrition, and varied by type of setting. To effectively tackle malnutrition, nutritional programs should be oriented to local needs.


Major adverse cardiovascular events of vascular endothelial growth factor tyrosine kinase inhibitors among patients with different malignancy: A systemic review and network meta-analysis.
Vascular endothelial growth factor tyrosine kinase inhibitors (VEGF-TKIs) are a common cancer treatment. However, the pharmacologic characteristics of VEGF-TKIs may influence cardiovascular risks. The relative risks of major adverse cardiovascular events (MACEs) associated with VEGF-TKIs are poorly understood. We searched PubMed, Embase, and ClinicalTrials.gov from inception until August 31, 2021, for phase II/III randomized controlled trials of 11 VEGF-TKIs (axitinib, cabozantinib, lenvatinib, pazopanib, ponatinib, ripretinib, regorafenib, sorafenib, sunitinib, tivozanib, and vandetanib). The endpoints were heart failure, thromboembolism, and cardiovascular death. The Mantel-Haenszel method was used to calculate the risk of VEGF-TKI among users by comparing to nonusers. Pairwise meta-analyses with a random-effects model was used to estimate the risks of the various VEGF-TKIs. We estimated ranked probability with a P-score and assessed credibility by employing the Confidence in Network Meta-Analysis framework. We identified 69 trials involving 30 180 patients with cancer. The highest risk of MACEs was associated with high-potency tivazonib (odds ratio [OR] 3.34), lenvatinib (OR 3.26), and axitinib (OR: 2.04), followed by low-potency pazopanib (OR: 1.79), sorafenib (OR: 1.77), and sunitinib (OR: 1.66). The risk of heart failure significantly increased in association with less selective sorafenib (OR: 3.53), pazopanib (OR: 3.10), and sunitinib (OR: 2.65). The risk of thromboembolism significantly increased in association with nonselective lenvatinib (OR: 3.12), sorafenib (OR: 1.54), and sunitinib (OR: 1.53). Higher potency (tivozanib, axitinib) and lower selectivity (sorafenib, vandetanib, pazopanib, sunitinib) were associated with a higher probability of heart failure. Low selectivity (lenvatinib, cabozantinib, sorafenib, sunitinib) was associated with a higher probability of thromboembolism. Higher-potency and lower-selectivity VEGF-TKIs may influence the risks of MACEs, heart failure, and thromboembolism. These findings may facilitate evidence-based decision-making in clinical practice.


On Laterally Perturbed Human Stance: Experiment, Model, and Control.
Understanding human balance is a key issue in many research areas. One goal is to suggest analytical models for the human balance. Specifically, we are interested in the stability of a subject when a lateral perturbation is being applied. Therefore, we conducted an experiment, laterally perturbing five subjects on a mobile platform. We observed that the recorded motion is divided into two parts. The legs act together as a first, the head-arms-trunk segment as a second rigid body with pelvis, and the ankle as hinge joints. Hence, we suggest using a planar double-inverted pendulum model for the analysis. We try to reproduce the human reaction utilizing torque control, applied at the ankle and pelvis. The fitting was realized by least square and nonlinear unconstrained optimization on training sets. Our model is not only able to fit to the human reaction, but also to predict it on test sets. We were able to extract and review key features of balance, like torque coupling and delays as outcomes of the aforementioned optimization process. Furthermore, the delays are well within the ranges typically for such compensatory motions, composed of reflex and higher level motor control.


Neural control of heart rate is an arrhythmia risk modifier in long QT syndrome.
The purpose of this study was to test the hypothesis that differences in autonomic responses might modify clinical severity in long QT syndrome type 1 (LQT1) patients, those with KCNQ1 mutations and reduced I(Ks), in whom the main arrhythmia trigger is sympathetic activation. Some long QT syndrome (LQTS) patients experience life-threatening cardiac arrhythmias, whereas others remain asymptomatic throughout life. This clinical heterogeneity is currently unexplained. In a South African LQT1 founder population segregating KCNQ1-A341V, we correlated major cardiac events to resting heart rate (HR) and to baroreflex sensitivity (BRS) on and off beta-adrenergic blockers (BB). In 56 mutation carriers (MCs), mean HR was lower among asymptomatic patients (p < 0.05). Among MCs with a QT interval corrected for heart rate <or=500 ms, those in the lower HR tertile were less likely to have suffered prior cardiac events (odds ratio [OR] 0.19, 95% confidence interval [CI] 0.04 to 0.79, p < 0.02). The BRS was lower among asymptomatic than symptomatic MCs (11.8 +/- 3.5 ms/mm Hg vs. 20.1 +/- 10.9 ms/mm Hg, p < 0.05). A BRS in the lower tertile was associated with a lower probability of being symptomatic (OR 0.13, 95% CI 0.02 to 0.96, p < 0.05). A similar trend was observed during BB. The MCs in the lower tertile for both HR and BRS were less frequently symptomatic than MCs with different patterns (20% vs. 76%, p < 0.05). Subjects with either ADRA2C-Del322-325 or homozygous for ADRB1-R389, 2 polymorphisms predicting enhanced adrenergic response, were more likely to have BRS values above the upper tertile (45% vs. 8%, p < 0.05). Lower resting HR and "relatively low" BRS are protective factors in KCNQ1-A341V carriers. A plausible underlying mechanism is that blunted autonomic responses prevent rapid HR changes, arrhythmogenic when I(Ks) is reduced. These findings help understanding phenotypic heterogeneity in LQTS and identify a physiological risk modifier, which is probably genetically determined.


Interleukin-7 enhances antimicrobial activity against Leishmania major in murine macrophages.
Recently, it has been shown that interleukin-7 (IL-7) is able to induce secretion of cytokines and tumoricidal activity by human monocytes. This study shows that treatment of murine macrophages infected with Leishmania major with IL-7 without any other stimulus reduced the percentage of infected cells, as well as the parasite burden per cell, in a dose-dependent manner to a limited degree (45% reduction of the number of amastigotes per 100 macrophages). Simultaneous treatment of macrophages with gamma interferon and IL-7 led to nearly complete (> 99%) elimination of amastigotes. Addition of anti-tumor necrosis factor alpha or N omega-monomethyl-L-arginine acetate reversed the leishmanicidal effects of IL-7, and production of nitric oxide was induced in the presence of IL-7.


Structural studies on rat prostatic binding protein. The primary structure of its glycosylated component C3.
The amino acid sequence of the glycosylated component C3 of rat prostatic binding protein has been determined. The peptides obtained by digestion of the S-carboxamidomethylated or S-aminoethylated glycoprotein with trypsin and Staphylococcus aureus protease were sequenced by manual Edman degradation. The alignment of the fragments was further established with overlapping peptides obtained by enzymic hydrolysis of the modified protein with chymotrypsin and thermolysin, and by chemical cleavage with cyanogen bromide. The glycopeptide C3 contains 77 amino acids corresponding to a molecular weight of 8653. the oligosaccharide chain is attached to the peptide by an N-glycosidic bond to asparagine-17. C3 is an acidic polypeptide due to the presence of ten acidic residues; its three cysteine residues are located at both extremities and in the middle of the molecule.


Fluorescence ratio imaging microscopy shows decreased access of vancomycin to cell wall synthetic sites in vancomycin-resistant Staphylococcus aureus.
A new method of fluorescence ratio imaging microscopy was used to compare the in vivo binding capacity and the access of a fluorescent derivative of vancomycin to the cell wall synthetic sites in isogenic pairs of vancomycin-susceptible and -resistant laboratory mutants and vancomycin-intermediate and -susceptible clinical isolates of Staphylococcus aureus. Live cells of resistant strains were found to bind approximately 1.5 times more antibiotic, but there was no correlation between the increased binding capacity and the MICs of the strains. In both susceptible and resistant bacteria, the subcellular sites of wall synthesis were localized to the division septa, but the rate of diffusion of drug molecules to these sites was reduced in resistant cells. The findings allow a reinterpretation of the mechanism of vancomycin resistance in which the path of vancomycin to its lethal target (lipid II) is considered to be through the division septum and therefore is dependent on the stage of the staphylococcal cell cycle.


Isolated acute knee injuries in the general population.
Knee injuries are common in the general population, yet most studies have concentrated on special populations. The purpose of this study was to describe the types of injuries, injury events, age, and sex distributions in a community population. The medical records of all residents of Olmsted County, Minnesota, 17 years of age or older who had a first visit for an isolated acute knee (acute was defined as occurring within 60 days of the physician visit) injury between January 1, 1993, and January 1, 1996, were abstracted. A total of 664 patients (4/1,000 community adults, 46% women and 54% men) presented to a physician for evaluation and treatment of an isolated acute knee injury. Injured men were on average younger than women and more likely to have an injury during a sports activity, whereas women's injuries were more likely to be the result of non-sports-related falls. Knee sprain or strain was the most common final diagnosis (36%). Approximately half (49%) of the patients had a single visit. The likelihood of orthopedic surgeon's care (37% overall) increased with injury severity and age of the patient. Overall, 12% of subjects had surgical treatment recommended. Knee injuries are common, often result in a single visit, seldom receive surgical intervention, and the majority are cared for entirely by generalist physicians.


[Capillary electrophoresis with field-enhanced stacking for determination of water-soluble active principles in Salvia miltiorrhiza var. miltiorrhiza f. alba].
A simple and sensitive capillary electrophoresis method with field-enhanced stacking concentration for the analysis of protocatechuic aldehyde, protocatechuic acid, danshensu, rosmarinic acid and salvianolic acid B in Salvia miltiorrhiza var. miltiorrhiza f. alba was developed. The separation was achieved with a fused-silica capillary (75 microm x 50.2 cm, effective length was 40 cm) and a running buffer 15 mmol x L(-1) borax (pH 10.0) containing 20% CH3 OH. The UV detection wavelength was 210 nm. The applied voltage was 28 kV, and the cartridge temperature was 25 degrees C. Water plug was introduced from the anode by 0.5 psi x 4 s before injection. Sample was injected by electrokinetic injection - 8 kV x 3 s. The linear range of protocatechuic aldehyde is 3.0-60.00 mg x L(-1) (R2 = 0.999 8); that of protocatechuic acid, danshensu, rosmarinic acid and salvianolic acid B are 1.0-20.00 mg x L(-1) (R2 are 0.999 1, 0.999 4, 0.998 9 and 0.999 8, respectively), and the limits of detection of five analyts are 0.55, 0.40, 0.25, 0.32, 0.38 microg x L(-1), respectively, Stacking factor is higher and precision is satisfactory. The recoveries ranges were from 97.3% to 99.8%. The proposed method was used to determine the protocatechuic aldehyde, protocatechuic acid, danshensu, rosmarinic acid and salvianolic acid B in S. miltiorrhiza var. miltiorrhiza f. alba. The proposed method is simple, rapid, accurate and high sensitivity, and can be used to control of the quality of S. miltiorrhiza var. miltiorrhiza f. alba.


Normal primitive haemopoietic progenitors are more frequent than their leukaemic counterpart in newly diagnosed patients with chronic myeloid leukaemia but rapidly decline with time.
We carried out studies to quantify Ph-negative progenitors both in steady state and during regeneration after chemotherapy and G-CSF in 23 newly diagnosed chronic myeloid leukaemia (CML) patients (group A) and in 14 individuals more than a year from diagnosis (nine in chronic and five in accelerated phase, group B). In steady-state bone marrow, Ph-negative long-term culture initiating cells (LTC-IC) and Ph-negative colony-forming-cells (CFC) were detected in 18/23 and 14/23 patients of group A versus 3/14 and 3/14 patients of group B (P<0.001 and P<0.02, respectively). The absolute number of mobilized Ph-negative progenitors was markedly higher in group A versus group B (P<0.02 for LTC-IC, P<0.003 for CFC). 12/16 newly diagnosed patients mobilized Ph-negative LTC-IC only and the yield was in the range of normal allogeneic donors. Overall the frequency of Ph-negative LTC-IC in the bone marrow predicted the yield of Ph-negative LTC-IC mobilized into peripheral blood (P<0.001). The bone marrow frequency of Ph-positive LTC-IC was considerably lower than the normal counterpart. Taken together, these findings suggest that normal progenitors are relatively well preserved in newly diagnosed CML patients, but tend to rapidly decline with time. This observation helps in the understanding of the pathogenesis of CML and has potential implications for autografting. The optimal time for a successful collection of Ph-negative circulating progenitors would appear to be soon after diagnosis.


Study and characterization of different developmental stages of Leishmania major.
The complete developmental cycle of Leishmania major in axenic culture was achieved by simply changing the temperature whether sudden from 22 degrees C to 37 degrees C or stepwise 22 degrees C, 29 degrees C and then 37 degrees C. The morphology by light microscopy, GPI isoenzyme pattern and PCR amplification of minicircle of kinetoplast DNA of the different stages were studied. The amastigotes obtained from the foot pads of mice were compared to those obtained from axenic culture. The GPI isoenzyme pattern and the PCR amplification products showed distinct differences between the promastigotes and the amastigotes. The amastigotes of the two sources also showed differences after temperature changes.


Effects of antimicrobial drugs on human neutrophil-microbe interactions.
Antimicrobial drugs may influence neutrophil-microbe interactions in several ways, and, conversely, neutrophils may interfere with the action(s) of antimicrobial drugs. Here, evidence for the existence of such effects is evaluated and attention drawn to the problems of in vitro experimentation in this area. The review is restricted to those studies that used human neutrophils, clinically achievable drug concentrations, and were well designed. Even so, it is noted that little attempt has been made to investigate underlying mechanisms. The effects of drugs on microbes, which influence neutrophil-microbe interactions, such as concentration and the post-antibiotic effect, are considered. The penetration of antimicrobial drugs into neutrophils and subsequent intracellular activity is discussed and contrasted with observations obtained using macrophages. Overall, neutrophil-microbe interactions are complex and difficult to dissect, and carefully designed experiments using closely defined conditions are required if meaningful results are to be obtained.


Pulmonary function tests and high resolution computed tomography of the lungs in patients with rheumatoid arthritis.
To compare the results of pulmonary function tests (PFTs) and high resolution computed tomography (HRCT) of the lungs in rheumatoid arthritis (RA) patients. Sixty eight patients (54 women, 14 men) fulfilling the revised criteria for RA were consecutively included in a transversal prospective study. Their mean age was 58.8 years (range: 35-82) and the mean duration of the disease was 12 years (range: 5-16). Rheumatoid factor was positive in 52 patients (76.5%). Fifty two patients (76.5%) were lifelong non-smokers. Detailed medical and drug histories were obtained. PFTs comprised spirometry and gas transfer measurements. Results for PFTs were expressed as percentage of predicted values for each individual adjusted for age, sex, and height. HRCT was undertaken with a Siemens Somatom Plus. A significant decrease of FEV1/ FVC, FEF25%, FEF50%, FEF75%, FEF25-75%, and TLCO was observed (p < 0.05) and 13.2% of the patients had a small airways involvement defined by a decrease of FEF25-75% below 1.64 SD. The most frequent HRCT findings were: bronchiectasis (30.5%), pulmonary nodules (28%), and air trapping (25%). The patients with small airways involvement had a high frequency of recurrent bronchitis (75% v 34%, p = 0.05) and bronchiectasis (71% v 23%, p = 0.019). The patients with bronchiectasis were characterised by low values of FEV1, FVC, FEF25-75%, and TLCO (p < 0.01), a high prevalence of small airways involvement (29% v 5%, p = 0.019), and a low prevalence of HLA DQA1 *0501 allele (14% v 33%, p < 0.05). This study suggests a significant association between small airways involvement on PFTs and bronchiectasis on HRCT in unselected RA patients.


Anti-diabetic effects of natural and modified 'Ganzhou' navel orange peel pectin on type 2 diabetic mice via gut microbiota.
Pectin, a kind of dietary fiber, has attracted much attention owing to its beneficial effect on human health in recent years. In this study, the effects of both 'Ganzhou' navel orange peel pectin (GOP) and modified GOP (MGOP) on type 2 diabetes (T2DM) were investigated. The results indicated that GOP and MGOP intervention had positive effects on T2DM in C57BL/6 mice. After modification, pectin can be changed into low methoxy pectin (LMP) and the content of GalA can increase, which endow MGOP with significant effects on improving lipid metabolism (TC, TG, and LDL-C decreased by 30.46%, 50%, and 37.56%, respectively, and HDL-C increased by 56%) and OGTT, further reducing insulin resistance (insulin decreased by 74.35%). In addition, MGOP was superior to GOP in improving oxidative stress (GSH and GSH-Px increased by 52.05% and 29.08% respectively, and MDA decreased by 84.02%), inhibiting inflammation and promoting SCFA synthesis. 16S rRNA analysis showed that MGOP changed the composition of intestinal microbiota in diabetic mice, decreased the abundance of Alistipes, Helicobacter and Oscillibacter, and increased the relative abundance of Dubosiella, Akkermansiaceae, and Atopobiaceae. The phenotypes of the gut microbiome also changed accordingly, which showed that MGOP significantly inhibited the growth of Gram-negative bacteria and potential pathogenic bacteria and reversed the related complications. Taken together, our findings revealed that MGOP intake regulated lipid metabolism and oxidative stress and improved the gut health of mice, with promising effects against T2DM and related complications.


Effects of dose-response of topical administration of nimodipine on cerebral vasospasm after subarachnoid hemorrhage in rabbits.
To explore the dose-response effects of topical administration of nimodipine on cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH) in rabbits. The CVS model was established by injection of fresh autologous nonheparinized arterial blood into the subtemporal area of basilar cisterns. The 24 CVS animals were randomly divided into 4 groups, group I (n=7): nimodipine original stock solution/normal saline=1/19 (0.01 mg/mL); group II (n=6): nimodipine original stock solution/normal saline=1/9 (0.02 mg/mL); group III (n=5): nimodipine original stock solution/normal saline=1/4 (0.04 mg/mL); and group IV (n=6) with no nimodipine, but 5% ethanol dissolved in normal saline as the control group. The operative area was administrated with nimodipine at different concentrations or alcohol-saline at 3 days after SAH. The blood flow velocity of middle cerebral artery was measured at 5, 15, 30, and 60 minutes after topical administration of nimodipine by transverse cerebellar diameter monitoring. Blood flow velocity of middle cerebral artery in group II (0.02 mg/mL) and in group III (0.04 mg/mL) significantly decreased at 60 and 15 minutes, respectively, after topical administration of nimodipine (P<0.05), and even more significantly at 30 and 60 minutes after topical administration of nimodipine in group III (0.04 mg/mL) (P<0.01). Topical administration of nimodipine at the concentrations of 1:5 (0.04 mg/mL) and 1:10 (0.02 mg/mL) significantly alleviates CVS after SAH, which indicates that topical administration of nimodipine may be useful for CVS of patients with SAH during surgical clip of intracranial aneurysms.


Utility of chromosomal chromogenic in situ hybridization as an alternative to flow cytometry and cytogenetics in the diagnosis of early partial hydatidiform moles: a validation study.
The introduction of p57 immunohistochemistry has aided the distinction between early complete moles (CMs) and hydropic abortus (HA), but no single technique has emerged for the distinction between early partial moles (PMs) and HA. Flow cytometry and cytogenetics have been used, but these require specialized equipment/expertise. The goal of this study is validation of chromosome in situ hybridization (CrISH), focusing on comparing the results to those obtained by cytogenetic methods. Archival paraffin blocks from molar and nonmolar gestations were retrieved. Sections were labeled with a chromosome 10 probe. Hybridization and visualization were performed using standard protocols. One hundred nuclei per sample were scored for the number ofsignals. Of 50 hydatidiform moles, 22 were PMs and 28 were CMs. The CMs showed 2 signals in 25 cases and 4 signals in 3 cases. The PMs showed 3 signals in 21 cases and 2 signals in 1 case. For the HAs there were 2 signals in 24 cases, and 1 case had 3 signals. Concordance between CrISH and flow cytometry studies for molar gestations was 95%. CrlSH is a highly effective adjunct in differentiating between PM and CM and between PM and HA. CrlSH is a simple, cost effective adjunct in evaluating molar gestations.


Evaluation of an inservice training program on dementia and wandering.
This study assessed the impact of an inservice training program in four nursing homes on nursing staff's knowledge of dementia, pacing/wandering behavior and management strategies, staff satisfaction, and their perceptions of work difficulty and quality of care. Additionally, unit-level behavioral observations of agitated behavior and the nature of resident interactions with staff members and other residents were also performed to assess whether changes in nursing staff's knowledge impacted the residents or interactions with the residents. Analyses revealed that quiz scores were significantly improved immediately following the inservice program as compared to pretest but returned to near pretest levels at the 1-month follow-up. Additionally, nursing staff reported that residents were allowed to pace/wander to a greater extent at follow-up compared to pretest. Methods for improving the effectiveness of inservice training are discussed.


Induction of fatty acid synthase and S14 gene expression by glucose, xylitol and dihydroxyacetone in cultured rat hepatocytes is closely correlated with glucose 6-phosphate concentrations.
It is now well established that the transcription of several genes belonging to the glycolytic and lipogenic pathway is stimulated in the presence of a high glucose concentration in adipocytes and hepatocytes. We have previously proposed that glucose 6-phosphate could be the signal metabolite that transduces the glucose effect. This proposal has recently been challenged and both an intermediate of the pentose phosphate pathway, xylulose 5-phosphate, and metabolites of the later part of glycolysis (3-phosphoglycerate and phosphoenolpyruvate) have been proposed. To discriminate between these possibilities, we have measured concomitantly, in primary cultures of adult rat hepatocytes, the expression of the fatty acid synthase (FAS) and S14 genes and the concentration of glucose metabolites. We have used various substrates entering at different steps of the glycolytic pathway (glucose, dihydroxyacetone) and the pentose phosphate pathway (xylitol). When compared with 5 mM glucose, 25 mM glucose induces a marked increase in both S14 and FAS gene expression, detectable as early as 2 h and peaking at 6 h. Increasing concentrations (1-5 mM) of xylitol and dihydroxyacetone in the presence of 5 mM glucose are also able to induce S14 and FAS gene expression progressively. Among the various glucose metabolites measured, glucose 6-phosphate, in contrast with xylulose 5-phosphate and metabolites of the lower part of glycolysis, is the only one that shows a clear-cut parallelism between its concentration and the degree of S14 and FAS gene expression. We conclude that glucose 6-phosphate is the most likely signal metabolite for the glucose-induced transcription of this group of genes.


EVIDENCE REQUIREMENTS FOR REIMBURSEMENT OF PHARMACEUTICALS ACROSS EUROPE.
The objective of this study was to compare evidence requirements for health technology assessment of pharmaceuticals by national agencies across Europe responsible for reimbursement decisions focusing specifically on relative effectiveness assessment. Evidence requirements from thirty-three European countries were requested and twenty-nine national agencies provided documents to review. Data were extracted from national documents (manufacturer's submission templates and associated guidance) into a purpose-made framework with categories covering information about the health condition, the technology, clinical effectiveness and safety. The level of detail in the required evidence varies considerably across countries. Some countries include specific questions while others request information under general headings. Some countries include all information in a single document, which may or may not include guidance on how to complete the template. Others have specific guidance documents or methods and process manuals that help with the completion of the submission templates. Despite differences in quantity and detail, the content of the evidence requirements is broadly similar. All countries ask for information on the health technology, target disease, and clinical effectiveness and safety. However, one country only requests clinical effectiveness information as part of cost-effectiveness analyses. We found twenty-six evidence requirements for which generic answers may apply across borders and nineteen in which countries requested nationally specific information. This work suggests that it would be possible to put together a minimum set of evidence requirements for HTA to support reimbursement decisions across Europe which could facilitate collaboration between jurisdictions.


[Pyourachal cyst. A diagnosis to keep in mind].
Urachal conditions are relatively exceptional, cystic formations being the most commonly reported. Urachal cysts remain silent and are not diagnosed except for the presence of associated complications such as: infection, lithiasis, re-permeabilization and malignant degeneration. This paper presents one patient with an infected urachal cyst (pyourachal) which required surgical exeresis. A literature revision is made of the clinical, diagnostic and therapeutic aspects. Emphasis is made on the relevance of a high index of suspicion considering its small incidence, the absence of specific clinical sings and the possibility of mimicking different abdominal pictures. Also the usefulness of ultrasound and computerized axial tomography in the diagnosis, and the need for surgical exeresis, including a vesical cup rim, are emphasised.


Do older adults' beliefs about their community mobility predict walking performance?
Among older adults, preserving community mobility (CM) is important for maintaining independent living. We explored whether perceptions of the environment and self-efficacy for CM (SE-CM) would predict walking performance for tasks reflecting CM. We hypothesized that perceptions of the environment and SE-CM would be additive predictors of walking performance on tasks reflecting the complexity of CM. Independent living older adults (N = 60) aged 64-85 completed six complex walking tasks (CWTs), SE-CM, and the environmental analysis of mobility questionnaire (EAMQ). Multiple regression analyses indicated that for each CWT, the EAMQ scales predicted walking performance (range: model R2Adj. = .078 to .139, p < .04). However, when SE-CM was added to the models, it was the sole significant predictor (p < .05). Contrary to our hypotheses, SE-CM was the best predictor in the additive models. SE-CM may be more correspondent to walking tests and thus a more sensitive predictor of CM walking performance.


Evidence for constriction of canine prenodal lymphatic vessels by vasoactive agents and carotid occlusion.
We measured pressure in a prenodal lymphatic in the canine forelimb during constant flow pump-perfusion of the brachial artery. We made bolus i.a. injections of 1.0 micrograms angiotensin II, norepinephrine, bombesin, or bradykinin, 20 micrograms 5-hydroxytryptamine (5HT), or occluded the carotid arteries. Norepinephrine, 5HT, or carotid occlusion produced regular rises in forelimb perfusion pressure and in lymphatic pressure. Angiotensin II increased forelimb arterial pressures but increased lymphatic pressure in only four experiments. Bombesin increased artery pressures but did not affect lymphatic pressure. Small vein pressure was increased by carotid occlusion, 5HT and norepinephrine. Increases in lymphatic pressure were coincident with increases in vein pressure but no related in magnitude. Bradykinin decreased forelimb arterial and venous pressures but did not affect lymphatic pressure. Either active constriction of lymphatic vessels or passive compression by movements of adjacent blood vessels could increase lymphatic pressure. These data do not preclude a passive component of pressure rise in the lymphatics nor do they support the concept. We conclude that active constriction of prenodal lymphatic vessels in the dog forelimb can occur in response to circulating vasoactive agents and bilateral carotid occlusion.


Pharmacological, pharmacokinetic, and clinical properties of benidipine hydrochloride, a novel, long-acting calcium channel blocker.
Benidipine is a dihydropyridine-derived calcium channel blocker developed in Japan, with several unique mechanisms of action, that is, triple calcium channels (L, N, and T) blocking action with a membrane approach. Benidipine has relatively high vascular selectivity and is expected to show protective effects on vascular endothelial cells. Renal protective effects of benidipine also have been shown in several basic and clinical studies. Moreover, anti-oxidative action and enhancing nitric oxide production have been noted with this drug, following its cardio-protective effects in patients with ischemic heart diseases. In fact, benidipine exerted a better prognostic effect than other calcium channel blockers in the therapy for patients with vasospastic angina. In addition, benidipine showed reliable antihypertensive, renoprotective effects if used in combination with angiotensin II type 1 receptor blockers (ARBs) when adequate anti-hypertensive effects are not achieved by ARBs alone, indicating that benidipine is an useful calcium channel blocker in combination therapy for hypertension. Benidipine was launched on the Japanese market 14 years ago, but few severe side effects have been reported, suggesting that this is a drug with established safety and long-acting pharmacological effects.


Severe injuries in football players. Influencing factors.
The aims of this prospective study were to analyze factors related to the occurrence of severe football injuries in players of different ages (14 to 42 years) and different skill levels (local teams to first league teams). In the Czech Republic, 398 players were followed up for 1 year, during which time they sustained 686 injuries. Of these, 113 (16.5%) were severe injuries. Ninety-seven severe injuries (86%) were able to be documented in detail. Trauma was the cause of 81.5% of the injuries and overuse was the cause of 18.5%. Joint sprains predominated (30%), followed by fractures (16%), muscle strains (15%), ligament ruptures (12%), meniscal tears and contusions (8%), and other injuries. Injuries to the knee were most prevalent (29%), followed by injuries to the ankle (19%) and spine (9%). More injuries occurred during games (59%) than in practice. Twenty-four percent of the injured players had suffered a previous injury of the same body part. Forty-six percent of injuries were caused by contact and 54% involved no body contact. Thirty-one percent of severe injuries were caused by foul play. From these results and the analysis of injuries in specific body parts, the following factors were determined to influence the occurrence of severe injuries: 1) personal factors (intrinsic): age of player, previous injuries, joint instability, abnormality of the spine, poor physical condition, poor football skills, or inadequate treatment and rehabilitation of injuries; 2) environmental factors (extrinsic): subjective exercise overload during practices and games, amount and quality of training, playing field conditions, equipment (wearing of shin guards and taping) and violations of existing rules (foul play).


Nivolumab plus ipilimumab versus sunitinib for first-line treatment of advanced renal cell carcinoma: extended 4-year follow-up of the phase III CheckMate 214 trial.
To report updated analyses of the phase III CheckMate 214 trial with extended minimum follow-up assessing long-term outcomes with first-line nivolumab plus ipilimumab (NIVO+IPI) versus (vs) sunitinib (SUN) in patients with advanced renal cell carcinoma (aRCC). Patients with aRCC with a clear cell component were stratified by International Metastatic Renal Cell Carcinoma Database Consortium risk and randomised to NIVO (3 mg/kg) plus IPI (1 mg/kg) every three weeks ×4 doses, followed by NIVO (3 mg/kg) every two weeks; or SUN (50 mg) once per day ×4 weeks (6-week cycle). Efficacy endpoints included overall survival (OS), progression-free survival (PFS) and objective response rate (ORR) per independent radiology review committee in patients with intermediate/poor-risk disease (I/P; primary), intent-to-treat patients (ITT; secondary) and in patients with favourable-risk disease (FAV; exploratory). Overall, 1096 patients were randomised (ITT: NIVO+IPI, n=550, SUN, n=546; I/P: NIVO+IPI, n=425, SUN, n=422; FAV: NIVO+IPI, n=125, SUN, n=124). After 4 years minimum follow-up, OS (HR; 95% CI) remained superior with NIVO+IPI vs SUN in ITT (0.69; 0.59 to 0.81) and I/P patients (0.65; 0.54 to 0.78). Four-year PFS probabilities were 31.0% vs 17.3% (ITT) and 32.7% vs 12.3% (I/P), with NIVO+IPI vs SUN. ORR remained higher with NIVO+IPI vs SUN in ITT (39.1% vs 32.4%) and I/P (41.9% vs 26.8%) patients. In FAV patients, the HRs (95% CI) for OS and PFS were 0.93 (0.62 to 1.40) and 1.84 (1.29 to 2.62); ORR was lower with NIVO+IPI vs SUN. However, more patients in all risk groups achieved complete responses with NIVO+IPI: ITT (10.7% vs 2.6%), I/P (10.4% vs 1.4%) and FAV (12.0% vs 6.5%). Probability (95% CI) of response ≥4 years was higher with NIVO+IPI vs SUN (ITT, 59% (0.51 to 0.66) vs 30% (0.21 to 0.39); I/P, 59% (0.50 to 0.67) vs 24% (0.14 to 0.36); and FAV, 60% (0.41 to 0.75) vs 38% (0.22 to 0.54)) regardless of risk category. Safety remained favourable with NIVO+IPI vs SUN. After long-term follow-up, NIVO+IPI continues to demonstrate durable efficacy benefits vs SUN, with manageable safety. ClinicalTrials.gov identifier: NCT02231749.


Arrhythmias in children with ventricular assist devices.
Children with decompensated heart failure are at high risk for arrhythmias, and ventricular assist device placement is becoming a more common treatment strategy. The impact of ventricular assist devices on arrhythmias and how arrhythmias affect the clinical course of this population are not well described. A single-centre retrospective analysis of children receiving a ventricular assist device between 1998 and 2011 was performed. In all, 45 patients received 56 ventricular assist devices. The median age at initial placement was 13 years (interquartile range 6-15). The median duration of support was 10 days (range 2-260). The aetiology of heart failure included cardiomyopathy, transplant rejection, myocarditis, and congenital heart disease. In all, 32 patients (71%) had an arrhythmia; 19 patients (42%) had an arrhythmia before ventricular assist device and eight patients (18%) developed new arrhythmias on ventricular assist device. Ventricular tachycardia was most common (25/32, 78%). There was no correlation between arrhythmia and risk of death or transplantation (p=0.14). Of the 15 patients who weaned from ventricular assist device, post-ventricular assist device arrhythmias occurred in nine (60%), with five (33%) having their first arrhythmia after weaning. Patients with ventricular dysfunction after ventricular assist device were more likely to have arrhythmias (p<0.02). Arrhythmias, especially ventricular, are common in children requiring ventricular assist device. They frequently persist for those able to wean from ventricular assist device.


Anthelmintic activity of cytochrome P450 inhibitors miconazole and clotrimazole: in-vitro effect on the liver fluke Opisthorchis felineus.
Discovery of drugs for the treatment of opisthorchiasis and schistosomiasis is a high priority. The basic metabolic cytochrome P450 (CYP) system in parasitic flatworms contains a single gene. CYP of the liver fluke Opisthorchis felineus, the causative agent of opisthorchiasis, is important for survival of the worm, so it may be a promising target for therapeutics against liver fluke infection. The aims of this study were: (i) to analyse in-vitro anthelmintic activity of various CYP inhibitors using standard motility and mortality assays against juvenile and adult O. felineus worms; and (ii) to characterize their anthelminthic effects. Azole inhibitors (ketoconazole, miconazole, triadimenol, clotrimazole and 4-phenyl imidazole) and other inhibitors of haem-containing enzymes (disulfiram, metyrapone, benzyl isothiocyanate, and ticlopidine) were tested. This study revealed that inhibitors of haem mono-oxygenase enzymes possess anthelmintic activity. The most effective anthelmintic agents against the newly excysted metacercariae (NEM) were the antifungal agents miconazole [concentration to reduce the response by 50% (IC50) 0.79 µM] and clotrimazole (IC50 1.25 µM), both approved by the US Food and Drug Administration. The activity of miconazole and clotrimazole was comparable to that for praziquantel (IC50 0.98 µM). In addition, 100% mortality was observed among NEM after 1 d of treatment with 10 µM miconazole, after 3 d of treatment with 10 µM clotrimazole, or after 7 d of treatment with 40 µM ketoconazole. When various CYP inhibitors were tested on adult worms, clotrimazole, miconazole and ketoconazole were found to be the most effective (IC50 13-20 µM). It is speculated that CYP may represent a promising drug target for combined treatment with other anthelmintic agents. The use of inhibitor-drug combinations may improve the action of standard anthelmintic agents.