European Academy of Paediatrics Research in Ambulatory Setting network (EAPRASnet): a multi-national general paediatric research network for better child health.
In 2008, the European Academy of Paediatrics launched a paediatric-based research network - EAPRASnet (European Academy of Paediatrics Research in Ambulatory Setting network). The network has recruited primary care and general paediatricians from European and Mediterranean countries. Every paediatrician joining the network has been asked to complete a recruitment survey. The aims of the survey were to characterize paediatrician's demographics, practice arrangements and patient's demographics, to define main incentives for research, and to learn what paediatricians view as unsolved issues that need to be studied. A total of 156 paediatricians from 19 countries were recruited with 144 completing the questionnaire (92%). Majority of respondents (89%) were general paediatricians for more than half of their time. Practice arrangement of 47% of paediatricians was solo practice, with 40% in group practice. Electronic medical records were being used by 72% of respondents. Over 70% of the paediatricians had more than 1000 patients under their clinical care, and patients younger than 6 years old contributed nearly half of the patient population. Areas of most interest for research were: quality of care indicators, communication with parents, obesity, attention deficit hyperactivity disorder and effective well child care. Main incentives for participation in a research project were interest in the topic (81%) and effort to improve quality of care (71%). Lack of time was the leading reported obstacle for research activity (72%). EAPRASnet is growing, and the network's structure, operation and funding are described. Methods for joining the network and the process of study development are presented. A core group of EAP general paediatricians are committed to research in their practices. The information gathered will serve for future planning of research projects in the EAPRASnet to harmonize and optimize the care given to children in the primary care setting in Europe.


Clostridium Difficile Infection from a Surgical Perspective.
The incidence and the severity of Clostridium difficile infection (CDI) have increased significantly over the last decade, especially in high-risk populations such as patients with inflammatory bowel disease (IBD). Surgeons must be able to both identify and minimize the risk of CDI in their own surgical patients and determine which CDI patients will benefit from surgery. We sought to define the risk factors, compare the treatment options, define the surgical indications, and identify factors that affect surgical outcomes for CDI based on the currently available literature. Antibiotic use, exposure to the C. difficile bacteria, IBD, and higher levels of co-morbidity are all risk factors for CDI. The majority of CDI can be treated with antibiotics. Severe or fulminant colitis, however, has a high potential for poor outcome, but experience and some data suggest a lower mortality rate with colectomy rather than with continued medical treatment. Open total abdominal colectomy with end ileostomy is typically the preferred surgical strategy. It is often difficult to determine which patients will fail medical management as some may not manifest clinical signs of severe infection. Surrogate parameters of failure of medical therapy include respiratory and/or renal insufficiency, age greater than 60 years, peripheral vascular disease, congestive heart failure, and coagulopathy, all of which have been associated with worse surgical outcomes. Evidence suggests that in appropriately selected patients, colectomy performed before the development of shock requiring vasopressors, respiratory failure, renal failure, multi-organ dysfunction, and mental status changes may reduce mortality of the most severe forms of colitis. For less severe or recurrent presentations, creation of a loop ileostomy with intra-operative colonic lavage, fecal microbiota transfer, and C. difficile vaccinations are being discussed but have only been studied in small case-controlled series. Prevention, containment, and non-surgical treatment are the cornerstone of management for CDI. However, the most severe forms with toxic colitis benefit from involvement of a surgical team. Swift open total abdominal colectomy with end ileostomy in patients with severe or fulminant C. difficile colitis has the best chance to reduce mortality if it is not delayed until shock, end organ damage, vasopressor requirement, mental status changes develop. Less aggressive approaches may be appropriate for milder and refractory forms but require further study before their applicability can be determined.


Vertigo and metabolic disorders.
Metabolic disorders are accepted by many authors as being responsible for balance disorders. Because of the importance of metabolic disorders in the field of labyrinthine dysfunction, we decided to assess the prevalence of carbohydrates, lipids and thyroid hormones disorders in our patients with vestibular diseases. The study evaluates the metabolic profile of 325 patients with vertigo who sought the Otolaryngology Department of the University of São Paulo in the Hospital das Clínicas da Universidade de São Paulo. The laboratory tests ordered according to the classical research protocol were: low-density lipoprotein cholesterol fraction, TSH, T3, T4 and fasting blood sugar level. The metabolic disorders found and the ones that were observed in the general population were compared. The high level of low-density lipoprotein cholesterol, the altered levels of thyroid hormones, the higher prevalence of diabetes mellitus were the most significant changes found in the group of study. The higher amount of metabolic disorders in patients with vertigo disease reinforces the hypothesis of its influence on the etiopathogenesis of cochleovestibular symptoms.


Polychlorinated biphenyl exposure and neuropsychological status among older residents of upper Hudson River communities.
Polychlorinated biphenyls (PCBs) may accelerate the cognitive and motor dysfunction found in normal aging, but few studies have examined these outcomes and PCB exposure among older adults. We evaluated neuropsychological status and low-level PCB exposure among older adults living along contaminated portions of the upper Hudson River in New York. A total of 253 persons between 55 and 74 years of age were recruited and interviewed, and provided blood samples for congener-specific PCB analysis. Participants also underwent a neuropsychological battery consisting of 34 tests capable of detecting subtle deficits in cognition, motor function, affective state, and olfactory function. After adjustment for potential confounders, the results indicated that an increase in serum total PCB concentration from 250 to 500 ppb (lipid basis) was associated with a 6.2% decrease in verbal learning, as measured by California Verbal Learning Test trial 1 score (p = 0.035), and with a 19.2% increase in depressive symptoms, as measured by the Beck Depression Inventory (p = 0.007). The results suggest that exposure to PCBs may be associated with some measures of memory and learning and depression among adults 55-74 years of age whose current body burdens are similar to those of the general population. Although the results are useful in delineating the neuropsychological effects of low-level exposure to PCBs, further studies of whether older men and women are a sensitive subpopulation are needed.


Simple concentration method enables the use of gargle and mouthwash instead of nasopharyngeal swab sampling for the diagnosis of COVID-19 by PCR.
Since its emergence in December 2019, SARS-CoV-2 is causing one of the most devastating pandemics in human history. Currently, the most important method for definitive diagnosis of COVID-19 is identification of SARS-CoV-2 RNA in nasopharyngeal swab samples by RT-PCR. Nasopharyngeal swab sampling is a discomforting procedure sometimes with adverse effects, which also poses a risk for infection for the personnel performing the sampling. We have developed a new method for concentrating biological samples, which enabled us to use gargle and mouthwash samples to be used in RT-PCR, for the diagnosis of COVID-19, as an alternative to nasopharyngeal swab samples. We have analyzed nasopharyngeal and gargle and mouthwash samples, before and after concentration, of 363 patients by RT-PCR for the presence of SARS-CoV-2. Among 114 patients in which SARS-CoV-2 was identified in at least one of their samples, the virus was identified in 76 (66.7%), 67 (58.8%), and 101 (88.6%) of nasopharyngeal swab, gargle, and mouthwash samples before and after concentration, respectively. When concentrated by our new method, gargle and mouthwash samples can be used instead of nasopharyngeal samples in identification of SARS-CoV-2 by RT-PCR, with the same or better sensitivity. Eliminating the need for nasopharyngeal sampling will save the patients from an invasive and painful procedure and will lower the risk of infection for the healthcare personnel taking the sample. This easy sampling procedure may decrease the workload of hospitals, shorten the turnaround time of obtaining test results, and thus enable rapid isolation of infected patients.


The impact of electronic meal ordering systems on hospital and patient outcomes: A systematic review.
Malnutrition is a serious clinical issue associated with adverse patient and hospital outcomes. Hospitals need to consider interventions that support the provision of optimal nutritional management and care for patients. Electronic meal ordering (EMO) systems provide an alternative to traditional paper-based meal ordering with the capacity to support appropriate orders, monitor nutritional status, and potentially improve clinical outcomes. This review aimed to identify the impact of EMO systems on hospital and patient outcomes. We sought quantitative evidence (peer-reviewed and grey literature) from studies evaluating EMO systems in healthcare facilities, published after 1999 and available in the English language. We identified 23 studies evaluating one of three distinct EMO system-supported models: spoken menu, room service, and self-service. While limited, the evidence indicated that EMO systems were associated with: improved patient satisfaction; decreased food waste; increased consumption; and, for spoken menus, more time with patients. There was no substantive evidence of impact on clinical outcomes. Whether EMO systems meet their potential to support nutritional monitoring and positively impact clinical outcomes remains unanswered within the evidence. Thus, policy makers and hospital management currently have a poor evidence base upon which to make decisions about the value of implementing EMO. Whether these systems can provide support and guidance to patients during meal ordering, improve order appropriateness and accuracy through compliance checking, identify patients in need of dietary education or those at risk of malnutrition are critical areas of focus for future research.


The hepatitis C virus E1 glycoprotein undergoes productive folding but accelerated degradation when expressed as an individual subunit in CHO cells.
Hepatitis C Virus E1E2 heterodimers are components of the viral spike. Although there is a general agreement on the necessity of the co-expression of both E1 and E2 on a single coding unit for their productive folding and assembly, in a previous study using an in vitro system we obtained strong indications that E1 can achieve folding in absence of E2. Here, we have studied the folding pathway of unescorted E1 from stably expressing CHO cells, compared to the folding observed in presence of the E2 protein. A DTT-resistant conformation is achieved by E1 in both situations, consistent with the presence of an E2-independent oxidative pathway. However, while the E1E2 heterodimer is stable inside cells, E1 expressed alone is degraded within a few hours. On the other hand, the oxidation and stability of individually expressed E2 subunits is dependent on E1 co-expression. These data are consistent with E1 and E2 assisting each other for correct folding via different mechanisms: E2 assists E1 by stabilizing a semi-native conformation meanwhile E1 drives E2 towards a productive folding pathway.


Stronger control of ATP/ADP by proton leak in pancreatic beta-cells than skeletal muscle mitochondria.
Pancreatic beta cells respond to rising blood glucose concentrations by increasing their oxidative metabolism, which leads to an increased ATP/ADP ratio, closure of K(ATP) channels, depolarization of the plasma membrane potential, influx of calcium and the eventual secretion of insulin. Such a signalling mechanism implies that the ATP/ADP ratio is flexible in beta cells (beta-cells), which is in contrast with other cell types (e.g. muscle and liver) that maintain a stable ATP/ADP poise while respiring at widely varying rates. To determine whether this difference in flexibility is accounted for by mitochondrial peculiarities, we performed a top-down metabolic control analysis to quantitatively assess how ATP/ADP is controlled in mitochondria isolated from rat skeletal muscle and cultured beta cells. We show that the ATP/ADP ratio is more strongly controlled (approx. 7.5-fold) by proton leak in beta cells than in muscle. The comparatively high importance of proton leak in beta cell mitochondria (relative to phosphorylation) is evidenced furthermore by its relatively high level of control over membrane potential and overall respiratory activity. Modular-kinetic analysis of oxidative phosphorylation reveals that these control differences can be fully explained by a higher relative leak activity in beta cell mitochondria, which results in a comparatively high contribution of proton leak to the overall respiratory activity in this system.


Nonparametric regression applied to quantitative structure-activity relationships
Several nonparametric regressors have been applied to modeling quantitative structure-activity relationship (QSAR) data. The simplest regressor, the Nadaraya-Watson, was assessed in a genuine multivariate setting. Other regressors, the local linear and the shifted Nadaraya-Watson, were implemented within additive models--a computationally more expedient approach, better suited for low-density designs. Performances were benchmarked against the nonlinear method of smoothing splines. A linear reference point was provided by multilinear regression (MLR). Variable selection was explored using systematic combinations of different variables and combinations of principal components. For the data set examined, 47 inhibitors of dopamine beta-hydroxylase, the additive nonparametric regressors have greater predictive accuracy (as measured by the mean absolute error of the predictions or the Pearson correlation in cross-validation trails) than MLR. The use of principal components did not improve the performance of the nonparametric regressors over use of the original descriptors, since the original descriptors are not strongly correlated. It remains to be seen if the nonparametric regressors can be successfully coupled with better variable selection and dimensionality reduction in the context of high-dimensional QSARs.


Occurrence of soil- and tick-borne fungi and related virulence tests for pathogenicity to Ixodes scapularis (Acari: Ixodidae).
Ixodes scapularis Say, the blacklegged tick, vectors Borrelia burgdorferi Johnson et al. 1984, the bacterium that causes Lyme disease, the most important vector-borne disease in the United States. Efforts to reduce I. scapularis populations are shifting toward the development of biological control methods. Currently, only a few entomopathogenic fungal species are considered virulent to ticks. We hypothesized that these species may not represent the most abundant local taxa that would be pathogenic to ticks in situ. To identify potential entomopathogenic fungi at a study site in Westchester County, New York, we sampled soils and ticks, extracted and amplified the internal transcribed spacer region of nuclear ribosomal DNA (nrDNA), and compared sequences with those in GenBank. Over three sampling periods from June 2007 to May 2008, 70 fungal taxa were isolated and identified from soils (48 taxa) and ticks (27 taxa; 5 taxa were found both in soil and on ticks) collected in this study, encompassing species in 25 different genera. In laboratory bioassays, 15 fungal taxa were found to be significantly virulent, although none of these were previously considered common pathogens of I. scapularis. Two species, Hypocrea lixii Patouillard 1891 and Penicillium soppii K. M. Zalessky 1927, were tested in field trials by spraying suspensions on forested plots. Mean tick mortality was 71% after treatment with H. lixii, 58% after treatment with P. soppii, and 32% in the control plots. The complete diversity of entomopathogenic fungal species at this site is yet to be defined, but, in general, such fungi appear to be more common in forest habitats where I. scapularis resides than previously thought. Examination of intact fungal communities can provide information that serves as the foundation for site-specific biocontrol programs.


The air transport of patients.
Air transport of patients by light aircraft and air ambulance is safe, fast and comfortable, and it is becoming increasingly popular. There are, however, specific problems, shortcomings and other details which must be dealt with effectively at various levels to ensure the best service for the patient.


Adjuvant radiation therapy for patients with type II endometrial carcinoma: impact on tumor recurrence and survival.
The optimal adjuvant treatment of type II endometrial carcinoma after hysterectomy remains controversial. The objective of this study was to determine the effect of adjuvant radiation therapy (RT) on recurrence-free survival (RFS), disease-specific survival (DSS), and overall survival in patients with early-stage type II endometrial carcinoma. In this institutional review board-approved study, our database of 1450 patients with endometrial cancer was reviewed. Seventy-nine surgically staged patients with 2009 International Federation of Gynecology and Obstetrics (FIGO) stages I and II serous and clear cell carcinoma were treated from 1991 to 2010. These patients were then divided into 2 groups; one group received adjuvant RT, and the other group included patients who did not receive adjuvant RT. The median age of the study cohort is 65 years, and the median follow-up is 47 months. Thirty-nine patients (49%) received adjuvant RT, and 40 patients did not. The 5-year RFS was significantly improved in patients who received RT (84% vs 58%; P = 0.002). Similarly, 5-year DSS was significantly improved in patients who received RT (87% vs 58%; P = 0.023) with a trend toward improved 5-year overall survival (74% vs 58%; P = 0.088). On multivariate analysis, lack of angiolymphatic invasion (P < 0.001 and P < 0.001), adjuvant RT (P < 0.001 and P = 0.004), and lack of lower uterine segment involvement (P = 0.007 and P = 0.009) were independent predictors of improved RFS and DSS, respectively. In the current study of surgically staged patients with type II endometrial carcinoma International Federation of Gynecology and Obstetrics stages I and II, adjuvant radiation therapy with or without chemotherapy resulted in a significant improvement in recurrence-free and disease-specific survival.


Hypothesis: systemic lupus erythematosus as a disease secondary to polyclonal lymphocyte activation.
Systemic lupus erythematosus (SLE) is characterized by the concomitance of B cell polyclonal activation and the production of a wide spectrum of autoantibodies showing a marked preference for nuclear antigens. We propose the hypothesis that the production of the most pathogenic antinuclear antibodies with high affinity and IgG isotype is driven by antigens liberated by the death of rapidly proliferating lymphoid cells. The disease could evolve in three steps: (1) polyclonal B cell activation and proliferation caused by unknown etiologic factors (virus?, mitogen? ...); (2) primary activation of natural antibody-producing clones leading to the appearance in the serum of large amounts of natural autoantibodies with public idiotype; and (3) secondary immunization against molecules synthesized in large quantities during lymphocyte proliferation, released by dying cells and subsequently presented in an immunogenic form to T cells. These molecules include nucleic acids (DNA, RNA), nucleic acid-binding proteins (histones, Ro, La), and parts of the RNA splicing machinery (RNP). Other autoantibody activities could relate to cross-reactivities or the binding properties of DNA-anti-DNA immune complexes.


Hybrid origins of honeybees from italy (Apis mellifera ligustica) and sicily (A. m. sicula).
The genetic variability of honeybee populations Apis mellifera ligustica, in continental Italy, and of A. m. sicula, in Sicily, was investigated using nuclear (microsatellite) and mitochondrial markers. Six populations (236 individual bees) and 17 populations (664 colonies) were, respectively, analysed using eight microsatellite loci and DraI restriction fragment length polymorphism (RFLP) of the cytochrome oxidase I (COI)-cytochrome oxidase II (COII) region. Microsatellite loci globally confirmed the southeastern European heritage of both subspecies (evolutionary branch C). However, A. m. ligustica mitochondrial DNA (mtDNA) appeared to be a composite of the two European (M and C) lineages over most of the Italian peninsula, and only mitotypes from the African (A) lineage were found in A. m. sicula samples. This demonstrates a hybrid origin for both subspecies. For A. m. ligustica, the most widely exported subspecies, this hybrid origin has long been obscured by the fact that in the main area of queen production (from which most of the previous ligustica bee samples originated) the M mitochondrial lineage is absent, whereas it is present almost everywhere else in Italy. This presents a new view of the evolutionary history of European honeybees. For instance, the Iberian peninsula was considered as the unique refuge for the M branch during the quaternary ice periods. Our results show that the Apennine peninsula played a similar role. The differential distribution of nuclear and mitochondrial markers observed in Italy seems to be a general feature of introgressed honeybee populations. Presumably, it stems from the social nature of the species in which both genome compartments are differentially affected by the two (individual and colonial) reproduction levels.


CuInS2 Photocathodes with Atomic Gradation-Controlled (Ta,Mo)x(O,S)y Passivation Layers for Efficient Photoelectrochemical H2 Production.
An atomic gradient passivation layer, (Ta,Mo)x(O,S)y, is designed to improve the charge transportation and photoelectrochemical activity of CuInS2-based photoelectrodes. We found that Mo spontaneously diffused to the a-TaOx layer during e-beam evaporation. This result indicates that the gradient profile of MoOx/TaOx is formed in the sublayer of (Ta,Mo)x(O,S)y. To understand the atomic-gradation effects of the (Ta,Mo)x(O,S)y passive layer, the composition and (photo)electrochemical properties have been characterized in detail. When this atomic gradient-passive layer is applied to CuInS2-based photocathodes, promising photocurrent and onset potential are seen without using Pt cocatalysts. This is one of the highest activities among reported CuInS2 photocathodes, which are not combined with noble metal cocatalysts. Excellent photoelectrochemical activity of the photoelectrode can be mainly achieved by (1) the electron transient time improved due to the conductive Mo-incorporated TaOx layer and (2) the boosted electrocatalytic activity by Mox(O,S)y formation.


Catalytic efficiency and sequence selectivity of a restriction endonuclease modulated by a distal manganese ion binding site.
Crystal structures of EcoRV endonuclease bound in a ternary complex with cognate duplex DNA and manganese ions have previously revealed an Mn(2+)-binding site located between the enzyme and the DNA outside of the dyad-symmetric GATATC recognition sequence. In each of the two enzyme subunits, this metal ion bridges between a distal phosphate group of the DNA and the imidazole ring of His71. The new metal- binding site is specific to Mn(2+) and is not occupied in ternary cocrystal structures with either Mg(2+) or Ca(2+). Characterization of the H71A and H71Q mutants of EcoRV now demonstrates that these distal Mn(2+) sites significantly modulate activity toward both cognate and non-cognate DNA substrates. Single-turnover and steady-state kinetic analyses show that removal of the distal site in the mutant enzymes increases Mn(2+)-dependent cleavage rates of specific substrates by tenfold. Conversely, the enhancement of non-cognate cleavage at GTTATC sequences by Mn(2+) is significantly attenuated in the mutants. As a consequence, under Mn(2+) conditions EcoRV-H71A and EcoRV-H71Q are 100 to 700-fold more specific than the wild-type enzyme for cognate DNA relative to the GTTATC non-cognate site. These data reveal a strong dependence of DNA cleavage efficiency upon metal ion-mediated interactions located some 20 A distant from the scissile phosphodiester linkages. They also show that discrimination of cognate versus non-cognate DNA sequences by EcoRV depends in part on contacts with the sugar-phosphate backbone outside of the target site.


Signatures of Microevolutionary Processes in Phylogenetic Patterns.
Phylogenetic trees are representations of evolutionary relationships among species and contain signatures of the processes responsible for the speciation events they display. Inferring processes from tree properties, however, is challenging. To address this problem, we analyzed a spatially-explicit model of speciation where genome size and mating range can be controlled. We simulated parapatric and sympatric (narrow and wide mating range, respectively) radiations and constructed their phylogenetic trees, computing structural properties such as tree balance and speed of diversification. We showed that parapatric and sympatric speciation are well separated by these structural tree properties. Balanced trees with constant rates of diversification only originate in sympatry and genome size affected both the balance and the speed of diversification of the simulated trees. Comparison with empirical data showed that most of the evolutionary radiations considered to have developed in parapatry or sympatry are in good agreement with model predictions. Even though additional forces other than spatial restriction of gene flow, genome size, and genetic incompatibilities, do play a role in the evolution of species formation, the microevolutionary processes modeled here capture signatures of the diversification pattern of evolutionary radiations, regarding the symmetry and speed of diversification of lineages.


Radiation oncology: past achievements and ongoing controversies.
With the development of megavoltage treatment and computerized treatment planning the quality and precision of radiation oncology has steadily improved. Likewise, these developments have contributed to better local control for some cancers; however, micrometastatic lesions beyond the radiation treatment field and ineffective systemic treatments for many malignancies hamper efforts at the most important oncological end point, survival. Major advances in cancer therapy are therefore likely to come with improved combined modality treatment representing integration of local modalities with the systemic. These advances, in our opinion, will come from biological developments that address the problems that the modern oncologist faces at the cellular level. The biological developments will incorporate modern molecular biology, continued probing for biochemical mechanisms, and an intensified effort to learn more about the complexities of human tumor physiology.


Human-Relevant Sensitivity of iPSC-Derived Human Motor Neurons to BoNT/A1 and B1.
The application of botulinum neurotoxins (BoNTs) for medical treatments necessitates a potency quantification of these lethal bacterial toxins, resulting in the use of a large number of test animals. Available alternative methods are limited in their relevance, as they are based on rodent cells or neuroblastoma cell lines or applicable for single toxin serotypes only. Here, human motor neurons (MNs), which are the physiological target of BoNTs, were generated from induced pluripotent stem cells (iPSCs) and compared to the neuroblastoma cell line SiMa, which is often used in cell-based assays for BoNT potency determination. In comparison with the mouse bioassay, human MNs exhibit a superior sensitivity to the BoNT serotypes A1 and B1 at levels that are reflective of human sensitivity. SiMa cells were able to detect BoNT/A1, but with much lower sensitivity than human MNs and appear unsuitable to detect any BoNT/B1 activity. The MNs used for these experiments were generated according to three differentiation protocols, which resulted in distinct sensitivity levels. Molecular parameters such as receptor protein concentration and electrical activity of the MNs were analyzed, but are not predictive for BoNT sensitivity. These results show that human MNs from several sources should be considered in BoNT testing and that human MNs are a physiologically relevant model, which could be used to optimize current BoNT potency testing.


Biallelic mutations in the death domain of PIDD1 impair caspase-2 activation and are associated with intellectual disability.
PIDD1 encodes p53-Induced Death Domain protein 1, which acts as a sensor surveilling centrosome numbers and p53 activity in mammalian cells. Early results also suggest a role in DNA damage response where PIDD1 may act as a cell-fate switch, through interaction with RIP1 and NEMO/IKKg, activating NF-κB signaling for survival, or as an apoptosis-inducing protein by activating caspase-2. Biallelic truncating mutations in CRADD-the protein bridging PIDD1 and caspase-2-have been reported in intellectual disability (ID), and in a form of lissencephaly. Here, we identified five families with ID from Iran, Pakistan, and India, with four different biallelic mutations in PIDD1, all disrupting the Death Domain (DD), through which PIDD1 interacts with CRADD or RIP1. Nonsense mutations Gln863* and Arg637* directly disrupt the DD, as does a missense mutation, Arg815Trp. A homozygous splice mutation in the fifth family is predicted to disrupt splicing upstream of the DD, as confirmed using an exon trap. In HEK293 cells, we show that both Gln863* and Arg815Trp mutants fail to co-localize with CRADD, leading to its aggregation and mis-localization, and fail to co-precipitate CRADD. Using genome-edited cell lines, we show that these three PIDD1 mutations all cause loss of PIDDosome function. Pidd1 null mice show decreased anxiety, but no motor abnormalities. Together this indicates that PIDD1 mutations in humans may cause ID (and possibly lissencephaly) either through gain of function or secondarily, due to altered scaffolding properties, while complete loss of PIDD1, as modeled in mice, may be well tolerated or is compensated for.


Tsga10 encodes a 65-kilodalton protein that is processed to the 27-kilodalton fibrous sheath protein.
We had previously reported the isolation of the testis-specific human gene Tsga10, which is not expressed in testes from two infertile patients. To study its role and function, we cloned the mouse homologue Mtsga10. Mtsga10 localizes to mouse chromosome 1, band B. This region is syntenic with human chromosome 2q11.2, where Tsga10 is located. We demonstrate that Mtsga10 mRNA is expressed in testis, but not in other adult tissues, and in several human fetal tissues and primary tumors. We uncovered that different species use different first exons and, consequently, different promoters. Using several antibodies, we discovered that, in mouse testis, Mtsga10 encodes a 65-kDa spermatid protein that appears to be processed to a 27-kDa protein of the fibrous sheath, a major sperm tail structure, in mature spermatozoa. Mtsga10 protein contains a putative myosin/Ezrin/radixin/moesin (ERM) domain. Transfection of fibroblasts with GFP-Mtsga10 fusion protein results in formation of short, thick filaments and deletion of the myosin/ERM domain abolished filament formation. Our results suggest the possibility that Tsga10 plays a role in the sperm tail fibrous sheath.


Health Literacy Requires Empowerment - Results of an Analysis of Patient Needs and the Demand for Qualification over a 10-Year Period.
Patients are expected to be increasingly competent about health issues, and patient self-determination is on the rise. This makes it ever more necessary to provide high quality information which is increasingly being offered to patients at all levels of the healthcare system. One example of a regionally available information service offered to patients is the Women's Academy, created 10 years ago, at the University Women's Hospital Tübingen in association with the Institute for Women's Health. The Women's Academy also serves as a forum and an outreach hub for the (cancer) centres of the Women's Hospital. The topics addressed include problematic areas in medicine and problems in coping with health and disease and additionally reflect the emergence of various patient interests and needs over time. The communication between physician and patient is central to the promotion of health literacy and individualised medicine. Regionally available services like the Women's Academy offer complementary benefits and are able to reach vulnerable groups of women who have little or no access to online information.


Understanding leptospirosis: application of state-of-the-art molecular typing tools with a One Health lens.
Leptospirosis is an archetypal One Health problem as described in the companion Currents in One Health article in the October 2022 issue of the Journal of the American Veterinary Medical Association by Sykes et al. A thorough understanding of leptospirosis requires a detailed analysis of the elaborate interplay among pathogenic leptospiral strains, host species, and the environment. Such an understanding is required to inform appropriate preventative measures including vaccine design, prophylaxis efforts, educational programs that help to reduce exposure to pathogenic spirochetes, as well as policy development. Because of the complex epidemiology of leptospirosis, a One Health approach as defined by the One Health Initiative Task Force is critical-an approach that calls for "the collaborative efforts of multiple disciplines working locally, nationally, and globally, to attain optimal health for people, animals and our environment." Over the last three decades, progressive advances in cutting-edge molecular typing techniques, as well as our ability to rapidly generate and share large amounts of sequence data through establishment and growth of databases, have been central to accelerating a One Health understanding of the epidemiology of leptospirosis. Nevertheless, our dependence on serotype information because of the serovar-specific nature of current vaccines means that laborious serotyping efforts continue. With the advent of new approaches such as mRNA vaccines that are based on lipopolysaccharide immunogens, sequence- and/or proteomics-based typing methods may replace these methods.


NMR studies of calcium-binding to mutant alpha-spectrin EF-hands.
The co-operative calcium binding mechanism of the two C-terminal EF-hands of human alphaII-spectrin has been investigated by site-specific mutagenesis and multi-dimensional NMR spectroscopy. To analyse the calcium binding of each EF-hand independently, two mutant structures (E33A and D69S) of wild type alpha-spectrin were prepared. According to NMR analysis both E33A and D69S were properly folded. The unmutated EF-hand in these mutants remained nearly intact and active in calcium binding, whereas the mutated EF-hand lost its affinity for calcium completely. The apparent calcium binding affinity of the E33A mutant was much lower compared to the D39S mutant (approximately 2470 microM and approximately 240 microM, respectively). When the chemical shift perturbations were followed upon calcium titration, a positive correlation between the D69S mutant and the binding of the first calcium ion to the wild type was revealed. These observations showed that the first EF-hand in spectrin binds the first calcium ion and thereby triggers a conformational change that allows the second calcium ion to bind to the other EF-hand.


[Eye care management in Russian Federation].
The article presents an analysis of Russian eye care performance indicators based on federal and sector statistics over the recent years provided by the Ministry of Health of the Russian Federation, that is the incidence of eye diseases, eye care equipment provision, inpatient and outpatient volumes. Legal acts of the Russian Federation on health system in general and eye care in particular were taken into consideration when preparing the section on organizational matters. Problems of human resources, efficiency of specialists' time management, hospital beds use, and administrative issues in particular regions and Russia as a whole are discussed.


Utilization of electrochemically generated ozone in the degradation and detoxication of benzo[a]pyrene.
The ability of electrochemically generated ozone (O3) to degrade and detoxify the polycyclic aromatic hydrocarbon (PAH) benzo[a]pyrene (BaP) was assessed utilizing the chick embryotoxicity screening test (CHEST) and Hydra attenuata bioassays. Aqueous solutions containing 10 microg/ml BaP and 0.5% (v/v) acetonitrile were subjected to ozonolysis for 0 to 30 min. Rapid degradation of BaP was evident by both gas chromatography/mass spectroscopy (GC/MS) and high-performance liquid chromatography (HPLC) analysis. HPLC fluorescence detection revealed no BaP shortly after 5 min of ozonolysis, while HPLC with PDA detection demonstrated continued reactions with ozone over the 30-min time course. As little as 2 min of O3 treatment afforded protection from BaP-induced mortality and toxicity (embryolethality and liver discoloration) in the chicken embryos. In the hydra bioassay, no toxicity was observed in the adult hydra until the ozonolysis products were reconstituted 100-fold from their initial post-ozonolysis concentrations. The results obtained from this study clearly demonstrate the potential application of electrochemically generated O3 for the detoxication and prevention of toxicity of BaP. Both CHEST and hydra assays predict that the ozonolysis products of BaP are less toxic than the parent compound.


Characteristics of Bacterial Strains with Desirable Flavor Compounds from Korean Traditional Fermented Soybean Paste (Doenjang).
To identify and analyze the characteristics of the microorganisms involved in the formation of the desirable flavor of Doenjang, a total of 179 strains were isolated from ninety-four Doenjang collected from six regions in South Korea, and fourteen strains were selected through a sensory evaluation of the aroma of each culture. The enzyme activities of amylase, protease and lipase was shown in the various strains. Bacillus sp.-K3, Bacillus sp.-K4 and Bacillus amyloliquefaciens-J2 showed relatively high protease activity, at 317.1 U, 317.3 U and 319.5 U, respectively. The Bacillus sp.-K1 showed the highest lipase activity at 2453.6 U. In the case of amylase, Bacillus subtilis-H6 showed the highest activity at 4105.5 U. The results of the PCA showed that Bacillus subtilis-H2, Bacillus subtilis-H3, and Bacillus sp.-K2 were closely related to the production of 3-hydroxy-2-butanone (23.51%~43.37%), and that Bacillus subtilis-H5 and Bacillus amyloliquefaciens-J2 were significantly associated with the production of phenethyl alcohol (0.39% and 0.37%). The production of peptides was observed to vary among the Bacillus cultures such as Val-Val-Pro-Pro-Phe-Leu and Pro-Ala-Glu-Val-Leu-Asp-Ile. These peptides are precursors of related volatile flavor compounds created in Doenjang via the enzymatic or non-enzymatic route; it is expected that these strains could be used to enhance the flavor of Doenjang.


Psychometric properties of the 8-item Morisky Medication Adherence Scale in patients taking warfarin.
There is no patient-reported medication adherence measure that has been validated in Singapore. This study aimed to validate the 8-item Morisky Medication Adherence Scale (MMAS) in patients taking warfarin in Singapore. A cross-sectional survey was conducted in a convenience sample of 151 patients taking warfarin at an anticoagulation clinic in 2011. Respondents completed the MMAS in English or Chinese depending on their preference. The MMAS had a Cronbach's alpha of 0.56 and good criterion-related validity as the scale scores were associated with warfarin refill rates (p = 0.04). Respondents with higher MMAS scores were found to have a higher percentage of International Normalised Ratios (INRs) within the therapeutic range (p = 0.01), higher adherence to diet recommendations (p = 0.02), and less perceived difficulty in taking all medications (p < 0.001); they were also more likely to take warfarin at the same time every day (p < 0.001). Confirmatory factor analysis showed that the eight items loaded onto one factor (RMSEA = 0.03). The sensitivity, specificity, positive predictive value and negative predictive value of the MMAS for identifying patients with poor INR control were 73.0%, 35.6%, 49.5% and 60.5%, respectively, using the time in the therapeutic INR range as the gold standard. This study shows that the 8-item MMAS has good validity and moderate reliability in patients taking warfarin. Future research is needed to investigate the scale's psychometric properties in other patient populations and clinical settings.


Enhancement of cyclic electron flow around PSI at high light and its contribution to the induction of non-photochemical quenching of chl fluorescence in intact leaves of tobacco plants.
Non-photochemical quenching (NPQ) of Chl fluorescence is a mechanism for dissipating excess photon energy and is dependent on the formation of a DeltapH across the thylakoid membranes. The role of cyclic electron flow around photosystem I (PSI) (CEF-PSI) in the formation of this DeltapH was elucidated by studying the relationships between O2-evolution rate [V(O2)], quantum yield of both PSII and PSI [Phi(PSII) and Phi(PSI)], and Chl fluorescence parameters measured simultaneously in intact leaves of tobacco plants in CO2-saturated air. Although increases in light intensity raised V(O2) and the relative electron fluxes through both PSII and PSI [Phi(PSII) x PFD and Phi(PSI) x PFD] only Phi(PSI) x PFD continued to increase after V(O2) and Phi(PSII) x PFD became light saturated. These results revealed the activity of an electron transport reaction in PSI not related to photosynthetic linear electron flow (LEF), namely CEF-PSI. NPQ of Chl fluorescence drastically increased after Phi(PSII) x PFD became light saturated and the values of NPQ correlated positively with the relative activity of CEF-PSI. At low temperatures, the light-saturation point of Phi(PSII) x PFD was lower than that of Phi(PSI) x PFD and NPQ was high. On the other hand, at high temperatures, the light-dependence curves of Phi(PSII) x PFD and Phi(PSI) x PFD corresponded completely and NPQ was not induced. These results indicate that limitation of LEF induced CEF-PSI, which, in turn, helped to dissipate excess photon energy by driving NPQ of Chl fluorescence.


Cytoadherence characteristics of Plasmodium falciparum isolates in Thailand using an in vitro human lung endothelial cells model.
Using an in vitro model of human lung endothelial cells, we studied different characteristics of Plasmodium falciparum isolates as potential factors for malaria severity in 2 Thai patient groups: 27 with complicated malaria and 42 with uncomplicated malaria. In regard to binding properties, no association existed between cytoadherence and rosette phenotypes (P = 0.1) and hypothrombocytemia increased the cytoadherence level (P = 0.007). Cytoadherence was significantly associated with malaria severity (P = 0.05) in contrast to rosette formation (P = 0.9). Intercellular adhesion molecule-1 and chondroitin-4-sulfate were major receptors of cytoadherence in those with complicated malaria compared with those with uncomplicated malaria (P < 10(-4)). Chondroitin-4-sulfate could act as a putative receptor for malaria complications in non-pregnant women. CD36 was the main receptor in patients with uncomplicated malaria (P < 10(-3)). Vascular cell adhesion molecule-1 and E-selectin played a minor role in 2 groups (P = 0.6). Qinghaosu derivatives were more efficient than other antimalarial drugs, but a positive correlation was observed between the 50% inhibitory concentrations of halofantrine and quinine and the number of adhesive parasitized red blood cells, suggesting their influence on cytoadherence.


The phosphate ester group in secondary metabolites.
Covering: 2000 to mid-2021The phosphate ester is a versatile, widespread functional group involved in a plethora of biological activities. Its presence in secondary metabolites, however, is relatively rare compared to other functionalities and thus is part of a rather unexplored chemical space. Herein, the chemistry of secondary metabolites containing the phosphate ester group is discussed. The text emphasizes their structural diversity, biological and pharmacological profiles, and synthetic approaches employed in the phosphorylation step during total synthesis campaigns, covering the literature from 2000 to mid-2021.


Doppler sonography in renal diseases.
Using Doppler sonography, the circulation of the kidney can now be studied in the living person in various physiologic or pathologic situations. Our understanding of renal hemodynamics, gleaned from experimental animal models as well as human studies, is being enhanced by Doppler examination and the clinical uses are expanding. Some early concepts, sometimes hastily postulated, are being questioned. The highlights of ongoing debates and the results of animal studies that further our understanding of Doppler examinations in patients are the subject of this review.


DCOne as an Allogeneic Cell-based Vaccine for Multiple Myeloma.
Multiple myeloma (MM) is characterized by progressive immune dysregulation, loss of myeloma-specific immunity, and an immunosuppressive milieu that fosters disease growth and immune escape. Accordingly, cancer vaccines that reverse tumor-associated immune suppression represent a promising therapeutic avenue of investigation. We examined the potential of an allogeneic cellular vaccine to generate immune responses against MM tumor cells. The DCOne vaccine is comprised of a human myeloid leukemia cell line differentiated into a fully functional dendritic cell, expressing a range of tumor-associated antigens that are also known targets in MM. We found that the myeloma-specific antigens expressed by the DCOne vaccine can traffic via extracellular vesicles to surrounding antigen-presenting cells, thus stimulating autologous T-cell responses. Indeed, coculture of peripheral blood mononuclear cells from patients with MM with the DCOne vaccine resulted in the expansion of activated CD8 T cells expressing interferon-γ and perforin, with no significant change in the percentage of CD4 T cells producing interleukin-10. Further, coculture of patient's tumor cells with peripheral blood mononuclear cells and DCOne induced cytotoxic T-lymphocyte-mediated killing of autologous MM cells. These findings demonstrate that the allogeneic DCOne vaccine can induce T-cell activation and myeloma-specific immunity via cross presentation of antigens by native antigen-presenting cells.


Computer-based interventions for sexual health promotion: systematic review and meta-analyses.
This systematic review was conducted to determine the effects of self-help interactive computer-based interventions (ICBIs) for sexual health promotion. We searched 40 databases for randomized controlled trials (RCTs) of computer-based interventions, defining 'interactive' as programmes that require contributions from users to produce personally relevant material. We conducted searches and analysed data using Cochrane Collaboration methods. Results of RCTs were pooled using a random-effects model with standardized mean differences for continuous outcomes and odds ratios (ORs) for binary outcomes, with heterogeneity assessed using the I(2) statistic. We identified 15 RCTs of ICBIs (3917 participants). Comparing ICBIs to minimal interventions, there were significant effects on sexual health knowledge (standardized mean difference [SMD] 0.72, 95% confidence interval [CI] 0.27-1.18); safer sex self-efficacy (SMD 0.17, 95% CI 0.05-0.29); safer-sex intentions (SMD 0.16, 95% CI 0.02-0.30); and sexual behaviour (OR 1.75, 95% CI 1.18-2.59). ICBIs had a greater impact on sexual health knowledge than face-to-face interventions did (SMD 0.36, 95% CI 0.13-0.58). ICBIs are effective tools for learning about sexual health, and show promising effects on self-efficacy, intention and sexual behaviour. More data are needed to analyse biological outcomes and cost-effectiveness.


Differential impact of the expression of the androgen receptor by age in estrogen receptor-positive breast cancer.
We evaluated the expression of the androgen receptor (AR) to determine its significance in breast cancer. AR expression levels were analyzed in 250 invasive breast cancers by immunohistochemistry and any association with the clinicopathological features was evaluated. AR expression was higher in estrogen receptor (ER)-positive cases than in ER-negative cases (P < 0.0001). AR expression was associated with ER level, and it increased with age in ER-positive cases. The cut-off value was determined to be 75% (Cancer Res. 2009;69:6131-6140), and AR expression was considered to be high in 155 (62%) cases. High AR expression significantly correlated with lower nuclear grade (P < 0.0001), ER and progesterone receptor (PR) positivity (P < 0.0001 and P = 0.0022), HER2 negativity (P = 0.0113), lower Ki67 index (P < 0.0001) and a longer disease-free survival (DFS) and distant metastasis-free survival (DMFS) (P = 0.0003 and 0.0107). This association between a high AR expression and a good DFS and DMFS was significant for ER-positive tumors (P < 0.0001 and P = 0.0018); however, no association existed between AR expression and prognosis for ER-negative tumors. In patients ≤51 years old, a high AR expression level significantly correlated with a better prognosis, but this was not significant in patients who were 50 or younger. Multivariate Cox hazard analyses revealed AR expression to be independently associated with a good prognosis in overall patients (HR 0.46, P = 0.0052) and in the ER-positive cohort (HR 0.34, P = 0.0009). AR expression is associated with a less aggressive phenotype and a good prognosis in patients with ER-positive breast cancer. This is considered to be a specific phenomenon for postmenopausal breast cancer patients.


The problems of hypertension in the elderly.
The cut off age for elderly person in India is 60-65 years, in the USA is 75-80 years and 6th Joint Committee on Detection, Evaluation and Treatment of High Blood Pressure (JNC-VI) has identified it as above 60 years. Elderly people may have (i) systolic-diastolic hypertension, (ii) isolated systolic hypertension or (iii) pseudohypertension. JNC-VI has classified hypertension in stage 1, stage 2 and stage 3 according to its severity. Hypertension is confirmed when BP measured on three separate occasions over 1-2 weeks and when consistently it is raised above 140/90 mm Hg. The management includes lifestyle modification and drug treatment. Lifestyle modification includes rationality of diet, regular exercise, stop smoking, stoppage of alcohol or moderation and yoga. Drugs commonly used are diureties and beta-blockers. Other antihypertensive drugs are calcium channel blockers, ACE-inhibitors, alpha-blockers and vasodilators.


Prescriptive Method for Optimizing Cost of Data Collection and Annotation in Machine Learning of Clinical Ultrasound.
Machine learning in medical ultrasound faces a major challenge: the prohibitive costs of producing and annotating clinical data. Optimizing the data collection and annotation will improve model training efficiency, reducing project cost and times. This paper prescribes a 2-phase method for cost optimization based on iterative accuracy/sample size predictions, and active learning for annotation optimization. Using public breast, fetal, and lung ultrasound datasets we can: Optimize data collection by statistically predicting accuracy for a desired dataset size; and optimize labeling efficiency using Active Learning, where predictions with lowest certainty were labelled manually using feedback. A practical case study on BUSI data was used to demonstrate the method prescribed in this work. With small data subsets, ~10%, dataset size vs. final accuracy relations can be predicted with diminishing results after 50% usage. Manual annotation was reduced by ~10% using active learning to focus the annotation. This led to cost reductions of 50%-66%, depending on requirements and initial cost model, on BUSI dataset with a negligible accuracy drop of 3.75% from theoretical maximums.Clinical Relevance- This work provides methodology to optimize dataset size and manual data labelling, this allows generation of cost-effective datasets, of interest to all, but particularly for financially limited trials and feasibility studies, Reducing the time burden on annotating clinicians.


ADBP-1 regulates an ADAR RNA-editing enzyme to antagonize RNA-interference-mediated gene silencing in Caenorhabditis elegans.
Small interfering RNAs (siRNAs) and microRNAs (miRNAs) mediate gene silencing through evolutionarily conserved pathways. In Caenorhabditis elegans, the siRNA/miRNA pathways are also known to affect transgene expression. To identify genes that regulate the efficiencies of the siRNA/miRNA pathways, we used the expression level of a transgene as an indicator of gene silencing and isolated a transgene-silencing mutant, adbp-1 (ADR-2 binding protein). The adbp-1 mutation caused transgene silencing in hypodermal and intestinal cells in a cell-autonomous manner, depending on the RNA interference (RNAi) machinery. The adbp-1 gene encodes a protein with no conserved domains that is localized in the nucleus. Yeast two-hybrid screening and co-immunoprecipitation analysis demonstrated that ADBP-1 physically interacts with ADR-2, an RNA-editing enzyme from the ADAR (adenosine deaminase acting on dsRNA) family. In the adbp-1 mutant, as previously shown in adr-2 mutants, A-to-I RNA editing was not detected, suggesting that ADBP-1 is required for the RNA-editing activity of ADR-2. We found that ADBP-1 facilitates the nuclear localization of ADR-2. ADBP-1 may regulate ADR-2 activity and the consequent RNA editing and thereby antagonize RNAi-mediated transgene silencing in C. elegans.


Overreplication of short DNA regions during S phase in human cells.
DNA replication origins (ORI) are regulatory regions from which the genome is replicated once every cell cycle. A widely used method for their identification in mammalian chromosomes relies on quantitative PCR of DNA nascent strands across candidate regions. We developed a new high-resolution PCR strategy to localize ORIs directly on total unfractionated human DNA. The increase in sensitivity provided by this approach has revealed that a short region of approximately 200-base-pair overlapping well-characterized replication origins undergoes several rounds of replication, coinciding with their specific time of activation during S phase. This process generates a population of discrete dsDNA fragments detectable as free molecules in preparations of total DNA in normally proliferating cells. Overreplicated regions have precise boundaries at the edge of the nucleosome-free gap that encompasses the transcription initiation sites of CpG island promoters. By itself, active transcription does not induce overreplication but does stimulate it at ORIs associated with promoters. The coincidence in time and space between the overproduction of short DNA fragments and ORI activity predicts the precise localization of thousands of ORIs in the human genome and uncovers a previously unnoticed step in the initiation of DNA replication.


Neuronal, dendritic, and vascular profiles of human temporal lobe epilepsy correlated with cellular physiology in vivo.
Partial complex seizures are known to arise from abnormal firing of neurons in cortex that has histologic abnormalities associated with tumors, infarcts, or neuron loss. The latter pathology of sclerosis is most frequently found in the hippocampus, and partial seizures from this region are focalized by direct electrical recordings and treated by anterior temporal lobectomy. Although we can link this hippocampal sclerosis to nearby hyperexcitability, the synaptic mechanisms involved in hippocampal seizure genesis are not yet known. We have used in vivo microelectrode recordings from hippocampal neurons and found rare instances of anomalous bursting patterns as well as coupled firing. Postinhibitory "rebound excitation" has also been recorded, supporting the concept that synchronized hippocampal outputs are important for seizure genesis. Immunocytochemistry of GAD-positive inhibitory interneurons indicates no significant loss in inhibition in the sclerotic hippocampus and a normal number of inhibitory interneurons in its output target, the presubiculum. The presubiculum, with its multi-layered cortex, may amplify and propagate seizures to other cortices. Golgi and electron microscopy of epileptic neurons have shown pre- and postsynaptic alterations that may contribute to seizure genesis. Finally, ultrastructural analysis of capillaries in sclerotic hippocampus indicates deficient plasma-tissue transport that may contribute to cell loss or may alter neuronal excitability.


Phosphorus extractability of soils amended with stockpiled and composted cattle manure.
Managing fertilizer applications to maintain soil P below environmentally unacceptable levels should consider the contribution of manure and synthetic fertilizer sources to soluble and extractable forms of P. Our objective was to evaluate soil and manure characteristics and application rates on P extractability in recently amended soils. Five soils of the U.S. southern High Plains were amended with beef cattle manures, composted beef manure, and inorganic fertilizers [Ca(H(2)PO(4))(2) or KH(2)PO(4)] at five rates and incubated under controlled conditions. Mehlich 3-, Olsen (NaHCO(3))-, Texas A&M extractant (TAM)-, and water-extractable P were determined for the soils after selected incubation periods. Except for TAM and some water extractions, P extractability as a function of total P applied was linear (P < 0.001) for a wide range of application rates. Mehlich-3, NaHCO(3), and water P extraction efficiencies of KH(2)PO(4)-amended soils averaged 22, 34, and 115% greater (P < or = 0.036), respectively, than efficiencies of soils amended with manures except for the Texline (calcareous) loam and Pullman clay loam soils. Phosphorus extraction efficiencies decreased with time for KH(2)PO(4)-amended soils (P < 0.05) but remained stable or increased for manure-amended soils during the 8-wk incubation period. Across all soils and manure sources, changes in water-extractable P per unit increase in Mehlich 3-, NaHCO(3)-, and TAM-extractable P averaged 100, 85, and 125% greater, respectively, for inorganic as compared with manure-amended soils. These source-dependent relationships limit the use of agronomic soil extractants to make correct inferences about water-extractable P and dissolved P in runoff.


Enhancing cochlear duct length estimation by incorporating second-turn parameters.
Estimating insertion depth, cochlear duct length (CDL), and other inner ear parameters is vital to optimizing cochlear implantation outcomes. Most current formulas use only the basal turn dimensions for CDL prediction. In this study, we investigated the importance of the second turn parameters in estimating CDL. Two experienced neuro-otologists blindly used segmentation software to measure (in mm) cochlear parameters, including basal turn diameter (A), basal turn width (B), second-turn diameter (A2), second-turn width (B2), CDL, first-turn length, and second-turn length (STL). These readings were taken from 33 computed tomography (CT) images of temporal bones from anatomically normal ears. We constructed regression models using A, B, A2, and B2 values fitted to CDL, two-turn length, and five-fold cross-validation to ensure model validity. CDL, A value, and STL were longer in males than in females. The mean B2/A2 ratio was 0.91 ± 0.06. Adding A2 and B2 values improved CDL prediction accuracy to 86.11%. Therefore, we propose a new formula for more accurate CDL estimation using A, B, A2, and B2 values. In conclusion, the findings of this study revealed a notable improvement in the prediction of two-turn length (2TL), and CDL by clinically appreciable margins upon adding A2 and B2 values to the prediction formulas.


Discovery of Potent Inhibitors of Streptococcus mutans Biofilm with Antivirulence Activity.
Dental caries is a bacterial infectious disease characterized by demineralization of the tooth enamel. Treatment of this disease with conventional antibiotics is largely ineffective as the cariogenic bacteria form tenacious biofilms that are resistant to such treatments. The main etiological agent for dental caries is the bacterium Streptococcus mutans. S. mutans readily forms biofilms on the tooth surface and rapidly produces lactic acid from dietary sucrose. Glucosyl transferases (Gtfs) secreted by S. mutans are mainly responsible for the production of exopolysaccharides that are crucial for the biofilm architecture. Thus, inhibiting S. mutans' Gtfs is an effective approach to develop selective biofilm inhibitors that do not affect the growth of oral commensals. Herein, we report a library of 90 analogs of the previously identified lead compound, G43, and exploration of its structure activity relationships (SAR). All compounds were evaluated for the inhibition of S. mutans biofilms and bacterial growth. Selected compounds from this library were further evaluated for enzyme inhibition against Gtfs using a zymogram assay and for growth inhibition against oral commensal bacterial species such as Streptococcus gordonii and Streptococcus sanguinis. This study has led to the discovery of several new biofilm inhibitors with enhanced potency and selectivity. One of the leads, III F1 , showed marked reduction in buccal, sulcal, and proximal caries scores in a rat model of dental caries.


Mortality Risk Associated With Long-acting Injectable Antipsychotics: A Systematic Review and Meta-analyses of Randomized Controlled Trials.
Long-acting injectable (LAI) antipsychotics (LAI-APs) have several advantages over oral medications, but deaths reported in Japan during the early post-marketing phase vigilance period have raised safety concerns. We conducted a series of meta-analyses to assess whether LAI-APs affect the mortality of patients with schizophrenia. Three categorical meta-analyses of randomized controlled trials (RCTs) were performed to compare all-cause death (primary outcome) and death due to suicide: individual and pooled LAI-APs vs placebo, individual and pooled LAI-APs vs oral antipsychotics (OAPs), and head-to-head comparisons of LAI-APs. The risk ratios (RRs) and 95% CIs were calculated. We identified 52 RCTs (53 comparisons; total participants = 17 416, LAI-APs = 11 360, OAP = 3910, and placebo = 2146; mean study duration [wk]: LAI-APs vs placebo = 28.9, LAI-APs vs OAPs = 64.5). Neither pooled nor individual LAI-APs (aripiprazole, fluphenazine, olanzapine, paliperidone, and risperidone) differed from the placebo regarding the incidences of all-cause death (pooled LAI-APs: RR = 0.64, P = .37) and death due to suicide (pooled LAI-APs: RR = 0.98, P = .98). However, in a subgroup meta-analysis of only short-duration RCTs (≤13wk), pooled LAI-APs exhibited a trend toward lower incidence of all-cause death than placebo (RR = 0.29, P = .08). Pooled LAI-APs (aripiprazole, fluphenazine, haloperidol, olanzapine, paliperidone, risperidone, and zuclopenthixol) did not differ from pooled OAPs regarding all-cause death (pooled LAI-APs: RR = 0.71, P = .30) and death due to suicide (pooled LAI-APs: RR = 0.94, P = .91). Individual LAI-APs and OAPs were associated with similar risks of death. Data for head-to-head comparisons of individual LAI-APs were insufficient. In conclusion, there was no significant difference between LAI-APs and placebo or OAPs regarding all-cause death and death due to suicide.


The effect of dietary fiber in tube-fed elderly patients.
Diarrhea is a common problem with potentially serious consequences for elderly tube-fed patients. A review of the literature indicates that diarrhea has been associated with the use of antibiotics, low blood albumin concentration, and a lack of fiber in the diet. This study documents the observed effect of dietary fiber on the incidence of diarrhea in 50 tube-fed patients. Over a 3-week period, patients who received fiber-containing formula had fewer liquid/loose stools than patients who received fiber-free formula. Nurses who care for elderly, tube-fed patients can help to improve outcomes by advocacy and interdisciplinary collaboration with registered dietitians and primary providers and by teaching assistive staff the importance of accurate recording of bowel function.


Knowledge, Attitude, and Practices Regarding COVID-19 Among Chronic Illness Patients at Outpatient Departments in Ho Chi Minh City, Vietnam.
The novel coronavirus disease (COVID-19) has become a major threat to human life around the world. This study aims to assess the knowledge, attitude, and practices regarding COVID-19 among people with chronic diseases at the outpatient departments in Ho Chi Minh City. A cross-sectional study was carried out between February and March 2020 using a convenience sampling strategy in three hospitals in Ho Chi Minh City (HCMC) via the use of a structured self-administered questionnaire. Factors relating to practices, prevalence ratio (PR), and 95% confidence interval were estimated by using the Poisson regression with robust options. P-value <0.05 was considered as statistically different. A total of 522 participants had a mean age of 51.5 ± 10.6 years. Most of them reported seeing information regarding the COVID-19 pandemic (93.7%) via television and social media (72.8% and 62.1%, respectively). Just over two-thirds of the participants (68.4%) answered with sufficient knowledge of COVID-19. Most respondents had a positive attitude toward COVID-19 (90.8%), although some misconceptions existed. Almost over three-fourths of them (77.2%) maintained good practices for prevention. The rate of good practices in those who had sufficient knowledge was 1.24 times greater than that among those who had insufficient knowledge (PR 1.24, 95% CI: 1.10-1.41, P<0.05). Also, the rate of good practices in males was lower than that of females (PR: 0.91, 95% CI: 0.83-0.99, P<0.05). There still exists an amount of insufficient knowledge and negative attitude regarding COVID-19, which may be barriers to good prevention practices among chronic illness patients. Education programs need to continue via television and social media and emphasize that people with chronic diseases are more likely to experience severe symptoms, including death from COVID-19. Additionally, management authorities should prolong specific policies to protect the more vulnerable in our community.


Intergroup conflict self-perpetuates via meaning: Exposure to intergroup conflict increases meaning and fuels a desire for further conflict.
We investigated whether violent conflict provides individuals with a sense of meaning that they are hesitant to let go of, thus contributing to the perpetuation of intergroup conflict. Across a wide variety of contexts, we found that making intergroup conflict salient increased the meaning people found in conflict and, in turn, increased support for conflict-perpetuating beliefs, ideologies, policies, and behaviors. These effects were detected among participants exposed to reminders of intergroup conflict (the American Revolutionary War and the U.S.-led campaign against ISIS; Studies 1A and 1B), participants living through actual intergroup conflict (the 2014 Israel-Gaza war; Study 2), and participants who perceived actual intergroup conflicts to be larger versus smaller in scope (the November 2015 Paris attacks; Studies 3 and 4). We also found that directly manipulating the perceived meaning in conflict (in the context of the 2014 NYC "hatchet attack"; Study 5) led to greater perceived meaning in life in general and thereby greater support for conflict escalation. Together, these findings suggest that intergroup conflict can serve as a source of meaning that people are motivated to hold on to. We discuss our findings in the context of the meaning making and threat compensation literatures, and consider their implications for perspectives on conflict escalation and resolution. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Bayes variable selection in semiparametric linear models.
There is a rich literature on Bayesian variable selection for parametric models. Our focus is on generalizing methods and asymptotic theory established for mixtures of g-priors to semiparametric linear regression models having unknown residual densities. Using a Dirichlet process location mixture for the residual density, we propose a semiparametric g-prior which incorporates an unknown matrix of cluster allocation indicators. For this class of priors, posterior computation can proceed via a straightforward stochastic search variable selection algorithm. In addition, Bayes factor and variable selection consistency is shown to result under a class of proper priors on g even when the number of candidate predictors p is allowed to increase much faster than sample size n, while making sparsity assumptions on the true model size.


Oenococcus oeni Exopolysaccharide Biosynthesis, a Tool to Improve Malolactic Starter Performance.
Oenococcus oeni is the lactic acid bacterium that most commonly drives malolactic fermentation (MLF) in wine. Though the importance of MLF in terms of wine microbial stability and sensory improvement is well established, it remains a winemaking step not so easy to control. O. oeni displays many adaptation tools to resist the harsh wine conditions which explain its natural dominance at this stage of winemaking. Previous findings showed that capsular polysaccharides and endogenous produced dextran increased the survival rate and the conservation time of malolactic starters. In this paper, we showed that exopolysaccharides specific production rates were increased in the presence of single stressors relevant to wine (pH, ethanol). The transcription of the associated genes was investigated in distinct O. oeni strains. The conditions in which eps genes and EPS synthesis were most stimulated were then evaluated for the production of freeze dried malolactic starters, for acclimation procedures and for MLF efficiency. Sensory analysis tests on the resulting wines were finally performed.


Randomized clinical trial: Effective gluten degradation by Aspergillus niger-derived enzyme in a complex meal setting.
The Aspergillus niger-derived prolyl endoprotease (AN-PEP) has previously been shown to degrade gluten in healthy subjects when added to an intragastrically infused meal. The current study investigated the efficacy of AN-PEP in a physiological meal setting. In this randomized placebo-controlled crossover study, 18 gluten-sensitive subjects consumed a porridge containing 0.5 g gluten together with two tablets either containing a high or low dose of AN-PEP, or placebo. Gastric and duodenal content was sampled over 180 minutes, and areas under the curve of gluten concentrations were calculated. The primary outcome, i.e. success rate of high dose AN-PEP defined as at least 50% gluten degradation compared to placebo in the duodenum, was achieved in 10 of 13 comparisons. In the stomach, gluten levels were reduced from 176.9 (median, interquartile range 73.5-357.8) to 22.0 (10.6-50.8, p = 0.001) in the high dose and to 25.4 μg × min/ml (16.4-43.7, p = 0.001) in the low dose. In the duodenum, gluten levels were reduced from 14.1 (8.3-124.7) in the placebo to 6.3 (3.5-19.8, p = 0.019) in the high dose and to 7.4 μg × min/ml in the low dose (3.8-12.0, p = 0.015). Thus even in a physiological meal setting, AN-PEP significantly degraded most gluten in the stomach before it entered the duodenum.


THE START-TO-RUN DISTANCE AND RUNNING-RELATED INJURY AMONG OBESE NOVICE RUNNERS: A RANDOMIZED TRIAL.
High body mass index is associated with an increased risk of running-related injury among novice runners. However, the amount of running participation plays a fundamental explanatory role in regards to running-related injury development. Therefore, the purpose of the present study was to investigate if the risk of running-related injury among obese novice runners (BMI 30-35) was different when the start-to-run distance was 3km per week instead of 6km per week. A start-to-run distance of 3km per week is associated with 20% fewer running-related injuries and significantly fewer symptoms of overuse injury than a start-to-run distance of 6km per week among obese novice runners. Randomized trial. Fifty-six obese novice runners with a body mass index between 30-35 were enrolled and randomized to receive one of the two following Interventions: (i) a 4-week running program with a start-to-run distance of 3km per week including three sessions with 1km running per session (n=29), or (ii) a 4-week running program with a start-to-run distance of 6km per week including three sessions with 2km running per session (reference group, n=27). In both programs, the weekly running distance was increased by 10% each week throughout the follow-up. The intention-to-treat analysis revealed a protective cumulative risk difference of -16.3% (95%CI: -43.8%; 11.3%, p=0.25) after four weeks. Importantly, some participants completed much more running than prescribed (n=5) and some never uploaded any training (n=15). Therefore, a supplementary per-protocol analysis was performed revealing a cumulative risk difference of -31.2% (95%CI: -57.0%; -5.2%, p=0.02) after four weeks. Furthermore, in the per-protocol analysis, the cumulative risk difference of overuse-injury symptoms was -47.8% (95%CI: -81.0%; -14.6%, p=0.01) after four weeks of running. A 3km reduction from 6km per week to 3km per week in the start-to-run distance appears to be associated with fewer running-related injuries and significantly fewer symptoms of overuse injury. 2b.


Clinical significance of singleton pregnancies complicated by placental abruption associated with histological chorioamnionitis.
The aim of this study was to determine the perinatal outcomes of placental abruption associated with the presence of histological chorioamnionitis. We reviewed the obstetric records of 96 singleton deliveries complicated by placental abruption after 22 weeks' gestation. Of these 96 cases, 37 cases (39%) were diagnosed as having histological chorioamnionitis in the placenta. The incidence of premature delivery, preterm rupture of the membranes and low birth weight in the cases of placental abruption with chorioamnionitis were higher than in cases without chorioamnionitis. However, there were no significant differences in the incidence of other outcomes, such as fetal demise, low Apgar score, or low umbilical artery pH, between the cases of placental abruption with and without histological chorioamnionitis. Although the incidence of prematurity in the cases of placental abruption with chorioamnionitis was higher than that in cases without chorioamnionitis, there were no significant differences in fetal and neonatal conditions between the abruption cases with and without chorioamnionitis.


[Career, trainee physicians].
Medicine offers trainee physicians many different possibilities to practice their vocation. Young doctors should try to assess their own motivation and skills as precisely as possible, especially regarding a more theoretical, more practical, more academic or a purely patient-oriented career path. However, different phases of life may bring about changes in priorities. The course for the direction of a career path is often set early on. This is particularly true during the early career phase when institutions, such as medical faculties, have the responsibility to provide authoritative consulting and good mentorship.


Neutron and cobalt-60 gamma irradiation produce similar changes in DNA supercoiling.
The effects of 60Co gamma-ray and d(20 MeV)Be neutron irradiation on DNA supercoiling have been studied using a nucleoid rewinding technique. Irradiation of viable CHO AA8 cells on ice with from 4 to 25 Gy of either radiation produced a similar resistance to rewinding of nuclear supercoils after treatment with ethidium bromide. The restitution from the effects of 12 Gy of either radiation was also similar, leaving no detectable residual damage. The discrepancy between these data and the reduced ability of neutrons to produce DNA breaks, as defined by the alkaline elution assay, is explained by the discontinuous deposition of dose associated with neutron irradiation. It is suggested from a microdosimetric analysis that the neutron radiation interacts with DNA at sites on average 5-10 times further apart than the interactions with gamma rays. The long DNA sequences which results after neutron irradiation are consequently eluted inefficiently during alkaline elution, giving a reported RBE of approximately 0.3. Restrictions in the rewinding of individual supercoils are not dependent on the interionization distance and thus give rise to an RBE of approximately 1. Furthermore, the complete removal of DNA damage, as measured by this technique, supports the hypothesis that neutron toxicity is associated with incorrect, not incomplete, rejoining of the DNA molecule.


Lysophospholipids as Predictive Markers of ST-Elevation Myocardial Infarction (STEMI) and Non-ST-Elevation Myocardial Infarction (NSTEMI).
The present study explored patterns of circulating metabolites and proteins that can predict future risk for ST-elevation myocardial infarction (STEMI) and non-ST-elevation myocardial infarction (NSTEMI). We conducted a prospective nested case-control study in northern Sweden in individuals who developed STEMI (N = 50) and NSTEMI (N = 50) within 5 years and individually matched controls (N = 100). Fasted plasma samples were subjected to multiplatform mass spectrometry-based metabolomics and multiplex protein analyses. Multivariate analyses were used to elucidate infarction-specific metabolite and protein risk profiles associated with future incident STEMI and NSTEMI. We found that altered lysophosphatidylcholine (LPC) to lysophosphatidylethanolamine (LPE) ratio predicted STEMI and NSTEMI events in different ways. In STEMI, lysophospholipids (mainly LPEs) were lower, whereas in NSTEMI, lysophospholipids (mainly LPEs) were higher. We found a similar response for all detected lysophospholipids but significant alterations only for those containing linoleic acid (C18:2, p < 0.05). Patients with STEMI had higher secretoglobin family 3A member 2 and tartrate-resistant acid phosphate type 5 and lower platelet-derived growth factor subunit A, which are proteins associated with atherosclerosis severity and plaque development mediated via altered phospholipid metabolism. In contrast, patients with NSTEMI had higher levels of proteins associated with inflammation and macrophage activation, including interleukin 6, C-reactive protein, chemerin, and cathepsin X and D. The STEMI risk marker profile includes factors closely related to the development of unstable plaque, including a higher LPC:LPE ratio, whereas NSTEMI is characterized by a lower LPC:LPE ratio and increased inflammation.


Health-related quality of life: a population based study from Slovenia.
Health status is represented by people's subjective assessment of their sense of well-being and ability to perform social roles and has been well accepted as a health indicator of different populations. The aim of this study was to determine health-related quality of life in Slovenian population. We performed a cross-sectional postal survey in a random stratified sample of 1,000 adult Slovenian inhabitants. The questionnaire consisted of the respondents' demographic data (sex, age, education level, employment status, living environment), self-reported chronic conditions, self-reported use of health services and EQ-5D instrument for measuring quality of life. The response rate was 41% (53.1% men, mean age 51.5 years). Respondents reported most problems in the pain dimension of EQ-5D (59.3%), following by mobility (30.4%), anxiety/depression (30.3%), daily activities (29.8%) and self-care (9.0%). At least one moderate problem was reported by 272 (66.3%) respondents. Independent factors, associated with problems in any EQ-5D dimension were primary and vocational education, older age, high blood pressure, rheumatic diseases, back problems, anxiety/depression, a visit to the emergency department in the past year, and a house visit from a family doctor in the past year. The present study showed that the health-related quality of life of the Slovenian inhabitants is lower than the one found in some other European countries. This finding is surprising and also worrying. Because we cannot find any perceptible reason for this observation, larger and prospective studies are needed to confirm those results and to determine the reasons for that.


Survival estimates for patients with homozygous sickle-cell disease in Jamaica: a clinic-based population study.
Information about life expectancy of patients with homozygous sickle-cell disease is needed for research and patient counselling. Our aim was to study two Jamaican populations, one clinic-based and one birth cohort and, by careful consideration of data quality and statistical analysis, to identify ways to increase the chances of obtaining valid and generalisable results. We investigated the survival experience of 3301 patients with homozygous sickle-cell disease attending the Jamaican sickle-cell clinic between Jan 1, 1987, and Dec 31, 1996. We applied and assessed a simulation technique for incorporating early life mortality using a birth cohort, and analysed the precision of this technique. Kaplan-Meier survival estimates are produced. 290 of the 3301 patients died. Median survival calculated with the excess mortality rate simulation data was 53 years (95% CI 49.3-57.0) for men and 58.5 (55.1-67.5) for women. Our simulation technique, with realistic assumptions based on empirical evidence, offers a new estimate of median survival for patients with homozygous sickle-cell disease. We present the precision of these survival estimates, which introduces an important level of uncertainty. The inherent biases of clinically ascertained populations of patients, and the assumptions underlying analysis techniques are crucial features of survival studies in sickle-cell disease, and can modify summary statistics substantially.


Breast-Feeding Analgesia in Infants: An Update on the Current State of Evidence.
To provide an updated synthesis of the current state of the evidence for the effectiveness of breast-feeding and expressed breast milk feeding in reducing procedural pain in preterm and full-term born infants. A systematic search of key electronic databases (PubMed, CINAHL, EMBASE) was completed. Of the 1032 abstracts screened, 21 were found eligible for inclusion. Fifteen studies reported on the use of breast-feeding or expressed breast milk in full-term infants and 6 reported on preterm infants. Direct breast-feeding was more effective than maternal holding, maternal skin-to-skin contact, topical anesthetics, and music therapy, and was as or more effective than sweet tasting solutions in full-term infants. Expressed breast milk was not consistently found to reduce pain response in full-term or preterm infants. Studies generally had moderate to high risk of bias. There is sufficient evidence to recommend direct breast-feeding for procedural pain management in full-term infants. Based on current evidence, expressed breast milk alone should not be considered an adequate intervention.


Evidence that enforced sunlight exposure can cause hyperthermia in cattle ingesting low levels of ergot of rye (Claviceps purpurea), when air temperature and humidity conditions are only moderate.
To determine the rectal temperature response of cattle, following the oral administration of ergot of rye (Claviceps purpurea), under pen conditions of enforced sunlight compared with those of enforced shade. Hereford cross steers were divided into two groups of 18. One group was dosed once, on a Monday morning, with finely ground rye grass ergots at a rate of 180 mg/kg body weight and held in the sun for 7 h each day until Friday afternoon. The other group was not dosed but was similarly held in the sun during the same period. Their rectal temperatures were measured early morning and mid afternoon, from Monday to Friday inclusive. The process was repeated for each group, but this time they were held in the shade. The four treatment options were run concurrently by conducting the experiments over 6 weeks and using 3 animals in each treatment group, each week. The thermic response over all weeks, of the ergot treated, sunlight exposed cattle, was deemed greater than for the other groups, based on the following four parameters. The increase in rectal temperature between early morning and mid afternoon, the size of the mid afternoon rectal temperature rise, the difference between the maximum mid afternoon rectal temperature recorded by an animal in the sun compared with that recorded by the same animal in the shade, and finally the number of animals in a treatment group that recorded rectal temperatures > 40.00 degrees C. The difference in the daily increase in body temperature between the ergot treated, sun exposed cattle and the ergot treated, shaded cattle, was greater than that observed between the sun exposed and shade restricted control cattle. Nine of 18 ergot treated and sun exposed cattle developed hyperthermia; no cattle in the other three groups did. Some sunlight exposed cattle, dosed with a low amount of ergot of rye, can experience a body temperature elevation above the normal range, even under mild ambient temperature and humidity conditions. Sunlight exposure causes an increase in body temperature in normal cattle that is probably attributable to solar radiant heat. Ergot ingestion causes an increase in body temperature in shaded cattle that is probably attributable to ambient temperature and humidity effects. This response can be increased by sun exposure and this increase is attributable to solar radiant heat effects and possibly also solar radiation. A response to radiant heat is consistent with the known physiological effects of ingested ergot alkaloids, but a response to solar radiation is not.


[Complex surgical treatment of recurrent post-injection abscess in a patient with systemic autoimmune disease].
The authors present complex surgical treatment of a patient with rheumatoid arthritis and persistent wound in the right gluteal region after previous surgical treatment of post-injection abscess. According to these data, active surgical treatment of wounds and purulent-necrotic lesions of any etiology and localization, augmented by modern agents for local and systemic therapy (including phage therapy) provide favorable functional and cosmetic results even in patients with systemic autoimmune diseases receiving glucocorticosteroids. It is important to assess the wound process considering not only clinical data, but also objective information of qualitative and quantitative microbiological and cytological examinations.


The fungal mycobiome promotes pancreatic oncogenesis via activation of MBL.
Bacterial dysbiosis accompanies carcinogenesis in malignancies such as colon and liver cancer, and has recently been implicated in the pathogenesis of pancreatic ductal adenocarcinoma (PDA)1. However, the mycobiome has not been clearly implicated in tumorigenesis. Here we show that fungi migrate from the gut lumen to the pancreas, and that this is implicated in the pathogenesis of PDA. PDA tumours in humans and mouse models of this cancer displayed an increase in fungi of about 3,000-fold compared to normal pancreatic tissue. The composition of the mycobiome of PDA tumours was distinct from that of the gut or normal pancreas on the basis of alpha- and beta-diversity indices. Specifically, the fungal community that infiltrated PDA tumours was markedly enriched for Malassezia spp. in both mice and humans. Ablation of the mycobiome was protective against tumour growth in slowly progressive and invasive models of PDA, and repopulation with a Malassezia species-but not species in the genera Candida, Saccharomyces or Aspergillus-accelerated oncogenesis. We also discovered that ligation of mannose-binding lectin (MBL), which binds to glycans of the fungal wall to activate the complement cascade, was required for oncogenic progression, whereas deletion of MBL or C3 in the extratumoral compartment-or knockdown of C3aR in tumour cells-were both protective against tumour growth. In addition, reprogramming of the mycobiome did not alter the progression of PDA in Mbl- (also known as Mbl2) or C3-deficient mice. Collectively, our work shows that pathogenic fungi promote PDA by driving the complement cascade through the activation of MBL.


[Cutaneous Melanoma (CM): Current Diagnosis and Treatment].
Cutaneous melanoma (CM) is the third most common cancer of the skin, but it is the neoplasia with the greatest impact on mortality. Its etiology is multifactorial and it has been reported that its prevalence has increased in the last two decades. In Mexico, CM ranks seventh in frequency among all malignancies and 80% of cases are in locally advanced stages. The prognosis depends on the stage. The prognostic factors with greatest impact in survival are nodal status, tumor thickness or Breslow depth, ulceration, and in thin melanomas (< 1 mm thickness, without ulceration and Clarck level III), the mitotic index. The diagnostic approach is of great importance to achieve adequate treatment. Adherence to global guidelines of treatment allows us to obtain the best rates of locoregional control, which is the first target to be achieved in patients with CM. The goal of this manuscript is to provide a synthesis of the most important aspects in the diagnosis and treatment of CM, based on current evidence obtained in the literature.


Comparison of whole blood dried on filter paper and serum for measurement of the temporal antibody response to avian infectious bronchitis virus by enzyme-linked immunosorbent assay.
Two methods for collecting specimens for measuring sequential antibody activity of infectious bronchitis virus (IBV) were compared. Whole blood was collected on filter-paper strips, dried for 2 hr at 37 C, and then stored in plastic bags at 4 C or eluted overnight and tested immediately. Eluates of whole blood were paired with serum samples and tested for IBV antibody activity by enzyme-linked immunosorbent assay (ELISA) at four weekly intervals. Both sampling methods yielded ELISA antibody levels that were detectable at 7 days postinfection (PI), peaked at 21 days PI, and then began to decline by 28 days PI. The paired samples showed no significant difference (P less than 0.05) between ELISA titers at any time tested. Whole blood dried on filter paper could be stored sealed in plastic bags at 4 C for at least 2 weeks with no appreciable loss of antibody titers. Virus-neutralizing antibodies, measured in serum only, were not detectable until 14 days PI but then continued to rise through 28 days PI. It was concluded that eluates of whole blood dried on filter paper may be used as an alternative to sera in ELISA for measuring IBV antibodies.


Rat spinal cord alpha 2-adrenoceptors are of the alpha 2A-subtype: comparison with alpha 2A- and alpha 2B-adrenoceptors in rat spleen, cerebral cortex and kidney using 3H-RX821002 ligand binding.
Binding of the alpha 2-adrenoceptor antagonist radioligand 3H-RX821002 was investigated in membranes from rat spinal cord, spleen, cerebral cortex and kidney. The ligand was found to bind to saturable binding sites with apparent uniform affinities within each tissue. Seven compounds, some of which have previously been reported to be selective for either alpha 2A- or alpha 2B-adrenoceptors, were used in competition with 3H-RX821002. By using computer modelling, competition curves generated for three of these compounds (ARC 239, prazosin and oxymetazoline) could be resolved into two site fits in the kidney, Kd's of the drugs being compatible with the notion that these sites corresponded to alpha 2A- and alpha 2B-adrenoceptors. Moreover, rauwolscine and yohimbine were found to be about 14 and 9-fold selective for alpha 2B-adrenoceptors in the kidney. In all other tissues studied drug competition curves were uniphasic and computer modelled into one site fits, drug Kd's being well correlated to those for the alpha 2A-adrenoceptor. In rat spinal cord 26 further drugs, which showed wide variation in structure, were evaluated in competition with 3H-RX821002. Of these compounds, competion curves of the agonists UK-14,304, (-) and (+) adrenaline were modelled into two site fits whereas those of the remaining compounds could be modelled only into one site fits. Since the high affinity site for UK-14,304, (-) and (+) adrenaline was eliminated when EDTA, Gpp(NH)p and 140 mM NaCl was present in the assay the heterogeniety observed in spinal cord was considered to be due to formation of high and low affinity conformations of the alpha 2-adrenoceptor for agonists. It is concluded that 3H-RX821002 is useful to label both alpha 2A- and alpha 2B-adrenoceptors in the rat. Moreover, the binding sites labelled by 3H-RX821002 in the spinal cord appear to consist of a single population of alpha 2-adrenoceptors of the alpha 2A-type.


Cyclic stretch induces proliferation and TGF-beta1-mediated apoptosis via p38 and ERK in ureteric bud cells.
We previously reported that p38 mitogen-activated protein kinase (p38) and phosphorylated ERK are upregulated in cyst epithelium of human renal dysplasia and obstructive uropathy in fetal lambs (Omori S, Fukuzawa R, Hida M, Awazu M. Kidney Int 61: 899-906, 2002; Omori S, Kitagawa H, Koike J, Fujita H, Hida M, Pringle KC, Awazu M. Kidney Int 73: 1031-1037, 2008). Dysplastic epithelium is characterized by proliferation, apoptosis, and upregulation of Pax2 and transforming growth factor (TGF)-beta1. In the present study, we investigated whether cyclic mechanical stretching of ureteric bud cells, a mimic of the hydrodynamic derangement after fetal urinary tract obstruction, reproduces events seen in vivo. Cyclic stretch activated p38 and ERK and upregulated Pax2 expression in a time-dependent manner in ureteric bud cells. Stretch-stimulated Pax2 expression was suppressed by a p38 inhibitor, SB203580, or a MEK inhibitor, PD98059. 5-Deoxyuridine incorporation was increased by stretch at 24 h, which was also abolished by SB203580 or PD98059. On the other hand, apoptosis was not induced at 24 h by stretch but was significantly increased at 48 h. TGF-beta1 secretion was increased by stretch at 24 h, which was inhibited by SB203580 or PD98059. Inhibition of p38 or ERK as well as anti-TGF-beta antibody abolished the stretch-induced apoptosis. Finally, exogenous TGF-beta1 induced apoptosis of ureteric bud cells, which was inhibited by SB203580 and PD98059. In conclusion, cyclic stretch induces Pax2 upregulation, proliferation, and TGF-beta1-mediated apoptosis, features characteristic of dysplastic epithelium, via p38 and ERK in ureteric bud cells.


Characterization of the beta-lactamase promoter of pBR322.
The beta-lactamase promoter of pBR322, derived from Tn3, has been characterized using several techniques. The transcription initiation site is located 35 base pairs from the translation initiation codon of beta-lactamase. The mRNA produced in vitro has a 5' pppGpA terminus. RNA polymerase bound at this start site protects a region from about -50 to +20 from DNase I cleavage using the footprinting technique. RNA polymerase binds rapidly to the beta-lactamase promoter. The half-time of association is less than one-half minute. The half-time of dissociation is approximately 6 hr. A study of the binding of RNA polymerase at different temperatures showed a large change between 11 degrees and 15 degrees C. Comparison of these parameters with those reported for other promoters is discussed.


Synthesis of an enzymatically active FLP recombinase in vitro: search for a DNA-binding domain.
We have used an in vitro transcription and translation system to synthesize an enzymatically active FLP protein. The FLP mRNA synthesized in vitro by SP6 polymerase is translated efficiently in a rabbit reticulocyte lysate to produce enzymatically active FLP. Using this system, we assessed the effect of deletions and tetrapeptide insertions on the ability of the respective variant proteins synthesized in vitro to bind to the FLP recognition target site and to carry out excisive recombination. Deletions of as few as six amino acids from either the carboxy- or amino-terminal region of FLP resulted in loss of binding activity. Likewise, insertions at amino acid positions 79, 203, and 286 abolished DNA-binding activity. On the other hand, a protein with an insertion at amino acid 364 retained significant DNA-binding activity but had no detectable recombination activity. Also, an insertion at amino acid 115 had no measurable effect on DNA binding, but recombination was reduced by 95%. In addition, an insertion at amino acid 411 had no effect on DNA binding and recombination. On the basis of these results, we conclude that this approach fails to define a discrete DNA-binding domain. The possible reasons for this result are discussed.