[Visual evoked potentials: methodologic problems--variations of stimulus parameters].
The evaluation of visual potentials (VEP) evoked by pattern reversal includes the latency and amplitude of P100. An increase in the mean brightness or contrast of the checkerboard stimulus results in an increase in amplitude and a decrease in latency of P100. A square edge subtense of 20' in checkerboard pattern or a spatial frequency of 3-4 cyc/degrees in stripe pattern produce maximum responses. Transient VEPs are recorded at low repetition rates (less than 2 Hz), steady state VEPs at temporal frequencies above 8 Hz. In the latter condition the VEP takes the character of a sinusoidal train of waves which, compared to transient stimulation allows for a better measurement of VEP amplitude. The latency of P100 increases with the age of the subject. The influence of the different stimulus parameters and recording techniques must be known and have to be kept constant in order to record reliable and reproducible VEPs.


Sensitivity and specificity of the high fluorescent lymphocyte count-gate on the Sysmex XE-5000 hematology analyzer for detection of peripheral plasma cells.
Plasma cells are one of the end products of the B-lymphocyte mediated immune response. These cells normally reside in the bone marrow or some peripheral lymphoid tissues. Increased numbers of plasma cells in the blood, usually indicates pathology such as infection, auto-immunity or haematological malignancy. Therefore, the ability to measure plasma cells (PCs) on an automated cell analyzer might be advantageous. The performance of the Sysmex XE-5000 leukocyte differential channel [high fluorescent lymphocyte count (HFLC) area] was evaluated for the ability to detect plasma cells in peripheral blood and compared to the detection of plasma cells by flow cytometric analyses. Our results show that the HFL count from the XE-5000 correlates (R(2)=0.8) with the number of PCs in peripheral blood, but detects PCs with moderate to good sensitivity (88.9%) and specificity (87.8%). The Sysmex XE-5000 is suitable for screening blood samples for the presence of elevated number of plasma cells in peripheral blood, but the actual quantification needs to be confirmed by flow cytometry.


Probing cytoskeletal pre-stress and nuclear mechanics in endothelial cells with spatiotemporally controlled (de-)adhesion kinetics on micropatterned substrates.
The mechanical properties of living cells reflect their propensity to migrate and respond to external forces. Both cellular and nuclear stiffnesses are strongly influenced by the rigidity of the extracellular matrix (ECM) through reorganization of the cyto- and nucleoskeletal protein connections. Changes in this architectural continuum affect cell mechanics and underlie many pathological conditions. In this context, an accurate and combined quantification of the mechanical properties of both cells and nuclei can contribute to a better understanding of cellular (dys-)function. To address this challenge, we have established a robust method for probing cellular and nuclear deformation during spreading and detachment from micropatterned substrates. We show that (de-)adhesion kinetics of endothelial cells are modulated by substrate stiffness and rely on the actomyosin network. We combined this approach with measurements of cell stiffness by magnetic tweezers to show that relaxation dynamics can be considered as a reliable parameter of cellular pre-stress in adherent cells. During the adhesion stage, large cellular and nuclear deformations occur over a long time span (>60 min). Conversely, nuclear deformation and condensed chromatin are relaxed in a few seconds after detachment. Finally, our results show that accumulation of farnesylated prelamin leads to modifications of the nuclear viscoelastic properties, as reflected by increased nuclear relaxation times. Our method offers an original and non-intrusive way of simultaneously gauging cellular and nuclear mechanics, which can be extended to high-throughput screens of pathological conditions and potential countermeasures.


Inflammatory Chemokines in Atherosclerosis.
Atherosclerosis is a long-term, chronic inflammatory disease of the vessel wall leading to the formation of occlusive or rupture-prone lesions in large arteries. Complications of atherosclerosis can become severe and lead to cardiovascular diseases (CVD) with lethal consequences. During the last three decades, chemokines and their receptors earned great attention in the research of atherosclerosis as they play a key role in development and progression of atherosclerotic lesions. They orchestrate activation, recruitment, and infiltration of immune cells and subsequent phenotypic changes, e.g., increased uptake of oxidized low-density lipoprotein (oxLDL) by macrophages, promoting the development of foam cells, a key feature developing plaques. In addition, chemokines and their receptors maintain homing of adaptive immune cells but also drive pro-atherosclerotic leukocyte responses. Recently, specific targeting, e.g., by applying cell specific knock out models have shed new light on their functions in chronic vascular inflammation. This article reviews recent findings on the role of immunomodulatory chemokines in the development of atherosclerosis and their potential for targeting.


Evidence for polyreactivity seen with monoclonal antibodies produced against type II collagen.
Specific type II collagen monoclonal antibodies are needed for the quantification of articular cartilage collagen. In this study we produced and characterized 29 type II collagen monoclonal antibodies. Hybridomas were generated from mice immunized with rat type II collagen, selected for high antibody production against type II collagen using ELISA. Antibodies from selected and cloned hybridoma cells were purified by affinity chromatography and their reactivity tested by ELISA against a panel of antigens including actin, thyroglobulin, and single stranded DNA, all of which have been used to characterize the 'naturally occurring antibodies.' It was found that many of the anti-type II collagen monoclonal antibodies reacted to more than one antigen. The monospecific antibodies had higher affinity to type II collagen than the antibodies which demonstrated multireactivity. Because of the prevalence of polyreactive anti-type II collagen antibodies, it is advisable to employ highly selective methodologies to isolate high affinity monospecific antibodies to type II collagen.


Evaluating experimental bias and completeness in comparative phosphoproteomics analysis.
Unraveling the functional dynamics of phosphorylation networks is a crucial step in understanding the way in which biological networks form a living cell. Recently there has been an enormous increase in the number of measured phosphorylation events. Nevertheless, comparative and integrative analysis of phosphoproteomes is confounded by incomplete coverage and biases introduced by different experimental workflows. As a result, we cannot differentiate whether phosphosites indentified in only one or two samples are the result of condition or species specific phosphorylation, or reflect missing data. Here, we evaluate the impact of incomplete phosphoproteomics datasets on comparative analysis, and we present bioinformatics strategies to quantify the impact of different experimental workflows on measured phosphoproteomes. We show that plotting the saturation in observed phosphosites in replicates provides a reproducible picture of the extent of a particular phosphoproteome. Still, we are still far away from a complete picture of the total human phosphoproteome. The impact of different experimental techniques on the similarity between phosphoproteomes can be estimated by comparing datasets from different experimental pipelines to a common reference. Our results show that comparative analysis is most powerful when datasets have been generated using the same experimental workflow. We show this experimentally by measuring the tyrosine phosphoproteome from Caenorhabditis elegans and comparing it to the tyrosine phosphoproteome of HeLa cells, resulting in an overlap of about 4%. This overlap between very different organisms represents a three-fold increase when compared to dataset of older studies, wherein different workflows were used. The strategies we suggest enable an estimation of the impact of differences in experimental workflows on the overlap between datasets. This will allow us to perform comparative analyses not only on datasets specifically generated for this purpose, but also to extract insights through comparative analysis of the ever-increasing wealth of publically available phosphorylation data.


Cross-cultural translation and validation of the Chinese Oxford Knee Score and the Activity and Participation Questionnaire.
To cross-culturally translate and validate the Chinese versions of the Oxford Knee Score (OKS) and the Activity and Participation Questionnaire (APQ) in patients with end-stage knee osteoarthritis who are also candidates for knee replacement. The Chinese version of the OKS and APQ was completed by standard forward-backward translation and adaption. The feasibility was validated by a pretest in 30 patients. The final version together with the Short Form-36 (SF-36), EQ-5D, and EQ visual analog scale were assessed in 150 patients, and the OKS and APQ were repeated in 30 patients after a 2-week interval. The psychometric properties of the OKS and APQ were evaluated for test-retest reliability using intraclass correlation coefficients (ICCs), internal consistency using Cronbach's α, and construct validity using Spearman's correlation analysis. All patients were able to understand and complete both the OKS and APQ without difficulty (i.e. no missing data). The ICCs were 0.959 for the OKS, 0.956 for the APQ for total scores, and >0.7 for each item. Cronbach's α was greater than 0.7, and the corrected item-total correlation was greater than 0.4 for each item of both questionnaires. The OKS and APQ showed better correlations with questions from the pain and function domains than with those from the mental status domains of the SF-36 and EQ-5D. No floor or ceiling effect was identified in either questionnaire. The Chinese versions of the OKS and APQ are easy to understand and complete and showed good reliability and validity. They can be used to assess patient-reported outcomes after undergoing knee replacement in mainland China.


Improving oral squamous cell carcinoma diagnosis and treatment with fluorescence molecular imaging.
Timely identification and complete removal of oral squamous cell carcinoma (OSCC) through surgery is crucial for effective treatment. However, current diagnostic methods that rely on physical abnormalities are not very informative and practical in clinical settings, leading to the late detection of oral cancer. Furthermore, no dependable intraoperative tools available for assessing surgical margins in real-time. Fluorescence imaging allows the visualization of biological processes occurring in the early stages of cancer, and as a result, small tumors can be detected at an early stage. Fluorescence imaging can effectively aid in assessing excised edges during surgery for OSCC as it possesses high sensitivity and spatial resolution. This review focuses on tongue cancer as a representation of OSCC and delves into various fluorescence techniques that can aid in early diagnosis and surgical guidance. The review also discusses the potential clinical applications of these techniques in the future.


Nurses Exploring the Spirituality of Their Patients With Cancer: Participant Observation on a Medical Oncology Ward.
Attention for spirituality should be an integral part of professionals' caregiving. Particularly, nurses caring for patients with cancer might have opportunities to give attention to this dimension. The aim of this study was to gain insight in the way and extent to which nurses during daily caregiving observe and explore spiritual issues of hospitalized patients with cancer. We performed an ethnographic study with participant observation. Data were collected in 2015 during 4 shifts at the medical oncology department of a university hospital. The researcher, a spiritual care provider (chaplain) wearing the same kind of uniform as the nurses, observed the nurses, participated in their actions, and interviewed them after the shift. Although the patients did send many implicit and explicit messages concerning spiritual issues, the nurses did not explore them. If noticed, 3 barriers for exploring spiritual issues were mentioned by the nurses: lack of time, conflict with their mindset, and being reserved to talk about such issues. During their daily caregiving to patients with a life-threatening illness, nurses have many opportunities to explore spiritual issues, but they do not often recognize them. If they do, they tend not to explore the spiritual issues. Communication training for nurses is necessary to develop skills for exploring the spiritual dimension in patients with cancer. In such training, attention to the misconception that such a conversation requires a lot of time and for recognizing signals from patients inviting an exploration of their concerns is necessary.


Septicemic listeriosis in wild hares from Saskatchewan, Canada.
The bacterium Listeria monocytogenes causes disease in a wide variety of mammals including rabbits and hares. We describe naturally acquired metritis and septicemic listeriosis in wild female hares from Saskatchewan, Canada. Between April 2012 and July 2013, two white-tailed jackrabbits (Lepus townsendii) and a snowshoe hare (Lepus americanus) were presented to the Veterinary Medical Centre at the Western College of Veterinary Medicine, Saskatoon, Saskatchewan, Canada with nonspecific neurologic signs. The hares were euthanized and autopsied. Necrotizing fibrinosuppurative metritis was present in all. Additional findings in individual hares included fetal maceration, multifocal necrotizing myocarditis, multifocal hepatic necrosis, and nonsuppurative encephalitis. Listeria monocytogenes was cultured from multiple tissues in each hare. Although listeriosis in pregnant domestic rabbits has been studied, this is the first detailed description in wild North American hares. The epidemiology of listeriosis, including prevalence and the role of environmental sources and coprophagy in transmission among hares, requires further investigation.


Small fenestra stapedectomy technique: reducing risk and improving hearing.
During the past 25 years many variations have emerged in stapedectomy, most of which centered around either a change in the prosthesis itself or in the type of oval window seal. The small fenestra stapedectomy technique (SFT) represents a change in surgical procedure rather than in prosthetic design. This technique offers the opportunity to improve hearing results while reducing risks in stapedectomy surgery. Four areas of significant improvement are seen in patients in whom the SFT was used: (1) improved hearing in the high frequencies of 2000, 4000, and 8000 Hz, (2) improved speech discrimination scores, (3) a significant reduction in the number of reported vestibular complaints, and (4) a reduction in the number of serious postoperative sensorineural hearing losses.


Total Synthesis and Structural Revision of Chaetoviridins A.
The first synthesis of the proposed structures of chaetoviridins A 1-4 has been achieved in 10 steps by controlling the syn- or anti-aldol side chain. The angular lactone has been regioselectively introduced by condensation of a chiral dioxin-4-one to cazisochromene. Comparison of the NMR and circular dichroism data of the synthesized and reported natural products led to the complete reassignment and renaming of the chaetoviridins.


Treating age-related macular degeneration: comparing the use of two drugs among medicare and veterans affairs populations.
While new biologics have revolutionized the treatment of age-related macular degeneration-the leading cause of severe vision loss among older adults-these new drugs have also raised concerns over the economic impact of medical innovation. The two leading agents are similar in effectiveness but vary greatly in price-up to $2,000 per injection for ranibizumab compared to $50 for bevacizumab. We examined the diffusion of these drugs in fee-for-service Medicare and Veterans Affairs (VA) systems during 2005-11, in part to assess the impact that differing financial incentives had on prescribing. Physicians treating Medicare patients have a direct financial incentive to prescribe the more expensive agent (ranibizumab), while VA physicians do not. Medicare injections of the more expensive ranibizumab peaked in 2007 at 47 percent. Beginning in 2009 the less expensive bevacizumab became the predominant therapy for Medicare patients, accounting for more than 60 percent of injections. For VA patients, the distribution of injections across the two drugs was relatively equal, particularly from 2009 to 2011. Our analysis indicates that there are opportunities in both the VA and Medicare to adopt more value-conscious treatment patterns and that multiple mechanisms exist to influence utilization.


Intranasal immunization with CpG oligodeoxynucleotides as an adjuvant dramatically increases IgA and protection against herpes simplex virus-2 in the genital tract.
Development of vaccines capable of preventing the transmission or limiting the severity of sexually transmitted viruses, such as HSV and HIV, will likely be dependent on the induction of potent long-lasting mucosal immune responses in the genital tract. Recently, synthetic oligodeoxynucleotides (ODN) containing immunostimulatory CpG motifs were shown to serve as potent adjuvants for the induction of mucosal immune responses. Here, we show that intranasal immunization with CpG ODN, plus recombinant glycoprotein B (rgB) of HSV-1, results in significantly elevated levels of specific anti-gB IgA Abs in vaginal washes that remained high throughout the estrous cycle. Additionally, dramatically elevated numbers of specific IgA Ab-secreting cells were present and persisted in the genital tract in response to intravaginal (IVAG) HSV-2 challenge. HSV-2-specific CTL were observed at moderate levels in the spleens of CpG or non-CpG ODN-immunized mice. In contrast, strong CTL responses were observed locally in the genital tissues of both groups following IVAG HSV-2 challenge. Interestingly, mice immunized intranasally with rgB plus CpG ODN, but not non-CpG ODN, were significantly protected following IVAG HSV-2 challenge. Measurement of virus in protected CpG-immunized mice revealed a log lower level of replication within the first few days after infection. In conclusion, these results indicate that intranasal immunization with CpG ODN plus protein mediates immunity in the female genital tract capable of protecting against a sexually transmitted pathogen.


Safety concern between autologous fat graft, mesenchymal stem cell and osteosarcoma recurrence.
Osteosarcoma is the most common malignant primary bone tumour in young adult treated by neo adjuvant chemotherapy, surgical tumor removal and adjuvant multidrug chemotherapy. For correction of soft tissue defect consecutive to surgery and/or tumor treatment, autologous fat graft has been proposed in plastic and reconstructive surgery. We report here a case of a late local recurrence of osteosarcoma which occurred 13 years after the initial pathology and 18 months after a lipofilling procedure. Because such recurrence was highly unexpected, we investigated the possible relationship of tumor growth with fat injections and with mesenchymal stem/stromal cell like cells which are largely found in fatty tissue. Results obtained in osteosarcoma pre-clinical models show that fat grafts or progenitor cells promoted tumor growth. These observations and results raise the question of whether autologous fat grafting is a safe reconstructive procedure in a known post neoplasic context.


Evidence for suppression of cellular growth in vitro and selection against the indigenous mouse X chromosome in A9 cell hybrids after microcell-mediated transfer of an X from other mammalian species.
Introduction of a human or Syrian hamster X chromosome (derived from BHK-191-5C cell hybrids) into tumorigenic mouse A9 cells via microcell fusion induced changes in cellular morphology and a retardation of cellular growth. The suppression of growth of the hybrids could be abolished, however, by daily changes of medium containing 20% serum. G-banding analysis showed the absence of a single, cytogenetically identifiable, indigenous X chromosome (marker Z) in two of four hybrid clones after an X chromosome was transferred from either hamster or human cells. All hybrids were tumorigenic when tested in nude mice. Together, these data suggest that the loss of the mouse X chromosome took place probably because of growth inhibitory effects imposed on hybrid cells due to the increase in X chromosome dosage. In addition, our results show a lack of association between the phenotype of cellular growth suppression in vitro and the phenotype of suppression of tumorigenicity in vivo.


MEDIATOR18 and MEDIATOR20 confer susceptibility to Fusarium oxysporum in Arabidopsis thaliana.
The conserved protein complex known as Mediator conveys transcriptional signals by acting as an intermediary between transcription factors and RNA polymerase II. As a result, Mediator subunits play multiple roles in regulating developmental as well as abiotic and biotic stress pathways. In this report we identify the head domain subunits MEDIATOR18 and MEDIATOR20 as important susceptibility factors for Fusarium oxysporum infection in Arabidopsis thaliana. Mutants of MED18 and MED20 display down-regulation of genes associated with jasmonate signaling and biosynthesis while up-regulation of salicylic acid associated pathogenesis related genes and reactive oxygen producing and scavenging genes. We propose that MED18 and MED20 form a sub-domain within Mediator that controls the balance of salicylic acid and jasmonate associated defense pathways.


A diffusing wave spectroscopy study of pharmaceutical emulsions for physical stability assessment.
Emulsions are broadly used in pharmaceutics either as intermediate products or as final dosage forms. Such disperse systems are only kinetically stabilized and therefore early detection of physical instability is highly desirable. This work employed diffusing wave spectroscopy (DWS) to study a series of model emulsions that were categorized, based on their composition, as either "simple" or "complex". DWS data were compared with results of droplet size imaging, apparent viscosity obtained by microfluidics, and near-infrared (NIR) analytical centrifugation. A mathematical model of the droplet mean square displacement (MSD) was modified by us regarding improved fitting of experimental data. Although the emulsions showed different types of instability like creaming and sedimentation, a good rank correlation was found between the DWS parameters and results from the comparative stability methods. Our findings indicate that DWS provides a highly attractive method for stability analysis of pharmaceutical emulsions because it requires only low sample volumes, is rapid and non-invasive. The proposed data modeling provides the means for a better understanding of emulsion microstructure that in turn will help designing quality into pharmaceutical dispersions.


[Experience in using integral rheography for evaluating the hemodynamic indices in emergency states in cardiology].
The work deals with the comparative assessment of integral rheography, a modern method for studying hemodynamic indices. As compared to the method of dye dilution and radiocardiography, integral rheography has proved to be sufficiently precise and at the same time has definite advantages as a method of express diagnosis in patients with acute cardiovascular diseases. As compared with the other method of indirect determination of the cardiac output, tetrapolar rheography, integral rheography is more suitable for being used in emergencies, particularly in cases with disorders of the rhythm of the cardiac activity.


Treatment of Pneumocystis carinii pneumonitis with trimethoprim-sulfamethoxazole.
Pneumocystis carinii pneumonitis (PCP) is fatal in 90 to 100% of the cases if no treatment is given. Trimethoprim-sulfamethoxazole (TMP-SMX) was used at one of two dosage levels in the treatment of 20 children with PCP and cancer. Of 14 patients treated with 20 mg TMP--100 mg SMX/kgd, 12 recovered and 2 died. Treatment of the fatal cases and one of the patients who recovered was supplemented with pentamidine. When six patients were treated with 4 to 7 mg TMP--20 to 35 mg SMX/kgd, four recovered and two died. Both fatal cases and one of the patients who recovered were also treated with pentamidine. There was no significant adverse effects from TMP-SMX.


Effects of veratrine and cocaine on cerebral carbohydrate-amino acid interrelations.
The rate of aerobic transformation of glucose-U-C(14) into radioactive amino acids by slices of rat-brain cortex is greatly influenced by the K(+)/Ca(++) ratio in the incubating medium. Protoveratrine has effects on the amino acid pattern resembling that due to an increase in the K(+)/Ca(++) ratio. These effects are antagonized by cocaine and may be correlated with the neurophysiological activities of these drugs.


Atovaquone maintenance therapy prevents reactivation of toxoplasmic encephalitis in a murine model of reactivated toxoplasmosis.
Acute therapy with pyrimethamine plus sulfadiazine is the treatment of choice for reactivated toxoplasmic encephalitis (TE). Acute therapy is followed by lifelong maintenance therapy (secondary prophylaxis) with the same drugs at lower dosages. The use of pyrimethamine plus sulfadiazine is hampered by severe side effects including allergic reactions and hematotoxicity. Alternative treatment regimens with pyrimethamine plus clindamycin or other antiparasitic drugs are less efficacious. Atovaquone nanosuspensions show excellent therapeutic effects for "acute" intravenous (i.v.) treatment of reactivated TE in a murine model. In the present study, the therapeutic efficacy of atovaquone for oral "maintenance" therapy was investigated. Mice with a targeted mutation in the interferon regulatory factor 8 gene were latently infected with Toxoplasma gondii, developed reactivated TE, and received acute i.v. therapy with atovaquone nanosuspensions. Mice were then treated orally with atovaquone suspension or other antiparasitic drugs to prevent relapse of TE. Maintenance therapy with atovaquone at daily doses of 50 or 100 mg/kg (body weight) protected mice against reactivated TE and death. This maintenance treatment was superior to standard therapy with pyrimethamine plus sulfadiazine. The latter combination was superior to the combination of pyrimethamine plus clindamycin. Inflammatory changes in the brain parenchyma and meninges, as well as parasite numbers, in the brains of mice confirmed the therapeutic efficacy of atovaquone for maintenance therapy. Atovaquone was detectable in sera, brains, livers, and lungs of infected mice by high-performance liquid chromatography and/or mass spectrometry. In conclusion, atovaquone appears to be superior to the standard maintenance therapy regimens in a murine model of reactivated TE. The therapeutic efficacy of atovaquone for maintenance therapy against TE should be further investigated in clinical trials.


Reducing the health effects of hot weather and heat extremes: from personal cooling strategies to green cities.
Heat extremes (ie, heatwaves) already have a serious impact on human health, with ageing, poverty, and chronic illnesses as aggravating factors. As the global community seeks to contend with even hotter weather in the future as a consequence of global climate change, there is a pressing need to better understand the most effective prevention and response measures that can be implemented, particularly in low-resource settings. In this Series paper, we describe how a future reliance on air conditioning is unsustainable and further marginalises the communities most vulnerable to the heat. We then show that a more holistic understanding of the thermal environment at the landscape and urban, building, and individual scales supports the identification of numerous sustainable opportunities to keep people cooler. We summarise the benefits (eg, effectiveness) and limitations of each identified cooling strategy, and recommend optimal interventions for settings such as aged care homes, slums, workplaces, mass gatherings, refugee camps, and playing sport. The integration of this information into well communicated heat action plans with robust surveillance and monitoring is essential for reducing the adverse health consequences of current and future extreme heat.


Disentangling the Influence of Environment, Host Specificity and Thallus Differentiation on Bacterial Communities in Siphonous Green Seaweeds.
Siphonous green seaweeds, such as Caulerpa, are among the most morphologically complex algae with differentiated algal structures (morphological niches). Caulerpa is also host to a rich diversity of bacterial endo- and epibionts. The degree to which these bacterial communities are species-, or even niche-specific remains largely unknown. To address this, we investigated the diversity of bacteria associated to different morphological niches of both native and invasive species of Caulerpa from different geographic locations along the Turkish coastline of the Aegean sea. Associated bacteria were identified using the 16S rDNA marker gene for three morphological niches, such as the endobiome, epibiome, and rhizobiome. Bacterial community structure was explored and deterministic factors behind bacterial variation were investigated. Of the total variation, only 21.5% could be explained. Pronounced differences in bacterial community composition were observed and variation was partly explained by a combination of host species, biogeography and nutrient levels. The majority of the explained bacterial variation within the algal holobiont was attributed to the micro-environments established by distinct morphological niches. This study further supports the hypothesis that the bacterial assembly is largely stochastic in nature and bacterial community structure is most likely linked to functional genes rather than taxonomy.


Infective endocarditis: a consumptive disease among the elderly.
The clinical presentation of infective endocarditis varies according to the etiologic agent and the host. In elderly individuals, infective endocarditis can be difficult to diagnose and poses a challenge for the physician. The course of subacute infective endocarditis is indolent, and the onset of cardiac structural lesion is slow and gradual. In elderly patients, anemia and weight loss are occasionally the only or the most striking symptoms. In such cases, the clinical reasoning process leads to a hypothesis of wasting syndrome or neoplastic disease, especially when there is no fever. We report the case of an elderly patient who had mitral insufficiency due to degenerative valve disease and presented with bacterial endocarditis due to Streptococcus mitis. The patient was not treated, because the diagnosis was not established in a timely manner. It is of note that the patient presented with marked weight loss and no fever. The autopsy revealed impairment of the mitral valve and left atrium due to endocarditis, as well as lung involvement due to chronic inhalation of smoke from biomass burning, such as that produced by wood-burning stoves.


Taste acuity in response to zinc supplementation in older Europeans.
Taste acuity declines with age and may be dependent upon Zn status. The aim of the present double-blind, randomised controlled intervention trial has been to determine taste acuity in response to Zn supplementation (placebo, or 15 or 30 mg Zn/d). Healthy older European adults aged 70-87 years were recruited within Italy (Rome) (n 108) and France (Grenoble) (n 91) to the European Commission-funded Zenith project. A signal detection theory approach was adopted for taste assessment. The data were converted to R indices and analysed by repeated-measures ANOVA controlling for baseline taste acuity as well as serum and erythrocyte Zn. Serum Zn increased post-intervention, indicating compliance with the intervention. Results differed across geographical region. Salt taste acuity was greater in response to Zn (30 mg) than placebo post-intervention among those recruited in Grenoble. There was no apparent change in acuity for sweet, sour or bitter taste in response to Zn. Supplemented Zn may have potential to enhance salt taste acuity in those over the age of 70 years. Further research is required to determine if enhanced salt taste acuity is reflected in the eating experiences of older individuals.


Sensory Afferent Renal Nerve Activated Gαi2 Subunit Proteins Mediate the Natriuretic, Sympathoinhibitory and Normotensive Responses to Peripheral Sodium Challenges.
We have previously reported that brain Gαi2 subunit proteins are required to maintain sodium homeostasis and are endogenously upregulated in the hypothalamic paraventricular nucleus (PVN) in response to increased dietary salt intake to maintain a salt resistant phenotype in rats. However, the origin of the signal that drives the endogenous activation and up-regulation of PVN Gαi2 subunit protein signal transduction pathways is unknown. By central oligodeoxynucleotide (ODN) administration we show that the pressor responses to central acute administration and central infusion of sodium chloride occur independently of brain Gαi2 protein pathways. In response to an acute volume expansion, we demonstrate, via the use of selective afferent renal denervation (ADNX) and anteroventral third ventricle (AV3V) lesions, that the sensory afferent renal nerves, but not the sodium sensitive AV3V region, are mechanistically involved in Gαi2 protein mediated natriuresis to an acute volume expansion [peak natriuresis (μeq/min) sham AV3V: 43 ± 4 vs. AV3V 45 ± 4 vs. AV3V + Gαi2 ODN 25 ± 4, p < 0.05; sham ADNX: 43 ± 4 vs. ADNX 23 ± 6, AV3V + Gαi2 ODN 25 ± 3, p < 0.05]. Furthermore, in response to chronically elevated dietary sodium intake, endogenous up-regulation of PVN specific Gαi2 proteins does not involve the AV3V region and is mediated by the sensory afferent renal nerves to counter the development of the salt sensitivity of blood pressure (MAP [mmHg] 4% NaCl; Sham ADNX 124 ± 4 vs. ADNX 145 ± 4, p < 0.05; Sham AV3V 125 ± 4 vs. AV3V 121 ± 5). Additionally, the development of the salt sensitivity of blood pressure following central ODN-mediated Gαi2 protein down-regulation occurs independently of the actions of the brain angiotensin II type 1 receptor. Collectively, our data suggest that in response to alterations in whole body sodium the peripheral sensory afferent renal nerves, but not the central AV3V sodium sensitive region, evoke the up-regulation and activation of PVN Gαi2 protein gated pathways to maintain a salt resistant phenotype. As such, both the sensory afferent renal nerves and PVN Gαi2 protein gated pathways, represent potential targets for the treatment of the salt sensitivity of blood pressure.


Thermoluminescence dosimetry of gamma rays using ceramic samples from Hiroshima and Nagasaki: a comparison with DS86 estimates.
This study reports gamma-ray doses measured using thermoluminescence (TL) dosimetry of atomic-bomb-exposed ceramic samples from Hiroshima and Nagasaki. Advances in the dosimetry of TL-sensitive minerals in the field of TL dating of archaeological and geological materials made it possible to measure a radiation dose of 10(-2) Gy. Ceramic samples such as tiles and bricks were collected from locations between 523 and 2,453 m from the hypocenter in Hiroshima and from between 731 and 1,565 m in Nagasaki. The gamma-ray dose estimates derived from these samples are compared with estimates from DS86. A significant discrepancy was noted between the doses measured for samples from Hiroshima and DS86 values. One possible cause of this discrepancy might be a secondary gamma-ray dose related to the neutron dose.


A pilot study of participatory and rapid implementation approaches to increase depression screening in primary care.
Most individuals with depression go unidentified and untreated. In 2016 the US Preventive Services Task Force released guidelines recommending universal screening in primary care to identify patients with depression and to link them to treatment. Feasible, acceptable, and effective strategies to implement these guidelines are needed. This three-phased study employed rapid participatory methods to design and test strategies to increase depression screening at Penn Medicine, a large health system with 90 primary care practices. First, researchers solicited ideas and barriers from stakeholders to increase screening using an innovation tournament-a crowdsourcing method that invites stakeholders to submit ideas to address a workplace challenge. Second, a panel of stakeholders and scientists deliberated over and ranked the tournament ideas. An instant runoff election was held to select the winning idea. Third, the research team piloted the winning idea in a primary care practice using rapid prototyping, an approach that quickly refines and iterates strategy designs. The innovation tournament yielded 31 ideas and 32 barriers from diverse stakeholders (12 primary care physicians, 10 medical assistants, 4 nurse practitioners, 2 practice managers, and 4 patient support assistants). A panel of 6 stakeholders and scientists deliberated on the ideas and voted for patient self-report (i.e., through tablet computers, text message, or an online patient portal) as the winning idea. The research team rapid prototyped tablets in one primary care practice with one physician over 5 five-hour shifts to examine the feasibility, acceptability, and effectiveness of the strategy. Most patients, the physician, and medical assistants found the tablets acceptable and feasible. However, patient support assistants struggled to incorporate them in their workflow and expressed concerns about scaling up the process. Depression screening rates were higher using tablets compared to usual care; follow-up was comparable between tablets and usual care. Rapid participatory methods engaged and amplified the voices of diverse stakeholders in primary care. These methods helped design an acceptable and feasible implementation strategy that showed promise for increasing depression screening in a primary care setting. The next step is to evaluate the strategy in a randomized controlled trial across primary care practices.


Cardiorespiratory and Immunologic Effects of Electronic Cigarettes.
Although e-cigarettes have become popular, especially among youth, the health effects associated with e-cigarette use remain unclear. This review discusses current evidence relating to the cardiovascular, pulmonary, and immunological effects of e-cigarettes. The use of e-cigarettes by healthy adults has been shown to increase blood pressure, heart rate, and arterial stiffness, as well as resistance to air flow in lungs. Inhalation of e-cigarette aerosol has been shown to elicit immune responses and increase the production of immunomodulatory cytokines in young tobacco-naïve individuals. In animal models, long-term exposure to e-cigarettes leads to marked changes in lung architecture, dysregulation of immune genes, and low-grade inflammation. Exposure to e-cigarette aerosols in mice has been shown to induce DNA damage, inhibit DNA repair, and promote carcinogenesis. Chronic exposure to e-cigarettes has also been reported to result in the accumulation of lipid-laden macrophages in the lung and dysregulation of lipid metabolism and transport in mice. Although, the genotoxic and inflammatory effects of e-cigarettes are milder than those of combustible cigarettes, some of the cardiorespiratory effects of the two insults are comparable. The toxicity of e-cigarettes has been variably linked to nicotine, as well as other e-cigarette constituents, operating conditions, and use patterns. The use of e-cigarettes in humans is associated with significant adverse cardiorespiratory and immunological changes. Data from animal models and in vitro studies support the notion that long-term use of e-cigarettes may pose significant health risks.


A double-blind, randomized, sham-controlled study of cranial electrotherapy stimulation as an add-on treatment for tic disorders in children and adolescents.
The aim of this study was to determine the efficacy and safety of cranial electrotherapy stimulation (CES) as an add-on treatment for TD. A randomized, double-blind, sham-controlled trial was conducted at an outpatient, single-center academic setting. A total of 62 patients aged 6-17 years with TD and lack of clinical response to 4 weeks' pharmacotherapy were enrolled. Patients were divided randomly into 2 groups and given 4 weeks' treatment, including 30 min sessions of active CES (500 μA-2 mA) or sham CES (lower than 100 μA) per day for 40 d on weekdays. Change in Yale Global Tic Severity Scale (YGTSS), Clinical Global Impression-severity of illness-severity (CGI-S) and Hamilton Anxiety Scale-14 items (HAMA-14) were performed at baseline, week 2, week 4. Adverse events (AEs) were also evaluated. 53 patients (34 males and 9 females) completed the trial, including 29 in the active CES group and 24 in the sham CES group. Both groups showed clinical improvement in tic severities compared to baseline respectively at week 4. Participants receiving active CES showed a reduction of 31.66 % in YGTSS score, compared with 23.96 % in participants in sham CES group, resulting in no significant difference between the two groups (t = 1.54, p = 0.13). Four-week's treatment of CES for children and adolescents with TD is effective and safe, but the improvement for tic severity may be related to placebo effect.


Upper body kinematics in patients with cerebellar ataxia.
Although abnormal oscillations of the trunk are a common clinical feature in patients with cerebellar ataxia, the kinematic behaviour of the upper body in ataxic patients has yet to be investigated in quantitative studies. In this study, an optoelectronic motion analysis system was used to measure the ranges of motion (ROMs) of the head and trunk segments in the sagittal, frontal and yaw planes in 16 patients with degenerative cerebellar ataxia during gait at self-selected speed. The data obtained were compared with those collected in a gender-, age- and gait speed-matched sample of healthy subjects and correlated with gait variables (time-distance means and coefficients of variation) and clinical variables (disease onset, duration and severity). The results showed significantly larger head and/or trunk ROMs in ataxic patients compared with controls in all three spatial planes, and significant correlations between trunk ROMs and disease duration and severity (in sagittal and frontal planes) and time-distance parameters (in the yaw plane), and between both head and trunk ROMs and swing phase duration variability (in the sagittal plane). Furthermore, the ataxic patients showed a flexed posture of both the head and the trunk during walking. In conclusion, our study revealed abnormal motor behaviour of the upper body in ataxic patients, mainly resulting in a flexed posture and larger oscillations of the head and trunk. The results of the correlation analyses suggest that the longer and more severe the disease, the larger the upper body oscillations and that large trunk oscillations may explain some aspects of gait variability. These results suggest the need of specific rehabilitation treatments or the use of elastic orthoses that may be particularly useful to reduce trunk oscillations and improve dynamic stability.


Escherichia coli amino acid auxotrophic expression host strains for investigating protein structure-function relationships.
A set of C43(DE3) and BL21(DE3) Escherichia coli host strains that are auxotrophic for various amino acids is briefly reviewed. These strains require the addition of a defined set of one or more amino acids in the growth medium, and have been specifically designed for overproduction of membrane or water-soluble proteins selectively labelled with stable isotopes, such as 2H, 13C and 15N. The strains described here are available for use and have been deposited into public strain banks. Although they cannot fully eliminate the possibility of isotope dilution and mixing, metabolic scrambling of the different amino acid types can be minimized through a careful consideration of the bacterial metabolic pathways. The use of a suitable auxotrophic expression host strain with an appropriately isotopically labelled growth medium ensures high levels of isotope labelling efficiency as well as selectivity for providing deeper insight into protein structure-function relationships.


An approach to immunotherapy using antibody to IgE in mast cell leukemia.
Passive immunotherapy was attempted in a patient with mast cell leukemia using antibody against IgE. The results of both in vitro and in vivo studies indicate that although receptor sites for IgE were retained by the malignant mast cell, a secretory defect was present characterized by the spontaneous release of histamine and an impaired secretory response to anti-IgE antibody. Anti-IgE antibody selectively and reproducibly reduced the number of circulating mast cells probably by facilitating their permanent uptake by the reticuloendothelial system. Tolerance was not achieved with high dose deaggregated sheep IgG, nor were we able to confirm the effectivity of immunochemotherapy based on linking chlorambucil to antibody directed against the tumor-associated "antigen" IgE.


Sitagliptin Induces Tolerogenic Human Dendritic Cells.
Sitagliptin, an anti-diabetic drug, is a dipeptidyl peptidase (DPP)-4/CD26 inhibitor with additional anti-inflammatory and immunomodulatory properties. In this study, we investigated for the first time the effect of sitagliptin on the differentiation and functions of human dendritic cells generated from monocytes (MoDCs) for 4 days using the standard GM-CSF/IL-4 procedure. LPS/IFN-γ treatment for an additional 24 h was used for maturation induction of MoDCs. Sitagliptin was added at the highest non-cytotoxic concentration (500 µg/mL) either at the beginning (sita 0d protocol) or after MoDC differentiation (sita 4d protocol). Sitagliptin impaired differentiation and maturation of MoDCs as judged with the lower expression of CD40, CD83, CD86, NLRP3, and HLA-DR, retention of CD14 expression, and inhibited production of IL-β, IL-12p70, IL-23, and IL-27. In contrast, the expression of CD26, tolerogenic DC markers (ILT4 and IDO1), and production of immunoregulatory cytokines (IL-10 and TGF-β) were increased. Generally, the sita 0d protocol was more efficient. Sitagliptin-treated MoDCs were poorer allostimulators of T-cells in MoDC/T-cell co-culture and inhibited Th1 and Th17 but augmented Th2 and Treg responses. Tolerogenic properties of sitagliptin-treated MoDCs were additionally confirmed by an increased frequency of CD4+CD25+CD127- FoxP3+ Tregs and Tr1 cells (CD4+IL-10+FoxP3-) in MoDC/T-cell co-culture. The differentiation of IL-10+ and TGF-β+ Tregs depended on the sitagliptin protocol used. A Western blot analysis showed that sitagliptin inhibited p65 expression of NF-kB and p38MAPK during the maturation of MoDCs. In conclusion, sitagliptin induces differentiation of tolerogenic DCs, and the effect is important when considering sitagliptin for treating autoimmune diseases and allotransplant rejection.


Turnover of tritiated-thymidine-labeled mononuclear cells in tuberculous lesions of rabbits: a comparison of primary dermal BCG lesions and those of reinfection.
Rabbits were infected or reinfected intradermally in multiple sites with BCG. One day before the BCG infection they were injected iv with a single pulse of tritiated thymidine (3HT). At various times thereafter, the BCG lesions were biopsied and evaluated for 3HT-labeled mononuclear cells (MN), which are mostly macrophages but include some lymphocytes. Periodically Old Tuberculin (OT) was infected intradermally, creating MN traps, which were biopsied and evaluated 1 day after their onset. These traps monitor the population of 3HT-labeled MN in the blood stream that readily enters sites of inflammation. During the first 5 days after the 3HT pulse, a higher percentage of labeled MN accumulated in tuberculin traps on rabbits reinfected with BCG than in traps on rabbits with primary infection. The high percentage of labeled MN in the former was probably due to the more intense inflammation cuased by preexisting tuberculin hypersensitivity. By 8 days after the 3HT pulse, a lower percentage of labeled MN had accumulated in BCG lesions of reinfection than in primary BCG lesions. Evidently the presence of delayed hypersensitivity increased the turnover of MN in tuberculous lesions. By 15 days after the 3HT pulse, the percentage of 3HT-labeled MN was much higher in BCG lesions than in 1-day traps, suggesting that such MN had accumulated in the lesions over a period of several days.


[Chemical accidents and catastrophes as a source of the greatest hazard to the environment and human health in Poland].
The author presents sources of the greatest chemical hazards to the environment in Poland, among which industry and transport occupy the first two places. The availability of data and information on chemical hazards to the environment and the human health are discussed. Legislation, organisation and competences of institutions designed to prevent and mitigate the outcomes of chemical disasters are outlined. It was found that in Poland a possible danger of chemical disasters with serious consequences for the people and environment does really exist. Certain industrial installations as well as road and railway transport belong to the major threats. Among chemicals, ammonia, chlorine, sulphur dioxide and phosgene are considered as most dangerous. In Poland the Ministry of Internal Affairs and the Ministry of Environmental Protection, Natural Resources and Forestry are legally responsible for prevention and mitigation of the outcomes of chemical disasters.


Functional characterization of the complement receptor type 1 and its circulating ligands in patients with schizophrenia.
Whereas the complement system alterations contribute to schizophrenia, complement receptors and regulators are little studied. We investigated complement receptor type 1 (CR1) expression on blood cells, the levels of circulating immune complexes (CIC) containing ligands of CR1, C1q complement protein and fragments of C3 complement protein (C1q-CIC, C3d-CIC), and CR1 C5507G functional polymorphism in schizophrenia patients and controls. We found an increased C1q-CIC level and CR1 expression on blood cells, elevated number of CR1 positive erythrocytes and reduced number of CR1 positive lymphocytes and monocytes in patients compared to controls. No difference in the levels of C3d-CIC between groups was observed. Higher CR1 expression on erythrocytes in CC genotype versus CG+GG for both groups was detected, whereas no difference was observed for other cell populations. Our results indicated that schizophrenia is associated with the increased CR1 expression and C1q-CIC level. Our study for the first time indicated that schizophrenia is associated with the increased CR1 expression and C1q-CIC level. Further studies in other ethnic groups are needed to replicate these findings.


Water-Compatible Cycloadditions of Oligonucleotide-Conjugated Strained Allenes for DNA-Encoded Library Synthesis.
DNA-encoded libraries of small molecules are being explored extensively for the identification of binders in early drug-discovery efforts. Combinatorial syntheses of such libraries require water- and DNA-compatible reactions, and the paucity of these reactions currently limit the chemical features of resulting barcoded products. The present work introduces strain-promoted cycloadditions of cyclic allenes under mild conditions to DNA-encoded library synthesis. Owing to distinct cycloaddition modes of these reactive intermediates with activated olefins, 1,3-dipoles, and dienes, the process generates diverse molecular architectures from a single precursor. The resulting DNA-barcoded compounds exhibit unprecedented ring and topographic features, related to elements found to be powerful in phenotypic screening.


Evaluation of periodontal status among saudi females with gestational diabetes and its relation to glucose and lipid homeostasis in ohud hospital, Al madina Al-munwarrah.
Due to the lack of pregnant based oral health studies in our country, we conducted this study to evaluate periodontal status among females with gestational diabetes in Ohud Hospital, Al Madinah Al-Munwarrah and to assess its effect on insulin sensitivity and lipid metabolism. This cross-sectional descriptive study was conducted from January 2008 till June 2008. The sample includes 250 Saudi females, 100 were pregnant with gestational diabetes (GDM), and 100 were pregnant without GDM and 50 were normal non pregnant females. The study cases were matched age and body mass index. All the participants were subjected to history taking, physical examination and assessment of their periodontal status. Laboratory tests include fasting blood sugar, insulin levels, Homeostasis Model Assessment Insulin Resistance (HOMA-IR) test and serum lipids. Serum levels of inflammatory markers (CRP, ESR, IL-1β, and TNF-α) also measured. The periodontal health status was assessed using the Community Periodontal Index of Treatment Needs (CPITN). Severe periodontal diseases were elicited in 37% of the pregnant females with GDM, 29% of the pregnant females without GDM and 14% of non pregnant normal control group. Females with GDM showed higher systolic and diastolic blood pressure, fasting insulin, HOMA-IR, serum lipids and serum inflammatory markers levels than the other groups. Serum fasting insulin levels, HOMA-IR, triglyceride levels, and serum inflammatory markers were higher in females with severe periodontal diseases. Females with recurrent GDM were found to have higher Body Mass Index (BMI), severe periodontal diseases, as well as higher HOMA-IR, than those presented for first time with GDM. Females with GDM; also shows significant positive correlation between CPITN scoring and patient age, HOMA-IR and inflammatory markers. The prevalence of periodontal disease is high among pregnant females especially those with GDM. Periodontal disease had important deleterious effects on carbohydrates and lipids metabolism. Strategies are needed to improve dental health care and to reduce periodontal diseases among pregnant females.


Japanese Honeybees (Apis cerana japonica Radoszkowski, 1877) May Be Resilient to Land Use Change.
Pollinators are being threatened globally by urbanisation and agricultural intensification, driven by a growing human population. Understanding these impacts on landscapes and pollinators is critical to ensuring a robust pollination system. Remote sensing data on land use attributes have previously linked honeybee nutrition to land use in the Western Honeybee (Apis mellifera L.). Here, we instead focus on the less commonly studied Apis cerana japonica-the Japanese Honeybee. Our study presents preliminary data comparing forage (honey and pollen) with land use across a rural-urban gradient from 22 sites in Kyushu, southern Japan. Honey samples were collected from hives between June 2018 and August 2019. Pollen were collected and biotyped from hives in urban and rural locations (n = 4). Previous studies of honey show substantial variation in monosaccharide content. Our analysis of A. cerana japonica honey found very little variation in glucose and fructose (which accounted for 97% of monosaccharides), despite substantial differences in surrounding forage composition. As expected, we observed temporal variation in pollen foraged by A. cerana japonica, likely dependent on flowering phenology. These preliminary results suggest that the forage and nutrition of A. cerana japonica may not be negatively affected by urban land use. This highlights the need for further comparative studies between A. cerana japonica and A. mellifera as it could suggest a resilience in pollinators foraging in their native range.


Chronic pain in patients with hemophilia: is it preventable?
: Current management of chronic pain in patients with hemophilia (PWH) focuses on pain relief with analgesics and symptom control. The clinical practice of managing chronic pain in PWH varies considerably across hemophilia treatment centers. Here, we aim to study the appropriate intervention of hemophilic arthropathy for prevention and treatment of chronic pain in PWH. Medline, Embase, Cochrane databases were searched for randomized controlled trials, and the European Hemophilia Therapy Standardization Board, The World Federation of Hemophilia, Nordic Hemophilia Guidelines, American Society of the International Pain Physicians and the Medical and Scientific Advisory Council guidelines were studied through November 2019 for chronic pain in PWH for a narrative review. We found no standardized approach for the prevention and management of chronic pain in PWH. Evidence suggests that prophylactic factor concentrate therapy, programmed exercise and educational intervention may help PWH manage their chronic pain.


Spatial coating inhomogeneity of highly reflective mirrors determined by cavity ringdown measurements.
The inhomogeneity of high-reflectivity mirror coatings is a potential error source in the application of the cavity ringdown technique. Here, the ringdown times for different transverse modes were recorded. Together with the observed spatial distribution of these modes the ringdown times can be used to approximately locate the position of coating defects. A simple model based on a weighted sum of Hermite-Gaussian mode functions is used to explain the experimental results.


Interleukin-1 as an injury signal mobilizes retinyl esters in hepatic stellate cells through down regulation of lecithin retinol acyltransferase.
Retinoids are mostly stored as retinyl esters in hepatic stellate cells (HSCs) through esterification of retinol and fatty acid, catalyzed by lecithin-retinol acyltransferase (LRAT). This study is designated to address how retinyl esters are mobilized in liver injury for tissue repair and wound healing. Initially, we speculated that acute inflammatory cytokines may act as injury signal to mobilize retinyl esters by down-regulation of LRAT in HSCs. By examining a panel of cytokines we found interleukin-1 (IL-1) can potently down-regulate mRNA and protein levels of LRAT, resulting in mobilization of retinyl esters in primary rat HSCs. To simulate the microenvironment in the space of Disse, HSCs were embedded in three-dimensional extracellular matrix, by which HSCs retaine quiescent phenotypes, indicated by up-regulation of LRAT and accumulation of lipid droplets. Upon IL-1 stimulation, LRAT expression went down together with mobilization of lipid droplets. Secreted factors from Kupffer cells were able to suppress LRAT expression in HSCs, which was neutralized by IL-1 receptor antagonist. To explore the underlying mechanism we noted that the stability of LRAT protein is not significantly regulated by IL-1, indicating the regulation is likely at transcriptional level. Indeed, we found that IL-1 failed to down-regulate recombinant LRAT protein expressed in HSCs by adenovirus, while transcription of endogenous LRAT was promptly decreased. Following liver damage, IL-1 was promptly elevated in a close pace with down-regulation of LRAT transcription, implying their causative relationship. After administration of IL-1, retinyl ester levels in the liver, as measured by LC/MS/MS, decreased in association with down-regulation of LRAT. Likewise, IL-1 receptor knockout mice were protected from injury-induced down-regulation of LRAT. In summary, we identified IL-1 as an injury signal to mobilize retinyl ester in HSCs through down-regulation of LRAT, implying a mechanism governing transition from hepatic injury to wound healing.


Multiscale conformational heterogeneity in staphylococcal protein a: possible determinant of functional plasticity.
The Staphylococcus aureus virulence factor staphylococcal protein A (SpA) is a major contributor to bacterial evasion of the host immune system, through high-affinity binding to host proteins such as antibodies. SpA includes five small three-helix-bundle domains (E-D-A-B-C) separated by conserved flexible linkers. Prior attempts to crystallize individual domains in the absence of a binding partner have apparently been unsuccessful. There have also been no previous structures of tandem domains. Here we report the high-resolution crystal structures of a single C domain, and of two B domains connected by the conserved linker. Both structures exhibit extensive multiscale conformational heterogeneity, which required novel modeling protocols. Comparison of domain structures shows that helix1 orientation is especially heterogeneous, coordinated with changes in side chain conformational networks and contacting protein interfaces. This represents the kind of structural plasticity that could enable SpA to bind multiple partners.


Reliability of corticomotor excitability in leg and thigh musculature at 14 and 28 days.
Alterations in corticomotor excitability are observed in a variety of patient populations, including the musculature surrounding the knee and ankle after joint injury. Active motor threshold (AMT) and motor-evoked-potential (MEP) amplitudes elicited through transcranial magnetic stimulation (TMS) are outcome measures used to assess corticomotor excitability and have been deemed reliable in upper-extremity musculature. However, there are few studies assessing the reliability of TMS measures in lower-extremity musculature. To determine the intersession reliability of AMT and MEP amplitudes over 14 and 28 d in the quadriceps and fibularis longus (FL). Descriptive laboratory study. University laboratory 20 able-bodied volunteers (10 men, 10 women; 22.35 ± 2.3 y, 1.71 ± 0.11 m, 73.61 ± 16.77 kg). AMT and MEP amplitudes were evaluated at 95%, 100%, 105%, 110%, 120%, 130%, and 140% of AMT in the dominant and nondominant quadriceps and FL. Interclass correlation coefficients (ICCs) were used to assess reliability for absolute agreement and internal consistency between baseline and 2 follow-up sessions at 14 and 28 d postbaseline. Each ICC was fit with the best-fit straight line or parabola to smooth out noise in the observations and best determine if a pattern existed in determining the most reliable MEP value. All muscles yielded strong ICCs between baseline and both time points for AMT. MEPs in both the quadriceps and FL produced varying degrees of reliability, with the greatest reliability demonstrated on day 28 at 130% and 140% of AMT in the quadriceps and FL, respectively. The dominant FL muscle showed a significant pattern; as TMS intensity increased, MEP reliability increased. TMS can be used to reliably identify corticomotor alterations after therapeutic interventions, as well as monitor disease progression.


Enhancing Protein Conformational Space Sampling Using Distance Profile-Guided Differential Evolution.
De novo protein structure prediction aims to search for low-energy conformations as it follows the thermodynamics hypothesis that places native conformations at the global minimum of the protein energy surface. However, the native conformation is not necessarily located in the lowest-energy regions owing to the inaccuracies of the energy model. This study presents a differential evolution algorithm using distance profile-based selection strategy to sample conformations with reasonable structure effectively. In the proposed algorithm, besides energy, the residue-residue distance is considered another measure of the conformation. The average distance errors of decoys between the distance of each residue pair and the corresponding distance in the distance profiles are first calculated when the trial conformation yields a larger energy value than that of the target. Then, the distance acceptance probability of the trial conformation is designed based on distance profiles if the trial conformation obtains a lower average distance error compared with that of the target conformation. The trial conformation is accepted to the next generation in accordance with its distance acceptance probability. By using the dual constraints of energy and distance in guiding sampling, the algorithm can sample conformations with lower energies and more reasonable structures. Experimental results of 28 benchmark proteins show that the proposed algorithm can effectively predict near-native protein structures.


Quantitative Analysis of Transnasal Anterior Skull Base Approach: Report of Technology for Intraoperative Assessment of Instrument Motion.
To develop a method to measure intraoperative surgical instrument motion. This model will be applicable to the study of surgical instrument kinematics including surgical training, skill verification, and the development of surgical warning systems that detect aberrant instrument motion that may result in patient injury. We developed an algorithm to automate derivation of surgical instrument kinematics in an endoscopic endonasal skull base surgery model. Surgical instrument motion was recorded during a cadaveric endoscopic transnasal approach to the pituitary using a navigation system modified to record intraoperative time-stamped Euclidian coordinates and Euler angles. Microdebrider tip coordinates and angles were referenced to the cadaver's preoperative computed tomography scan allowing us to assess surgical instrument kinematics over time. A representative cadaveric endoscopic endonasal approach to the pituitary was performed to demonstrate feasibility of our algorithm for deriving surgical instrument kinematics. Technical feasibility of automatically measuring intraoperative surgical instrument motion and deriving kinematics measurements was demonstrated using standard navigation equipment.


Ethical issues in using social media for health and health care research.
The dramatic growth of social media in recent years has not gone unnoticed in the health sector. Media such as Facebook and Twitter are increasingly being used to disseminate information among health professionals and patients but, more recently, are being seen as a source of data for surveillance and research, for example by tracking public concerns or capturing discourses taking place outside traditional media outlets. This raises ethical issues, in particular the extent to which postings are considered public or private and the right to anonymity of those posting on social media. These issues are not clear cut as social media, by their nature, blur the boundary between public and private. There is a need for further research on the beliefs and expectations of those using social media in relation to how their material might be used in research. In contrast, there are areas where the ethical issues are more clear cut, such as when individuals are active participants in research, where traditional considerations apply.


Energy use strategies and implications for fire risk amongst low-income households.
Despite near universal access to electricity in Cape Town, usage of informal electrical connections and nonelectric energy sources remains high and pose significant fire risk to such households. This research set out to examine the energy sources being utilised by low-income households in Lwandle, Nomzamo and Asanda Village to understand the factors that influence these energy use choices and what implications these energy choices have for fire risk. This research utilised a mixture of qualitative and quantitative data collection methods including focus group sessions with residents and a household survey to collect information on household energy use strategies, perceptions of safety and accessibility of energy sources and experiences of energy-related fires from residents residing in different types of dwellings. The research observed that despite high access to electricity, household utilisation is constrained by economic and physical factors. Consequently, they are forced to resort to employing an energy stacking approach, alternating between electric and nonelectric energy sources, which include usage of cheaper yet potentially hazardous energy sources such as paraffin (kerosene), candles, firewood, coal and gas to meet their daily energy needs. A potential consequence of this energy stacking approach employed by households to meet their energy needs is that the majority of households continue to face the risk of a dwelling fire caused by nonelectric energy sources. Whereas nonelectric energy sources were both perceived and experienced by residents as the main cause of dwelling fires in the study site, electricity was found to contribute to a number of dwelling fires, with a slight increase in the number of fires caused by electric sources observed over the last few years.


Current concepts of intraepithelial neoplasia in the uterine cervix (CIN).
The unifying concept of precancerous events in the epithelia of the uterine cervix is reviewed in the light of epidemiologic and morphologic observations. The role of human papillomavirus (HPV) infection as a factor in the genesis of precancerous states and carcinoma of the uterine cervix is assessed. Although cytologic, histologic, ultrastructural and molecular biologic data support the concept of HPV as an oncogenic virus, the precise mechanisms linking the infection with cancer are not known at this time. A hypothesis of the presumed sequence of events is presented.


Roles of sulfite oxidoreductase and sulfite reductase in improving desulfurization by Rhodococcus erythropolis.
Rhodococcus erythropolis has been widely studied for desulfurization. However, activity levels required for commercial application have not been achieved. A major limitation of the current work in biodesulfurization is inadequate information regarding sulfur metabolism generally, and in particular the metabolism of the sulfur obtained from dibenzothiophene (DBT) metabolism via the 4S pathway. In this work, we have investigated the possible routes taken by the sulfur from DBT to convert into biomass or other metabolites. We propose two alternate hypotheses. In the first, we hypothesize that the cell can convert via sulfite reductase (SR) the sulfite from the metabolism of DBT into sulfide that can be assimilated into biomass. However, in the process, it may convert any excess sulfite into extracellular sulfate via sulfite oxidoreductase (SOR) to avoid the toxic effects of sulfite. In the second, we speculate that the cell cannot assimilate the sulfite directly into biomass via SR. It must first use SOR to produce extracellular sulfate, and then recapture that sulfate into biomass via SR. Thus, either way, we propose that SOR and SR activities, in addition to dsz genes and cofactors, may be critical in increasing desulfurization levels significantly. In particular, we suggest that the simultaneous increase in SOR activity and decrease in SR activity can enable increased desulfurization activity.


[Immunoexpression of p53 and cyclin D1 in adenomas of the gallbladder].
Gallbladder adenomas are infrequent neoplasms whose relation to adenocarcinoma is not well understood. It has been suggested that adenomas and adenocarcinomas follow different molecular pathways. This is a comparative, cross-sectional study in which we compared p53 and D1 cyclin expression in adenomas and adenocarcinomas of the gallbladder. We included 12 cases in each group. Expression of p53 occurred in 83.3% of adenocarcinomas and in 16.6% of adenomas (p = 0.003). D1 cyclin was expressed in a similar number of adenomas and adenocarcinomas. Our results support the hypothesis that p53 is an important step in the pathogenesis of adenocarcinomas but not of adenomas of the gallbladder. D1 cyclin is apparently a common pathway involved in the genesis of both tumors.


Serendipitous SAD phasing of an echinomycin-(ACGTACGT)2 bisintercalation complex.
A new crystal form was obtained for the complex between (ACGTACGT)2 and echinomycin and X-ray data were collected to 1.6 A. The structure was phased by the SAD method based on a single unexpected anomalous scatterer that could be identified as a mixture of nickel and zinc by measurements of the anomalous scattering at different wavelengths. This cation is coordinated by two guanines from two different duplexes and four water molecules. The structure resembles previously reported crystal structures of DNA-echinomycin complexes, except that three of the eight base pairs flanking the echinomycin bisintercalator sites have the Watson-Crick rather than the Hoogsteen configuration. Hoogsteen binding was found for the corresponding base pairs of the crystallographically independent duplex, indicating that the two configurations are very close in energy.


Seroprevalence of Louping Ill virus (LIV) antibodies in sheep submitted for post mortem examination in the North West of Ireland in 2011.
Blood samples were collected opportunistically at routine post mortem examination from 199 sheep which came from 152 flocks. The location of each submitting flock was mapped. Sera were tested using a goose blood haemagglutination inhibition assay for louping ill virus. There was an animal level prevalence of 8.5%, and a flock level prevalence of 9.8%. The greatest proportion of seropositive animals was identified among the animals older than 24 months of age. The elevation of the land associated with positive flocks was greater than that of negative flocks. Lesions of non-suppurative meningoencephalitis were observed in three of the 199 animals.


Hepatocellular 22Na+ uptake: effect of oleate.
Whether cellular oleate uptake is Na+ coupled remains controversial. Our present studies document that hepatocellular [3H]oleate uptake is unaltered by isomotic substitution of Na+ with K+, Li+, or sucrose in Hanks' HEPES buffer. In parallel studies 22Na+ uptake was significantly (P less than 0.02) increased by concentrations of alanine that yielded [3H]-alanine uptakes greater than 24% of basal 22Na+ uptake when both were expressed as nanomoles per minute per 10(6) hepatocytes. Although [3H]glutamine uptake exceeded this threshold, maximal specific [3H]taurocholate uptake did not. Consistent with the observations with alanine, addition of glutamine, but not taurocholate, to the incubations resulted in a significant increase in 22Na+ uptake. The ionophore monensin increased uptake of 22Na+ under all conditions. Both in the absence and in the presence of HCO3- (4 and 25 mM), specific [3H]oleate uptake was sufficient to elicit a readily detectable effect on 22Na+ uptake, if Na+ and oleate were cotransported. However, addition of 1 mM oleate did not affect 22Na+ uptake in the absence of HCO3- (5.4 +/- 0.6 vs. control 5.4 +/- 0.4 nmol.min-1.10(6) cells-1) as well as in the presence of 4mM HCO3- (10.4 +/- 1.8 vs. control 9.9 +/- 1.1) and 25 mM HCO3- (10 +/- 1.3 vs. control 10.8 +/- 0.5). These data indicate that the predominant component of hepatocellular oleate uptake is not directly associated with Na+ influx. They do not exclude, however, a more complex or indirect link.


A measure of palliative care in nursing homes.
Efforts to improve care for nursing home residents stand to be enhanced by measures to assess the degree to which staff provide palliative care. As the incidence of death in nursing homes increases with the aging population, the gap in measurement must be addressed. To that end, we report the development and psychometric testing of a nursing home palliative care survey. The purpose of this study was to evaluate the psychometric properties of the Palliative Care Survey (PCS) for use in nursing homes. Psychometric evaluation of the instrument was completed in two phases. Phase 1 focused on individual item analyses and subsequent revision or deletion of items, and Phase 2 evaluated evidence for reliability and validity. Phase 1 included 26 nursing homes and staff (n=717), and Phase 2 included 85 nursing homes and staff (n=2779). Data were analyzed using item-total correlations, Cronbach's alpha, confirmatory factor analysis, and analysis of variance. Support was obtained for a 51-item PCS made up of two constructs, Palliative Care Practice and Palliative Care Knowledge. The PCS measures the extent to which the nursing home staff engage in palliative care practices and have knowledge consistent with good end-of-life care. Both practice and knowledge are an essential foundation to providing good end-of-life care to nursing home residents. Efforts to improve care for the dying in nursing homes have been slowed by an absence of measurement tools that capture care processes, a gap that the PCS reported here helps fill.


The contribution of visceral adipose tissue to splanchnic cortisol production in healthy humans.
Cortisol is regenerated from cortisone by 11beta-hydroxysteroid dehydrogenase type 1 (11HSD1), amplifying glucocorticoid action in adipose tissue and liver. 11HSD1 inhibitors are being developed for type 2 diabetes and may be most effective in obesity, where adipose 11HSD1 is increased. However, the magnitude of regeneration of cortisol in different tissues in humans is unknown, hindering understanding of the pathophysiological and therapeutic importance of 11HSD1. In eight healthy men, we infused 9,11,12,12-(2)H4-cortisol and measured tracer enrichment in the hepatic vein as an indicator of total splanchnic cortisol generation. Oral cortisone (25 mg) was then given to measure first-pass hepatic cortisol generation. In steady state, splanchnic cortisol production was 45 +/- 11 nmol/min when arterialized plasma cortisone concentration was 92 +/- 7 nmol/l. Extrapolation from hepatic cortisol generation after oral cortisone suggested that, at steady state, the liver contributes 15.2 nmol/min and extrahepatic splanchnic tissue contributes 29.8 nmol/min to the total splanchnic cortisol production. We conclude that tissues draining into the portal vein, including visceral adipose tissue, contribute substantially to the regeneration of cortisol. Thus, in addition to free fatty acids and adipokines, the portal vein delivers cortisol to the liver, and inhibition of 11HSD1 in visceral adipose tissue may indeed be valuable in ameliorating insulin resistance in obesity.


Effect of PDE5 inhibition on coronary hemodynamics in pacing-induced heart failure.
Inhibition of phosphodiesterase type 5 (PDE5) can relax systemic and coronary vessels by causing accumulation of cGMP. Both the endothelial dysfunction with decreased nitric oxide production and increased natriuretic peptide levels in congestive heart failure (CHF) have the potential to alter cGMP production, thereby influencing the response to PDE5 inhibition. Consequently, this study examined the effects of PDE5 inhibition with sildenafil in dogs with CHF produced by rapid ventricular pacing. CHF resulted in decreases of left ventricular (LV) systolic pressure, coronary blood flow, and the maximal first time derivative of LV pressure (LV dP/dt(max)) at rest and during treadmill exercise compared with normal, whereas resting LV end-diastolic pressure increased from 10 +/- 1.4 to 23 +/- 1.4 mmHg. Sildenafil (2 and 10 mg/kg per os) caused a 5- to 6-mmHg decrease of aortic pressure (P < 0.05), with no change of heart rate, LV systolic pressure, or LV dP/dt(max). Sildenafil caused no change in coronary flow or myocardial oxygen consumption in animals with CHF at rest or during exercise. In contrast to findings in normal animals, sildenafil did not augment endothelium-dependent coronary vasodilation in response to acetylcholine in animals with CHF. Furthermore, Western blotting showed decreased PDE5 protein expression in myocardium from failing hearts. These findings demonstrate that PDE5 contributes little to regulation of coronary hemodynamics in CHF.


Fully automated system for the gas chromatographic characterization of polar biopolymers based on thermally assisted hydrolysis and methylation.
Pyrolysis-gas chromatography (Py-GC) is a powerful tool for the detailed compositional analysis of polymers. A major problem of Py-GC is that polar (bio)polymers yield polar pyrolyzates which are not easily accessible to further GC characterization. In the present work, a newly developed fully automated procedure for thermally assisted hydrolysis and methylation (THM) of biopolymers is described. Drying of the sample, addition of the reagent, incubation and pyrolysis are performed inside the liner of a programmable temperature vaporizer injector. The new system not only allows efficient analysis of large series of samples, but also allows automated optimization of the experimental parameters based on an experimental design approach. The performance of the automated THM-procedure was evaluated by performing THM-GC of a poly(acrylic acid)-poly(maleic anhydride) copolymer (PAA/PMAH) and several polysaccharides. The optimized THM-procedure was applied for the structural characterization and differentiation of several lignins and hydroxypropylmethyl-celluloses. It was also applied to proteins. Here myoglobin and cytochrome c were used as the model compounds. Both conventional GC-mass spectrometry (MS) and comprehensive two-dimensional gas chromatography (GCxGC)-time-of-flight (TOF) MS were used for separation and identification of the species formed. The information obtained can aid in structure elucidation of polar biopolymers as well as in providing detailed compositional information which can be used to differentiate structurally similar biopolymers.


Antimicrobial Susceptibility Patterns and Clinical Parameters in 208 Dogs with Positive Urine Cultures (2012-2014).
Urinary tract infections (UTI) occur commonly in dogs, and gram-negative enteric bacteria are the most prevalent pathogens. Clinical parameters, urinalysis, and urine culture and sensitivity results were retrieved from the medical records of 208 dogs with positive urine cultures over a 3 yr period at the Louisiana State University Veterinary Teaching Hospital. Several groups were defined including dogs presented for primary care versus referred cases; simple UTI, complicated UTI, and pyelonephritis; dogs pretreated with antimicrobials; and dogs having an indwelling catheter in place prior to sampling. Nearly 80% of dogs had complicated UTI. Of all dogs, 70% had no documented clinical signs of lower urinary tract disease (LUTD), with 68% of them showing hematuria and/or pyuria. Based on clinical signs or urinalysis, 19% of all dogs had no evidence of lower UTI. In dogs without LUTD signs the most common comorbidities were immunosuppressive treatment and severely restricted mobility (23%). Chronic recurring UTI were present in 19% of dogs with LUTD signs. Distribution of bacterial species was comparable with the existing literature and not significantly different between clinical subgroups. Isolates from dogs pretreated with antimicrobials showed decreased susceptibility to enrofloxacin. The prevalence of multidrug-resistant Escherichia coli and Staphylococcus spp. was moderate (29%).


Autoantibodies to leukocyte alphaMbeta2 integrin glycoproteins in HIV infection.
HIV infection is often associated with polyclonal B-cell activation, autoantibodies, and clinically evident autoimmune disease. Because neutropenia and anti-neutrophil autoantibodies are common clinical features of HIV disease, we studied a series of HIV+ patients to determine whether anti-alphaMbeta2 integrin (MAC-1) specific anti-neutrophil autoantibodies occur in HIV disease, as we have shown to occur in patients with immune neutropenia not associated with HIV. Two new assays specific for anti-alphaMbeta2 IgG were developed to carry out these studies: an ELISA method using affinity-purified alphaMbeta2 integrin protein, and a flow cytometry method using subclones of the 293 human fetal kidney cell line, stably transfected with cDNAs for the alphaM and/or beta2 integrin subunits. In studies of the sera of 20 untreated HIV+ individuals, anti-alphaMbeta2 activity was detected in 9 (45%) by one or the other of these assays and in 5 (25%) by both assays. Seven of the 20 HIV+ study subjects had unexplained neutropenia, and of these, 6 (86%) were positive for anti-alphaMbeta2 autoantibodies. Our findings indicate that anti-alphaMbeta2 integrin autoantibodies are frequent in HIV+ individuals, particularly when unexplained neutropenia is also present, and raise the possibility that these autoantibodies may have a role in the acquired neutrophil dysfunction and increased risk of nonopportunistic bacterial infections observed in HIV disease.


Glycation Interferes with the Expression of Sialyltransferases and Leads to Increased Polysialylation in Glioblastoma Cells.
Glioblastoma (GBM) is a highly aggressive brain tumor that often utilizes aerobic glycolysis for energy production (Warburg effect), resulting in increased methylglyoxal (MGO) production. MGO, a reactive dicarbonyl compound, causes protein alterations and cellular dysfunction via glycation. In this study, we investigated the effect of glycation on sialylation, a common post-translational modification implicated in cancer. Our experiments using glioma cell lines, human astrocytes (hA), and primary glioma samples revealed different gene expressions of sialyltransferases among cells, highlighting the complexity of the system. Glycation has a differential effect on sialyltransferase expression, upregulating ST8SIA4 in the LN229 and U251 cell lines and decreasing the expression in normal hA. Subsequently, polysialylation increased in the LN229 and U251 cell lines and decreased in hA. This increase in polysialylation could lead to a more aggressive phenotype due to its involvement in cancer hallmark processes such as immune evasion, resistance to apoptosis, and enhancing invasion. Our findings provide insights into the mechanisms underlying GBM aggressiveness and suggest that targeting glycation and sialylation could be a potential therapeutic strategy.


A retrospective study of the incidence and causes of failed spinal anesthetics in a university hospital.
One hundred sequential spinal anesthetic procedures were reviewed retrospectively to study specifically the incidence and causes of spinal anesthesia. Variables examined included the patient population, the technical aspects of performing subarachnoid tap and subsequent blockade, and the level of training of the anesthetists. We found a 17% incidence of spinal failure, defined as the need to use general anesthesia during the surgical procedure. Failure was found to be significantly associated with a lack of free flow of cerebral spinal fluid, the use of tetracaine without epinephrine, and an increased administration of intravenous supplementation. Forty-one% of the failures represented errors in judgement, either in not properly anticipating the duration of surgery or injecting local anesthetic solution in the absence of a free flow of cerebral spinal fluid. An incidental finding was the lack of documentation in many of the variables examined. We attribute the high incidence of failed spinal anesthesia mainly to technical reasons, most of them avoidable. The use of local and regional anesthesia requires considerable technical skills and demands a precise and total understanding of regional anatomic relationships. With the decreasing use of regional anesthesia in our operating rooms, only those regional anesthesia techniques that require minimum dexterity, such as spinal and epidural anesthesia, continue to be utilized widely; and even these techniques, safe as they are, are being poorly taught.


Walk Score® and the prevalence of utilitarian walking and obesity among Ontario adults: A cross-sectional study.
Evidence from large, population-based studies about the association between neighbourhood walkability and the prevalence of obesity is limited. The study population consisted of 106,337 people aged 20 or older living in urban and suburban Ontario, who participated in the National Population Health Survey and the Canadian Community Health Survey from 1996/1997 to 2008. Based on their postal code, individuals were grouped into one of five walkability categories, ranging from very car-dependent to "Walker's Paradise," according to the Street Smart Walk Score®, a composite measure of neighbourhood walkability. Logistic regression models, adjusted for demographic, socioeconomic and lifestyle characteristics, were used to estimate odds ratios relating neighbourhood walkability to overweight/obesity and physical activity. Compared with residents of "Walker's Paradise" areas, those in very car-dependent areas had significantly higher odds of being overweight or obese. Despite similar levels of leisure physical activity among residents of all walkability areas, those in "Walker's Paradise" areas reported more utilitarian walking and weighed, on average, 3.0 kg less than did those in very car-dependent areas. Living in a low-walkability area is associated with a higher prevalence of overweight/obesity. Neighbourhood walkability is related to the frequency of utilitarian walking.


Enhanced protective immunity against Baylisascaris schroederi infection in mice through a multi-antigen cocktail vaccine approach.
Baylisascaris schroederi is among the most severe intestinal nematodes affecting giant pandas. Developing effective and secure vaccines can be used as a novel strategy for controlling repeated roundworm infection and addressing drug resistance. In our previous study, three recombinant antigens (rBsHP2, rBsGAL, and rBsUP) exhibited promising effects against B. schroederi infection in the mice model. This study extends the findings by formulating four-form cocktail vaccines (GAL+UP, HP2+UP, GAL+HP2, and GAL+HP2+UP) using three B. schroederi recombinant antigens to improve protection in mice further. Additionally, the protective differences after immunizing mice with different doses of cocktail antigens (150 μg, 100 μg, and 50 μg) were analyzed. Administration of rBs(GAL+UP), rBs(HP2+UP), rBs(GAL+HP2), and rBs(GAL+HP2+UP) significantly reduced liver and lung lesions, along with a decrease in L3 larvae by 83.7%, 82.1%, 76.4%, and 75.1%, respectively. These vaccines induced a Th1/Th2 mixed immunity, evidenced by elevated serum antibody levels (IgG, IgG1, IgG2a, IgE, and IgA) and splenocyte cytokines [interferon gamma (IFN-γ), interleukin (IL)-5, and IL-10]. Furthermore, varying cocktail vaccine dosages did not significantly affect protection. The results confirm that a 50 μg rBs(GAL+UP) dosage holds promise as a better candidate vaccine combination against B. schroederi infection, providing a basis for developing the B. schroederi vaccine.


Functional comparison of the mouse DC-SIGN, SIGNR1, SIGNR3 and Langerin, C-type lectins.
The mouse (m) DC-SIGN family consists of several homologous type II transmembrane proteins located in close proximity on chromosome 8 and having a single carboxyl terminal carbohydrate recognition domain. We first used transfected non-macrophage cell lines to compare the polysaccharide and microbial uptake capacities of three of these lectins--DC-SIGN, SIGNR1 and SIGNR3--to another homologue mLangerin. Each molecule shares a potential mannose-recognition EPN-motif in its carbohydrate recognition domain. Using an anti-Tag antibody to follow Tag-labeled transfectants, we found that each molecule could be internalized, although the rates differed. However, mDC-SIGN was unable to take up FITC-dextran, FITC-ovalbumin, zymosan or heat-killed Candida albicans. The other three lectins showed distinct carbohydrate recognition properties, assessed by blocking FITC-dextran uptake at 37 degrees C and by mannan binding activity at 4 degrees C. Furthermore, only SIGNR1 was efficient in mediating the capture by transfected cells of Gram-negative bacteria, such as Escherichia coli and Salmonella typhimurium, while none of the lectins tested were competent to capture Gram-positive bacteria, Staphylococcus aureus. Interestingly, transfectants with SIGNR1 lacking the cytoplasmic domain were capable of binding FITC-zymosan in a manner that was abolished by EDTA or mannan, but not laminarin. In addition, resident peritoneal CD11b+ cells expressing SIGNR1 bound zymosan at 4 degrees C in concert with a laminarin-sensitive receptor. Therefore these homologous C-type lectins have distinct recognition patters for microbes despite similarities in the carbohydrate recognition domains.


Examination of the association between male gender and preterm delivery.
To examine possible reasons why a male fetus constitutes a risk factor for preterm delivery. Retrospective study of deliveries from hospital database in a UK teaching hospital. The population comprised all deliveries >23 weeks over an 11-year period, excluding multiples, terminations and pregnancies with major abnormalities including indeterminate gender. Obstetric variables and outcomes were initially compared in male and female babies for preterm births in different gestation bands, extreme (<28 weeks), severe (29-32 weeks) and moderate (33-36 weeks). For each, the odds ratios with 95% confidence intervals for preterm delivery were calculated. Then, using binary logistic regression with adjusted odds ratios with 95% confidence intervals, putative causal pathways that might explain the male excess were tested. 75,725 deliveries occurred, of which 4003 (5.3%) were preterm. Males delivered preterm more frequently (OR 1.13, 95% CI 1.06-1.20). This was due to spontaneous (OR 1.30, 95% CI 1.19-1.42) but not iatrogenic (OR 0.96, 95% CI 0.87-1.05) preterm birth. There was an increased risk of pre eclampsia among preterm females. Although males were larger, and male pregnancies were more frequently nulliparous and affected by some other obstetric complications (abruption, urinary tract infection), these did not account for their increased risk. Any effect of growth restriction could not be properly determined. Being male carries an increased risk of spontaneous but not iatrogenic preterm birth. The reasons behind this remain obscure.


Management of Adults with Newly Diagnosed Atrial Fibrillation with and without CKD.
Atrial fibrillation (AF) is highly prevalent in CKD and is associated with worse cardiovascular and kidney outcomes. Limited data exist on use of AF pharmacotherapies and AF-related procedures by CKD status. We examined a large "real-world" contemporary population with incident AF to study the association of CKD with management of AF. We identified patients with newly diagnosed AF between 2010 and 2017 from two large, integrated health care delivery systems. eGFR (≥60, 45-59, 30-44, 15-29, <15 ml/min per 1.73 m2) was calculated from a minimum of two ambulatory serum creatinine measures separated by ≥90 days. AF medications and procedures were identified from electronic health records. We performed multivariable Fine-Gray subdistribution hazards regression to test the association of CKD severity with receipt of targeted AF therapies. Among 115,564 patients with incident AF, 34% had baseline CKD. In multivariable models, compared with those with eGFR >60 ml/min per 1.73 m2, patients with eGFR 30-44 (adjusted hazard ratio [aHR] 0.91; 95% CI, 0.99 to 0.93), 15-29 (aHR, 0.78; 95% CI, 0.75 to 0.82), and <15 ml/min per 1.73 m2 (aHR, 0.64; 95% CI, 0.58-0.70) had lower use of any AF therapy. Patients with eGFR 15-29 ml/min per 1.73 m2 had lower adjusted use of rate control agents (aHR, 0.61; 95% CI, 0.56 to 0.67), warfarin (aHR, 0.89; 95% CI, 0.84 to 0.94), and DOACs (aHR, 0.23; 95% CI, 0.19 to 0.27) compared with patients with eGFR >60 ml/min per 1.73 m2. These associations were even stronger for eGFR <15 ml/min per 1.73 m2. There was also a graded association between CKD severity and receipt of AF-related procedures (vs eGFR >60 ml/min per 1.73 m2): eGFR 30-44 ml/min per 1.73 (aHR, 0.78; 95% CI, 0.70 to 0.87), eGFR 15-29 ml/min per 1.73 m2 (aHR, 0.73; 95% CI, 0.61 to 0.88), and eGFR <15 ml/min per 1.73 m2 (aHR, 0.48; 95% CI, 0.31 to 0.74). In adults with newly diagnosed AF, CKD severity was associated with lower receipt of rate control agents, anticoagulation, and AF procedures. Additional data on efficacy and safety of AF therapies in CKD populations are needed to inform management strategies.


The karrikin signaling regulator SMAX1 controls Lotus japonicus root and root hair development by suppressing ethylene biosynthesis.
An evolutionarily ancient plant hormone receptor complex comprising the α/β-fold hydrolase receptor KARRIKIN INSENSITIVE 2 (KAI2) and the F-box protein MORE AXILLARY GROWTH 2 (MAX2) mediates a range of developmental responses to smoke-derived butenolides called karrikins (KARs) and to yet elusive endogenous KAI2 ligands (KLs). Degradation of SUPPRESSOR OF MAX2 1 (SMAX1) after ligand perception is considered to be a key step in KAR/KL signaling. However, molecular events which regulate plant development downstream of SMAX1 removal have not been identified. Here we show that Lotus japonicus SMAX1 is specifically degraded in the presence of KAI2 and MAX2 and plays an important role in regulating root and root hair development. smax1 mutants display very short primary roots and elongated root hairs. Their root transcriptome reveals elevated ethylene responses and expression of ACC Synthase 7 (ACS7), which encodes a rate-limiting enzyme in ethylene biosynthesis. smax1 mutants release increased amounts of ethylene and their root phenotype is rescued by treatment with ethylene biosynthesis and signaling inhibitors. KAR treatment induces ACS7 expression in a KAI2-dependent manner and root developmental responses to KAR treatment depend on ethylene signaling. Furthermore, in Arabidopsis, KAR-induced root hair elongation depends on ACS7 Thus, we reveal a connection between KAR/KL and ethylene signaling in which the KAR/KL signaling module (KAI2-MAX2-SMAX1) regulates the biosynthesis of ethylene to fine-tune root and root hair development, which are important for seedling establishment at the beginning of the plant life cycle.