Review of the etiologic heterogeneity of the oculo-auriculo-vertebral spectrum (Hemifacial Microsomia).
Hemifacial microsomia is a congenital asymmetry of the lower face that may be associated with other cranial and extracranial anomalies. The variability of its severity, and wide range of anomalies that have been reported with it in some cases has resulted in these composite manifestations being given a number of names, including oculo-auriculo-vertebral spectrum (OAVS). Etiology is often stated to be a perturbation of embryonic blood flow in the developing region, although other factors may also play a role in some cases. Depending on what is considered to be minimum criteria for affected classification, what is often to be presumed to be a sporadic event in a family may be the more severe manifestation of a familial condition. Etiological factors are clearly heterogeneous, the investigation of which is confounded by not only the lack of a refined affected phenotype, but also the apparent influence of genetic factors in some instances that directly influence phenotype perhaps through alteration of mesodermal development, or indirectly through increased susceptibility to vascular disruption. Future studies likely to advance knowledge in this area will need to incorporate an analysis of who may be minimally affected in families, so that advances in genotyping will have greater power to distinguish genetic factors that may influence OVAS through interaction with environmental factors in particular families. The same genetic-environmental factors and or etiological mechanisms may then be investigated in apparently sporadic cases.


Naturally occurring tuberculosis in a squirrel monkey and a cebus monkey.
Tuberculosis was detected in a Brazilian squirrel monkey (Saimiri sciureus) and in a cebus monkey (Cebus apella). Both of these animals were housed in the same room with a rhesus monkey (Macaca mulatta) subsequently found to be tuberculous. All 3 of the monkeys died within a 4-month period.


Transcriptional regulation of an endoglucanase and a cellodextrinase gene in Ruminococcus flavefaciens FD-1.
A gene which encodes a 35 kDa protein with both carboxymethylcellulase and xylanase activity was cloned from Ruminococcus flavefaciens FD-1 and the nucleotide sequence determined. The FD-1 gene, celE, and the celA gene, which encodes a cellodextrinase, were used as probes to analyse transcription in R. flavefaciens grown under different conditions. Transcription of both genes was induced when cellulose was added to cells growing in cellobiose. This induction continued after cellulose depletion and after cell division had ceased. Transcription of both genes was also induced by cellotriose, although the effect was not as pronounced as induction by cellulose and was greater for the celA gene than for the celE gene.


All-cause mortality and multimorbidity in older adults: The role of social support and loneliness.
To determine whether the effect of multimorbidity on time to mortality is modified by level of social support and loneliness in a representative sample of 2113 participants aged 60+. Vital status was ascertained through national registers or by asking participants' relatives. Baseline variables included number of illnesses, self-perceived social support (Oslo social support scale) and loneliness (UCLA loneliness scale). Kaplan-Meier survival curves were used to estimate the time to death by multimorbidity, social support and loneliness. Adjusted cox proportional hazards regression models were conducted to explore interactions between multimorbidity and social support and loneliness. Multimorbidity was associated with low probability of survival, whereas high loneliness and low social support were not related with time to death. Only the interaction multimorbidity∗social support was significant. Participants with low social support and 2 chronic diseases, compared with none, presented lower probability of survival (HR=2.43, 95%CI=1.14-5.18, p<0.05), whereas the effect of multimorbidity, in comparison with not having chronic conditions, did not affect mortality if participants had high social support. For participants with low social support, there were no differences between having one, two or more than two diseases. When there is high social support, the probability of death is significantly lower if one or two chronic diseases are present, compared with more than two. These findings indicate that having a supportive social environment increases the survival of people with physical illnesses, especially those with one or two. For those with more than two illnesses, survival remains unchanged regardless of the level of social support and other protective factors should be explored in future research. Geriatric health professionals are encouraged to evaluate social relationships and stimulate support given by relatives, friends or neighbors.


Examining risk behavior and syringe coverage among people who inject drugs accessing a syringe services program: A latent class analysis.
Injection drug use (IDU) remains a significant public health problem. IDU has been associated closely with the opioid crisis; driving overdose, HIV, and Hepatitis C (HCV) infection nationwide. Syringe services programs (SSPs) remain pivotal evidence-based interventions to reduce harm and engage subgroups of people who inject drugs (PWID). This study aims to provide policy considerations from the IDEA SSP, the first legal SSP in the state of Florida. We performed a latent class analysis on patterns of substance use among participants (N = 982) newly enrolled in a syringe services program (SSP). Associations between classes of substance use and sociodemographic variables, risky injection and sex behaviors, HIV/HCV status and syringe coverage were analyzed using the R3STEP and BCH 3-step procedures in latent class regression. We found a three-class solution: Heroin-Dominant class (73.9%), Methamphetamine-Dominant class (9.5%) and Heroin/Cocaine class (16.6%). Compared to Heroin-Dominant class, the Heroin/Cocaine class were more likely to report homelessness, sharing works, unprotected sex, public injection, and to be HCV positive. Compared to both Heroin-Dominant and Heroin/Cocaine classes, the Methamphetamine-Dominant class were more likely to be male, Hispanic, gay or bisexual orientation, HIV positive, to report unprotected sex and sex with PWID. In addition, the lowest and highest syringe coverage were among those in the Heroin/Cocaine and Methamphetamine-Dominant classes, respectively. Existing interventions among this population to mitigate infectious disease risk, such as SSPs, can be a used to engage differing PWID populations. However, multi-component, targeted preventive interventions and need-based syringe distribution policies are required to further reduce HIV and HCV risk among various PWID populations.


Rio1 promotes rDNA stability and downregulates RNA polymerase I to ensure rDNA segregation.
The conserved protein kinase Rio1 localizes to the cytoplasm and nucleus of eukaryotic cells. While the roles of Rio1 in the cytoplasm are well characterized, its nuclear function remains unknown. Here we show that nuclear Rio1 promotes rDNA array stability and segregation in Saccharomyces cerevisiae. During rDNA replication in S phase, Rio1 downregulates RNA polymerase I (PolI) and recruits the histone deacetylase Sir2. Both interventions ensure rDNA copy-number homeostasis and prevent the formation of extrachromosomal rDNA circles, which are linked to accelerated ageing in yeast. During anaphase, Rio1 downregulates PolI by targeting its subunit Rpa43, causing PolI to dissociate from the rDNA. By stimulating the processing of PolI-generated transcripts at the rDNA, Rio1 allows for rDNA condensation and segregation in late anaphase. These events finalize the genome transmission process. We identify Rio1 as an essential nucleolar housekeeper that integrates rDNA replication and segregation with ribosome biogenesis.


Detection of Earth-rotation Doppler shift from Suomi National Polar-Orbiting Partnership Cross-Track Infrared Sounder.
The Cross-Track Infrared Sounder (CrIS) on the Suomi National Polar-Orbiting Partnership Satellite is a Fourier transform spectrometer and provides a total of 1305 channels for sounding the atmosphere. Quantifying the CrIS spectral accuracy, which is directly related to radiometric accuracy, is crucial for improving its data assimilation in numerical weather prediction. In this study, a cross-correlation method is used for detecting the effect of Earth-rotation Doppler shift (ERDS) on CrIS observations. Based on a theoretical calculation, the ERDS can be as large as about 1.3 parts in 10(6) (ppm) near Earth's equator and at the satellite scan edge for a field of regard (FOR) of 1 or 30. The CrIS observations exhibit a relative Doppler shift as large as 2.6 ppm for a FOR pair of 1 and 30 near the equator. The variation of the ERDS with latitude and scan position detected from CrIS observations is similar to that derived theoretically, which indicates that the spectral stability of the CrIS instrument is very high. To accurately calibrate CrIS spectral accuracy, the ERDS effect should be removed. Since the ERDS is easily predictable, the Doppler shift is correctable in the CrIS spectra.


In vitro evaluation of T-Scan®III through study of the sensels.
To evaluate the validity and reliability of the T-Scan®III sensors system by measuring force and time in sensels. In order to evaluate the validity, known forces were applied to 31 sensels selected at random of a T-Scan® III sensor. In order to evaluate the reliability, the existing agreement between two different sensors was measured. Both validity and reliability were assessed using Lin's concordance correlation coefficient (CCC). Using the McBride's scale, for the total group of sensels, by measuring force, a poor level of validity was obtained (Lin's CCC:0.698); when measuring time, an almost perfect agreement was obtained (Lin's CCC: 0.9995). In contrast, the reliability level was poor (Lin's CCC: 0.099). The T-Scan®III system neither shows an appropriate degree of validity when estimating absolute force, nor presents an adequate level of reliability when measuring relative force; however, it presents a high degree of validity when quantifying times.


Estimating the burden of disease from water, sanitation, and hygiene at a global level.
We estimated the disease burden from water, sanitation, and hygiene at the global level taking into account various disease outcomes, principally diarrheal diseases. The disability-adjusted life year (DALY) combines the burden from death and disability in a single index and permits the comparison of the burden from water, sanitation, and hygiene with the burden from other risk factors or diseases. We divided the world's population into typical exposure scenarios for 14 geographical regions. We then matched these scenarios with relative risk information obtained mainly from intervention studies. We estimated the disease burden from water, sanitation, and hygiene to be 4.0% of all deaths and 5.7% of the total disease burden (in DALYs) occurring worldwide, taking into account diarrheal diseases, schistosomiasis, trachoma, ascariasis, trichuriasis, and hookworm disease. Because we based these estimates mainly on intervention studies, this burden is largely preventable. Other water- and sanitation-related diseases remain to be evaluated. This preliminary estimation of the global disease burden caused by water, sanitation, and hygiene provides a basic model that could be further refined for national or regional assessments. This significant and avoidable burden suggests that it should be a priority for public health policy.


Antiatherosclerotic effects of alfalfa meal ingestion in chicks: a biochemical evaluation.
Feeding Alfalfa seed extract to chicks resulted in significant reduction of total cholesterol, phospholipid, triglyceride, LDL-Cholesterol and VLDL-cholesterol. Simultaneously, an increased in the HDL-cholesterol/total cholesterol ratio occurred with reduction in total cholesterol and phospholipid contents of liver and (ventricular) muscle of the heart. Results obtained with alfalfa seed extract ingestion were evaluated with a standard drug, compound, clofibrate.


Overactivation of the Pupillary Response to Emotional Information in Short- and Long-Term Alcohol Abstinent Patients.
The aim of the study was to compare emotional information processing in patients with severe alcohol use disorder in short-term abstinence (<1 month) and long-term abstinence (at least 6 months to 9 years) with control participants. We studied the variation in pupil diameter during the presentation of pictures of human interactions associated with positive, negative or neutral valences. Overall, the results of the short-term abstinent group revealed greater pupil dilation regardless of the valence of the pictures while the pupillary response of long-term abstainers did not differ from the control group. More specifically, according to each valence, the pupil response to neutral pictures was greater for both patient groups than for controls. For the long-term abstainers, a negative correlation was found between the length of abstinence and the pupillary response to emotional stimuli. In long-term abstainers group, the high activation by neutral stimuli suggests however some difficulties in the processing of nonemotional stimuli, considered emotional ones and may constitute a potential relapse factor or the maintenance of addiction.


Personality traits in women with endometrial or cervical cancer.
The objective of the study was to describe the personality characteristics of women with endometrial (n = 30) or cervical cancer (n = 30) and to test the hypothesis of diagnosis-specific personality profiles. The Karolinska Scales of Personality were administered at the first interview with the women before the initiation of treatment. Significant differences occurred both between the two clinical groups and in comparisons with healthy controls. Whereas the personality profile of the cervical group was almost identical to the controls, the endometrial group scored significantly lower on aggression scales and higher on social desirability and socialization. The findings of a normal personality profile in cervical cancer patients allow us to abandon some of the existing misconceptions and encourage us to consider personality factors in the management of those patients.


[Disease management programmes in Germany: first reactions by patients suffering from diabetes].
The performance of health delivery for chronic diseases in Germany has been considered deficient for a long time and in need of improvement. The launch of Disease Management Programmes (DMP) for type 2 diabetes in the summer of 2002 can be seen as an attempt to optimise health care for the chronically ill. The aim of our study was to find out at an early stage of DMP development which factors, personal or otherwise, would have an influence on whether diabetic patients know of, and are willing to participate in, such a programme. The data of 679 diabetic patients, collected between late 2002 and the end of 2003, presented here have been extracted from a written follow-up study to the three baseline surveys S1 (1984/85), S2 (1989/90) und S3 (1994/95) in Augsburg as part of the WHO-MONICA project with more than 13,000 participants. The study population is characterised by a rather high age (mean: 67.4 years), a long duration of the disease (mean: 9.8 years) as well as a high intensity of contacts with physicians. Only 28 % of the diabetic patients had heard of the DMP, but a majority of the participants (54 %) stated their readiness to participate in DMP once offered. Higher age, low education as well as no participation in diabetes education have been identified in logistic regression models as impeding factors both for the knowing of and the participation in DMP. As a consequence, promotion of the DMP should be directed to those sections of the diabetic population outside the effective reach of the traditional diabetes care organisations.


Failure of pyruvate to salvage myocardium after prolonged ischemia.
Thrombolytic therapy for acute coronary occlusion may be more effective if combined with substrate-enhanced reperfusion. In this study, we examined the utility of pyruvic acid, an important metabolic substrate, in salvaging ischemic myocardium. Twenty-six anesthetized dogs underwent 3 h of circumflex coronary occlusion followed by 90 min of reperfusion with administration of intracoronary pyruvate or vehicle. To test the sensitivity of the model in detecting differences in infarct size, eight additional dogs underwent coronary occlusion of shorter duration (45 min), an intervention that is known to reduce infarct size. Collateral perfusion to the ischemic zone during coronary occlusion was similar in experimental and control groups. Whereas a shorter duration of occlusion (45 min) decreased the infarct-to-risk area ratio by 54% compared with a longer duration of occlusion (90 min), neither early (15 min prior to occlusion) nor late (3 h after occlusion) onset of intracoronary infusion of pyruvate shifted the infarct-risk relationship (control: y = 74x - 8.7, r = 0.99; early infusion: y = 0.76x - 9.5, r = 0.85; late infusion: y = 0.58x - 5.5, r = 0.79). The failure of intracoronary administration of pyruvate to limit infarct size raises questions as to its potential clinical utility in the setting of acute myocardial ischemia.


An antibiofilm coating of 5-aryl-2-aminoimidazole covalently attached to a titanium surface.
Biofilms, especially those formed by Staphylococcus aureus, play a key role in the development of orthopedic implant infections. Eradication of these infections is challenging due to the elevated tolerance of biofilm cells against antimicrobial agents. In this study, we developed an antibiofilm coating consisting of 5-(4-bromophenyl)-N-cyclopentyl-1-octyl-1H-imidazol-2-amine, designated as LC0024, covalently bound to a titanium implant surface (LC0024-Ti). We showed in vitro that the LC0024-Ti surface reduces biofilm formation of S. aureus in a specific manner without reducing the planktonic cells above the biofilm, as evaluated by plate counting and fluorescence microscopy. The advantage of compounds that only inhibit biofilm formation without affecting the viability of the planktonic cells, is that reduced development of bacterial resistance is expected. To determine the antibiofilm activity of LC0024-Ti surfaces in vivo, a biomaterial-associated murine infection model was used. The results indicated a significant reduction in S. aureus biofilm formation (up to 96%) on the LC0024-Ti substrates compared to pristine titanium controls. Additionally, we found that the LC0024-Ti substrates did not affect the attachment and proliferation of human cells involved in osseointegration and bone repair. In summary, our results emphasize the clinical potential of covalent coatings of LC0024 on titanium implant surfaces to reduce the risk of orthopedic implant infections. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1908-1919, 2019.


Multivariate time-to-event models for studies of recurrent childhood diseases.
Standard Poisson regression analyses of infectious disease incidence may not be optimal when covariates change over time. When a longitudinal study is of sufficient duration so that multiple disease episodes per person may occur, within-subject correlation may be present and require special statistical consideration. Seasonality of disease incidence is a common feature of field studies of respiratory and diarrhoeal diseases in children. Accurate analysis may require keeping the children aligned with respect to the same baseline hazard function. These data features often are ignored in the analysis of such studies. Methods for accounting for multiple events are discussed. We propose the use of a counting process model to retain the alignment with respect to season. This model is supplemented by a simple bootstrap procedure to account for the within-child correlation. The bootstrap technique is illustrated with data from a randomized trial of the effects of vitamin A supplementation on childhood morbidity. Standard errors for the main effect of vitamin A are larger using the bootstrap, indicating the presence of positive correlation. Time-to-event models can be useful in the analysis of studies of childhood morbidity, especially for situations with seasonality, waning of effect, or multiple events. The bootstrap provides an appropriate, straightforward method for handling within-child correlation in such settings.


Thirty Years of Global Deep Brain Stimulation: "Plus ça change, plus c'est la même chose"?
The advent of deep brain stimulation (DBS) of the subthalamic nucleus (STN) for Parkinson's disease 30 years ago has ushered a global breakthrough of DBS as a universal method for therapy and research in wide areas of neurology and psychiatry. The literature of the last three decades has described numerous concepts and practices of DBS, often branded as novelties or discoveries. However, reading the contemporary publications often elicits a sense of déjà vu in relation to several methods, attributes, and practices of DBS. Here, we review various applications and techniques of the modern-era DBS and compare them with practices of the past. Compared with modern literature, publications of the old-era functional stereotactic neurosurgery, including old-era DBS, show that from the very beginning multidisciplinarity and teamwork were often prevalent and insisted upon, ethical concerns were recognized, brain circuitries and rational for brain targets were discussed, surgical indications were similar, closed-loop stimulation was attempted, evaluations of surgical results were debated, and controversies were common. Thus, it appears that virtually everything done today in the field of DBS bears resemblance to old-time practices, or has been done before, albeit with partly other tools and techniques. Movement disorders remain the main indications for modern DBS as was the case for lesional surgery and old-era DBS. The novelties today consist of the STN as the dominant target for DBS, the tremendous advances in computerized brain imaging, the sophistication and versatility of implantable DBS hardware, and the large potential for research. Many aspects of contemporary DBS bear strong resemblance to practices of the past. The dominant clinical indications remain movement disorders with virtually the same brain targets as in the past, with one exception: the STN. Other novel brain targets - that are so far subject to DBS trials - are the pedunculopontine nucleus for gait freezing, the anteromedial internal pallidum for Gilles de la Tourette and the fornix for Alzheimer's disease. The major innovations and novelties compared to the past concern mainly the unmatched level of research activity, its high degree of sponsorship, and the outstanding advances in technology that have enabled multimodal brain imaging and the miniaturization, versatility, and sophistication of implantable hardware. The greatest benefit for patients today, compared to the past, is the higher level of precision and safety of DBS, and of all functional stereotactic neurosurgery.


Picomolar-scale compound-specific isotope analyses.
We report modifications to compound-specific isotope analyses (CSIA) to enable high-precision isotopic analyses of picomoles of carbon for intact organic molecules. This sample size is two orders of magnitude below the amounts required for commercial systems. The greatly enhanced sensitivity of this system expands molecular isotope studies and applications previously prohibited by low concentrations and small samples. We utilize the resolving power and low volumetric flow rates of narrow-bore capillary gas chromatography to improve sample transfer efficiency while maintaining narrow peak widths. Post-column peak broadening is minimized using a micro-fluidic valve for solvent diversion, capillary combustion reactor, narrow-bore capillary transfer lines, and cryogenic water trap. The mass spectrometer was fitted with collector amplifiers configured to 25 ms response times and a data logger board with firmware capable of rapid data acquisition. Carbon dioxide gas was introduced directly into the ion source to evaluate the dynamic range of the system and accuracy and precision of carbon isotope ratio (δ13 C value) measurements. The accuracy and precision for combusted compounds were evaluated using a suite of n-alkanes. For ≥30 pmol carbon introduced directly into the ion source, the mean difference between the measured and expected δ13 C values is 0.03‰ (1σ, n = 57) and the standard deviation of replicate measurements is 0.11‰ (1σ). The CO2 peak widths generated by the exponential dilution flask were 250 ms and the peak widths produced by combusting n-alkanes were ca 500 ms, less than 25% the width of conventional gas chromatography peaks. For a mixture of 15 n-alkanes (n-C16 to n-C30 ), the accuracy is 0.3‰ (1σ) and precision is 0.9‰ (1σ) for replicate δ13 C measurements with 100 pmol carbon per compound on column. The pico-CSIA method described here offers improved chromatographic resolution and reduces sample size requirements by two orders of magnitude. These advances significantly broaden the available analytical window for CSIA in research areas frequently hindered by sample size limitations, such as forensics, paleoclimate, astrobiology, and biochemistry.


Role of Microelectrode Recording in Deep Brain Stimulation of the Pedunculopontine Nucleus: A Physiological Study of Two Cases.
Deep brain stimulation (DBS) of the pedunculopontine nucleus (PPN) has been reported to improve gait disturbances in Parkinson's disease (PD); however, there are controversies on the radiological and electrophysiological techniques for intraoperative and postoperative confirmation of the target and determination of optimal stimulation parameters. We investigated the correlation between the location of the estimated PPN (ePPN) and neuronal activity collected during intraoperative electrophysiological mapping to evaluate the role of microelectrode recording (MER) in identifying the effective stimulation site in two PD patients. Bilateral PPN DBS was performed in two patients who had suffered from levodopa refractory gait disturbance. They had been implanted previously with DBS in the internal globus pallidus and the subthalamic nucleus, respectively. The PPN was determined on MRI and identified by intraoperative MER. Neuronal activity recorded was analyzed for mean discharge rate, bursting, and oscillatory activity. The effects were assessed by clinical ratings for motor signs before and after surgery. The PPN location was detected by MER. Groups of neurons characterized by tonic discharges were found 9-10 mm below the thalamus. The mean discharge rate in the ePPN was 19.1 ± 15.1 Hz, and 33% of the neurons of the ePPN responded with increased discharge rate during passive manipulation of the limbs and orofacial structures. PPN DBS with bipolar stimulation at a frequency range 10-30 Hz improved gait disturbances in both patients. Although PPN DBS provided therapeutic effects post-surgery in both cases, the effects waned after a year in case 1 and three years in case 2. Estimation of stimulation site within the PPN is possible by combining physiological guidance using MER and MRI findings. The PPN is a potential target for gait disturbances, although the efficacy of PPN DBS may depend on the location of the electrode and the stimulation parameters.


T-cell receptor V beta gene expression differs in tumor-infiltrating lymphocytes within primary and metastatic melanoma.
Expression of T-cell receptor (TCR) gene rearrangements in tumor-infiltrating lymphocytes (TILs) within primary and metastatic melanoma specimens was studied. In order to analyze TCR gene transcription in TILs within these tissues, we analyzed reverse transcribed complementary DNA from mRNA directly from tissues using the polymerase chain reaction. The polymerase chain reaction-amplified products were confirmed by dot or Southern blot hybridization with C alpha or C beta oligoprobes. First, we investigated the diversity of TCR V alpha and V beta gene usage in human malignant melanoma patients with multiple metastasis. We found in one patient, bearing multiple skin lesions, that the patterns of TCR V alpha and V beta repertoires in different sites of the skin (leg and chest wall) were almost the same. However, in another patient with skin and brain melanomas, different TCR repertoires were presented. Next, we examined the usage of murine TCR V beta genes in TILs within the primary and metastatic sites (liver, lung, and brain) of C57BL/6 mice bearing B16-F10 murine melanoma. The population of TILs in each primary and metastatic site expressed from one to four TCR V beta genes. In each metastatic site, the profile of TCR V beta gene expression was different. A different TCR V beta usage in TILs distributed within metastases of various organs may reflect differences in tumor antigenicity at these sites or may be due to differential homing patterns to these tumors.


Discrimination between adenocarcinoma and normal pancreatic ductal fluid by proteomic and glycomic analysis.
Sensitive and specific biomarkers for pancreatic cancer are currently unavailable. The high mortality associated with adenocarcinoma of the pancreatic epithelium justifies the broadest possible search for new biomarkers that can facilitate early detection or monitor treatment efficacy. Protein glycosylation is altered in many cancers, leading many to propose that glycoproteomic changes may provide suitable biomarkers. In order to assess this possibility for pancreatic cancer, we have performed an in-depth LC-MS/MS analysis of the proteome and MS(n)-based characterization of the N-linked glycome of a small set of pancreatic ductal fluid obtained from normal, pancreatitis, intraductal papillary mucinous neoplasm (IPMN), and pancreatic adenocarcinoma patients. Our results identify a set of seven proteins that were consistently increased in cancer ductal fluid compared to normal (AMYP, PRSS1, GP2-1, CCDC132, REG1A, REG1B, and REG3A) and one protein that was consistently decreased (LIPR2). These proteins are all directly or indirectly associated with the secretory pathway in normal pancreatic cells. Validation of these changes in abundance by Western blotting revealed increased REG protein glycoform diversity in cancer. Characterization of the total N-linked glycome of normal, IPMN, and adenocarcinoma ductal fluid clustered samples into three discrete groups based on the prevalence of six dominant glycans. Within each group, the profiles of less prevalent glycans were able to distinguish normal from cancer on this small set of samples. Our results emphasize that individual variation in protein glycosylation must be considered when assessing the value of a glycoproteomic marker, but also indicate that glycosylation diversity across human subjects can be reduced to simpler clusters of individuals whose N-linked glycans share structural features.


Survival after valve replacement for aortic stenosis: implications for decision making.
Recommendations for aortic valve replacement in severe aortic stenosis are based primarily on the presence of symptoms. However, the onset of symptoms is often insidious, potentially leading to delayed intervention and suboptimal results. Identifying factors that reduce the survival of patients undergoing aortic valve replacement could lead to revised treatment guidelines and improved outcomes. We conducted a single-center observational clinical study of 3049 patients with aortic stenosis who underwent native aortic valve replacement with a single type of bioprosthesis. The primary end point was all-cause mortality from the date of operation. Multivariable analysis of risk factors for death was performed in the multiphase hazard function domain. The presence of severe left ventricular hypertrophy at operation, which preceded symptoms in 17% of patients, was associated with decreased survival. This effect was magnified by the severity of aortic stenosis (P = .02) and use of small prostheses (P = .01). The presence of left ventricular dysfunction reduced survival (P = .0003). Although older age was a risk factor for death (P < .0001), elderly patients had survival comparable to their age, race, and sex-matched cohorts, whereas younger patients had worse than expected survival that was further diminished with insertion of a small prosthesis (P = .01). To optimize survival, earlier aortic valve replacement should be considered even in asymptomatic patients before severe left ventricular hypertrophy or dysfunction develops. In younger patients, the largest possible prosthesis should be implanted to minimize residual gradient; in elderly patients, complex operations just to insert larger prostheses should be avoided.


The low ionic strength form of the sodium salt of poly(dm5C-dG) is a B DNA.
The low-salt Z form of poly(dm5C-dG) reported in the literature was shown to be due to minute amounts of bivalent cations. If the polymer was purified by dialysis against 1 mM EDTA, poly(dm5C-dG) showed the normal B form. No change in this B form was observed up to 0.8 M NaCl. Very small amounts of Mg2+ or Ca2+ (10(-6) to 10(-5) M) induced the characteristic Z form CD spectrum, which could easily and rapidly be reverted to B-DNA by addition of EDTA.


Limited reporting of major harms in studies of initial combination antiretroviral therapy: a systematic review.
Risk-benefit assessment of combination antiretroviral therapy (cART) requires consideration of all potential serious harms. Studies of initial cART may permit identification of associations between particular regimens and uncommon harms, but only if comprehensively reported in the public domain. Study-based, systematic review of published initial cART studies (in adult patients) for completeness of serious harms reporting. Electronic databases, abstracts, and regulatory/sponsor reports were searched (1 January 1996 - 31 December 2012). Reporting of pre-specified harms - deaths, new/recurrent AIDS events, serious non-AIDS events (2010 INSIGHT classification) and serious adverse events (SAEs) - were assessed as the proportion of studies providing data (reporting frequency). Pharmaceutical sponsors were approached for unreported data. 103 studies (86% randomized, 54% industry-sponsored) were included. Deaths, AIDS events, serious non-AIDS events and SAEs were reported for 85 (83%), 55 (53%), 26 (25%) and 43 (42%) studies, respectively. Deaths were better reported for academic than industry-sponsored studies (91 vs. 75%; P = 0.03); the converse applied for SAEs (26 vs. 55%; P = 0.002). SAEs were better reported for randomized than cohort studies (46 vs. 14%; P = 0.03), and for phase 3 than phase 2 or 4 studies (58 vs. 32 and 29%, respectively; P = 0.02). SAE reporting increased over time [ρ = 0.704, P = 0.002 (Spearman)]. Unreported data acquired for 34 (61%) of 56 industry-sponsored studies improved ascertainment in these studies to between 82 and 100% (P < 0.001). Public domain reporting of serious harms for initial cART studies is limited. Insufficient data exist to determine if particular ART drugs/regimens are associated with most serious harms.


Inhibition of Influenza A Virus by Human Infant Saliva.
Innate antiviral factors in saliva play a role in protection against respiratory infections. We tested the anti-influenza virus activities of saliva samples taken from human infants, 1-12 months old, with no history of prior exposure to influenza. In contrast to the inhibitory activity we observed in mouse and ferret saliva, the activity of human infant saliva was complex, with both sialic acid-dependent and independent components, the proportion of which differed between individuals. Taken as a whole, we showed that the major anti-influenza activity of infant saliva is acquired over the first year of life and is associated with sialic acid-containing molecules. The activity of sialic acid-independent inhibitors was lower overall, more variable between individuals, and less dependent on age. The results show that the saliva of very young infants can provide a degree of protection against influenza, which may be critical in the absence of adaptive immunity.


Effects of carbofuran and deltamethrin on acetylcholinesterase activity in brain and muscle of the common carp.
This work investigated the effect from exposure to insecticides carbofuran and deltamethrin on acetylcholinesterase (AChE) activity in the brain and muscle of common carp (Cyprinus carpio). Both pesticides were evaluated through two separate experiments, and carp were exposed in a semi-static system to three different concentrations of carbofuran (10, 50, and 100 μg/L) and deltamethrin (0.08, 0.4, and 0.8 μg/L) during a month with sampling times at 0, 4, 15, and 30 days (n = 7 from each aquarium). AChE activity was significantly inhibited in both organs of carps exposed to carbofuran at all sampling times depending on dose and time, reaching inhibition values of 73.5 and 67.1%, in brain and muscle tissues respectively, after 30 days with the highest concentration. On the contrary, AChE activity was not significantly affected after deltamethrin exposure at all concentrations and times of the assay. This study shows that the measurement of brain and muscle AChE activity in Cyprinus carpio is a useful biomarker of carbamates exposure and/or effects, but has no application with pyrethroids.


Differences in respiratory function, depressive symptoms and quality of life between patients with hereditary motor and sensory neuropathy and myotonic dystrophy undergoing maintenance rehabilitation.
Hereditary motor and sensory neuropathy (HMSN) and myotonic dystrophy (MD) are chronic neuromuscular diseases that cause progressive muscular impairment and impact patient's quality of life. Conflicting findings in existing literature underscore the need for focussed research on specific health aspects in these patients. The aim of the study was to explore the differences in respiratory function, depressive symptoms and quality of life between patients with HMSN and MD undergoing maintenance rehabilitation. Our prospective observational study included 62 HMSN patients (median age 53.5, range 19-79 years; 38 women) and 50 MD patients (median age 54.0, range 18-77 years; 34 women) undergoing maintenance rehabilitation. They performed respiratory function tests (vital capacity, forced vital capacity, forced expiratory volume and peak expiratory flow) and respiratory muscle strength tests (maximum inspiratory pressure, maximum expiratory pressure and sniff nasal inspiratory force). The Center for Epidemiological Studies-Depression Scale was used to evaluate depression, and their health-related quality of life was assessed using the Quality of Life in Genetic Neuromuscular Disease Questionnaire. Using logistic regression, we examined group-difference in presence of depressive symptoms while accounting for age, symptoms duration, and marital status. Multiple linear regression was used to assess the difference in quality-of-life scores, adjusting for age, gender, and symptoms' duration. The HMSN group achieved statistically significantly better results than the MD group on all respiratory measures (about 17% absolute difference in the respiratory function measures and 30% relative difference in the respiratory muscle strength measures on average). Both groups exhibited a high level of depression symptoms (HMSN 24%, MD 44%; estimated adjusted odds-ratio MD vs. HMSN 1.9, 95% CI 0.8-4.5, P = 0.127). We did not find a statistically significant difference between the groups regarding quality-of-life domains, though a trend towards better quality-of-life among the HMSN patients could be observed. The implication for future practice is that the MD patients would potentially benefit the most from targeted respiratory-rehabilitation interventions, and both groups could benefit from focussed mental-health interventions.


Patterns of bovine dairy submissions to the California Veterinary Diagnostic Laboratory System.
Dairy cattle submissions to the California Veterinary Diagnostic Laboratory System (CVDLS) were analyzed to determine submitter statistics. Eligible submissions were those received July 1, 1987 through December 31, 1989 for reasons other than regulatory brucellosis serology. A comprehensive list frame of California dairies was constructed from Brucella Ring Test information and served as the comparison population for the study. Analyses were performed based on geographic location, herd size, proximity to a CVDLS laboratory, and frequency of submission. Thirty-nine percent of the 2,490 California dairies in the reference population had submitted specimens greater than or equal to 1 time to the CVDLS during the study period. Twenty-three percent of the reference population had submitted greater than or equal to 2 times. Specimens were more likely to be submitted from larger herds than smaller herds. Larger dairies also submitted specimens more frequently. Dairies in the northern part of the state were more likely to submit specimens and submitted more frequently than southern herds when herd size was accounted for in the analysis. Mean submission rate (+/- SD) for the 970 submitting dairies was 1.4 (+/- 1.8) submissions/year. Forty-six percent of the dairies accounted for 80% of submissions, whereas only 14% of dairies accounted for 50% of all diagnostic dairy submissions.


Multiscale, multimodal analysis of tumor heterogeneity in IDH1 mutant vs wild-type diffuse gliomas.
Glioma is recognized to be a highly heterogeneous CNS malignancy, whose diverse cellular composition and cellular interactions have not been well characterized. To gain new clinical- and biological-insights into the genetically-bifurcated IDH1 mutant (mt) vs wildtype (wt) forms of glioma, we integrated data from protein, genomic and MR imaging from 20 treatment-naïve glioma cases and 16 recurrent GBM cases. Multiplexed immunofluorescence (MxIF) was used to generate single cell data for 43 protein markers representing all cancer hallmarks, Genomic sequencing (exome and RNA (normal and tumor) and magnetic resonance imaging (MRI) quantitative features (protocols were T1-post, FLAIR and ADC) from whole tumor, peritumoral edema and enhancing core vs equivalent normal region were also collected from patients. Based on MxIF analysis, 85,767 cells (glioma cases) and 56,304 cells (GBM cases) were used to generate cell-level data for 24 biomarkers. K-means clustering was used to generate 7 distinct groups of cells with divergent biomarker profiles and deconvolution was used to assign RNA data into three classes. Spatial and molecular heterogeneity metrics were generated for the cell data. All features were compared between IDH mt and IDHwt patients and were finally combined to provide a holistic/integrated comparison. Protein expression by hallmark was generally lower in the IDHmt vs wt patients. Molecular and spatial heterogeneity scores for angiogenesis and cell invasion also differed between IDHmt and wt gliomas irrespective of prior treatment and tumor grade; these differences also persisted in the MR imaging features of peritumoral edema and contrast enhancement volumes. A coherent picture of enhanced angiogenesis in IDHwt tumors was derived from multiple platforms (genomic, proteomic and imaging) and scales from individual proteins to cell clusters and heterogeneity, as well as bulk tumor RNA and imaging features. Longer overall survival for IDH1mt glioma patients may reflect mutation-driven alterations in cellular, molecular, and spatial heterogeneity which manifest in discernable radiological manifestations.


Chemical ecology of the palm weevilRhynchophorus palmarum (L.) (Coleoptera: Curculionidae): Attraction to host plants and to a male-produced aggregation pheromone.
Attraction to host plants by adultRhynchophorus palmarum (L.) palm weevils was studied in the field and in the laboratory. Chemical analysis revealed the presence of ethanol and ethyl-acetate in stems of coco palms and in pineapple fruits and of pentane, hexanal, and isopentanol in coco stems. In the olfactometer, the first two compounds and isoamyl-acetate were attractive to the insects and the last three compounds, although not attractive by themselves, increased attractiveness when mixed with the first two compounds. Mixtures of these compounds, in proportions similar to the one occurring in attractive plant tissue, were as attractive as natural coconut tissue. In the field, the chemical compounds, either presented alone or as a mixture, did not attract the weevil. Males produce an aggregation pheromone when smelling ethyl-acetate. Rhynchophorol, 2(E)-6-methyl-2-hepten-4-ol, the known active component of the aggregation pheromone, attracts weevils in the olfactometer and in the field only if plant tissue, ethyl-acetate, or the above-mentioned odor mix are present. We propose that a complex mix of ethanol, ethyl-acetate, pentane, hexanal, isolamyl-acetate, and/or isopentanol serve as a short-range orientation cue to fresh wounds on the plant and that additional host odors, attracting weevils from a distance, have still to be discovered. Rhynchophorol can be considered to be a Synergist, having an anemotactic action at a distance. We recommend the use of retention traps baited with rhynchophorol, ethyl-acetate, and sugar cane as an alternative control method for the pest.


Eight-week, multimodal exercise counteracts a progress of chemotherapy-induced peripheral neuropathy and improves balance and strength in metastasized colorectal cancer patients: a randomized controlled trial.
Physical activity is supposed to decrease mortality of colorectal cancer (CRC) and is suggested to reduce side-effects of the disease and its treatment. However, the knowledge about the influence of exercise interventions on patients suffering from CRC and metastasized CRC (mCRC) is still sparse. One frequently observed side effect in mCRC is chemotherapy-induced peripheral neuropathy (CIPN). This randomized controlled trial investigated the influence of a supervised exercise program on CIPN in mCRC. Thirty patients (stage IV) undergoing outpatient palliative treatment including a median of 23.5 chemotherapy cycles of various regimens were randomly assigned to an intervention or control group (IG, n = 17; CG, n = 13). The IG participated in an eight-week supervised exercise program, including endurance, resistance and balance training (2×/week for 60 min) whereas the CG received written standard recommendations to obtain physical fitness. CIPN was assessed using the FACT/GOG-NTX questionnaire. Moreover, endurance capacity (6MWT), strength (h1RM) and balance (GGT-Reha) were evaluated before (t 0) and after (t 1) the intervention as well as after 4 weeks follow-up (t2). Neuropathic symptoms remained stable in the IG over time, while CIPN significantly worsened in the CG from t 0 to t 1 and t 0 to t 2. In contrast to the CG, the IG significantly improved in strength and balance function. Changes in CIPN correlated with changes in balance. This is the first investigation showing positive effects of a multimodal exercise program on CIPN, balance and strength on mCRC patients in a palliative setting, thereby consequently increasing patients` quality of life. The results support earlier findings stating a positive influence of balance exercise on CIPN.


High-energy chemistry of formamide: a simpler way for nucleobase formation.
The formation of nucleobases from formamide during a high-energy density event, i.e., the impact of an extraterrestrial body into the planetary atmosphere, was studied by irradiation of formamide ice and liquid samples with a high-power laser in the presence of potential catalysts. FTIR spectroscopy, time-resolved emission spectroscopy, and GC-MS were subsequently used to monitor the dissociation of this molecule into stable molecular fragments (HCN, H2O, HNCO, H2, CO, and NH3) and unstable species (HNC, •CN, and •NH). The kinetic and thermodynamic models of the high-energy density event molecular dynamics have been suggested together with the reaction routes leading from the dissociation products to the nucleobases. In addition, using theoretical calculations, we propose a simple new reaction pathway for the formation of both pyrimidine and purine nucleobases involving •CN radical chemistry.


Relating physico-chemical properties of frozen green peas (Pisum sativum L.) to sensory quality.
The acceptability of frozen green peas depends on their sensory quality. There is a need to relate physico-chemical parameters to sensory quality. In this research, six brands of frozen green peas representing product sold for retail and caterer's markets were purchased and subjected to descriptive sensory evaluation and physico-chemical analyses (including dry matter content, alcohol insoluble solids content, starch content, °Brix, residual peroxidase activity, size sorting, hardness using texture analysis and colour measurements) to assess and explain product quality. The sensory quality of frozen green peas, particularly texture properties, were well explained using physico-chemical methods of analysis notably alcohol insoluble solids, starch content, hardness and °Brix. Generally, retail class peas were of superior sensory quality to caterer's class peas although one caterer's brand was comparable to the retail brands. Retail class peas were sweeter, smaller, greener, more moist and more tender than the caterer's peas. Retail class peas also had higher °Brix, a(*) , hue and chroma values; lower starch, alcohol insoluble solids, dry matter content and hardness measured. The sensory quality of frozen green peas can be partially predicted by measuring physico-chemical parameters particularly °Brix and to a lesser extent hardness by texture analyser, alcohol insoluble solids, dry matter and starch content.


Misrepresentations of the Tuskegee Study of Untreated Syphilis.
The Tuskegee Study of Untreated Syphilis (TSUS) is the subject of medical research, healthcare, ethics, race relations and fictitious media portrayals (e.g., movie, play). Unraveling misrepresentations about the TSUS is important because prevailing views continue to contribute to alleged mistrust of the medical system by the black community and compromised doctor-patient and researcher-participant relationships. A comparative analysis of standard TSUS information was conducted and included evidenced-based alternative information in an effort to: (a) contextualize the TSUS with accurate medical, public health and historical information and (b) balance claims of racism, nontreatment and denial of treatment. An article in the Winter 2003 Radcliffe Quarterly misrepresented the images of two historical African-American health institutions, the John A. Andrew Memorial Hospital and the Tuskegee Veterans Administration Hospital. A critical analysis challenges these pictorial misrepresentations and documents, in part, a continuing and prevailing pattern of biased misrepresentations in discussions about the TSUS. The National Medical Association (NMA) and the Journal of the National Medical Association have a history of advocacy for African-American health, a record of combating racial discrimination and a quest for truth about the status of the black community. As past NMA historical notables have performed through their advancement, service and leadership, health professionals now have the opportunity, obligation and information to educate the American community about the TSUS with evidenced-based information.


Uteroglobin: a potential novel tumor suppressor and molecular therapeutic for prostate cancer.
Currently, there are very few diagnostic or therapeutic strategies targeted at controlling tumor growth and progression towards metastasis. Uteroglobin (UG) is a naturally occurring, small, stable, secretory protein that is normally expressed by most cells of epithelial origin but is known to be lost during the progression of prostate, lung, and uterine cancers to invasive malignancy. Uteroglobin -/- knockout mice appear to be extremely cancer prone. Both pharmacological and transgenic reconstitution of recombinant human UG (rhUG) to prostate, lung, and endometrial tumor cell lines markedly inhibits their invasiveness and antagonizes the neoplastic phenotype. In preliminary studies, rhUG inhibited angiogenesis in the ex vivo rat aorta model and showed antitumor activity against human prostate tumor cells (PC-3) in the chick chorioallantoic membrane assay, reducing both tumor volume and vascularity. A recent in vivo pilot study showed that twice daily dosing with rhUG resulted in a statistically significant increase in survival without evidence of toxicity in severe combined immunodeficient mice challenged with a PC-3 cell metastasizing tumor. Thus, rhUG may slow the progression of cancer by inhibiting both tumor cell invasiveness and tumor angiogenesis. It therefore holds the potential to serve as a new weapon in the arsenal of cytostatic, antimetastatic, adjuvant treatment for cancer. In this paper, we will briefly discuss the therapeutic potential of uteroglobin-based strategies for managing prostate cancer.


Leaf vein patterning is regulated by the aperture of plasmodesmata intercellular channels.
To form tissue networks, animal cells migrate and interact through proteins protruding from their plasma membranes. Plant cells can do neither, yet plants form vein networks. How plants do so is unclear, but veins are thought to form by the coordinated action of the polar transport and signal transduction of the plant hormone auxin. However, plants inhibited in both pathways still form veins. Patterning of vascular cells into veins is instead prevented in mutants lacking the function of the GNOM (GN) regulator of auxin transport and signaling, suggesting the existence of at least one more GN-dependent vein-patterning pathway. Here we show that in Arabidopsis such a pathway depends on the movement of auxin or an auxin-dependent signal through plasmodesmata (PDs) intercellular channels. PD permeability is high where veins are forming, lowers between veins and nonvascular tissues, but remains high between vein cells. Impaired ability to regulate PD aperture leads to defects in auxin transport and signaling, ultimately leading to vein patterning defects that are enhanced by inhibition of auxin transport or signaling. GN controls PD aperture regulation, and simultaneous inhibition of auxin signaling, auxin transport, and regulated PD aperture phenocopies null gn mutants. Therefore, veins are patterned by the coordinated action of three GN-dependent pathways: auxin signaling, polar auxin transport, and movement of auxin or an auxin-dependent signal through PDs. Such a mechanism of tissue network formation is unprecedented in multicellular organisms.


Investigation of the attitudes of university students to discrimination of the elderly.
This study was conducted to determine the attitudes of university students studying in different fields toward discrimination of the elderly. This descriptive study was conducted with students who were still studying in the 2015-2016 period. A sample size of 416 students was determined by the stratified sampling method, and students were selected by simple random sampling. Data were collected using an identifying information form and an Age Discrimination Attitude Scale (ADAS) by face-to-face interview. Statistical analysis was performed using the program SPSS 20.0. The mean total ADAS score of students was 67.7±6.0. The total ADAS scores and the scores of male students on limiting the life of the elderly was significantly higher than those of female students (p<0.05). It was determined that university students studying in different fields have a positive attitude toward the elderly. Action must be taken to remove discrimination of the elderly, and policies must be developed to increase social sensitivity.


VEP optimal channel selection using genetic algorithm for neural network classification of alcoholics.
In this letter, neural networks (NNs) classify alcoholics and nonalcoholics using features extracted from visual evoked potential (VEP). A genetic algorithm (GA) is used to select the minimum number of channels that maximize classification performance. GA population fitness is evaluated using fuzzy ARTMAP (FA) NN, instead of the widely used multilayer perceptron (MLP). MLP, despite its effective classification, requires long training time (on the order of 10(3) times compared to FA). This causes it to be unsuitable to be used with GA, especially for on-line training. It is shown empirically that the optimal channel configuration selected by the proposed method is unbiased, i.e., it is optimal not only for FA but also for MLP classification. Therefore, it is proposed that for future experiments, these optimal channels could be considered for applications that involve classification of alcoholics.


Effects of Renal Impairment and Hemodialysis on the Pharmacokinetics and Safety of the Glecaprevir and Pibrentasvir Combination in Hepatitis C Virus-Negative Subjects.
Hepatitis C virus (HCV) infection is an independent risk factor for developing chronic renal impairment and end-stage renal disease. Limited treatment options are available for HCV genotype 2, 3, 5, and 6 infections in patients with an estimated glomerular filtration rate (eGFR) of <30 ml/min. Glecaprevir and pibrentasvir are active against all six major HCV genotypes, are primarily excreted in the bile, and have minimal renal elimination. Therefore, combined treatment with these direct-acting antivirals may be useful for patients with HCV infection and chronic kidney disease. A phase 1, multicenter, open-label study evaluated the effects of renal impairment on the pharmacokinetics and safety of glecaprevir-pibrentasvir. In substudy 1, 38 subjects with stage 2 to 5 chronic kidney disease who were not on dialysis or who had normal renal function received single doses of the combination of 300 mg glecaprevir and 120 mg pibrentasvir. In substudy 2, 8 subjects requiring hemodialysis received single doses of the combination of 300 mg glecaprevir and 120 mg pibrentasvir under dialysis and nondialysis conditions. Regression analyses demonstrated increased glecaprevir and pibrentasvir plasma exposures, as determined by the area under the curve, with decreasing renal function, up to 56% and 46%, respectively, in subjects with an eGFR of <15 ml/min/1.73 m2 In dialysis-dependent subjects, glecaprevir and pibrentasvir exposures were similar (≤18% difference) when study drugs were administered before hemodialysis or on a nondialysis day. Adverse events were mostly mild, with the most common being self-limited fatigue (3 subjects). The study findings support the clinical evaluation of glecaprevir-pibrentasvir without dose adjustment in HCV-infected subjects with renal impairment. (This study has been registered at ClinicalTrials.gov under registration number NCT02442258.).


Assessing Patient Participation Burden Based on Protocol Design Characteristics.
Although a number of studies have quantitatively measured investigative site burden to administer increasingly complex protocol designs, robust scholarly research has not been performed to quantify the burden that patients face as participants in clinical trials. This paper presents the results of a cross-sectional pilot study conducted by the Tufts Center for the Study of Drug Development and ZS Associates among nearly 600 patients via an online survey conducted between February and March 2019. Respondents rated the perceived burden of 60 commonly administered protocol procedures. The association and relationship between overall patient burden-derived from aggregating mean perceived burden ratings for individual procedures-and performance (eg, screen failure and retention rates, clinical trial cycle times) for a cross-sectional sample of 137 individual protocols was assessed. Descriptive statistics, significance tests, and univariate analyses were performed. Strong positive, statistically significant associations were observed between a composite measure of patient burden and protocol-specific design and performance measures, most notably study visits above the tolerable mean and the study conduct duration from first patient first visit to last patient last visit. The study results suggest a new and viable approach to optimize protocol design and improve patient engagement.


Preparing Lamellae from Vitreous Biological Samples using a Dual-Beam Scanning Electron Microscope for Cryo-Electron Tomography.
Presented here is a protocol for preparing cryo-lamellae from plunge-frozen grids of Plasmodium falciparum-infected human erythrocytes, which could easily be adapted for other biological samples. The basic principles for preparing samples, milling, and viewing lamellae are common to all instruments and the protocol can be followed as a general guide to on-grid cryo-lamella preparation for cryo-electron microscopy (cryoEM) and cryo-electron tomography (cryoET). Electron microscopy grids supporting the cells are plunge-frozen into liquid nitrogen-cooled liquid ethane using a manual or automated plunge freezer, then screened on a light microscope equipped with a cryo-stage. Frozen grids are transferred into a cryo-scanning electron microscope equipped with a focused ion beam (cryoFIB-SEM). Grids are routinely sputter coated prior to milling, which aids dispersal of charge build-up during milling. Alternatively, an e-beam rotary coater can be used to apply a layer of carbon-platinum to the grids, the exact thickness of which can be more precisely controlled. Once inside the cryoFIB-SEM an additional coating of an organoplatinum compound is applied to the surface of the grid via a gas injection system (GIS). This layer protects the front edge of the lamella as it is milled, the integrity of which is critical for achieving uniformly thin lamellae. Regions of interest are identified via SEM and milling is carried out in a step-wise fashion, reducing the current of the ion beam as the lamella reaches electron transparency, in order to avoid excessive heat generation. A grid with multiple lamellae is then transferred to a transmission electron microscope (TEM) under cryogenic conditions for tilt-series acquisition. A robust and contamination-free workflow for lamella preparation is an essential step for downstream techniques, including cellular cryoEM, cryoET, and sub-tomogram averaging. Development of these techniques, especially for lift-out and milling of high-pressure frozen samples, is of high-priority in the field.


Staple suturing of the colon above the peritoneal reflection.
We review the few reports in the literature involving the results of stapled colonic anastomoses. Our personal experience with 88 consecutive stapled colonic anastomoses performed above the peritoneal reflection is analyzed. There were no mortalities and no evidence of anastomotic leaks in our series. With proper attention to the usual tenets of good colonic surgery and knowledge of the limitations of the stapling instruments, stapling colonic anastomoses is a safe technique.


Characterization of Higher Order Structural Changes of a Thermally Stressed Monoclonal Antibody via Mass Spectrometry Footprinting and Other Biophysical Approaches.
Characterizing changes in the higher order structure (HOS) of monoclonal antibodies upon stressed conditions is critical to gaining a better understanding of the product and process. One single biophysical approach may not be best suited to assess HOS comprehensively; thus, the synergy from multiple, complementary approaches improves characterization accuracy and resolution. In this study, we employed two mass spectrometry (MS )-based footprinting techniques, namely, fast photochemical oxidation of proteins (FPOP)-MS and hydrogen-deuterium exchange (HDX)-MS, supported by dynamic light scattering (DLS), differential scanning calorimetry (DSC), circular dichroism (CD), and nuclear magnetic resonance (NMR) to study changes to the HOS of a mAb upon thermal stress. The biophysical techniques report a nuanced characterization of the HOS in which CD detects no changes to the secondary or tertiary structure, yet DLS measurements show an increase in the hydrodynamic radius. DSC indicates that the stability decreases, and chemical or conformational changes accumulate with incubation time according to NMR. Furthermore, whereas HDX-MS does not indicate HOS changes, FPOP-MS footprinting reveals conformational changes at residue resolution for some amino acids. The local phenomena observed with FPOP-MS indicate that several residues show various patterns of degradation during thermal stress: no change, an increase in solvent exposure, and a biphasic response to solvent exposure. All evidences show that FPOP-MS efficiently resolves subtle structural changes and novel degradation pathways upon thermal stress treatment at residue-level resolution.


Machine-assisted interpolation algorithm for semi-automated segmentation of highly deformable organs.
Accurate and robust auto-segmentation of highly deformable organs (HDOs), for example, stomach or bowel, remains an outstanding problem due to these organs' frequent and large anatomical variations. Yet, time-consuming manual segmentation of these organs presents a particular challenge to time-limited modern radiotherapy techniques such as on-line adaptive radiotherapy and high-dose-rate brachytherapy. We propose a machine-assisted interpolation (MAI) that uses prior information in the form of sparse manual delineations to facilitate rapid, accurate segmentation of the stomach from low field magnetic resonance images (MRI) and the bowel from computed tomography (CT) images. Stomach MR images from 116 patients undergoing 0.35T MRI-guided abdominal radiotherapy and bowel CT images from 120 patients undergoing high dose rate pelvic brachytherapy treatment were collected. For each patient volume, the manual delineation of the HDO was extracted from every 8th slice. These manually drawn contours were first interpolated to obtain an initial estimate of the HDO contour. A two-channel 64 × 64 pixel patch-based convolutional neural network (CNN) was trained to localize the position of the organ's boundary on each slice within a five-pixel wide road using the image and interpolated contour estimate. This boundary prediction was then input, in conjunction with the image, to an organ closing CNN which output the final organ segmentation. A Dense-UNet architecture was used for both networks. The MAI algorithm was separately trained for the stomach segmentation and the bowel segmentation. Algorithm performance was compared against linear interpolation (LI) alone and against fully automated segmentation (FAS) using a Dense-UNet trained on the same datasets. The Dice Similarity Coefficient (DSC) and mean surface distance (MSD) metrics were used to compare the predictions from the three methods. Statistically significance was tested using Student's t test. For the stomach segmentation, the mean DSC from MAI (0.91 ± 0.02) was 5.0% and 10.0% higher as compared to LI and FAS, respectively. The average MSD from MAI (0.77 ± 0.25 mm) was 0.54 and 3.19 mm lower compared to the two other methods. Only 7% of MAI stomach predictions resulted in a DSC < 0.8, as compared to 30% and 28% for LI and FAS, respectively. For the bowel segmentation, the mean DSC of MAI (0.90 ± 0.04) was 6% and 18% higher, and the average MSD of MAI (0.93 ± 0.48 mm) was 0.42 and 4.9 mm lower as compared to LI and FAS. Sixteen percent of the predicted contour from MAI resulted in a DSC < 0.8, as compared to 46% and 60% for FAS and LI, respectively. All comparisons between MAI and the baseline methods were found to be statistically significant (p-value < 0.001). The proposed MAI algorithm significantly outperformed LI in terms of accuracy and robustness for both stomach segmentation from low-field MRIs and bowel segmentation from CT images. At this time, FAS methods for HDOs still require significant manual editing. Therefore, we believe that the MAI algorithm has the potential to expedite the process of HDO delineation within the radiation therapy workflow.


Schrödinger filtering: a precise EEG despiking technique for EEG-fMRI gradient artifact.
In EEG data acquired in the presence of fMRI, gradient-related spike artifacts contaminate the signal following the common preprocessing step of average artifact subtraction. Spike artifacts compromise EEG data quality since they overlap with the EEG signal in frequency, thereby confounding frequency-based inferences on activity. As well, spike artifacts can inflate or deflate correlations among time series, thereby confounding inferences on functional connectivity. We present Schrödinger filtering, which uses the Schrödinger equation to decompose the spike-containing input. The basis functions of the decomposition are localized and pulse-shaped, and selectively capture the various input peaks, with the spike components clustered at the beginning of the spectrum. Schrödinger filtering automatically subtracts the spike components from the data. On real and simulated data, we show that Schrödinger filtering (1) simultaneously accomplishes high spike removal and high signal preservation without affecting evoked activity, and (2) reduces spurious pairwise correlations in spontaneous activity. In these regards, Schrödinger filtering was significantly better than three other despiking techniques: median filtering, amplitude thresholding, and wavelet denoising. These results encourage the use of Schrödinger filtering in future EEG-fMRI pipelines, as well as in other spike-related applications (e.g., fMRI motion artifact removal or action potential extraction).


Spatial Transcriptomics Resolve an Emphysema-specific Lymphoid Follicle B Cell Signature in COPD.
Within chronic obstructive pulmonary disease (COPD), emphysema is characterized by a significant yet partially understood B cell immune component. To characterize the transcriptomic signatures from lymphoid follicles (LFs) in ever-smokers without COPD and COPD patients with varying degrees of emphysema. Lung sections from 40 COPD patients and ever-smokers were used for LF proteomic and transcriptomic spatial profiling. Formalin and OCT-fixed lung samples obtained from biopsies or lung explants, were assessed for LF presence. Emphysema measurements were obtained from clinical chest CT scans. High confidence transcriptional (HCT) target intersection analyses were conducted to resolve emphysema-induced transcriptional networks. Overall, 115 LFs from ever-smokers and GOLD 1-2 and GOLD 3-4 patients were analyzed. No LFs were found in never-smokers. Differential gene expression analysis revealed significantly increased expression of LF assembly and B cell markers genes in subjects with severe emphysema. HCT analysis revealed activation of abnormal B cell activity signature in LFs (q-value: 2.56E-111). LFs from GOLD 1-2 COPD patients with emphysema showed significantly increased expression of genes associated with antigen presentation, inflammation, and B cell activation and proliferation. LFs from GOLD 1-2 COPD patients without emphysema showed an anti-inflammatory profile. The extent of centrilobular emphysema was significantly associated with genes involved in B cell maturation and antibody production. Protein-RNA network analysis showed that LFs in emphysema have a unique signature skewed towards chronic B cell activation. An off-targeted B cell activation within LFs is associated with autoimmune-mediated emphysema pathogenesis.


The non-linear relationship between globalization, financial development and energy consumption: Evidence from BRICS economies.
In the era of globalization, financial development plays a key role in socioeconomic and environmental development. However, its adverse consequences on human life, environmental hazards, and high energy consumption cannot be ignored. Thus, this study investigates the non-linear relationship between globalization, financial development, and energy consumption for BRICS economies. In doing so, we have applied second-generation tests to identify cross-sectional dependence in the data. Cross-sectional augmented Dickey-Fuller (CADF) and Cross-sectional Im-Pesaran Shin (CIPS) have been performed to find the stationary level of variables. The long-term equilibrium link between the investigated variables has been established in continuance using the Westerlund Cointegration test. The Dynamic Seemingly Unrelated Regression (DSUR) indicates that U-shaped relationships exist for financial development and globalization with energy consumption. Conversely, there is an inverted U-shaped relationship exist between economic growth and energy consumption in BRICS. The Dumitrescu-Hurlin panel causality test findings show that a unidirectional link runs from energy consumption to financial development, economic growth to energy consumption, and globalization towards energy usage. Important policy implications have also been discussed.


Vibrational and configurational heat capacity of poly(vinyl acetate) from dynamic measurements.
The complex heat capacity C(p) (*) of poly(vinyl acetate) has been measured at 20.95 mrads modulation frequency during the cooling as well as on heating at 24, 8, and 2 Kh and during cooling at 0.5 Kh. The study is complemented with (the rate-dependent) C(p,app) measured during cooling and heating at 60, 24, and 8 Kh. At low temperatures, the real component of C(p) (*) yields the unrelaxed C(p) or C(p,vib), the vibrational part of C(p). It is found to be indistinguishable from C(p,glass) and lies on a line extrapolated to its equilibrium melt's temperature. At T near T(g),DeltaC(p)(=C(p,melt)-C(p,glass)) shows no detectable contribution from C(p,vib). The finding conflicts with a modified entropy theory calculation [E. A. DiMarzio and F. Dowell, J. Appl. Phys. 50, 6061 (1979)], which had predicted that approximately 27% of DeltaC(p) of poly(vinyl acetate) at T near T(g) is vibrational in origin and the remainder configurational. At T<T(g), the real component of C(p) (*) varies more slowly with T than C(p,app).


G-quadruplex RNA motifs influence gene expression in the malaria parasite Plasmodium falciparum.
G-quadruplexes are non-helical secondary structures that can fold in vivo in both DNA and RNA. In human cells, they can influence replication, transcription and telomere maintenance in DNA, or translation, transcript processing and stability of RNA. We have previously showed that G-quadruplexes are detectable in the DNA of the malaria parasite Plasmodium falciparum, despite a very highly A/T-biased genome with unusually few guanine-rich sequences. Here, we show that RNA G-quadruplexes can also form in P. falciparum RNA, using rG4-seq for transcriptome-wide structure-specific RNA probing. Many of the motifs, detected here via the rG4seeker pipeline, have non-canonical forms and would not be predicted by standard in silico algorithms. However, in vitro biophysical assays verified formation of non-canonical motifs. The G-quadruplexes in the P. falciparum transcriptome are frequently clustered in certain genes and associated with regions encoding low-complexity peptide repeats. They are overrepresented in particular classes of genes, notably those that encode PfEMP1 virulence factors, stress response genes and DNA binding proteins. In vitro translation experiments and in vivo measures of translation efficiency showed that G-quadruplexes can influence the translation of P. falciparum mRNAs. Thus, the G-quadruplex is a novel player in post-transcriptional regulation of gene expression in this major human pathogen.


Evaluation of comprehensive rehabilitation services for elderly homebound patients with arthritis and orthopedic disability.
Fifty-seven elderly homebound patients with arthritis and orthopedic disabilities were randomized to a goal-oriented outreach rehabilitation program or to usual treatment. Although 64% of patient goals were met, there were no overall significant differences in functional scores, institutionalization, or contentment between treatment and control periods. Twenty-three patients had maintained clinical improvement at the end of the study and some patients were dramatically improved with simple measures. The program's marginal costs were modest and consisted primarily of expenses associated with therapist's visits. The total costs of assistive devices and home modifications amounted to $1,902. Twenty-five percent of the homebound population could benefit from such services but the actual number who would partake is small.


Inhibition of the Proliferation of Human Lung Fibroblasts by Prostacyclin Receptor Agonists is Linked to a Sustained cAMP Signal in the Nucleus.
Idiopathic pulmonary fibrosis is a chronic and progressive fibrotic lung disease, and current treatments are limited by their side effects. Proliferation of human lung fibroblasts in the pulmonary interstitial tissue is a hallmark of this disease and is driven by prolonged ERK signalling in the nucleus in response to growth factors such as platelet-derived growth factor (PDGF). Agents that increase cAMP have been suggested as alternative therapies, as this second messenger can inhibit the ERK cascade. We previously examined a panel of eight Gαs-cAMP-coupled G protein-coupled receptors (GPCRs) endogenously expressed in human lung fibroblasts. Although the cAMP response was important for the anti-fibrotic effects of GPCR agonists, the magnitude of the acute cAMP response was not predictive of anti-fibrotic efficacy. Here we examined the reason for this apparent disconnect by stimulating the Gαs-coupled prostacyclin receptor and measuring downstream signalling at a sub-cellular level. MRE-269 and treprostinil caused sustained cAMP signalling in the nucleus and complete inhibition of PDGF-induced nuclear ERK and fibroblast proliferation. In contrast, iloprost caused a transient increase in nuclear cAMP, there was no effect of iloprost on PDGF-induced ERK in the nucleus, and this agonist was much less effective at reversing PDGF-induced proliferation. This suggests that sustained elevation of cAMP in the nucleus is necessary for efficient inhibition of PDGF-induced nuclear ERK and fibroblast proliferation. This is an important first step towards understanding of the signalling events that drive GPCR inhibition of fibrosis.


The airway antigen sampling system: respiratory M cells as an alternative gateway for inhaled antigens.
In this study, we demonstrated a new airway Ag sampling site by analyzing tissue sections of the murine nasal passages. We revealed the presence of respiratory M cells, which had the ability to take up OVA and recombinant Salmonella typhimurium expressing GFP, in the turbinates covered with single-layer epithelium. These M cells were also capable of taking up respiratory pathogen group A Streptococcus after nasal challenge. Inhibitor of DNA binding/differentiation 2 (Id2)-deficient mice, which are deficient in lymphoid tissues, including nasopharynx-associated lymphoid tissue, had a similar frequency of M cell clusters in their nasal epithelia to that of their littermates, Id2(+/-) mice. The titers of Ag-specific Abs were as high in Id2(-/-) mice as in Id2(+/-) mice after nasal immunization with recombinant Salmonella-ToxC or group A Streptococcus, indicating that respiratory M cells were capable of sampling inhaled bacterial Ag to initiate an Ag-specific immune response. Taken together, these findings suggest that respiratory M cells act as a nasopharynx-associated lymphoid tissue-independent alternative gateway for Ag sampling and subsequent induction of Ag-specific immune responses in the upper respiratory tract.


Weak Higher-Order Interactions in Macroscopic Functional Networks of the Resting Brain.
Interactions among different brain regions are usually examined through functional connectivity (FC) analysis, which is exclusively based on measuring pairwise correlations in activities. However, interactions beyond the pairwise level, that is, higher-order interactions (HOIs), are vital in understanding the behavior of many complex systems. So far, whether HOIs exist among brain regions and how they can affect the brain's activities remains largely elusive. To address these issues, here, we analyzed blood oxygenation level-dependent (BOLD) signals recorded from six typical macroscopic functional networks of the brain in 100 human subjects (46 males and 54 females) during the resting state. Through examining the binarized BOLD signals, we found that HOIs within and across individual networks were both very weak regardless of the network size, topology, degree of spatial proximity, spatial scales, and whether the global signal was regressed. To investigate the potential mechanisms underlying the weak HOIs, we analyzed the dynamics of a network model and also found that HOIs were generally weak within a wide range of key parameters provided that the overall dynamic feature of the model was similar to the empirical data and it was operating close to a linear fluctuation regime. Our results suggest that weak HOI may be a general property of brain's macroscopic functional networks, which implies the dominance of pairwise interactions in shaping brain activities at such a scale and warrants the validity of widely used pairwise-based FC approaches.SIGNIFICANCE STATEMENT To explain how activities of different brain areas are coordinated through interactions is essential to revealing the mechanisms underlying various brain functions. Traditionally, such an interaction structure is commonly studied using pairwise-based functional network analyses. It is unclear whether the interactions beyond the pairwise level (higher-order interactions or HOIs) play any role in this process. Here, we show that HOIs are generally weak in macroscopic brain networks. We also suggest a possible dynamical mechanism that may underlie this phenomenon. These results provide plausible explanation for the effectiveness of widely used pairwise-based approaches in analyzing brain networks. More importantly, it reveals a previously unknown, simple organization of the brain's macroscopic functional systems.


Development and validation of sensitive and rapid CRISPR/Cas12-based PCR method to detect hazelnut in unlabeled products.
Hazelnut, one of the most popular tree nuts, is widely found in processed food and even very small amounts can trigger severe allergic reactions in susceptible people. Herein, we developed a sensitive and rapid method based on CRISPR and qPCR capable of detecting low-abundance hazelnut in processed food. The assay, known as CRISPR-based nucleic acid test method (Crinac) can detect 1 % of hazelnut in a mixture and allows the species to be identified in a complex processed sample. The detection process can be completed within 60 min. Contributed to amplification via PCR and CRISPR/Cas12a, enables end-fluorescence measurement for the quantification of hazelnut, thus reducing assay time and eliminating the need for costly real-time fluorescence PCR instruments. The assay based on CRISPR/Cas12 and PCR has potential as a sensitive and reliable analytical tool for the detection of food authenticity.


Hollow Cu2MoS4 nanoparticles loaded with immune checkpoint inhibitors reshape the tumor microenvironment to enhance immunotherapy for pancreatic cancer.
Pancreatic ductal adenocarcinoma (PDAC) is a fatal disease that responds poorly to single-drug immunotherapy with PD-L1 (CD274) inhibitors. Here, we prepared mesoporous nanomaterials Cu2MoS4 (CMS)/PEG loaded with PD-L1 inhibitor BMS-1 and CXCR4 inhibitor Plerixafor to form the nanodrug CMS/PEG-B-P. In vitro experiments, CMS/PEG-B-P have a more substantial inhibitory effect on the expression of PD-L1 and CXCR4 as well as to promote the apoptosis of pancreatic cancer cells KPC and suppressed KPC cell proliferation were detected by flow cytometry, qPCR and Western blotting (WB). Promotes the release of the cytotoxic substance reactive oxygen species (ROS) and the production of the immunogenic cell death (ICD) marker calreticulin (CRT) in KPC cells. CMS/PEG-B-P was also detected to have a certain activating effect on mouse immune cells, dendritic cells (mDC) and macrophage RAW264.7. Subcutaneous tumorigenicity experiments in C57BL/6 mice verified that CMS/PEG-B-P had an inhibitory effect on the growth of tumors and remodeling of the tumor immune microenvironment, including infiltration of CD4+ and CD8+ T cells and polarization of macrophages, as well as reduction of immunosuppressive cells. Meanwhile, CMS/PEG-B-P was found to have different effects on the release of cytokines in the tumor immune microenvironment, including The levels of immunostimulatory cytokines INF-γ and IL-12 are increased and the levels of immunosuppressive cytokines IL-6, IL-10 and IFN-α are decreased. In conclusion, nanomaterial-loaded immune checkpoint inhibitor therapies can enhance the immune response and reduce side effects, a combination that shows great potential as a new immunotherapeutic approach. STATEMENT OF SIGNIFICANCE: Pancreatic ductal adenocarcinoma (PDAC) is a fatal disease that has a low response to single-drug immunotherapy with PD-L1 (CD274) inhibitors. We preared PEG-modified mesoporous nanomaterials Cu2MoS4 (CMS) loaded with PD-L1 inhibitor BMS-1 and CXCR4 inhibitor Plerixafor to form the nanodrug CMS/PEG-B-P. Our study demonstrated that Nanomaterial-loaded immune checkpoint inhibitor therapies can enhance the immune response and reduce side effects, a combination that shows great potential as a new immunotherapeutic approach.


Accumulation of radiolabelled platelets and fibrin on the carotid artery of rabbits after angioplasty: effects of heparin and dipyridamole.
We investigated the dynamic accumulation of platelets and fibrin after balloon injury of the carotid arteries in rabbits in vivo. In addition, effects of heparin and dipyridamole treatment were also tested. Autologous (99m)Tc-labelled platelet and (123)I-labelled fibrin accumulation was measured at one minute intervals for 4 hours following balloon injury of the carotid artery. Platelet accumulation occurred rapidly, with an approximately 125% increase occurring within 30 min after injury. There was no further activity for up to 4 hours. This accumulation could be inhibited with an intravenous infusion of PGI2 (500 ng/kg/hr). Fibrin accumulation occurred slowly and continuously over the 4 hour measurement period. Injection of an anti-fibrin antibody inhibited fibrin accumulation. Heparin (25 U/kg/hr for 4 hrs) administration resulted in a significant 82 +/- 19% and 68 +/- 13% reduction in platelet and fibrin accumulation, respectively. This dose of heparin was associated with a 2-fold prolongation of the aPTT. Dipyridamole (0.45 mg/kg/hr for 4 hrs) resulted in a 46 +/- 12% and 70 +/- 25% reduction of platelet and fibrin accumulation, respectively. Thus, we demonstrated that the dynamics of platelet and fibrin accumulation following balloon injury in rabbits are very different. The vessel wall continues to be thrombogenic for fibrin up to 4 hours after injury even though platelet accumulation has ceased after one hour. We conclude that the local thrombotic events following balloon injury are complex and that not only platelets but also fibrin is important in regulating responses to injury.


Novel lineages of Giardia intestinalis identified by genetic analysis of organisms isolated from dogs in Australia.
Infection of suckling mice with Giardia trophozoites recovered from the intestines of 11 dogs autopsied in Central and Southern Australia in each case produced an established isolate. In contrast, only 1 isolate was obtained by inoculation of faecal cysts. The organisms grew poorly in comparison with isolates from humans or non-canine animal hosts. Light microscopy revealed that the trophozoites had median bodies with the 'claw hammer' appearance typical of G. intestinalis (syn. G. duodenalis, G. lamblia) but that they differed in shape and nuclear morphology from axenic isolates of human or canine origin. Allozymic analysis of electrophoretic data representing 26 loci and phylogenetic analysis of nucleotide sequences obtained from DNA amplified from the glutamate dehydrogenase locus showed that the 11 isolates examined from Australian dogs were genetically distinct from all isolates of G. intestinalis that have been established previously from humans and animals, and also from G. muris. Both analytical methods placed 10 of the Australian canine isolates into a unique genetic lineage (designated Assemblage C) and the eleventh into a deep-rooted second branch (designated Assemblage D), each well separated from the 2 lineages (Assemblages A and B) of G. intestinalis that encompass all the genotypes known to infect humans. In contrast, 4 axenic isolates derived from dogs in Canada and Europe (the only other isolates to have been established from dogs) have genotypes characteristic of genetic Assemblages A or B. The findings indicate that the novel Giardia identified in these rural Australian dogs have a restricted host range, possibly confined to canine species. The poor success rate in establishing Giardia from dogs in vitro suggests, further, that similar genotypes may predominate as canine parasites world-wide. The absence of such organisms among isolates of Giardia that have been established from humans by propagation in suckling mice indicates that they are unlikely to infect humans. However, infection of humans by those dog-derived genotypes that grow in vitro cannot be excluded.


[Morphological study of brainstem auditory evoked potentials. The effect of the position of the reference electrode].
In view of the discrepancy between the siting of electrodes and the effects on BAEP, we made a morphological study of 30 healthy volunteers, 15 men and 15 women. In each person we obtained the BAEP with the active electrode on the ear lobe ipsilateral to the stimulated ear and the reference ear successively at four different sites: vertex (Cz), high frontal (Fz), earlobe contralateral to the stimulated ear and the spinous process of the seventh cervical vertebra (CV7). The I, III and V waves were present in all cases and tests, followed in frequency by II, IV and VII. In Cz and Fz the first five deflections were almost always seen, and these are the most useful for neurodiagnosis. The waves of greatest amplitude were 1 in both sexes in the ear lobe and CV7 for Fz and Cz in men and V for women. Regarding the complex morphological variants, the greatest incidence in the III IV V complex was observed in Fz in both sexes. In CV7 there was a greater range of variants, as found by other workers, but with greater morphological separation of waves IV and V. They were infrequent in waves I II and VI VII and least common in Fz and Cz. We consider that the BAEP obtained with reference to Cz and Fz are ideal for interpretation, since there is greater constancy, global amplitude and morphological clarity of the first five deflections and less morphological complexity of the other waves.


The influence of parenting style on health related behavior of children: findings from the ChiBS study.
Exploring associations between parenting behavior and children's health related behavior including physical activity, sedentary behavior, diet and sleep. We recruited 288 parents and their children (6-12y old). Children's weight and height were measured. Fat percentage was determined by air displacement plethysmography. Parents reported socio-demographic data, sleep information, physical activity and sedentary behavior of their child and completed the Comprehensive General Parenting Questionnaire (CGPQ) and a Food Frequency Questionnaire. Children completed the Dutch Eating Behavior Questionnaire. Associations between parenting behavior (CGPQ) and children's health related behavior were assessed with univariate and multiple regression analyses. A small positive correlation was found between sweet food consumption frequency and "coercive control" (r = 0.139) and a small negative correlation between fruit and vegetables consumption frequency and "overprotection" (r = -0.151). Children consumed more frequently soft drinks when their parents scored lower on "structure" (r = -0.124) and higher on "overprotection" (r = 0.123); for the light soft drinks separately, a small positive correlation with "behavioral control" was found (r = 0.172). A small negative correlation was found between "emotional eating" and "structure" (r = -0.172) as well as "behavioral control" (r = -0.166). "Coercive control" was negatively correlated with the child's sleep duration (r = -0.171). After correction for confounding factors, the following significant associations were found: (1) a small negative association between "structure" and soft drinks consumption (β = -0.17 for all soft drinks and -0.22 for light soft drinks), (2) a small positive association between "behavioral control" and light soft drinks (β = 0.34), (3) a small positive association of "nurturance" and "coercive control" with sedentary behavior (β = 0.16 for both parent constructs) and (4) a small negative association between the parenting construct "coercive control" and sleep duration (β = -0.23). The significant but small associations between parenting constructs and the investigated variables suggest that different aspects of parenting style play an important role in the genesis of the health related behavior of children. Overall, our findings suggest that health professionals should encourage parents to apply the more positive parenting constructs i.e., more "structure" and "behavioral control", and less "coercive control". They could, for instance, supervise and manage their child's activities and help their child to achieve certain goals.


FAS 116 and 117: the implementation process.
The Financial Accounting Standards Board finalized and issued two new statements in June 1993: "Financial Statements of Not-for-Profit Organizations" (FAS 117) and "Accounting for Contributions Received and Contributions Made" (FAS 116). The statements will become effective for fiscal years beginning after December 15, 1994. Until a revised audit guide is issued, however, several factors will need to be carefully considered when implementing the two standards.


Differential Processing of Isolated Object and Multi-item Pop-Out Displays in LIP and PFC.
Objects that are highly distinct from their surroundings appear to visually "pop-out." This effect is present for displays in which: (1) a single cue object is shown on a blank background, and (2) a single cue object is highly distinct from surrounding objects; it is generally assumed that these 2 display types are processed in the same way. To directly examine this, we applied a decoding analysis to neural activity recorded from the lateral intraparietal (LIP) area and the dorsolateral prefrontal cortex (dlPFC). Our analyses showed that for the single-object displays, cue location information appeared earlier in LIP than in dlPFC. However, for the display with distractors, location information was substantially delayed in both brain regions, and information first appeared in dlPFC. Additionally, we see that pattern of neural activity is similar for both types of displays and across different color transformations of the stimuli, indicating that location information is being coded in the same way regardless of display type. These results lead us to hypothesize that 2 different pathways are involved processing these 2 types of pop-out displays.


Dimericursones A and B: two unprecedented hexacyclic dimeric diterpenoids from the root barks of Jatropha curcas.
Dimericursones A and B (1 and 2), two unprecedented hexacyclic dimeric diterpenoids, were obtained from the root barks of Jatropha curcas. Their structures were elucidated by extensive spectroscopic analysis, electronic circular dichroism calculations, and single-crystal X-ray diffraction. Dimericursone B (2) showed significant inhibition on nitric oxide production of lipopolysaccharide-induced RAW264.7 macrophages with IC50 values of 5.65 μM.


Electroreduction of aromatic oximes: diprotonation, adsorption, imine formation, and substituent effects.
Aromatic oximes are reduced in aqueous solution in a four-electron process. The reducible species in the pH range 5-8 is a diprotonated form of the oxime. This species is generated in the course of electrolysis in the vicinity of the electrode surface from the adsorbed neutral form of the oxime. The reduction is initiated by a cleavage of the N-O bond. The diprotonation facilitates the reduction process by the preformation of OH2+ as a good leaving group and by a positive charge on the azomethine nitrogen. Diprotonation has been proven based on shapes of i = f(pH) plots, by observed shifts of half-wave potentials with pH and by comparison with the reduction of nitrones. Some observed deviations from theoretical i = f(pH) plots were attributed to the role of adsorption on the rate of protonation. Adsorption is also responsible for dips on some of the i-E curves. Adsorption plays a role at concentrations as low as 1 x 10(-5) M, when the electrode surface is still not fully covered. This indicates that catalyzed protonation occurs on islets of adsorbed materials. At pH 2-5 the studied oximes in the vicinity of the electrode are predominately present in a protonated form, which is less strongly adsorbed. In this pH range the protonation takes place in a homogeneous reaction layer of the electrode. It yields a monoprotonated form, which is reduced. The separation of two two-electron waves observed for some oximes in acidic media serves as an experimental proof of the formation of imines as reduction intermediates. This separation is caused by the differences in pKa values of protonated forms of oximes and imines. The effects of substituents in the para position on the benzene ring are characterized by correlation with the Hammett substituent constant sigmax. This has been proven at pH 1.5 for substituted benzaldehyde oximes and at pH 5.0 for substituted acetophenone oximes.