[Tuberculosis in the indigenous population of Peru 2008].
To identify the indigenous inhabitants affected by tuberculosis (TB) in Peru during the year 2008. Descriptive observational study, performed from August to December 2009, including all indigenous patients affected by tuberculosis that were including in the Control Program during the year 2008 in the 25 regions of Peru. We identified 702 indigenous patients with tuberculosis. The ethnical groups that have most patients were the Quechua group (417/702; 59.4%) and the Amazonic indigenous (201/702, 28.6%). Out of the Amazonic, more than 60% belonged to the Ashaninka (Campas), Shipibo and Matsiguenga groups. In third place, we found the Aymara natives, who had 84/702 (11.97%) of cases of tuberculosis. It is important to mention that the distribution of the cases of multidrugresistant tuberculosis (MDR TB) involves five departments (nine cases of MDR TB), being the greater number of cases of MDR TB in patients previously treated (6/9), and only 3 cases were primary MDR TB, belonging to the quechua group. High incidence rates of tuberculosis in indigenous population have been found, which raises the need of further research in order to guarantee the correct gathering of information in ethnic groups in order to have more and better evidence about the situation of tuberculosis in the indigenous population of Peru.


Harnessing proteasome dynamics and allostery in drug design.
The proteasome is the essential protease that is responsible for regulated cleavage of the bulk of intracellular proteins. Its central role in cellular physiology has been exploited in therapies against aggressive cancers where proteasome-specific competitive inhibitors that block proteasome active centers are very effectively used. However, drugs regulating this essential protease are likely to have broader clinical usefulness. The non-catalytic sites of the proteasome emerge as an attractive alternative target in search of highly specific and diverse proteasome regulators. Crystallographic models of the proteasome leave the false impression of fixed structures with minimal molecular dynamics lacking long-distance allosteric signaling. However, accumulating biochemical and structural observations strongly support the notion that the proteasome is regulated by precise allosteric interactions arising from protein dynamics, encouraging the active search for allosteric regulators. Here, we discuss properties of several promising compounds that affect substrate gating and processing in antechambers, and interactions of the catalytic core with regulatory proteins. Given the structural complexity of proteasome assemblies, it is a painstaking process to better understand their allosteric regulation and molecular dynamics. Here, we discuss the challenges and achievements in this field. We place special emphasis on the role of atomic force microscopy imaging in probing the allostery and dynamics of the proteasome, and in dissecting the mechanisms involving small-molecule allosteric regulators. New small-molecule allosteric regulators may become a next generation of drugs targeting the proteasome, which is critical to the development of new therapies in cancers and other diseases.


Seasonal occurrence and production effects of the biting louse Damalinia limbata on Angora goats and 2 treatment options.
Adult biting lice Damalinia limbata (Gervais 1844) were most numerous on Angora goats in southwestern Free State Province from November to May with the smallest numbers present during August. Nymphs were most numerous from January to March with the smallest numbers present during July. The largest proportion of adult lice was present on the ventral surface of goats during winter, when population numbers were lowest, and the largest proportion of nymphs during summer, when population numbers were highest. Infestation did not affect the body weight of goats, but adversely affected both the quantity and quality of mohair produced. A single treatment with deltamethrin, applied as a pour-on along the mid-line of the backs of the goats, effectively controlled infestation and resulted in a significant increase in mohair production compared with untreated goats. Deltamethrin sprayed along the sides of the animals was equally effective in controlling infestation.


A multi-year longitudinal study of water quality parameters in four salmon-bearing and recreational streams on mount hood, Oregon.
Four streams-Clear Fork, Lost Creek, Camp Creek and Still Creek-in northwestern Oregon's Sandy River Basin were monitored for temperature, dissolved oxygen levels, and fecal bacterial concentrations in a multi-year analysis examining stream health for recreational users and anchor habitat for Pacific Salmon. Temperatures were recorded using micro -T temperature loggers at 15 locations, during 22 July - 5 September 2006, 2 July - 4 September 2007, 20 June - 7 September 2008, 23 June - 9 September 2009, and 2 July -9 September 2010. The Seven-Day Average Maximum water temperature (7-DAM) of 13°C was used as a reference value for the biological limit governing suitable salmonid spawning and egg incubation conditions. The maximum 7-DAM temperatures occurred on different dates and all streams neared or exceeded the 13°C standard at least once each summer. Dissolved oxygen levels were measured at weekly or longer intervals in 2006, 2007, 2008, and 2009. Dissolved oxygen levels fell below the 9.0 ppm standard for Clear Fork on almost half the sampling dates in 2006, 2007, and 2009. Concentrations of the bacterial genus Enterococcus were measured as an indicator of fecal contamination. Samples were collected at 15 sites along the four streams. Weekly samples were collected during a 9 week period from July - September 2007, an 11 week period from June - September 2008, and an 11 week period from June - September 2009. Enterococcus counts exceeded the federal recommended national criterion value of 61 colony forming units (CFU) per 100 mL every year in Camp Creek and occasionally elsewhere, with exceedances trending towards late summer.


Operational performance of an STD control programme in Mwanza Region, Tanzania.
To describe important details of the design and operational features of the Mwanza sexually transmitted diseases (STD) control programme. To assess the feasibility of the intervention, the distribution of STD syndromes observed, the clinical effectiveness of syndromic STD case management, the utilisation of STD services by the population, and the quality of syndromic STD services delivered at rural health units. The intervention was integrated into rural primary healthcare (PHC) units. It comprised improved STD case management using the syndromic approach, facilitated by a regional programme office which ensured the training of health workers, a reliable supply of effective drugs, and regular support supervision. Five studies were performed to evaluate operational performance: (i) a survey of register books to collect data on patients presenting with STDs and reproductive tract infections (RTIs) to rural health units with improved STD services, (ii) a survey of register books from health units in communities without improved services, (iii) a survey of register books from referral clinics, (iv) a home based cross sectional study of STD patients who did not return to the intervention health units for follow up, (v) a cross sectional survey of reported STD treatment seeking behaviour in a random cohort of 8845 adults served by rural health units. During the 2 years of the Mwanza trial, 12,895 STD syndromes were treated at the 25 intervention health units. The most common syndromes were urethral discharge (67%) and genital ulcers (26%) in men and vaginal discharge (50%), lower abdominal tenderness (33%), and genital ulcers (13%) in women. Clinical treatment effectiveness was high in patients from whom complete follow up data were available, reaching between 81% and 98% after first line treatment and 97%-99% after first, second, and third line treatment. Only 26% of patients referred to higher levels of health care had presented to their referral institutions. During the trial period, data from the cohort showed that 12.8% of men and 8.6% of women in the intervention communities experienced at least one STD syndrome. Based on various approaches, utilisation of the improved health units by symptomatic STD patients in these communities was estimated at between 50% and 75%. During the first 6 months of intervention attendance at intervention units increased by 53%. Thereafter, the average attendance rate was about 25% higher than in comparison communities. Home visits to 367 non-returners revealed that 89% had been free of symptoms after treatment, but 28% became symptomatic again within 3 months of treatment. 100% of these patients reported that they had received treatment, but only 74% had been examined, only 57% had been given health education, and only 30% were offered condoms. Patients did not fully recall which treatment they had been given, but possibly only 63% had been treated exactly according to guidelines. This study demonstrated that it is feasible to integrate effective STD services into the existing PHC structure of a developing country. Improved services attract more patients, but additional educational efforts are needed to further improve treatment seeking behaviour. Furthermore, clear treatment guidelines, a reliable drug supply system, and regular supervision are critical. All efforts should be made to treat patients on the spot, without delay, as referral to higher levels of care led to a high number of dropouts. The syndromic approach to STD control should be supported by at least one reference clinic and laboratory per country to ensure monitoring of prevalent aetiologies, of the development of bacterial resistance, and of the effectiveness of the syndromic algorithms in use.


Carrier-mediated active transport of BQ-123, a peptidic endothelin antagonist, into rat hepatocytes.
The hepatic uptake mechanisms and pharmacokinetics of BQ-123, an anionic cyclopentapeptidic endothelin ET(A) receptor antagonist, were studied in rats. Elimination of BQ-123 from plasma after intravenous injection of the compound was very rapid as evidenced by high total body clearance (CLtot, 50 ml/min/kg), which is comparable with hepatic blood flow rate. Within 1 hr after injection, 86% of the dose was excreted as its intact form in the bile. BQ-123 was taken up extensively by isolated rat hepatocytes both Na(+)-dependently and Na(+)-independently. The Na(+)-dependent system transported BQ-123 with higher affinity than did the Na(+)-independent system (Km; 6 and 12 microM, respectively), but its capacity was lower (Vmax; 140 and 390 pmol/min/10(6) cells, respectively). Both uptake systems were found to be active transport systems because of explicit inhibition by metabolic inhibitors. BQ-485, an anionic linear tripeptide, strongly inhibited BQ-123 uptake with Ki values of 1.6 and 2.5 microM for Na(+)-dependent and Na(+)-independent systems, respectively, whereas BQ-587, a cationic cyclopentapeptide, inhibited BQ-123 uptake only slightly. Considering this in vitro finding and the low in vivo CLtot of BQ-587 together, the carrier systems for BQ-123 seem to recognize negatively charged substances selectively. Both Na(+)-dependent and Na(+)-independent uptake of BQ-123 were competitively inhibited by a bile acid (taurocholate) and an organic anion (dibromosulfophthalein). The Ki values were comparable with the Km values of taurocholate and dibromosulfophthalein transport, which suggests that the Na(+)-dependent system corresponds to the bile acid transporter and that the Na(+)-independent system corresponds to the organic anion transporter.


Polypeptide components of the apamin receptor associated with a calcium activated potassium channel.
Photoaffinity labeling of rat brain membranes with [125I]ANPAA-apamin incorporated radioactivity into polypeptides of 86 and 59 kDa and occasionally a more weakly labeled component of 45 kDa. These polypeptides were immunoprecipitated with anti-apamin antibodies and treated with glycosidases. Neither the 86 nor the 59 kDa polypeptide appeared to be N-glycosylated. Partial proteolytic mapping of affinity labeled polypeptides with chymotrypsin or V8 protease generated an identical pattern. These results suggest that the 59 and 45 kDa components are not additional subunits of an oligomeric protein but result from cleavage of the 86 kDa polypeptide.


Anisotropic hinge model for polarization-mode dispersion in installed fibers.
We discuss a generalized waveplate hinge model to characterize anisotropic effects associated with the hinge model of polarization-mode dispersion in installed systems. In this model, the action of the hinges is a random time-dependent rotation about a fixed axis. We obtain the probability density function of the differential group delay and the outage probability of an individual wavelength band using a combination of importance sampling and the cross-entropy method, and we then compute the noncompliant capacity ratio by averaging over wavelength bands. The results show that there are significant differences between the outage statistics predicted by isotropic and anisotropic hinge models.


Effectiveness of inactivated influenza vaccine in autoimmune rheumatic diseases treated with disease-modifying anti-rheumatic drugs.
The effectiveness of inactivated influenza vaccine in people with autoimmune rheumatic disease (AIRDs) is not known. We investigated whether the influenza vaccine is effective in preventing respiratory morbidity, mortality and all-cause mortality in AIRD patients. Adults with AIRDs treated with DMARDs prior to 1 September of each year between 2006 and 2009, and 2010 and 2015 were identified from the Clinical Practice Research Datalink. Exposure and outcome data were extracted. Data from multiple seasons were pooled. Propensity score (PS) for vaccination was calculated. Cox-proportional hazard ratios (HRs) and 95% CIs were calculated, and were (i) adjusted, (ii) matched for PS for vaccination. Data for 30 788 AIRD patients (65.7% female, 75.5% with RA, 61.1% prescribed MTX) contributing 125 034 influenza cycles were included. Vaccination reduced risk of influenza-like illness [adjusted HR (aHR) 0.70], hospitalization for pneumonia (aHR 0.61) and chronic obstructive pulmonary disease exacerbations (aHR 0.67), and death due to pneumonia (aHR 0.56) on PS-adjusted analysis in the influenza active periods (IAPs). The associations were of similar magnitude and remained statistically significant on PS-matched analysis except for protection from influenza-like illness, which became non-significant. Sub-analysis restricted to pre-IAP, IAP and post-IAP did not yield evidence of residual confounding on influenza-like illness and death due to pneumonia. Vaccination reduced risk of all-cause mortality, although IAP-restricted analysis demonstrated residual confounding for this outcome. Influenza vaccine associates with reduced risk of respiratory morbidity and mortality in people with AIRDs. These findings call for active promotion of seasonal influenza vaccination in immunosuppressed people with AIRDs by healthcare professionals.


Too poor to live? A case study of vulnerability and maternal mortality in Burkina Faso.
This paper examines the concept of vulnerability in the context of maternal morbidity and mortality in Burkina Faso, an impoverished country in West Africa. Drawing on a longitudinal cohort study into the consequences of life-threatening or 'near miss' obstetric complications, we provide an in-depth case study of one woman's experience of such morbidity and its aftermath. We follow Kalizeta's trajectory from her near miss and the stillbirth of her child to her death from pregnancy-related hypertension after a subsequent delivery less than two years later, in order to examine the impact of severe and persistent illness and catastrophic health expenditure on her health and on her family's everyday life. Kalizeta's case illustrates how vulnerability in health emerges and is maintained or exacerbated over time. Even where social arrangements are supportive, structural impediments, including unaffordable and inadequate healthcare, can severely limit individual resilience to mitigate the negative social and economic consequences of ill health.


The effect of surface colour on the formation of marine micro and macrofouling communities.
The effect of substratum colour on the formation of micro- and macro fouling communities was investigated. Acrylic tiles, painted either black or white were covered with transparent sheets in order to ensure similar surface properties. All substrata were exposed to biofouling at 1 m depth for 40 d in the Marina Bandar al Rowdha (Muscat, Sea of Oman). Studies were conducted in 2010 over a time course of 5, 10 and 20 d, and in 2012 samples were collected at 7, 14 and 21 d. The densities of bacteria on the black and white substrata were similar with the exception of day 10, when the black substrata had a higher abundance than white ones. Pyrosequencing via 454 of 16S rRNA genes of bacteria from white and black substrata revealed that Alphaproteobacteria and Firmicutes were the dominant groups. SIMPER analysis demonstrated that bacterial phylotypes (uncultured Gammaproteobacteria, Actibacter, Gaetbulicola, Thalassobius and Silicibacter) and the diatoms (Navicula directa, Navicula sp. and Nitzschia sp.) contributed to the dissimilarities between communities developed on white and black substrata. At day 20, the highest amount of chlorophyll a was recorded in biofilms developed on black substrata. SIMPER analysis showed that Folliculina sp., Ulva sp. and Balanus amphitrite were the major macro fouling species that contributed to the dissimilarities between the communities formed on white and black substrata. Higher densities of these species were observed on black tiles. The results emphasise the effect of substratum colour on the formation of micro and macro fouling communities; substratum colour should to be taken into account in future studies.


SMARCB1-mediated SWI/SNF complex function is essential for enhancer regulation.
SMARCB1 (also known as SNF5, INI1, and BAF47), a core subunit of the SWI/SNF (BAF) chromatin-remodeling complex, is inactivated in nearly all pediatric rhabdoid tumors. These aggressive cancers are among the most genomically stable, suggesting an epigenetic mechanism by which SMARCB1 loss drives transformation. Here we show that, despite having indistinguishable mutational landscapes, human rhabdoid tumors exhibit distinct enhancer H3K27ac signatures, which identify remnants of differentiation programs. We show that SMARCB1 is required for the integrity of SWI/SNF complexes and that its loss alters enhancer targeting-markedly impairing SWI/SNF binding to typical enhancers, particularly those required for differentiation, while maintaining SWI/SNF binding at super-enhancers. We show that these retained super-enhancers are essential for rhabdoid tumor survival, including some that are shared by all subtypes, such as SPRY1, and other lineage-specific super-enhancers, such as SOX2 in brain-derived rhabdoid tumors. Taken together, our findings identify a new chromatin-based epigenetic mechanism underlying the tumor-suppressive activity of SMARCB1.


Instrumental recording and biomonitoring of ambient ozone in the Greek countryside.
Among eight commercial Greek varieties of tobacco (Nicotiana tabacum L.) tested for their ozone-sensitivity levels, the Zichnomirodata (KK6/5) variety was found to be the most sensitive, although less sensitive than the well-known super-sensitive Bel-W3. Besides qualitative differences in the appearance of macroscopic symptoms these two varieties can be used simultaneously as a reliable pair of ozone bioindicators. The occurrence of ozone in the Greek countryside was surveyed by biomonitoring in 14 rural regions over the country and by a simultaneous biomonitoring and instrumental recording of ozone concentrations at a single remote side (Pournaria, Arcadia). Phytotoxic symptoms were observed mainly on the leaves of Bel-W3 and occasionally on those of Zichnomirodata varieties, suggesting that ozone levels were high enough to affect at least sensitive species. The instrumental monitoring (during a total period of 912 h) revealed maximum hourly O3 concentration 62 ppb, while the thresholds of 30, 40 and 50 ppb were exceeded for 40%, 20% and 6% of the recording period, respectively. The accumulated exposure over 40 ppb (AOT40) for the daylight hours over the 38 monitored days was 680 ppb h.


Pathogenic potential of antibodies to the GABAB receptor.
GABAB receptor (GABABR) autoantibodies have been detected in the serum of immunotherapy-responsive patients with autoimmune encephalitis. This study aimed to investigate the effect of immunoglobulin G (IgG) from a patient with GABABR antibodies on primary neuronal cultures and acute slices of entorhinal cortex. Primary hippocampal neuronal cultures were incubated with serum immunoglobulin from patients with GABABR or AMPA receptor (AMPAR) antibodies for up to 72 h to investigate their effect on receptor surface expression. Whole-cell patch-clamp recordings from layer III pyramidal cells of the medial entorhinal cortex were used to examine the effect on neuronal activity. GABABR surface expression was unaltered by incubation with GABABR antibodies. By contrast, after 24 h application of AMPAR antibodies, AMPARs were undetectable. However, acute application of GABABR IgG decreased both the duration of network UP states and the spike rate of pyramidal cells in the entorhinal cortex. GABABR antibodies do not appear to affect GABABRs by internalization but rather reduce excitability on the medial temporal lobe networks. This unusual mechanism of action may be exploited in rational drug development strategies.


Antioxidant, antimicrobial and neutrophil-modulating activities of herb extracts.
The present study provides a comprehensive data on the antioxidant, antimicrobial and neutrophil-modulating activities of extracts from six medicinal plants--blackberry (Rubus fruticosus) leaves, chokeberry (Aronia melanocarpa) leaves, hawthorn (Crataegus monogyna) leaves, lady's mantle (Alchemilla glabra) aerial parts, meadowsweet (Filipendula ulmaria) aerial parts and raspberry (Rubus idaeus) leaves. In order to analyze the antioxidant activity of the herbs, several methods (ORAC, TRAP, HORAC and inhibition of lipid peroxidation) were used. Blackberry leaves and meadowsweet extracts revealed the highest antioxidant activities via all methods. All extracts studied blocked almost completely the opsonized zymosan particle-activated ROS production by neutrophils from human whole blood. On the other hand, the effect of extracts on phorbol myristate acetate-activated ROS production was much milder and even nonsignificant in the case of chokeberry leaves. This latter result suggests that extracts (apart from their antioxidative activity) interfere with the signaling cascade of phagocyte activation upstream of the protein kinase C activation. The antimicrobial activity of the investigated extracts against 11 human pathogens was investigated using three different methods. Meadowsweet and blackberry leaves extracts had the highest antimicrobial effect and the lowest minimal inhibiting concentrations (MICs) against the microorganisms tested.


The value of emotion: how does episodic prospection modulate delay discounting?
Humans often show impatience when making intertemporal choice for monetary rewards, preferring small rewards delivered immediately to larger rewards delivered after a delay, which reflects a fundamental psychological principle: delay discounting. However, we propose that episodic prospection humans can vividly envisage exerts a strong and broad influence on individuals' delay discounting. Specifically, episodic prospection may affect individuals' intertemporal choice by the negative or positive emotion of prospection. The present study explored how episodic prospection modulated delay discounting by emotion. Study 1 showed that participants were more inclined to choose the delayed but larger rewards when they imaged positive future events than when they did not image events; Study 2 showed that participants were more inclined to choose the immediate but smaller rewards when they imaged negative future events than when they did not image events; In contrast, study 3 showed that choice preferences of participants when they imaged neutral future events were the same as when they did not image events. By manipulating the emotion valence of episodic prospection, our findings suggested that positive emotion made individuals tend to choose delayed rewards, while negative emotion made individuals tend to choose immediate rewards. Only imaging events with neutral emotion did not affect individuals' choice preference. Thus, the valence of imaged future events' emotion might play an important role in individuals' intertemporal choice. It is possible that the valence of emotion may affect the changed direction (promote or inhibit) of individuals' delay discounting, while the ability to image future events affects the changed degree of individuals' delay discounting.


3-hydroxykynurenine and other Parkinson's disease biomarkers discovered by metabolomic analysis.
Parkinson's disease (PD) biomarkers are needed to enhance therapeutics research and to understand PD pathogenesis. Methods that simultaneously measure hundreds of small molecular-weight compounds-metabolomic analysis-"fingerprint" disease-specific alterations in individual compounds or metabolic pathways. Beyond a nontargeted search for PD biomarkers, we hypothesized that PD cerebrospinal fluid would show increased formation of the excitotoxin 3-hydroxykynurenine and diminished concentration of the antioxidant glutathione. Cerebrospinal fluid was collected at <4 hours postmortem from 48 pathologically-verified PD subjects and 57 comparably-aged controls. Assays involved ultra-high-performance liquid and gas chromatography linked to mass spectrometry. We used univariate techniques to determine fold-changes in concentrations of biochemicals; false-discovery rates were calculated to exclude spurious findings. Data was modeled using a Support Vector Machine for analyzing compounds selected by Welch's t test. Classification accuracy was determined by cross-validation. Of 243 structurally-identified biochemicals,19 compounds differentiated PD from controls at a 20% false-discovery level. In PD, mean 3-hydroxykynurenine concentration was increased by one-third, and mean oxidized glutathione was decreased by 40% (for each, P < .01). Four of the 19 compounds differentiating PD from controls were N-acetylated amino acids, suggesting a generalized alteration in N-acetylation activity. The Support Vector Machine classification model distinguished between groups at 83% sensitivity and 91% specificity for the learning data, and at 65% and 79% from cross-validation. In this study, the first for metabolomic profiling of PD cerebrospinal fluid, we found several novel biomarkers and offer new directions for recognizing disease-specific biochemical indicators. The findings support involvement of excitotoxicity and oxidative stress in the pathogenesis of PD.


Intrafractional prostate motion during online image guided intensity-modulated radiotherapy for prostate cancer.
Intrafractional motion consists of two components: (1) the movement between the on-line repositioning procedure and the treatment start and (2) the movement during the treatment delivery. The goal of this study is to estimate this intrafractional movement of the prostate during prostate cancer radiotherapy. Twenty-seven patients with prostate cancer and implanted fiducials underwent a marker match procedure before a five-field IMRT treatment. For all fields, in-treatment images were obtained and then processed to enable automatic marker detection. Combining the subsequent projection images, five positions of each marker were determined using the shortest path approach. The residual set-up error (RSE) after kV-MV based prostate localization, the prostate position as a function of time during a radiotherapy session and the required margins to account for intrafractional motion were determined. The mean RSE and standard deviation in the antero-posterior, cranio-caudal and left-right direction were 2.3±1.5 mm, 0.2±1.1 mm and -0.1±1.1 mm, respectively. Almost all motions occurred in the posterior direction before the first treatment beam as the percentage of excursions>5 mm was reduced significantly when the RSE was not accounted for. The required margins for intrafractional motion increased with prolongation of the treatment. Application of a repositioning protocol after every beam could decrease the 1cm margin from CTV to PTV by 2 mm. The RSE is the main contributor to intrafractional motion. This RSE after on-line prostate localization and patient repositioning in the posterior direction emphasizes the need to speed up the marker match procedure. Also, a prostate IMRT treatment should be administered as fast as possible, to ensure that the pre-treatment repositioning efforts are not erased by intrafractional prostate motion. This warrants an optimized workflow with the use of faster treatment techniques.


Adjuvant chemoradiation therapy for adenocarcinoma of the distal pancreas.
This study was designed to examine the effect of adjuvant 5-FU-based chemoradiation therapy (CRT) after distal pancreatectomy for adenocarcinoma of the distal pancreas. All patients underwent curative resection for adenocarcinoma of the distal pancreas between December 1985 and June 2006. Patients who received adjuvant CRT were compared with those who underwent surgery alone. A Kaplan-Meier estimate of the survival curve was used to determine estimates of the median survival and proportion alive at 1 and 2 years; log-rank tests were used to make comparisons between groups. A total of 123 patients underwent distal pancreatectomy; 29 patients were excluded for distant metastases at the time of surgery (n = 12, 10%) or before adjuvant therapy (n = 11, 9%), death within 2 months of surgery (n = 2, 2%), or if CRT treatment status was unknown (n = 4, 3%). Of the remaining 94 patients, 72% received adjuvant 5-FU-based CRT and 28% underwent surgery alone. Overall median survival was 16.2 (95% confidence interval (CI), 13.1-18.9) months. The groups were similar with respect to tumor size, nodal status, and margin status. There was no significant difference in overall survival between patients treated with adjuvant CRT versus surgery alone (p = 0.23). An exploratory subgroup analysis suggested a potential survival benefit of adjuvant CRT in patients with lymph node metastases (16.7 vs. 12.1 months, p < 0.01). Adjuvant CRT did not increase survival compared with surgery alone; however, patients with node-positive disease appear to benefit from adjuvant CRT.


OVOL2, an Inhibitor of WNT Signaling, Reduces Invasive Activities of Human and Mouse Cancer Cells and Is Down-regulated in Human Colorectal Tumors.
Activation of WNT signaling promotes the invasive activities of several types of cancer cells, but it is not clear if it regulates the same processes in colorectal cancer (CRC) cells, or what mechanisms are involved. We studied the expression and function of OVOL2, a member of the Ovo family of conserved zinc-finger transcription factors regulated by the WNT signaling pathway, in intestinal tumors of mice and human beings. We analyzed the expression of OVOL2 protein and messenger RNA in CRC cell lines and tissue arrays, as well as CRC samples from patients who underwent surgery at Xiamen University in China from 2009 to 2012; clinical information also was collected. CRC cell lines (SW620) were infected with lentivirus expressing OVOL2, analyzed in migration and invasion assays, and injected into nude mice to assess tumor growth and metastasis. Tandem affinity purification was used to purify the OVOL2-containing complex from CRC cells; the complex was analyzed by liquid chromatography, tandem mass spectrometry, and immunoprecipitation experiments. Gene promoter activities were measured in luciferase reporter assays. We analyzed mice with an intestine-specific disruption of Ovol2 (Ovol2(flox/+) transgenic mice), as well as Apc(min/+) mice; these mice were crossed and analyzed. Analysis of data from patients indicated that the levels of OVOL2 messenger RNA were significantly lower in colon carcinomas than adenomas, and decreased significantly as carcinomas progressed from grades 2 to 4. Immunohistochemical analysis of a tissue array of 275 CRC samples showed a negative association between tumor stage and OVOL2 level. Overexpression of OVOL2 in SW620 cells decreased their migration and invasion, reduced markers of the epithelial-to-mesenchymal transition, and suppressed their metastasis as xenograft tumors in nude mice; knockdown of OVOL2 caused LS174T cells to transition from epithelial to mesenchymal phenotypes. OVOL2 bound T-cell factor (TCF)4 and β-catenin, facilitating recruitment of histone deacetylase 1 to the TCF4-β-catenin complex; this inhibited expression of epithelial-to-mesenchymal transition-related genes regulated by WNT, such as SLUG, in CRC cell lines. OVOL2 was a downstream target of WNT signaling in LS174T and SW480 cells. The OVOL2 promoter was hypermethylated in late-stage CRC specimens from patients and in SW620 cells; hypermethylation resulted in OVOL2 down-regulation and an inability to inhibit WNT signaling. Disruption of Ovol2 in Apc(min/+) mice increased WNT activity in intestinal tissues and the formation of invasive intestinal tumors. OVOL2 is a colorectal tumor suppressor that blocks WNT signaling by facilitating the recruitment of histone deacetylase 1 to the TCF4-β-catenin complex. Strategies to increase levels of OVOL2 might be developed to reduce colorectal tumor progression and metastasis.


Gene copy number detection in animal studies.
Sensitive methods for the quantification of DNA fragments can be used to study an individual's genetic constitution for duplicated regions of the genome or to determine the relative proportion of different DNA fragments in heterogeneous samples such as pooled DNA from different individuals or in samples in which a fraction of the chromosomes carry a mutation. Here, we describe how we are using Pyrosequencing for this purpose. In Subheading 3., we describe a sensitive method that can be used to quantify the relative proportion of X- and Y-carrying sperm after sperm sorting in cattle. We also discuss our method for determining the copy numbers at a duplicated locus. This method has been applied to study genetic variation at the KIT locus in pigs, which have a major effect on coat color variation in this species.


Molecular analysis of Salmonella enteritidis by pulsed-field gel electrophoresis and ribotyping.
A total of 61 isolates of Salmonella enteritidis were analyzed by the techniques of pulsed-field gel electrophoresis (PFGE) and ribotyping. Twenty-three of the isolates were from Zurich, Switzerland, and 38 isolates were from the University Hospital, Kuala Lumpur, Malaysia. Five of the Malaysian isolates were hospital-related outbreak strains and were shown to be indistinguishable by PFGE analysis following digestion with three different restriction endonucleases, XbaI (5'-TCTAGA-3'), SpeI (5'-ACTAGT-3'), and AvrII (5'-CCTAGG-3'). The PFGE pattern of an isolate from a suspected carrier staff nurse was found to be identical to those of the hospital outbreak isolates. These isolates were also indistinguishable by ribotyping with SmaI and SphI. The same single PFGE pattern was also detected in 29 of 32 sporadic isolates of S. enteritidis. Four closely related ribotypes were detected among these 29 isolates. Similarly, outbreak-related strains from Switzerland showed close genetic identity by PFGE and ribotyping. Strains obtained from poultry showed more variations in their PFGE patterns and ribotypes, although the patterns were still closely related. In addition, SphI ribotypes A and D among the Swiss strains correlated with phage types 4 and 8, respectively. No correlation of phage types with PFGE pattern was noted. Both PFGE and ribotyping indicate that the S. enteritidis strains circulating in Malaysia and Switzerland are very similar and may be clonally related. Comparison of the PFGE patterns with the ribotypes for 23 Swiss and 16 Malaysian isolates showed that there was a 69% concordance in the grouping of isolates. We conclude that the close genetic similarity observed between epidemiologically unrelated and outbreak-related isolates of S. enteritidis suggests that both PFGE and ribotyping are of limited value in the epidemiological analysis of these particular isolates, possibly because of the highly clonal nature of pathogenic strains of S. enteritidis.


The Association Between In Vitro Propiconazole Sensitivity and Field Efficacy of Five New England Sclerotinia homoeocarpa Populations.
Dollar spot (Sclerotinia homoeocarpa) is a major turfgrass disease requiring fungicide application to maintain acceptable conditions for golf. A 2-year field experiment was conducted to determine the association between field efficacy of propiconazole and in vitro fungicide sensitivity of isolates from five S. homoeocarpa populations. Four golf courses with prior propiconazole exposure (Hartford Golf Club, Hickory Ridge Country Club, Shuttle Meadow Country Club, and Wintonbury Hills Golf Club), and a baseline site with no prior propiconazole exposure (Joseph Troll Turf Research Facility) were chosen as field sites. Experimental plots at each site received the following treatments at 21-day intervals: untreated, propiconazole (0.44, 0.88, 1.32, and 1.76 kg a.i. ha-1), and chlorothalonil (8.18 kg a.i. ha-1). S. homoeocarpa isolates were sampled at three time points during 2009 and 2010: initial (directly before fungicide treatment), 7 days after treatment (DAT), and 21 days after the last treatment. Isolates sampled from dollar spot infection centers at 7 DAT (2009 and 2010) were considered to exhibit "practical field resistance". In parallel, S. homoeocarpa isolates from each site were assayed for in vitro sensitivity to propiconazole by determining relative mycelium growth percentages (RMG%) on potato dextrose agar amended with propiconazole at a discriminatory concentration of 0.1 μg a.i. ml-1. S. homoeocarpa isolates from the four exposed populations displayed significantly higher RMG% values than the baseline population. In general, field efficacy at all propiconazole rates tested was lower at the four locations with prior propiconazole exposure when compared with the baseline population. Increased RMG% values on the propiconazole discriminatory concentration 0.1 μg a.i. ml-1 were associated with decreased relative control values for all propiconazole rates in 2009 and 2010. Results suggest RMG values above 50% at the propiconazole discriminatory concentration of 0.1 μg a.i. ml-1 may be a suitable threshold for detection of S. homoeocarpa isolates that cause practical DMI field resistance.


Quantifying the economic impact of government and charity funding of medical research on private research and development funding in the United Kingdom.
Government- and charity-funded medical research and private sector research and development (R&D) are widely held to be complements. The only attempts to measure this complementarity so far have used data from the United States of America and are inevitably increasingly out of date. This study estimates the magnitude of the effect of government and charity biomedical and health research expenditure in the United Kingdom (UK), separately and in total, on subsequent private pharmaceutical sector R&D expenditure in the UK. The results for this study are obtained by fitting an econometric vector error correction model (VECM) to time series for biomedical and health R&D expenditure in the UK for ten disease areas (including 'other') for the government, charity and private sectors. The VECM model describes the relationship between public (i.e. government and charities combined) sector expenditure, private sector expenditure and global pharmaceutical sales as a combination of a long-term equilibrium and short-term movements. There is a statistically significant complementary relationship between public biomedical and health research expenditure and private pharmaceutical R&D expenditure. A 1% increase in public sector expenditure is associated in the best-fit model with a 0.81% increase in private sector expenditure. Sensitivity analysis produces a similar and statistically significant result with a slightly smaller positive elasticity of 0.68. Overall, every additional £1 of public research expenditure is associated with an additional £0.83-£1.07 of private sector R&D spend in the UK; 44% of that additional private sector expenditure occurs within 1 year, with the remainder accumulating over decades. This spillover effect implies a real annual rate of return (in terms of economic impact) to public biomedical and health research in the UK of 15-18%. When combined with previous estimates of the health gain that results from public medical research in cancer and cardiovascular disease, the total rate of return would be around 24-28%. Overall, this suggests that government and charity funded research in the UK crowds in additional private sector R&D in the UK. The implied historical returns from UK government and charity funded investment in medical research in the UK compare favourably with the rates of return achieved on investments in the rest of the UK economy and are greatly in excess of the 3.5% real annual rate of return required by the UK government to public investments generally.


Probiotics in health maintenance and disease prevention.
Probiotic microflora display numerous health benefits beyond providing basic nutritional value. They cooperatively maintain a delicate balance between the gastrointestinal tract and immune system. When this balance is disrupted, disease and inflammation result. Inflammation and over stimulation of the immune system by pathogenic bacteria are competitively inhibited by mucosal adherence of normal beneficial microflora. A healthy gastrointestinal tract with adequate mucus production and appropriate bacterial colonization prevents the overgrowth of pathogenic bacteria, modulates disease processes, and prevents widespread inflammatory disorders. The understanding of the function of probiotics in the maintenance of health and their importance in preventing disease serves to enhance the overall health of patients. With increasing understanding that beneficial microbes are required for health maintenance and disease prevention, probiotics may be commonly used as a therapeutic tool by health care practitioners in the not-too-distant future. This review presents a review of probiotics in health maintenance and disease prevention.


Return to Sport and Athletic Function in an Active Population After Primary Arthroscopic Labral Reconstruction of the Hip.
Labral reconstruction has been advocated as an alternative to debridement for the treatment of irreparable labral tears, showing favorable short-term results. However, literature is scarce regarding outcomes and return to sport in the nonelite athletic population. To report minimum 1-year clinical outcomes and the rate of return to sport in athletic patients who underwent primary hip arthroscopy with labral reconstruction in the setting of femoroacetabular impingement syndrome and irreparable labral tears. Case series; Level of evidence, 4. Data were prospectively collected and retrospectively analyzed for patients who underwent an arthroscopic labral reconstruction between August 2012 and December 2017. Patients were included if they identified as an athlete (high school, college, recreational, or amateur); had follow-up on the following patient-reported outcomes (PROs): modified Harris Hip Score (mHHS), Nonarthritic Hip Score (NAHS), Hip Outcome Score-Sport Specific Subscale (HOS-SSS), and visual analog scale (VAS); and completed a return-to-sport survey at 1 year postoperatively. Patients were excluded if they underwent any previous ipsilateral hip surgery, had dysplasia, or had prior hip conditions. The proportions of patients who achieved the minimal clinically important difference (MCID) and patient acceptable symptomatic state (PASS) for mHHS and HOS-SSS were calculated. Statistical significance was set at P = .05. There were 32 (14 females) athletes who underwent primary arthroscopic labral reconstruction during the study period. The mean age and body mass index of the group were 40.3 years (range, 15.5-58.7 years) and 27.9 kg/m2 (range, 19.6-40.1 kg/m2), respectively. The mean follow-up was 26.4 months (range, 12-64.2 months). All patients demonstrated significant improvement in mHHS, NAHS, HOS-SSS, and VAS (P < .001) at latest follow-up. Additionally, 84.4% achieved MCID and 81.3% achieved PASS for mHHS, and 87.5% achieved MCID and 75% achieved PASS for HOS-SSS. VAS pain scores decreased from 4.4 to 1.8, and the satisfaction with surgery was 7.9 out of 10. The rate of return to sport was 78%. At minimum 1-year follow-up, primary arthroscopic labral reconstruction, in the setting of femoroacetabular impingement syndrome and irreparable labral tears, was associated with significant improvement in PROs in athletic populations. Return to sport within 1 year of surgery was 78%.


Contiguous hydrophobic and charged surface patches in short helix-constrained peptides drive cell permeability.
Most protein-protein interactions occur inside cells. Peptides can inhibit protein-protein interactions but tend not to enter cells. We systematically compare cell permeability for 8-12 residue model peptides with helix-inducing lactam/hydrocarbon linkers between amino acid sidechains. Cell uptake increases when hydrophobic residues and lactam linkers (i, i + 4) form a contiguous hydrophobic surface patch. Uptake increases further when both hydrophobic and positively charged (but not neutral or negative) residues are clustered into like surface patches. Amphipathicity alone is however insufficient for cell uptake of acyclic sequences. Changing the linker from lactam to hydrocarbon further increases uptake, but also promotes cell lysis. Helicity, positive charge and amphipathicity together promote cell permeability. Most known bioactive helical peptides do not optimally cluster residues for amphipathicity and so are likely unoptimised for cell uptake.


Clostridium Difficile, Colitis, and Colonoscopy: Pediatric Perspective.
Review tests available for detection of Clostridium difficile (C. Diff) induced disease, including when such tests should be done in children and how they should be interpreted. Multiple tests are available for detecting disease due to C. diff. These include colonoscopy and stool analysis. Colonoscopy with biopsy is the most sensitive test for detecting the presence of colitis. The toxins produced by the C. diff. (toxin A, toxin B, and binary toxin) are the agents that cause injury and disease. Only toxin producing C. diff. Strains will cause disease. Binary toxin by itself is not thought to produce disease. Binary toxin causes disease in humans when present with toxin A and B producing bacteria, and has been implicated with fulminant life threatening disease. Stool analyses vary in sensitivity and specificity depending on the assay used. The presence of toxin producing strains of C diff. in the stool does not equate with disease. The presence of a toxin-producing bacteria or toxins (A or B) only equates with disease if diarrhea or a diseased colon (toxic megacolon, ileus, and sepsis) is present. Nucleic acid amplification testing (NAAT), when used in the stool from patients with diarrhea, appears to be the most efficient study to detect the gene that encodes for toxin A and B and thus to diagnose C. diff.-induced disease. Infants have a high carriage rate of C. diff. and are believed not to develop disease from it or its toxins. Infants should not be tested for C. difficile. The NAAT is most specific when done on patients with diarrhea with liquid stools. Testing for C. difficile should only be done on patients with diarrhea. One can assume that a patient who has no diarrhea and is not ill does not have C. diff.-induced disease. Treatment should be limited to patients with diarrhea who test positive for C. diff. toxin (A or B) or toxin-producing bacteria. Direct testing for binary toxin is not commercially available. Binary toxin is only thought to cause disease in humans when C. diff. toxin (A and B)-producing bacteria are present.


Ubiquitin Chain Analysis by Parallel Reaction Monitoring.
Assessment of the global profile of ubiquitin chain topologies within a proteome is of interest to answer a wide range of biological questions. The protocol outlined here takes advantage of the di-glycine (-GG) modification left after the tryptic digestion of ubiquitin incorporated in a chain. By quantifying these topology-characteristic peptides the relative abundance of each ubiquitin chain topology can be determined. The steps required to quantify these peptides by a parallel reaction monitoring experiment are reported taking into consideration the stabilization of ubiquitin chains. Preparation of heavy controls, cell lysis, and digestion are described along with the appropriate mass spectrometer setup and data analysis workflow. An example data set with perturbations in ubiquitin topology is presented, accompanied by examples of how optimization of the protocol can affect results. By following the steps outlined, a user will be able to perform a global assessment of the ubiquitin topology landscape within their biological context.


Measurement of human exposure to biologically relevant fractions of inhaled aerosols.
Aerosol sampling has evolved with changes in technology and our understanding of the importance of assessing the biologically relevant fractions of the total aerosol. During the past decade there has been international agreement on the definitions for the inhalable, thoracic, and respirable fractions and instruments have been developed to collect samples according to these conventions. These measurement techniques are now well established in the workplace and are increasingly being applied to assessments of non-occupational exposure, as the practical difficulties in obtaining samples over 24 hours have been solved. It is argued that multiple aerosol size fractions should be measured for hazardous substances--such as inorganic lead--in which inhaled material may either be absorbed in the alveoli (respirable) or cleared from the ciliated airways in the lung to the gut and then absorbed (thoracic and extrathoracic). Such measurements should improve the evaluation of the risk for inhaled lead, particularly for non-occupational exposure of children. Also, passive aerosol sampling techniques may enable measurements of non-occupational exposure to be made over several weeks and this would also help improve the reliability of the risk evaluation for aerosol toxicants with a long biological half life. However, further development of these techniques is required if they are to be practicable and biologically relevant.


Vaccination recommendations for adult patients with autoimmune inflammatory rheumatic diseases.
The number of individuals with autoimmune inflammatory rheumatic diseases (AIIRDs) treated with immunosuppressive drugs is increasing steadily. The variety of immunosuppressive drugs and, in particular, biological therapies is also rising. The immunosuppressants, as well as the AIIRD itself, increase the risk of infection in this population. Thus, preventing infections by means of vaccination is of utmost importance. New Swiss vaccination recommendations for AIIRD patients were initiated by the Swiss Federal Office of Public Health and prepared by a working group of the Federal Commission for Vaccination Issues as well as by consultation of international experts. A literature search was performed in electronic databases (Cochrane, Medline, PubMed, Embase). In addition, unpublished literature was identified through a targeted website search of relevant organisations and international conferences dealing with vaccination, infectious diseases and rheumatology. Although data are scarce, the following main points were retrieved from the literature. Inactivated vaccines are safe, but their immunogenicity may be reduced in AIIRD patients, especially if they are under immunosuppressive therapy. Rituximab and abatacept appear to reduce significantly immune responses after vaccination. Live vaccines are generally contraindicated under immunosuppressive therapy owing to safety concerns. Specific exceptions, as well as time intervals for the administration of live vaccines after interruption of an immunosuppressive therapy, have been formulated in this article. More evidence regarding the immunogenicity and safety of vaccinations in AIIRD patients under various therapies is needed. Vaccination recommendations should be updated on a regular basis, as more scientific data will become available.


Prooxidant activity of transferrin and lactoferrin.
Acceleration of the autoxidation of Fe2+ by apotransferrin or apolactoferrin at acid pH is indicated by the disappearance of Fe2+, the uptake of oxygen, and the binding of iron to transferrin or lactoferrin. The product(s) formed oxidize iodide to an iodinating species and are bactericidal to Escherichia coli. Toxicity to E. coli by FeSO4 (10(-5) M) and human apotransferrin (100 micrograms/ml) or human apolactoferrin (25 micrograms/ml) was optimal at acid pH (4.5-5.0) and with logarithmic phase organisms. Both the iodinating and bactericidal activities were inhibited by catalase and the hydroxyl radical (OH.) scavenger mannitol, whereas superoxide dismutase was ineffective. NaCl at 0.1 M inhibited bactericidal activity, but had little or no effect on iodination. Iodide increased the bactericidal activity of Fe2+ and apotransferrin or apolactoferrin. The formation of OH.was suggested by the formation of the OH.spin-trap adduct (5,5-dimethyl-1-pyroline N-oxide [DMPO]/OH)., with the spin trap DMPO and the formation of the methyl radical adduct on the further addition of dimethyl sulfoxide. (DMPO/OH).formation was inhibited by catalase, whereas superoxide dismutase had little or no effect. These findings suggest that Fe2+ and apotransferrin or apolactoferrin can generate OH.via an H2O2 intermediate with toxicity to microorganisms, and raise the possibility that such a mechanism may contribute to the microbicidal activity of phagocytes.


Utility of the functional activities questionnaire for distinguishing mild cognitive impairment from very mild Alzheimer disease.
Current criteria for mild cognitive impairment (MCI) require "essentially intact" performance of activities of daily living (ADLs), which has proven difficult to operationalize. We sought to determine how well the Functional Activities Questionnaire (FAQ), a standardized assessment of instrumental ADLs, delineates the clinical distinction between MCI and very mild Alzheimer disease (AD). We identified 1801 individuals in the National Alzheimer's Coordinating Center Uniform Data Set with MCI (n=1108) or very mild AD (n=693) assessed with the FAQ and randomized them to the development or test sets. Receiver-operator curve (ROC) analysis of the development set identified optimal cut-points that maximized the sensitivity and specificity of FAQ measures for differentiating AD from MCI and were validated with the test set. ROC analysis of total FAQ scores in the development set produced an area under the curve of 0.903 and an optimal cut-point of 5/6, which yielded 80.3% sensitivity, 87.0% specificity, and 84.7% classification accuracy in the test set. Bill paying, tracking current events, and transportation (P's<0.005) were the FAQ items of greatest diagnostic utility. These data suggest that the FAQ exhibits adequate sensitivity and specificity when used as a standardized assessment of instrumental ADLs in the diagnosis of AD versus MCI.


Cross-Polarized Surface-Enhanced Infrared Spectroscopy by Fano-Resonant Asymmetric Metamaterials.
Plasmonic metamaterials have overcome fundamental limitations in conventional optics by their capability to engineer material resonances and dispersions at will, holding great promise for sensing applications. Recent demonstrations of metamaterial sensors, however, have mainly relied on their resonant nature for strong optical interactions with molecules, but few examples fully exploit their functionality to manipulate the polarization of light. Here, we present cross-polarized surface-enhanced infrared absorption (SEIRA) by the Fano-resonant asymmetric metamaterial allowing for strong background suppression as well as significant field enhancement. The metamaterial is designed to exhibit the controlled Fano resonance with the cross-polarization conversion property at 1730 cm-1, which spectrally overlaps with the C=O vibrational mode. In the cross-polarized SEIRA measurement, the C=O mode of poly(methyl methacrylate) molecules is clearly observed as a distinct dip within a Fano-resonant transmission peak of the metamaterial. The vibrational signal contrast is then improved based on the cross-polarized detection scheme where only the light interacting with the metamaterial-molecular coupled system is detected by totally eliminating the unwanted background light. Our metamaterial approach achieves the zeptomole sensitivity with a large signal-to-noise ratio in the far-field measurement, paving the way toward the realization of ultrasensitive IR inspection technologies.


Effects of dietary omega-3 polyunsaturated fatty acids on brain gene expression.
Polyunsaturated fatty acids (PUFA) are essential structural components of the central nervous system. Their role in controlling learning and memory has been well documented. A nutrigenomic approach with high-density microarrays was used to reveal brain gene-expression changes in response to different PUFA-enriched diets in rats. In aged rats fed throughout life with PUFA-enriched diets, genes with altered expressions included transthyretin, alpha-synuclein, and calmodulins, which play important roles in synaptic plasticity and learning. The effect of perinatal omega-3 PUFA supply on gene expression later in life also was studied. Several genes showed similar changes in expression in rats fed omega-3-deficient diets in the perinatal period, regardless of whether they or their mothers were fed omega-3 PUFA-sufficient diets after giving birth. In this experiment, among the down-regulated genes were a kainate glutamate receptor and a DEAD-box polypeptide. Among the up-regulated genes were a chemokine-like factor, a tumor necrosis factor receptor, and cytochrome c. The possible involvement of the genes with altered expression attributable to different diets in different brain regions in young and aged rats and the possible mode of regulatory action of PUFA also are discussed. We conclude that PUFA-enriched diets lead to significant changes in expression of several genes in the central nervous tissue, and these effects appear to be mainly independent of their effects on membrane composition. The direct effects of PUFA on transcriptional modulators, the downstream developmentally and tissue-specifically activated elements might be one of the clues to understanding the beneficial effects of the omega-3 PUFA on the nervous system.


Gas-phase thermometry using delayed-probe-pulse picosecond coherent anti-Stokes Raman scattering spectra of H2.
We report the development and application of a simple theoretical model for extracting temperatures from picosecond-laser-based coherent anti-Stokes Raman scattering (CARS) spectra of H2 obtained using time-delayed probe pulses. This approach addresses the challenges associated with the effects of rotational-level-dependent decay lifetimes on time-delayed probing for CARS thermometry. A simple procedure is presented for accurate temperature determination based on a Boltzmann distribution using delayed-probe-pulse vibrational CARS spectra of H2; this procedure requires measurement at only a select handful of probe-pulse delays and requires no assumptions about sample environment.


Comparison of Rock Varnish Bacterial Communities with Surrounding Non-Varnished Rock Surfaces: Taxon-Specific Analysis and Morphological Description.
Rock varnish is a thin layer of Fe and Mn oxyhydroxides with embedded clay minerals that contain an increased Mn/Fe ratio compared to that of the Earth's crust. Even if the study of rock varnish has important implications in several fields, the composition of epilithic bacterial communities and the distribution of taxa on varnish surfaces are still not wholly described. The aim of this study was (i) to identify the bacterial taxa which show the greatest variation between varnish and non-varnish environments, collected from the same rock, and (ii) to describe the morphology of epilithic communities through scanning electron microscopy (SEM). Triplicate samples of rock surfaces with varnish and triplicate samples without varnish were collected from five sites in Matsch Valley (South Tyrol, Italy). The V4 region of 16S rRNA gene was analyzed by Illumina sequencing. Fifty-five ubiquitous taxa have been examined to assess variation between varnish and non-varnish. Cyanobacteria, Chloroflexi, Proteobacteria along with minor taxa such as Solirubrobacterales, Conexibaxter, and Rhodopila showed significant variations of abundance, diversity, or both responding to the ecology (presence/absence of varnish). Other taxa, such as the genus Edaphobacter, showed a more marked spatial variation responding to the sampling site. SEM images showed a multitude of bacterial morphologies and structures involved in the process of attachment and creation of a suitable environment for growth. The features emerging from this analysis suggest that the highly oxidative Fe and Mn-rich varnish environment favors anoxigenic autotrophy and establishment of highly specialized bacteria.


Relative importance of tree genetics and microhabitat on macrofungal biodiversity on coarse woody debris.
Understanding the contribution of genetic variation within foundation species to community-level pattern and diversity represents the cornerstone of the developing field of community genetics. We assessed the relative importance of intraspecific genetic variation, spatial variation within a forest and microhabitat variation on a macrofungal decay community developing on logs of the Australian forest tree, Eucalyptus globulus. Uniform logs were harvested from trees from eight geographic races of E. globulus growing in a 15-year-old genetic trial. Logs were placed as designed grids within a native E. globulus forest and after 3 years of natural colonisation the presence of 62 macrofungal taxa were recorded from eight microhabitats on each log. The key factor found to drive macrofungal distribution and biodiversity on structurally uniform coarse woody debris was log-microhabitat, explaining 42% of the total variation in richness. Differences between log-microhabitats appeared to be due to variation in aspect, substrate (bark vs wood) and area/time of exposure to colonisation. This findings demonstrates the importance of considering fine-scale (within substrate) variation in the conservation and management of macrofungal biodiversity, an area that has received little previous attention. While a number of recent studies have demonstrated that the genetics of foundation tree species can influence dependent communities, this was not found to be the case for the early log decay community associated with E. globulus. Despite genetic variation in wood and bark properties existing within this species, there was no significant effect of tree genetics on macrofungal community richness or composition. This finding highlights the variation that may exist among guilds of organisms in their response to genetic variation within foundation species, an important consideration in a promising new area of research.


Mitogenetic structure of brown bears (Ursus arctos L.) in northeastern Europe and a new time frame for the formation of European brown bear lineages.
We estimated the phylogenetic relationships of brown bear maternal haplotypes from countries of northeastern Europe (Estonia, Finland and European Russia), using sequences of mitochondrial DNA (mtDNA) control region of 231 bears. Twenty-five mtDNA haplotypes were identified. The brown bear population in northeastern Europe can be divided into three haplogroups: one with bears from all three countries, one with bears from Finland and Russia, and the third composed almost exclusively of bears from European Russia. Four haplotypes from Finland and European Russia matched exactly with haplotypes from Slovakia, suggesting the significance of the current territory of Slovakia in ancient demographic processes of brown bears. Based on the results of this study and those from the recent literature, we hypothesize that the West Carpathian Mountains have served either as one of the northernmost refuge areas or as an important movement corridor for brown bears of the Eastern lineage towards northern Europe during or after the last ice age. Bayesian analyses were performed to investigate the temporal framework of brown bear lineages in Europe. The molecular clock was calibrated using Beringian brown bear sequences derived from radiocarbon-dated ancient samples, and the estimated mutation rate was 29.8% (13.3%-47.6%) per million years. The whole European population and Western and Eastern lineages formed about 175,000, 70,000 and 25,000 years before present, respectively. Our approach to estimating the time frame of brown bear evolution demonstrates the importance of using an appropriate mutation rate, and this has implications for other studies of Pleistocene populations.


A Bayesian framework for cue integration in multistable grouping: Proximity, collinearity, and orientation priors in zigzag lattices.
Integration of proximity and good continuation cues is analyzed as a probabilistic inference problem in contour grouping. A Bayesian framework was tested in a multistable dot lattice experiment. In rectangular lattices, distance ratio and global orientation of rows and columns were manipulated. Discollinearity was introduced by imposing zigzag in one orientation, by either fixed or stochastic displacement of elements. Results indicate that proximity and good continuation are generally treated as independent sources of information, added to prior orientation log-odds to produce the odds of grouping percepts. Distance likelihood is well captured by a power law, and discollinearity likelihoods by generalized Laplace distributions, with higher kurtosis for stochastic zigzag. While observers prefer vertical over horizontal orientations, the exact prior distribution is idiosyncratic. Perceptual grouping along cardinal axes is less affected by distance, but more by discollinearity, than along oblique orientations. Results are qualitatively and quantitatively compared to ecological statistics of contours (J. H. Elder & R. M. Goldberg, 2002). The potential of hierarchically extended Bayes models for a better understanding of principles in cue integration is discussed.


A novel approach to the determination of clinical decision thresholds.
Our objective was to determine the test and treatment thresholds for common acute primary care conditions. We presented 200 clinicians with a series of web-based clinical vignettes, describing patients with possible influenza, acute coronary syndrome (ACS), pneumonia, deep vein thrombosis (DVT) and urinary tract infection (UTI). We randomly varied the probability of disease and asked whether the clinician wanted to rule out disease, order tests or rule in disease. By randomly varying the probability, we obtained clinical decisions across a broad range of disease probabilities that we used to create threshold curves. For influenza, the test (4.5% vs 32%, p<0.001) and treatment (55% vs 68%, p=0.11) thresholds were lower for US compared with Swiss physicians. US physicians had somewhat higher test (3.8% vs 0.7%, p=0.107) and treatment (76% vs 58%, p=0.005) thresholds for ACS than Swiss physicians. For both groups, the range between test and treatment thresholds was greater for ACS than for influenza (which is sensible, given the consequences of incorrect diagnosis). For pneumonia, US physicians had a trend towards higher test thresholds and lower treatment thresholds (48% vs 64%, p=0.076) than Swiss physicians. The DVT and UTI scenarios did not provide easily interpretable data, perhaps due to poor wording of the vignettes. We have developed a novel approach for determining decision thresholds. We found important differences in thresholds for US and Swiss physicians that may be a function of differences in healthcare systems. Our results can also guide development of clinical decision rules and guidelines.


High glucose increases diacylglycerol mass and activates protein kinase C in mesangial cell cultures.
We showed previously that glomerular mesangial cells displayed increased fibronectin, laminin, and type IV collagen synthesis and mRNA levels when grown in medium containing 30 mM glucose compared with those cells grown in 10 mM glucose [S. H. Ayo, R. A. Radnik, W. F. Glass II, J. A. Garoni, E. R. Rampt, D. R. Appling, and J. I. Kreisberg. Am. J. Physiol. 260 (Renal Fluid Electrolyte Physiol. 29): F185-F191, 1990]. However, total protein synthesis and actin mRNA were unchanged. In this report, we show that an increase in medium glucose concentration resulted in an increase in diacylglycerol (DAG) mass and transiently increased protein kinase C (PKC) activity as assessed by the translocation of PKC from the soluble to the particulate fraction. Effects of increased glucose on DAG were evident at 30 min and were maintained through 1 wk of growth in medium containing 30 mM glucose. Although total PKC activity (i.e., soluble plus particulate fractions) did not change with high-glucose treatment, the percent activity associated with the particulate fraction (i.e., activated PKC) increased significantly after 60 min in RPMI 1640 medium with 30 mM glucose. The distribution of PKC returned to control values by 24 h. High glucose did not stimulate phosphoinositide hydrolysis, as evidenced by the absence of an increase in the water-soluble inositol phosphates, indicating that DAG was not generated through the action of a phosphoinositide-specific phospholipase C. Cells treated with the cell-permeable DAG analogue 1-oleoyl-2-acetyl glycerol to activate PKC displayed approximately two-fold increases of fibronectin, laminin, and type IV collagen mRNA levels after normalization against actin.(ABSTRACT TRUNCATED AT 250 WORDS)


Oxytocin enhances the experience of attachment security.
Repeated interactions between infant and caregiver result in either secure or insecure relationship attachment patterns, and insecure attachment may affect individual emotion-regulation and health. Given that oxytocin enhances social approach behavior in animals and humans, we hypothesized that oxytocin might also promote the subjective experience of attachment security in humans. Within a 3-week interval, 26 healthy male students classified with an insecure attachment pattern were invited twice to an experimental session. At the beginning of each experiment, a single dose of oxytocin or placebo was administered intranasally, using a double-blind, placebo-controlled within-subject design. In both conditions, subjects completed an attachment task based on the Adult Attachment Projective Picture System (AAP). Thirty-two AAP picture system presentations depicted attachment-related events (e.g. illness, solitude, separation, and loss), and were each accompanied by four prototypical phrases representing one secure and three insecure attachment categories. In the oxytocin condition, a significant proportion of these insecure subjects (N=18; 69%) increased in their rankings of the AAP prototypical "secure attachment" phrases and decreased in overall ranking of the "insecure attachment" phrases. In particular, there was a significant decrease in the number of subjects ranking the pictures with "insecure-preoccupied" phrases from the placebo to the oxytocin condition. We find that a single dose of intranasally administered oxytocin is sufficient to induce a significant increase in the experience of attachment security in insecurely attached adults.


Effects of chronic alpha-adrenergic receptor blockade on peripheral nerve conduction, hypoxic resistance, polyols, Na(+)-K(+)-ATPase activity, and vascular supply in STZ-D rats.
The effects of alpha-receptor blockade on nerve conduction, hypoxic resistance, ouabain-sensitive Na(+)-K(+)-ATPase, nerve polyols, and capillary density were examined in streptozocin-induced diabetic (STZ-D) rats. Nondiabetic and untreated diabetic control groups were used. Diabetes duration was 2 mo. There were two treated diabetic groups. A "prevention" group received 5 mg/kg prazosin for 2 mo from the induction of diabetes. A "reversal" group was untreated for the 1st mo and was given prazosin for the subsequent month. Conduction was measured in motor nerves supplying tibialis anterior and gastrocnemius muscles and sensory saphenous nerve. Diabetes resulted in 15-29% reductions in conduction velocity (P less than 0.01). In the prevention group, conduction deficits were minimal compared with untreated diabetes (P less than 0.01). In the reversal group, motor conduction was also substantially improved, although sensory conduction was not significantly affected. In vitro measurement of sciatic nerve hypoxic resistance revealed a 49% increase in the time taken for compound action potential amplitude to reach half its initial value with diabetes (P less than 0.01). This was largely prevented by prazosin treatment (P less than 0.01), although treatment had a lesser effect in the reversal group. Treatment had no effect on nerve polyol levels or Na(+)-K(+)-ATPase activity. Functional improvements with prazosin were probably based on increased vasa nervorum perfusion. There was a 20% elevation of endoneurial capillary density (P less than 0.01) in both prevention and reversal groups. We conclude that vascular factors play an important role in the etiology of experimental diabetic neuropathy, and functional changes may be corrected by chronic vasodilator treatment.


Distribution and diversity of Verrucomicrobia methanotrophs in geothermal and acidic environments.
Recently, methanotrophic members of the phylum Verrucomicrobia have been described, but little is known about their distribution in nature. We surveyed methanotrophic bacteria in geothermal springs and acidic wetlands via pyrosequencing of 16S rRNA gene amplicons. Putative methanotrophic Verrucomicrobia were found in samples covering a broad temperature range (22.5-81.6°C), but only in acidic conditions (pH 1.8-5.0) and only in geothermal environments, not in acidic bogs or fens. Phylogenetically, three 16S rRNA gene sequence clusters of putative methanotrophic Verrucomicrobia were observed. Those detected in high-temperature geothermal samples (44.1-81.6°C) grouped with known thermoacidiphilic 'Methylacidiphilum' isolates. A second group dominated in moderate-temperature geothermal samples (22.5-40.1°C) and a representative mesophilic methanotroph from this group was isolated (strain LP2A). Genome sequencing verified that strain LP2A possessed particulate methane monooxygenase, but its 16S rRNA gene sequence identity to 'Methylacidiphilum infernorum' strain V4 was only 90.6%. A third group clustered distantly with known methanotrophic Verrucomicrobia. Using pmoA-gene targeted quantitative polymerase chain reaction, two geothermal soil profiles showed a dominance of LP2A-like pmoA sequences in the cooler surface layers and 'Methylacidiphilum'-like pmoA sequences in deeper, hotter layers. Based on these results, there appears to be a thermophilic group and a mesophilic group of methanotrophic Verrucomicrobia. However, both were detected only in acidic geothermal environments.


Canonical Notch signaling is dispensable for adult steady-state and stress myelo-erythropoiesis.
Although an essential role for canonical Notch signaling in generation of hematopoietic stem cells in the embryo and in thymic T-cell development is well established, its role in adult bone marrow (BM) myelopoiesis remains unclear. Some studies, analyzing myeloid progenitors in adult mice with inhibited Notch signaling, implicated distinct roles of canonical Notch signaling in regulation of progenitors for the megakaryocyte, erythroid, and granulocyte-macrophage cell lineages. However, these studies might also have targeted other pathways. Therefore, we specifically deleted, in adult BM, the transcription factor recombination signal-binding protein J κ (Rbpj), through which canonical signaling from all Notch receptors converges. Notably, detailed progenitor staging established that canonical Notch signaling is fully dispensable for all investigated stages of megakaryocyte, erythroid, and myeloid progenitors in steady state unperturbed hematopoiesis, after competitive BM transplantation, and in stress-induced erythropoiesis. Moreover, expression of key regulators of these hematopoietic lineages and Notch target genes were unaffected by Rbpj deficiency in BM progenitor cells.


Modeling competitive adsorption of mixtures of volatile organic compounds in a fixed-bed of beaded activated carbon.
A two-dimensional mathematical model was developed to study competitive adsorption of n-component mixtures in a fixed-bed adsorber. The model consists of an isotherm equation to predict adsorption equilibria of n-component volatile organic compounds (VOCs) mixture from single component isotherm data, and a dynamic adsorption model, the macroscopic mass, energy and momentum conservation equations, to simulate the competitive adsorption of the n-components onto a fixed-bed of adsorbent. The model was validated with experimentally measured data of competitive adsorption of binary and eight-component VOCs mixtures onto beaded activated carbon (BAC). The mean relative absolute error (MRAE) was used to compare the modeled and measured breakthrough profiles as well as the amounts of adsorbates adsorbed. For the binary and eight-component mixtures, the MRAE of the breakthrough profiles was 13 and 12%, respectively, whereas, the MRAE of the adsorbed amounts was 1 and 2%, respectively. These data show that the model provides accurate prediction of competitive adsorption of multicomponent VOCs mixtures and the competitive adsorption isotherm equation is able to accurately predict equilibrium adsorption of VOCs mixtures.


Does Losing 5-7% of Prediabetic Body Weight from a Diabetes Prevention Program decrease Cardiovascular Risks?
According to the Centers for Disease Control and Prevention (CDC), one-third of adults have prediabetes (i.e., at risk for developing type 2 diabetes), a leading risk factor for cardiovascular disease. The Diabetes Prevention Program (DPP) focuses on lifestyle modifications to help participants lose 5-7% of their body weight and prevent Type 2 Diabetes. The purpose of this community-based pilot study was to investigate how successful completion of the DPP might be associated with decreases in body weight and atherosclerotic cardiovascular disease (ASCVD) risks. Single-site, prospective cohort study. The DPP was implemented at the Farmington Village Family Practice Clinic and delivered virtually via Zoom from January 2020 through December 2020. During the first six months, participants met weekly for one hour. In the remaining six months, monthly sessions were held for one hour. Each session began with a private weigh-in followed by a uniquely designed lesson plan. A total of 14 prediabetic patients, based on hemoglobin A1c (A1c), fasting blood glucose levels, or diabetic risk calculator scores, were enrolled. For analyses, data concerning body mass index (BMI), smoking status, anti-hypertensive medications, age, race, sex, A1c, fasting blood glucose, total cholesterol, and high-density lipoprotein (HDL) levels were measured at baseline, six and 12 months. These parameters were used to calculate composite ASCVD risk percentages based on the 2013 Risk Calculator from the American Heart Association/American College of Cardiology. Using a series of Wilcoxon Matched Signed Rank Pair T test procedures, initial base-to six-month analyses showed a statistically significant improvement in ASCVD risk scores (p < 0.01), BMI (p < 0.01), HDL (p < 0.01), estimated weekly minutes of physical activity (p =< 0.01), and total cholesterol (p = 0.048) levels. In addition, base-to-12-month differences for ASCVD, BMI, HDL and physical activity outcomes remained statistically significant. After completion of the DPP program, both initial (base to six month) as well as follow up (base to 12-month) statistically significant improvements in ASCVD, HDL, BMI, physical activity levels, and total cholesterol were observed. These pilot study results are promising and consistent with the reduction of cardiovascular risk factors. These findings support the value of a structured, evidence-based educational curriculum focused on nonpharmacologic intervention to decrease weight loss and ASCVD risk scores for prediabetes adults.


Type I hypersensitivity reaction as a complication of lepa.
Adverse drug reaction is defined as response to a drug which is noxious and unintended, and which occurs at doses normally used in man for the prophylaxis, diagnosis or therapy of disease, or for the modification of physiological functions. Type I hypersensitivity reaction is known as anaphylactic reaction which is due to immediate immunoglobulin E-mediated reaction. It is characterized by symptoms such as fever nausea, back pain, angiodema, rash, flushing, etc. Lepa generally refers to the application of pastes formed by mixing powder of herbs with water, milk, etc., and liquids. Complementary and alternative medicines are frequently used by the general population. Many people consider them to be without side effects. Ayurvedic treatment involves Shodhana (biopurification), Shaman (pacification), Bahya (external therapy), and Abhyantara karma's (internal therapy) for treating different diseases. One such bahya karma or external therapy is lepa. Even though lepa is said as "Aadhya Upakrama," undue hypersensitivity is observed in many patients. A 60-year-old woman had an adverse reaction to lepa after being administered as an external medication. The observations were erythema, eruptions, and itching. Such case of hypersensitivity is discussed in the present study.


Little change in global drought over the past 60 years.
Drought is expected to increase in frequency and severity in the future as a result of climate change, mainly as a consequence of decreases in regional precipitation but also because of increasing evaporation driven by global warming. Previous assessments of historic changes in drought over the late twentieth and early twenty-first centuries indicate that this may already be happening globally. In particular, calculations of the Palmer Drought Severity Index (PDSI) show a decrease in moisture globally since the 1970s with a commensurate increase in the area in drought that is attributed, in part, to global warming. The simplicity of the PDSI, which is calculated from a simple water-balance model forced by monthly precipitation and temperature data, makes it an attractive tool in large-scale drought assessments, but may give biased results in the context of climate change. Here we show that the previously reported increase in global drought is overestimated because the PDSI uses a simplified model of potential evaporation that responds only to changes in temperature and thus responds incorrectly to global warming in recent decades. More realistic calculations, based on the underlying physical principles that take into account changes in available energy, humidity and wind speed, suggest that there has been little change in drought over the past 60 years. The results have implications for how we interpret the impact of global warming on the hydrological cycle and its extremes, and may help to explain why palaeoclimate drought reconstructions based on tree-ring data diverge from the PDSI-based drought record in recent years.


Development and the Baldwin effect.
Baldwin's classic hypothesis states that behavioral plasticity can speed evolution by (a) smoothing the fitness landscape and (b) indirect genetic assimilation of acquired characteristics. This latter phase demands a strong correlation between genotype and phenotype space. But the natural world shows signs of this correlation at only a very coarse level, since the intervening developmental process greatly complicates the mapping from genetics to physiology and ethology. Hence, development appears to preclude a strong Baldwin effect. However, by adding a simple developmental mechanism to Hinton and Nowlan's classic model of the Baldwin effect, and by allowing evolution to determine the proper balance between direct and indirect mapping of genome to phenotype, this research reveals several different effects of development on the Baldwin effect, some promoting and others inhibiting. Perhaps the most interesting result is an evolved cooperation between direct blueprints and indirect developmental recipes in searching for unstructured and partially structured target patterns in large, needle-in-the-haystack fitness landscapes.


Topical antimicrobial therapy: Current status and challenges.
Topically applied antibacterial agents are widely used. Opinions regarding the clinical efficacy of topical antibiotics are conflicting, and for most indications, alternative oral therapies are available. Topical application has many potential advantages over systemic therapy that includes high and sustained concentrations of drug directly at the infected site, low quantity of antibiotic needed, better compliance, fewer systemic side effects and potentially less chance of antimicrobial resistance. Despite these advantages, an important concern has been the difficulty in monitoring antibiotic dosage and duration of therapy. Most topical preparations are applied on sites with pre-existing normal bacterial flora, and the detrimental effect of antibiotic on the 'good' bacteria is difficult to control. Unnecessary exposure of the resident microflora to high drug levels may select drug-resistant phenotypes. The number of antibiotics available and the quality and composition of the formulations recommended for topical drug delivery are improving. Their role in the prevention and treatment of locally invasive infections is established for many clinical conditions. However, there is still a lacuna in the availability of pharmacokinetic (PK) knowledge of these topical preparations and translation of the same to clinical practice. In addition, reporting the clinical outcome following the use of these agents and its analysis considering the recently proposed epidemiological cut-off value-based cut-offs are also areas which merit further research. In this review, we highlight the clinical utility and the PK aspects of topical antimicrobials in various infections. We also discuss the limitations of the current antimicrobial susceptibility testing (AST) protocols and new methods for AMST for topical agents.


Genetic and pharmacologic inhibition of mTORC1 promotes EMT by a TGF-β-independent mechanism.
Epithelial-to-mesenchymal transition (EMT) is a transdifferentiation process that converts epithelial cells into highly motile mesenchymal cells. This physiologic process occurs largely during embryonic development but is aberrantly reactivated in different pathologic situations, including fibrosis and cancer. We conducted a siRNA screening targeted to the human kinome with the aim of discovering new EMT effectors. With this approach, we have identified mTOR complex 1 (mTORC1), a nutrient sensor that controls protein and lipid synthesis, as a key regulator of epithelial integrity. Using a combination of RNAi and pharmacologic approaches, we report here that inhibition of either mTOR or RPTOR triggers EMT in mammary epithelial cells. This EMT was characterized by the induction of the mesenchymal markers such as fibronectin, vimentin, and PAI-1, together with the repression of epithelial markers such as E-cadherin and ZO-3. In addition, mTORC1 blockade enhanced in vivo migratory properties of mammary cells and induced EMT independent of the TGF-β pathway. Finally, among the transcription factors known to activate EMT, both ZEB1 and ZEB2 were upregulated following mTOR repression. Their increased expression correlated with a marked reduction in miR-200b and miR-200c mRNA levels, two microRNAs known to downregulate ZEB1 and ZEB2 expression. Taken together, our findings unravel a novel function for mTORC1 in maintaining the epithelial phenotype and further indicate that this effect is mediated through the opposite regulation of ZEB1/ZEB2 and miR-200b and miR-200c. Furthermore, these results suggest a plausible etiologic explanation for the progressive pulmonary fibrosis, a frequent adverse condition associated with the therapeutic use of mTOR inhibitors.


Rotation is visualisation, 3D is 2D: using a novel measure to investigate the genetics of spatial ability.
Spatial abilities-defined broadly as the capacity to manipulate mental representations of objects and the relations between them-have been studied widely, but with little agreement reached concerning their nature or structure. Two major putative spatial abilities are "mental rotation" (rotating mental models) and "visualisation" (complex manipulations, such as identifying objects from incomplete information), but inconsistent findings have been presented regarding their relationship to one another. Similarly inconsistent findings have been reported for the relationship between two- and three-dimensional stimuli. Behavioural genetic methods offer a largely untapped means to investigate such relationships. 1,265 twin pairs from the Twins Early Development Study completed the novel "Bricks" test battery, designed to tap these abilities in isolation. The results suggest substantial genetic influence unique to spatial ability as a whole, but indicate that dissociations between the more specific constructs (rotation and visualisation, in 2D and 3D) disappear when tested under identical conditions: they are highly correlated phenotypically, perfectly correlated genetically (indicating that the same genetic influences underpin performance), and are related similarly to other abilities. This has important implications for the structure of spatial ability, suggesting that the proliferation of apparent sub-domains may sometimes reflect idiosyncratic tasks rather than meaningful dissociations.


Nomogram predicting clinical outcomes in breast cancer patients treated with neoadjuvant chemotherapy.
The aim of this study was to combine clinical pathologic variables that are associated with pathologic completer response (pCR) and relapse-free survival (RFS) after neoadjuvant chemotherapy into prediction nomograms. A total of 370 stage II or III breast cancer patients who received neoadjuvant docetaxel/doxorubicin chemotherapy were enrolled in this study. We developed the nomograms using logistic regression model for pCR and Cox proportional hazard regression model for RFS. The nomogram for pCR based on initial tumor size, estrogen receptor (ER), human epidermal growth factor receptor 2, and Ki67 had good discrimination performance (AUROC = 0.830). Multivariate Cox model identified age less than 35, initial clinical stage, pathologic stage, ER, Ki67 as prognostic factors, and the nomogram for RFS was developed based on these covariates. The concordance index for the second nomogram was 0.781, and calibration was also good. We developed nomograms based on clinical and pathologic characteristics to predict the probability of pCR and RFS for patients receiving neoadjuvant docetaxel/doxorubicin chemotherapy.


Short-term safety evaluation of processed calcium montmorillonite clay (NovaSil) in humans.
NovaSil clay (NS) provides significant protection from the adverse effects of aflatoxins (AFs) in multiple animal species by decreasing bioavailability from the gastrointestinal tract. It is postulated that NS clay can be safely added to human diets to diminish exposure and health risks from AF contaminated food. To determine the safety and tolerance of NS in humans and establish dosimetry protocols for long-term efficacy studies, a randomized and double-blinded phase I clinical trial was conducted. Volunteers (20-45 yr in age), were clinically screened for confirmation of their health status. Fifty subjects (23 males and 27 females) were randomly divided into two groups: The low-dose group received nine capsules containing 1.5 g/day, and the high-dose group received nine capsules containing 3.0 g/day for a period of 2?wk. NS capsules were manufactured in the same color and size and were distributed to each participant three times a day at designated sites where follow-up was taken to record any side effects and complaints. Blood and urine samples were collected before and after the study for laboratory analysis. All participants completed the trial and compliance was 99.1%. Mild GI effects were reported in some participants. Symptoms included abdominal pain (6%, 3/50), bloating (4%, 2/50), constipation (2%, 1/50), diarrhea (2%, 1/50), and flatulence (8%, 4/50). No statistical significance was found between the two groups for these adverse effects (p > 0.25). No significant differences were shown in hematology, liver and kidney function, electrolytes, vitamins A and E, and minerals in either group. These results demonstrate the relative safety of NS clay in human subjects and will serve as a basis for long-term human trials in populations at high risk for aflatoxicosis.


Maternal supplementation with cobalt sources, folic acid, and rumen-protected methionine and its effects on molecular and functional correlates of the immune system in neonatal Holstein calves.
Calves born to multiparous Holstein cows fed during the last 30 d of pregnancy 2 different cobalt sources [cobalt glucoheptonate (CoPro) or cobalt pectin (CoPectin)], folic acid (FOA), and rumen-protected methionine (RPM) were used to study neonatal immune responses after ex vivo lipopolysaccharide (LPS) challenge. Groups were (n = 12 calves/group) CoPro, FOA+CoPro, FOA+CoPectin, and FOA+CoPectin+RPM. Calves were weighed at birth and blood collected at birth (before colostrum), 21 d of age, and 42 d of age (at weaning). Growth performance was recorded once a week during the first 6 wk of age. Energy metabolism, inflammation, and antioxidant status were assessed at birth through various plasma biomarkers. Whole blood was challenged with 3 µg/mL of LPS or used for phagocytosis and oxidative burst assays. Target genes evaluated by real-time quantitative PCR in whole blood samples were associated with immune response, antioxidant function, and 1-carbon metabolism. The response in mRNA abundance in LPS challenged versus nonchallenged samples was assessed via Δ = LPS challenged - LPS nonchallenged samples. Phagocytosis capacity and oxidative burst activity were measured in neutrophils and monocytes, with data reported as ratio (percentage) of CD14 to CH138A-positive cells. Data including all time points were subjected to ANOVA using PROC MIXED in SAS 9.4 (SAS Institute Inc.), with Treatment, Sex, Age, and Treatment × Age as fixed effects. A 1-way ANOVA was used to determine differences at birth, with Treatment and Sex as fixed effects. Calf birth body weight and other growth parameters did not differ between groups. At birth, plasma haptoglobin concentration was lower in FOA+CoPro compared with CoPro calves. We detected no effect for other plasma biomarkers or immune function due to maternal treatments at birth. Compared with CoPro, in response to LPS challenge, whole blood from FOA+CoPectin and FOA+CoPectin+RPM calves had greater mRNA abundance of intercellular adhesion molecule 1 (ICAM1). No effect for other genes was detectable. Regardless of maternal treatments, sex-specific responses were observed due to greater plasma concentrations of haptoglobin, paraoxonase, total reactive oxygen metabolites, nitrite, and β-carotene in female versus male calves at birth. In contrast, whole blood from male calves had greater mRNA abundance of IRAK1, CADM1, and ITGAM in response to LPS challenge at birth. The longitudinal analysis of d 0, 21, and 42 data revealed greater bactericidal permeability-increasing protein (BPI) mRNA abundance in whole blood from FOA+CoPectin versus FOA+CoPro calves, coupled with greater abundance in FOA+CoPro compared with CoPro calves. Regardless of maternal treatments, most genes related to cytokines and cytokine receptors (IL1B, IL10, TNF, IRAK1, CXCR1), toll-like receptor pathway (TLR4, NFKB1), adhesion and migration (ICAM1, ITGAM), antimicrobial function (MPO), and antioxidant function (GPX1) were downregulated over time. Phagocytosis capacity and oxidative burst activity in both neutrophils and monocytes did not differ due to maternal treatment. Regardless of maternal treatments, we observed an increase in the percentage of neutrophils capable of phagocytosis and oxidative burst activity over time. Overall, these preliminary assessments suggested that maternal supplementation with FOA and Co combined with RPM had effects on a few plasma biomarkers of inflammation at birth and molecular responses associated with inflammatory mechanisms during the neonatal period.


Maintaining polarization in polarimetric multiphoton microscopy.
Polarimetric measurements in multiphoton microscopy can reveal information about the local molecular order of a sample. However, the presence of a dichroic through which the excitation beam propagates will generally scramble its polarization. We propose a simple scheme whereby a second properly-oriented compensation dichroic is used to negate any alteration regardless of the wavelength and the initial polarization. We demonstrate how this robust and rapid approach simplifies polarimetric measurements in second-harmonic generation, two-photon excited fluorescence and coherent anti-Stokes Raman scattering. Illustration of the polarization maintaining strategy with the compensating dichroic oriented such that its s- and p-axes are interchanged with these of the primary dichroic.