[Contact inhibition of desmin expression in rat liver Ito cells in vitro].
The presence of desmin is used to identify the Ito cells in rat liver and to evaluate the purity of separated and cultured cells. The present study was conducted to evaluate the expression of cytoskeletal (with special emphasis on desmin) and intercellular matrix proteins in cultured rat liver Ito cells. The Ito cells were isolated by enzymatic digestion of rat liver and purified by single-step Nycodenz gradient. During the first week of culturing, the Ito cells proliferated and all of them expressed desmin and vimentin. By the end of the first week, part of them were seen to transdifferentiate into myofibroblasts. These cells expressed alpha-smooth muscle actin. The Ito cells and myofibroblasts were stained with anti-laminin, and anti-collagen I and III antibodies. The expression of desmin in myofibroblasts was less than in untransformed (stellate shaped) Ito cells. By the end of the second week the cells developed a dense monolayer and lost desmin. We suggest that repression of desmin in the cablestone-like monolayer of cells may be due to cell-to-cell contact inhibition.


The relationship between methamphetamine and popper use and risk of HIV seroconversion in the multicenter AIDS cohort study.
The association between methamphetamine use and HIV seroconversion for men who have sex with men (MSM) was examined using longitudinal data from the Multicenter AIDS Cohort Study. Seronegative (n = 4003) men enrolled in 1984 to 1985, 1987 to 1991, and 2001 to 2003 were identified. Recent methamphetamine and popper use was determined at the current or previous visit. Time to HIV seroconversion was the outcome of interest. Covariates included race/ethnicity, cohort, study site, educational level, number of sexual partners, number of unprotected insertive anal sexual partners, number of unprotected receptive anal sexual partners, insertive rimming, cocaine use at the current or last visit, ecstasy use at the current or last visit, any needle use since the last visit, Center for Epidemiologic Study of Depression symptom checklist score >16 since the last visit, and alcohol consumption. After adjusting for covariates, there was a 1.46 (95% confidence interval [CI]: 1.12 to 1.92) increased relative hazard of HIV seroconversion associated with methamphetamine use. The relative hazard associated with popper use was 2.10 (95% CI: 1.63 to 2.70). The relative hazard of HIV seroconversion increased with the number of unprotected receptive anal sexual partners, ranging from 1.87 (95% CI: 1.40 to 2.51) for 1 partner to 9.32 (95% CI: 6.21 to 13.98) for 5+ partners. The joint relative hazard for methamphetamine and popper use was 3.05 (95% CI: 2.12 to 4.37). There was a significant joint relative hazard for methamphetamine use and number of unprotected receptive anal sexual partners of 2.71 (95% CI: 1.81 to 4.04) for men with 1 unprotected receptive anal sexual partner, which increased in a dose-dependent manner for >1 partners. Further examination of the mechanisms underlying the synergism of drug use and sexual risk behaviors on rates of HIV seroconversion is necessary for the development of new targeted HIV prevention strategies for non-monogamous drug-using MSM.


Tight coupling of metabolic oscillations and intracellular water dynamics in Saccharomyces cerevisiae.
We detected very strong coupling between the oscillating concentration of ATP and the dynamics of intracellular water during glycolysis in Saccharomyces cerevisiae. Our results indicate that: i) dipolar relaxation of intracellular water is heterogeneous within the cell and different from dilute conditions, ii) water dipolar relaxation oscillates with glycolysis and in phase with ATP concentration, iii) this phenomenon is scale-invariant from the subcellular to the ensemble of synchronized cells and, iv) the periodicity of both glycolytic oscillations and dipolar relaxation are equally affected by D2O in a dose-dependent manner. These results offer a new insight into the coupling of an emergent intensive physicochemical property of the cell, i.e. cell-wide water dipolar relaxation, and a central metabolite (ATP) produced by a robustly oscillating metabolic process.


pH-activated nanoparticles for controlled topical delivery of farnesol to disrupt oral biofilm virulence.
Development of effective therapies to control oral biofilms is challenging, as topically introduced agents must avoid rapid clearance from biofilm-tooth interfaces while targeting biofilm microenvironments. Additionally, exopolysaccharides-matrix and acidification of biofilm microenvironments are associated with cariogenic (caries-producing) biofilm virulence. Thus, nanoparticle carriers capable of binding to hydroxyapatite (HA), saliva-coated HA (sHA), and exopolysaccharides with enhanced drug release at acidic pH were developed. Nanoparticles are formed from diblock copolymers composed of 2-(dimethylamino)ethyl methacrylate (DMAEMA), butyl methacrylate (BMA), and 2-propylacrylic acid (PAA) (p(DMAEMA)-b-p(DMAEMA-co-BMA-co-PAA)) that self-assemble into ∼21 nm cationic nanoparticles. Nanoparticles exhibit outstanding adsorption affinities (∼244 L-mmol(-1)) to negatively charged HA, sHA, and exopolysaccharide-coated sHA due to strong electrostatic interactions via multivalent tertiary amines of p(DMAEMA). Owing to hydrophobic cores, nanoparticles load farnesol, a hydrophobic antibacterial drug, at ∼22 wt %. Farnesol release is pH-dependent with t1/2 = 7 and 15 h for release at pH 4.5 and 7.2, as nanoparticles undergo core destabilization at acidic pH, characteristic of cariogenic biofilm microenvironments. Importantly, topical applications of farnesol-loaded nanoparticles disrupted Streptococcus mutans biofilms 4-fold more effectively than free farnesol. Mechanical stability of biofilms treated with drug-loaded nanoparticles was compromised, resulting in >2-fold enhancement in biofilm removal under shear stress compared to free farnesol and controls. Farnesol-loaded nanoparticles effectively attenuated biofilm virulence in vivo using a clinically relevant topical treatment regimen (2×/day) in a rodent dental caries disease model. Strikingly, treatment with farnesol-loaded nanoparticles reduced both the number and severity of carious lesions, while free farnesol had no effect. Nanoparticle carriers have great potential to enhance the efficacy of antibiofilm agents through multitargeted binding and pH-responsive drug release due to microenvironmental triggers.


Sarcomatoid lesions of the urogenital tract.
Spindle-cell lesions other than sarcomas are relatively rare occurrences along the urogenital tract but include both benign and malignant processes. Inflammatory pseudotumors may occur anywhere along the urothelial tract; they may follow a surgical procedure or appear de novo. Malignant tumors may be classified as sarcomatoid carcinomas or carcinosarcomas according to their degree of differentiation, the latter containing obvious mesenchymal components. Generally both arise in a background of high-grade carcinoma and are of a high pathological stage at the time of initial presentation. For this reason they have a poor prognosis. Because the deferential diagnosis of spindle-cell neoplasms include both benign and malignant lesions, great care must be taken in making the correct diagnosis.


PDMS-based turbulent microfluidic mixer.
Over the past decade, homogeneous mixing in microfluidic devices has been a critical challenge, because of the inherently low flow rates in microfluidic channels. Although several mixer designs have been suggested to achieve efficient mixing, most of them involve intricate structures requiring a series of laborious fabrication processes. Operation at high flow rates can greatly enhance mixing by induction of turbulence, but devices that can resist such a high pressure drop to induce turbulence in microfluidic channels are difficult to fabricate, especially for commonly used poly(dimethylsiloxane) (PDMS)-based microfluidic devices. We have developed a Y-shaped, turbulent microfluidic mixer made of PDMS and a glass substrate by strong bonding of the substrates to a nanoadhesive layer deposited via initiated chemical vapor deposition. The high bonding strength of the nanoadhesive layer enables safe operation of the PDMS/glass turbulent microfluidic mixer at a total water flow rate of 40 mL min(-1), corresponding to a Reynolds number, Re, of ~4423, one of the highest values achieved in a microfluidic channel. The turbulence generated as a result of the high Re allows rapid mixing of the input fluids on contact. Image analysis showed that mixing started as soon as the fluids were introduced into the mixer. The experimental results matched the numerical predictions well, demonstrating that convective mixing was dominant as a result of turbulence induced in the microfluidic channel. Using the turbulent microfluidic mixer, we have demonstrated high throughput formation of emulsions with narrower size distribution. It was shown that as the flow rate increases inside the microfluidic channel, the size distribution of resulting emulsions decreases owing to the increase in the turbulent energy dissipation. The turbulent microfluidic mixer developed in this work not only enables rapid mixing of streams, but also increases throughputs of microfluidic reactors.


IMPACT-Scot 2 report on COVID-19 in hip fracture patients.
The primary aim was to determine the influence of COVID-19 on 30-day mortality following hip fracture. Secondary aims were to determine predictors of COVID-19 status on presentation and later in the admission; the rate of hospital acquired COVID-19; and the predictive value of negative swabs on admission. A nationwide multicentre retrospective cohort study was conducted of all patients presenting with a hip fracture to 17 Scottish centres in March and April 2020. Demographics, presentation blood tests, COVID-19 status, Nottingham Hip Fracture Score, management, length of stay, and 30-day mortality were recorded. In all, 78/833 (9.4%) patients were diagnosed with COVID-19. The 30-day survival of patients with COVID-19 was significantly lower than for those without (65.4% vs 91%; p < 0.001). Diagnosis of COVID-19 within seven days of admission (likely community acquired) was independently associated with male sex (odds ratio (OR) 2.34, p = 0.040, confidence interval (CI) 1.04 to 5.25) and symptoms of COVID-19 (OR 15.56, CI 6.61 to 36.60, p < 0.001). Diagnosis of COVID-19 made between seven and 30 days of admission to hospital (likely hospital acquired) was independently associated with male sex (OR 1.73, CI 1.05 to 2.87, p = 0.032), Nottingham Hip Fracture Score ≥ 7 (OR 1.91, CI 1.09 to 3.34, p = 0.024), pulmonary disease (OR 1.68, CI 1.00 to 2.81, p = 0.049), American Society of Anesthesiologists (ASA) grade ≥ 3 (OR 2.37, CI 1.13 to 4.97, p = 0.022), and length of stay ≥ nine days (OR 1.98, CI 1.18 to 3.31, p = 0.009). A total of 38 (58.5%) COVID-19 cases were probably hospital acquired infections. The false-negative rate of a negative swab on admission was 0% in asymptomatic patients and 2.9% in symptomatic patients. COVID-19 was independently associated with a three times increased 30-day mortality rate. Nosocomial transmission may have accounted for approximately half of all cases during the first wave of the pandemic. Identification of risk factors for having COVID-19 on admission or acquiring COVID-19 in hospital may guide pathways for isolating or shielding patients respectively. Length of stay was the only modifiable risk factor, which emphasizes the importance of high-quality and timely care in this patient group. Cite this article: Bone Joint J 2021;103-B(5):888-897.


Age-Specific Effects of Visual Feature Binding.
Temporary binding of visual features enables objects to be stored and maintained in the visual working memory as a singular structure, irrespective of its inherent complexity. Although working memory capacity is reduced in aging, previous behavioral studies suggest that binding is preserved. Using event-related brain potentials (ERPs), we tested whether stimulus encoding is different in younger (N = 26, mean age = 28.5) and older (N = 22; mean age = 67.4) participants in a change detection task. The processing costs of binding were defined by the difference between feature-alone (color or shape) and feature-binding (color-shape) conditions. The behavioral data revealed that discrimination ability was reduced in the feature-binding condition, and that this effect was more attenuated in older participants. A corresponding ERP effect was not found in early components related to visual feature detection and processing (posterior N1 and frontal P2). However, the late positive complex (LPC) was more often expressed in the feature-binding condition, and the increase in amplitude was more pronounced in older participants. The LPC can be related to attentional allocation processes which might support the maintenance of the more complex stimulus representation in the binding task. However, the selective neural overactivation in the encoding phase observed in older participants does not prevent swap errors in the subsequent retrieval phase.


Unexpected diversity of aryl hydrocarbon receptors in non-mammalian vertebrates: insights from comparative genomics.
Ligand-activated receptors are well-known targets of environmental chemicals that disrupt endocrine signaling. Genomic approaches are providing new opportunities to understand the comparative biology and molecular evolution of these receptors. One example of this is the aryl hydrocarbon receptor (AHR), a basic-helix-loop-helix (bHLH)-Per-Arnt-Sim (PAS) transcription factor through which planar aromatic hydrocarbons cause altered gene expression and toxicity. In contrast to humans and other mammals, which possess a single AHR, teleosts such as the Atlantic killifish (Fundulus heteroclitus) have at least two AHRs (AHR1 and AHR2). Analysis of sequenced genomes has revealed additional, unexpected AHR diversity in non-mammalian vertebrates, including the chicken Gallus gallus (three predicted AHR genes), bony fishes such as the pufferfish Takifugu (formerly Fugu) rubripes (five AHR genes) and zebrafish Danio rerio (three AHR genes), and cartilaginous fishes such as the spiny dogfish Squalus acanthias (three AHR genes). In contrast, invertebrates appear to possess single AHRs that do not bind typical ligands of vertebrate AHRs. We suggest that AHR diversity in vertebrates arose through both gene and whole-genome duplications combined with lineage-specific gene loss, and that sensitivity to the developmental toxicity of planar aromatic hydrocarbons may have had its origin in the evolution of the ligand-binding capacity of the AHR in the chordate lineage. Comparative molecular and genomic studies are providing new insights into AHR diversity and function in non-mammalian species, revealing additional complexity in mechanisms by which environmental chemicals interfere with receptor-dependent signaling.


"Calories in, calories out" and macronutrient intake: the hope, hype, and science of calories.
One of the central tenets in obesity prevention and management is caloric restriction. This perspective presents salient features of how calories and energy balance matter, also called the "calories in, calories out" paradigm. Determinants of energy balance and relationships to dietary macronutrient content are reviewed. The rationale and features of the carbohydrate-insulin hypothesis postulate that carbohydrate restriction confers a metabolic advantage. According to this model, a large amount of fat intake is enabled without weight gain. Evidence concerning this possibility is detailed. The relationship and application of the laws of thermodynamics are then clarified with current primary research. Strong data indicate that energy balance is not materially changed during isocaloric substitution of dietary fats for carbohydrates. Results from a number of sources refute both the theory and effectiveness of the carbohydrate-insulin hypothesis. Instead, risk for obesity is primarily determined by total calorie intake.


Glycan array on aluminum oxide-coated glass slides through phosphonate chemistry.
A new type of glycan array covalently or noncovalently attached to aluminum oxide-coated glass (ACG) slides has been developed for studies of enzymatic reactions and protein binding. To prepare the noncovalent array, glycans with a polyfluorinated hydrocarbon (-C(8)F(17)) tail are spotted robotically onto the ACG slide surface containing a layer of polyfluorinated hydrocarbon terminated with phosphonate. After incubation and washing, the noncovalent array can be characterized by MS-TOF via ionization/desorption at a low laser energy without addition of matrix. A representative cellotetraose array was developed to study the activity and specificity of different cellulases and to differentiate the exo- and endoglucanase activities. To prepare the covalent array, glycans with a phosphonic acid tail were synthesized and spotted robotically onto the ACG slide surface. After incubation, the slides can be used directly for quantitative protein binding analysis. Compared to the preparation of glycan arrays on glass slides and other surfaces, this method of arraying using phosphonic acid reacting with ACG is more direct, convenient, and effective and represents a new platform for the high-throughput analysis of protein-glycan interactions.


Phase II trial evaluating the clinical and biologic effects of bevacizumab in unresectable hepatocellular carcinoma.
To determine the clinical and biologic effects of bevacizumab, an anti-vascular endothelial growth factor (VEGF) monoclonal antibody, in unresectable hepatocellular carcinoma (HCC). Adults with organ-confined HCC, Eastern Cooperative Oncology Group performance status of 0 to 2, and compensated liver disease were eligible. Patients received bevacizumab 5 mg/kg (n = 12) or 10 mg/kg (n = 34) every 2 weeks until disease progression or treatment-limiting toxicity. The primary objective was to determine whether bevacizumab improved the 6-month progression-free survival (PFS) rate from 40% to 60%. Secondary end points included determining the effects of bevacizumab on arterial enhancement and on plasma cytokine levels and the capacity of patients' plasma to support angiogenesis via an in vitro assay. The study included 46 patients, of whom six had objective responses (13%; 95% CI, 3% to 23%), and 65% were progression free at 6 months. Median PFS time was 6.9 months (95% CI, 6.5 to 9.1 months); overall survival rate was 53% at 1 year, 28% at 2 years, and 23% at 3 years. Grade 3 to 4 adverse events included hypertension (15%) and thrombosis (6%, including 4% with arterial thrombosis). Grade 3 or higher hemorrhage occurred in 11% of patients, including one fatal variceal bleed. Bevacizumab was associated with significant reductions in tumor enhancement by dynamic contrast-enhanced magnetic resonance imaging and reductions in circulating VEGF-A and stromal-derived factor-1 levels. Functional angiogenic activity was associated with VEGF-A levels in patient plasma. We observed significant clinical and biologic activity for bevacizumab in nonmetastatic HCC and achieved the primary study end point. Serious bleeding complications occurred in 11% of patients. Further evaluation is warranted in carefully selected patients.


Differences in lipopolysaccharide- and lipoteichoic acid-induced cytokine/chemokine expression.
To investigate differences in cytokine/chemokine release in response to lipoteichoic acid (LTA) or lipopolysaccharide (LPS) and contributing cellular mechanisms, in order to improve understanding of the pathogenesis of sepsis. Levels of cytokines/chemokines were measured in plasma and peritoneal lavage fluid of 10-week-old male mice (C57/B16) following intraperitoneal injection of LTA or LPS (250 µg), and in supernatants of murine J774.2 cells, immortalised blood monocytes, or isolated human monocytes treated with LTA or LPS (0-10 µg/ml). The role of cytokine/chemokine messenger RNA (mRNA) stability versus nuclear factor-kappaB (NF-κB) and activator protein-1 (AP-1) in mediating cytokine/chemokine release in J774 cells was also assessed. In mice, plasma levels of keratinocyte-derived chemokine (KC), macrophage inflammatory protein (MIP)-2, interleukin (IL)-10, interferon (IFN)-γ and tumour necrosis factor-alpha (TNF-α) and peritoneal lavage fluid levels of KC, MIP-2 and TNF-α increased significantly 1 h after LPS. Only KC and MIP-2 levels increased 1 h after LTA. LPS-treated (10 μg/ml) J774 cells released MIP-2, IL-10, IFN-γ and TNF-α but not KC (24 h), whereas cells treated with 10 μg/ml LTA released only MIP-2. LPS-stimulated human monocytes released IL-10 and IL-8 (24 h); by contrast, LTA-treated cells released only IL-8. LPS and LTA activated NF-κB and AP-1 in J774 cells. The protein synthesis inhibitor cycloheximide abolished LPS-induced IL-10 mRNA expression and increased LTA- and LPS-induced mRNA for MIP-2 in J774 cells. LTA and LPS, at clinically relevant concentrations, induced differential cytokine/chemokine release in vitro and in vivo, via effects distal to activation of NF-κB/AP-1 that might include chromatin remodelling or mRNA stability.


Effect of acidic and basic isoferritins on in vitro growth of human granulocyte-monocyte progenitors.
Acidic isoferritins have been previously found to be highly potent inhibitors of hematopoietic progenitors at concentrations of 10(-16) to 10(-18) mol/L, and it has been suggested that acidic isoferritin inhibitory activity plays a role in the regulation of normal hematopoiesis and also in the pathogenesis of leukemia. To characterize the ferritin species that affect the in vitro growth of human colony-forming unit-granulocyte-macrophage (CFU-GM), we tested different preparations of basic (L-subunit-rich) and acidic (H-subunit-rich) isoferritins. Three preparations of human liver (basic) ferritin did not show any effects on CFU-GM growth at concentrations up to 10(-9) mol/L, irrespective of the degree of glycosylation. Acidic isoferritins were purified both from HeLa cells and human heart. HeLa cell ferritin did not affect in vitro colony formation. One of two preparations of human heart ferritin, containing 5% glycosylated ferritin, showed a mean inhibition of 26% +/- 8% of the control at 10(-9) mol/L (P less than .02), whereas the other preparation, which contained no glycosylated ferritin, did not show any effect of CFU-GM growth. A preparation enriched for glycosylated acidic isoferritins from human heart was found to produce a mean inhibition of 32% +/- 11% of the control at 10(-9) mol/L (P less than .01), whereas another one was ineffective. A significant part of the inhibitory activity was removed by preincubation with the monoclonal antibody 2A4 directed against human heart ferritin. The present findings indicate that basic isoferritins, ie, the predominant ferritin type in human blood, have no effect on the growth of human CFU-GM, and this is in keeping with indirect clinical evidence. Inhibition of colony formation may be obtained by some preparations of acidic isoferritins that are rich in H subunits and bind to concanavalin A. The mechanism(s) responsible for this are not clear, but the effective concentrations are higher than those found in human blood both under normal conditions and in leukemia. At present, the physiologic significance of the observed inhibitory activity is uncertain.


Twelve tips for how institutional ethnography (IE) is conducted in health professions education research (HPE).
Institutional ethnography (IE), a term coined by sociologist Dorothy Smith, explores the nuances of institutions and their complex relationships in sociology. IE is an approach to studying and analysing social organization, and it provides a more holistic understanding of 'invisible' relationships that govern institutions and how those relationships interact with each other. Health sciences researchers in patient care, patient experience, and allied health professionals have recently become more interested in the use of this methodology and how to incorporate it into their research. However, in health professions education (HPE) there is little use of IE. We hypothesize this may be because of limited practical knowledge of this methodology. This paper serves as an introduction to the use of IE in HPE, describing the differences between IE and traditional ethnographies, recognizing the common pitfalls when utilising IE, and incorporating texts into IE. While ethnographies may be daunting to researchers less familiar with these approaches, the tips in this paper will provide an introduction and help educators and researchers successfully navigate the use of IE in health profession scholarship and education.


Prevalence and factors associated with long COVID in adults from Southern Brazil: findings from the PAMPA cohort.
Most COVID-19 survivors have reported experiencing persistent symptoms after the infection - these types of cases are known as long COVID. Since Brazil was an epicenter of the COVID-19 pandemic, a high burden of long COVID is expected. This study aimed to identify the prevalence and factors associated with long COVID in adults in Southern Brazil, analyzing data from the PAMPA cohort. Participants filled out a self-reported online questionnaire in June 2022. This study only included subjects who tested positive for COVID-19. Long COVID was defined by any symptoms that persisted for at least three months after the SARS-CoV-2 infection. Poisson's regression models with robust variance were used to identify factors associated with long COVID; and results were reported as prevalence ratios (PR) and respective 95% confidence intervals (95%CI). A total of 1,001 participants (77.4% women, mean age [SD] = 38.3 [11.9] years) were analyzed. The prevalence of long COVID among these patients was 77.4% (95%CI: 74.7; 79.9). The likelihood of long COVID was higher in unvaccinated participants (PR = 1.23, 95%CI: 1.06; 1.42), in those with chronic conditions (PR = 1.13, 95%CI: 1.04; 1.24), and in those who were hospitalized due to the COVID-19 infection (PR = 1.24, 95%CI: 1.16; 1.32). This prevalence was also higher in women (PR = 1.21, 95%CI: 1.09; 1.33) than in men. Physical activity was associated with a reduced likelihood of fatigue, neurological complications, coughing, and headaches as persistent symptoms after a COVID-19 infection. It was found that three out of four adults in Southern Brazil experienced long COVID. Public policies aiming to reduce the burden of long COVID must be prioritized, especially in groups that are at higher risk of developing this harmful condition.


Racial and ethnic disparities and perceptions of health care: does health plan type matter?
To examine whether racial and ethnic differences in the distribution of individuals across types of health plans explain differences in satisfaction and trust with their physicians. Data were derived from the 1998-1999 Community Tracking Household and Followback Studies and consisted of a nationwide sample of adults (18 years and older). The data were collected by telephone survey. Surveys were administered in English and Spanish. The response rate for the Household Survey was 63 percent, and the match rate for the Followback Survey was 59 percent. Multivariate analyses used regression methods to detect independent effects of respondent race and ethnicity on satisfaction and trust with physician, while controlling for enrollment in different types of health plans. Racial and ethnic minorities are more likely than whites to have lower levels of trust and satisfaction with their physician. The most prominent differences occurred within the Latino and Native American/Asian American/Pacific Islander/Other ("Other") populations. Plan type does not mitigate the relationship between race/ethnicity and trust and satisfaction for the overall adult population. Disparate levels of trust and satisfaction exist within ethnic and minority populations, even when controlling for the distribution of individuals across types of health plans. The results demonstrate a need to better understand the health care-related factors that drive disparate trust and satisfaction.


Diagnosis of human immunodeficiency virus infection by immunoassay using a molecularly cloned and expressed virus envelope polypeptide. Comparison to Western blot on 2707 consecutive serum samples.
To detect human immunodeficiency virus (HIV) antibodies in a simple enzyme-linked immunoassay (CBre3-EIA), we used an Escherichia coli-expressed polypeptide antigen, representing the carboxy-terminal third of the external membrane glycoprotein gene fused with the amino-terminal half of the transmembrane glycoprotein gene. Over a 3-month period, 2707 consecutive serum samples referred for confirmatory testing for human T-lymphotrophic virus type III (HTLV-III) antibodies were evaluated by both Western blot and CBre3-EIA. On a single determination for each sample, the CBre3-EIA was found to have an estimated sensitivity (99.9%) and specificity (99.1%) similar or superior to the more cumbersome Western blot method. This study shows that all HIV-seropositive subjects have antibodies to the virus envelope protein; no other virus antigens are required for construction of highly sensitive immunoassays.


Extracting sequence features to predict protein-DNA interactions: a comparative study.
Predicting how and where proteins, especially transcription factors (TFs), interact with DNA is an important problem in biology. We present here a systematic study of predictive modeling approaches to the TF-DNA binding problem, which have been frequently shown to be more efficient than those methods only based on position-specific weight matrices (PWMs). In these approaches, a statistical relationship between genomic sequences and gene expression or ChIP-binding intensities is inferred through a regression framework; and influential sequence features are identified by variable selection. We examine a few state-of-the-art learning methods including stepwise linear regression, multivariate adaptive regression splines, neural networks, support vector machines, boosting and Bayesian additive regression trees (BART). These methods are applied to both simulated datasets and two whole-genome ChIP-chip datasets on the TFs Oct4 and Sox2, respectively, in human embryonic stem cells. We find that, with proper learning methods, predictive modeling approaches can significantly improve the predictive power and identify more biologically interesting features, such as TF-TF interactions, than the PWM approach. In particular, BART and boosting show the best and the most robust overall performance among all the methods.


Synaptic transmission at N-methyl-D-aspartate receptors in the proximal retina of the mudpuppy.
The effects of excitatory amino acid analogues and antagonists on retinal ganglion cells were studied using intracellular recording in the superfused mudpuppy eyecup preparation. Aspartate, glutamate, quisqualate (QA), kainate (KA) and N-methylaspartate (NMA) caused depolarization and decreased input resistance in all classes of ganglion cells. The order of sensitivity was QA greater than or equal to KA greater than NMA greater than aspartate greater than or equal to glutamate. All of these agonists were effective when transmitter release was blocked with 4 mM-Co2+ or Mn2+, indicating that they acted at receptor sites on the ganglion cells. At a concentration of 250 microM, 2-amino-5-phosphonovalerate (APV) blocked the responses of all ganglion cells to NMA, but not to QA or KA, indicating that NMA acts at different receptor sites from QA or KA. Responses to bath-applied aspartate and glutamate were reduced slightly or not at all in the presence of APV, indicating that they were acting mainly at non-NMDA (N-methyl-D-aspartate) receptors. In all ganglion cells 250 microM-APV strongly suppressed the sustained responses driven by the 'on'-pathway but not those driven by the 'off'-pathway. In most on-off ganglion cells the transient excitatory responses at 'light on' and 'light off' were not reduced by 500 microM-APV. APV-resistant transient excitatory responses were also present in some on-centre ganglion cells. APV did not block the transient inhibitory responses in any class of ganglion cells. At concentrations which blocked the sustained responses of ganglion cells, APV did not affect the sustained responses of bipolar cells, indicating that it acted at sites which were post-synaptic to bipolar cells. The simplest interpretation of these results is that the transmitter released by depolarizing bipolar cells acts at NMDA receptors on sustained depolarizing amacrine and ganglion cells. It may act at non-NMDA receptors at synapses which produce transient excitatory responses, but this could not be proved. The transmitter released by hyperpolarizing bipolar cells does not appear to act at NMDA receptors on any post-synaptic cells.


Radioimmunoassay of serum myoglobin in neuromuscular diseases.
Radioimmunoassay of serum myoglobin was performed in 85 patients with muscular symptoms. Elevated levels were found in 93% of patients with myogenic myopathy, in 54% with myasthenia gravis and in 50% with neurogenic myopathy. All 11 patients with polymyositis had elevated myoglobin concentrations. In six of seven patients with polymyositis, who were followed up with repeated determinations, a clear relationship between myoglobin levels and clinical course was found. In general serum myoglobin seemed to be a more sensitive indicator of muscle disease than creatine kinase.


The Hirsute woman: challenges in evaluation and management.
To review the etiology, pathogenesis, diagnostic approach, and management of hirsutism. We discuss the clinical course of hirsutism and provide our recommendations on the various treatment options available. Hirsutism is a common clinical problem characterized by the presence of increased terminal hair growth in androgen-dependent areas of the skin. The development of hirsutism depends on the presence of the pilosebaceous unit, which is genetically determined, as well as the presence of the androgen receptor and intracellular 5α-reductase activity, which converts testosterone to its more active metabolite, dihydrotestosterone. A detailed history and physical examination and the following laboratory tests can diagnose most causes of hirsutism: early-morning follicular phase measurement of total testosterone, testosterone not bound to sex hormone-binding globulin, dehydroepiandrosterone sulfate, 17-hydroxyprogesterone, prolactin, and thyrotropin levels. Oral contraceptive preparations may be effective monotherapy for mild hirsutism. For the treatment of more severe hirsutism, oral contraceptive pills combined with spironolactone are as effective as oral contraceptive pills containing cyproterone acetate, which are not available in the United States. Because of teratogenicity, spironolactone should be used with caution in premenopausal women when it is administered without an oral contraceptive pill. Metformin is an alternative therapy for hirsutism in women with polycystic ovary syndrome who have other indications for metformin use. Metformin is not as effective as antiandrogens for the management of hirsutism. The use of glucocorticoids, finasteride, or flutamide is not recommended. Hirsutism can be evaluated with a detailed history and physical examination and a limited number of hormonal tests. Serious disorders presenting as hirsutism are rare and can be excluded with the recommended evaluation. Treatment is targeted at reducing the production and bioavailability of testosterone, as well as blocking target tissue androgen action.


Dioxin exposure and age of pubertal onset among Russian boys.
Animal data demonstrate associations of dioxin, furan, and polychlorinated biphenyl (PCB) exposures with altered male gonadal maturation. It is unclear whether these associations apply to human populations. We investigated the association of dioxins, furans, PCBs, and corresponding toxic equivalent (TEQ) concentrations with pubertal onset among boys in a dioxin-contaminated region. Between 2003 and 2005, 499 boys 8-9 years of age were enrolled in a longitudinal study in Chapaevsk, Russia. Pubertal onset [stage 2 or higher for genitalia (G2+) or testicular volume (TV) > 3 mL] was assessed annually between ages 8 and 12 years. Serum levels at enrollment were analyzed by the Centers for Disease Control and Prevention, Atlanta, Georgia, USA. We used Cox proportional hazards models to assess age at pubertal onset as a function of exposure adjusted for potential confounders. We conducted sensitivity analyses excluding boys with pubertal onset at enrollment. The median (range) total serum TEQ concentration was 21 (4-175) pg/g lipid, approximately three times higher than values in European children. At enrollment, boys were generally healthy and normal weight (mean body mass index, 15.9 kg/m2), with 30% having entered puberty by G2+ and 14% by TV criteria. Higher dioxin TEQs were associated with later pubertal onset by TV (hazard ratio = 0.68, 95% confidence interval, 0.49-0.95 for the highest compared with the lowest quartile). Similar associations were observed for 2,3,7,8-tetrachlorodibenzo-p-dioxin and dioxin concentrations for TV but not G2+. Results were robust to sensitivity analyses. Findings support an association of higher peripubertal serum dioxin TEQs and concentrations with later male pubertal onset reflected in delayed testicular maturation.


Characterization of the Biodistribution and Systemic Absorption of TT-173, a New Hemostatic Agent of Recombinant Human Tissue Factor, Using Radiolabeling with 18F.
TT-173 is the first topical hemostatic agent based on tissue factor. To prevent thromboembolic events and intravascular coagulation it is necessary to rule out the systemic absorption of new bioactive hemostats. Here, we radiolabeled TT-173 with [18F]SBF to characterize its systemic absorption and biodistribution. A group of rats were administered intravenously with [18F]TT-173 and were subjected to a positron emission tomography study. A second group of animals received the [18F]TT-173 topically over a skin lesion in the flank. Topical absorption was quantified and the biodistribution patterns observed were compared. After topical application, low amounts of [18F]TT-173 were absorbed and distributed without relevant accumulation in any organ. On the other hand, after intravenous injection, [18F]TT-173 accumulated in lungs, liver and spleen, consistent with intravascular clot formation and the posterior capillary trapping and phagocytosis by the reticuloendothelial system. In both cases, a substantial concentration of radioactive product was found in urine consistent with renal excretion of degradation products of [18F]TT-173. After topical application, [18F]TT-173 did not show a significant systemic accumulation in animal organs. Minor radioactive concentration found in lungs, liver and spleen likely corresponds with fragments of the product without procoagulant activity. Radiolabeling with [18F]SFB enables the characterization of systemic absorption and biodistribution of new topical hemostats like TT-173.


Children's use of visual summary statistics for material categorization.
Although adults' ability to recognize materials from complex natural images has been well characterized, we still know very little about the development of material perception. When do children exhibit adult-like abilities to categorize materials? What visual features do they use to do so as a function of age and material category? In the present study, we attempted to address both of these issues in two experiments that we administered to school-age children (5-10 years old) and adults. In both tasks, we asked our participants to categorize natural materials (metal, stone, water, and wood) using original images of these materials as well as synthetic images made with the Portilla-Simoncelli algorithm. By including synthetic images in our stimulus set, we were able to assess both how material categorization develops during childhood and how visual summary statistics are recruited for material perception across age groups. We observed that when asked to provide category labels for individual images (Experiment 1), young children were disproportionately bad at categorizing some materials after they were synthesized, suggesting material-specific changes in information use over the course of development. However, when asked to match real and synthetic images according to material category without labeling (Experiment 2), these effects were weakened. We conclude that while children have adult-like abilities to encode and compare images based on summary statistics, the mapping between summary statistics and category labels undergoes prolonged development during childhood.


Single-institution outcomes in the first year of pancreas transplantation from brain-dead donors.
Under a revision to the law in 2010, the number of pancreas transplantations from brain-dead donors has been increasing in Japan. We started a new Pancreatic Transplant Program at Fujita Health University Hospital in September 2012. A total of 11 cases of pancreas transplantation from brain-dead donors performed at Fujita Health University Hospital were analyzed in terms of the background characteristics of the donors and recipients and the outcomes. The mean age of the recipients was 45.2 years, and all recipients had a long-term history of diabetes (mean: 32.5 years). In the simultaneous pancreas and kidney transplantation (SPK) cases, the patients also had a long history of hemodialysis (mean: 8.0 years). Although the average donor age was 42.5 years, 90% of the donors were marginal donors, defined according to the following factors: (1) >45 years old, (2) death from cardiovascular disease, (3) episodes of cardiac arrest, (4) use of high doses of catecholamines. The pancreatic graft survival rate was 100%, although 1 patient required a small amount of insulin to maintain euglycemia. In addition, the kidney graft survival rate was also 100% in the SPK cases. The new Pancreatic Transplant Program at Fujita Health University has provided excellent outcomes for type 1 diabetic patients.


The first immunoglobulin-like domain of HveC is sufficient to bind herpes simplex virus gD with full affinity, while the third domain is involved in oligomerization of HveC.
The human herpesvirus entry mediator C (HveC/PRR1) is a member of the immunoglobulin family used as a cellular receptor by the alphaherpesviruses herpes simplex virus (HSV), pseudorabies virus, and bovine herpesvirus type 1. We previously demonstrated direct binding of the purified HveC ectodomain to purified HSV type 1 (HSV-1) and HSV-2 glycoprotein D (gD). Here, using a baculovirus expression system, we constructed and purified truncated forms of the receptor containing one [HveC(143t)], two [HveC(245t)], or all three immunoglobulin-like domains [HveC(346t)] of the extracellular region. All three constructs were equally able to compete with HveC(346t) for gD binding. The variable domain bound to virions and blocked HSV infection as well as HveC(346t). Thus, all of the binding to the receptor occurs within the first immunoglobulin-like domain, or V-domain, of HveC. These data confirm and extend those of Cocchi et al. (F. Cocchi, M. Lopez, L. Menotti, M. Aoubala, P. Dubreuil, and G. Campadelli-Fiume, Proc. Natl. Acad. Sci. USA 95:15700, 1998). Using biosensor analysis, we measured the affinity of binding of gD from HSV strains KOS and rid1 to two forms of HveC. Soluble gDs from the KOS strain of HSV-1 had the same affinity for HveC(346t) and HveC(143t). The mutant gD(rid1t) had an increased affinity for HveC(346t) and HveC(143t) due to a faster rate of complex formation. Interestingly, we found that HveC(346t) was a tetramer in solution, whereas HveC(143t) and HveC(245t) formed dimers, suggesting a role for the third immunoglobulin-like domain of HveC in oligomerization. In addition, the stoichiometry between gD and HveC appeared to be influenced by the level of HveC oligomerization.


Folic acid supplementation and neural tube defect recurrence prevention.
It is well established that women who have had a pregnancy affected by a neural tube defect (NTD) have an elevated risk of a subsequent NTD-affected pregnancy and that a high dose (4 mg/day) of folic acid taken around the time of conception prevents most recurrences of NTDs. We reviewed the literature to identify studies that quantify the reduction in risk if women with a prior-NTD affected pregnancy consistently take folic acid before and during a subsequent pregnancy and the effectiveness of NTD recurrence prevention programs in increasing the percentage of women who consistently consume folic acid supplements. A meta-analysis of randomized trials of folic acid for the prevention of recurrent NTDs indicates a 69% reduction in recurrence risk if analyzed on an intention-to-treat basis and an 87% reduction among those women who took supplements prior to the beginning of pregnancy. Observational studies report reductions in recurrence risk of 85% to 100% among women taking folic acid prior to subsequent pregnancies. The percentage of women who take folic acid prior to a subsequent pregnancy has been reported to vary from 33% to 85%, varying with the demographic background and the intensity of folic acid counseling efforts. Targeted folic acid information and counseling provided to women with an NTD-affected pregnancy has been demonstrated to substantially reduce the risk of recurrent NTDs and is feasible to implement on a public health basis.


Drug resistant tuberculosis in sub-Saharan Africa: an estimation of incidence and cost for the year 2000.
Rwanda. To evaluate the tuberculosis (TB) drug resistance in Rwanda on smear positive sputa, collected prospectively at the start of the National TB programme, before the start of any treatment or retreatment. To adapt the scenarios of Schulzer et al (1992) to the data from Rwanda, in order to obtain an estimation of the number of drug resistant and multi-drug resistant (MDR) TB cases expected by the year 2000. A total of 298 specimens (236 randomly selected new cases and 62 retreated cases), collected between January 1991 and June 1993, were sent to Belgium in 1% cetylpyridinium chloride. Drug resistance was determined using the proportion method. MDR, i.e. resistance to at least rifampicin (R) and isoniazid (H), was observed in 3 (1.3%) out of 236 new cases and in 4 (6.5%) out of 62 treated cases. For new cases, single drug resistance to H, R and ethambutol (E) was 3%, 0.4% respectively; for treated cases it was 14.5%, 1.6% and 6.5 respectively. Based on the estimate of the size of the TB problem in sub-Saharan Africa by the year of 2000 (Schulzer), we calculated that the region should expect between 15,543 and 223, 417 cases of MDR, all forms combined (between 2.3 and 32.7 per 100,000 inhabitants), by the end of the century. The results from Rwanda during the period studied do not appear dramatic. However, for some other developing countries, they may just represent the tip of the iceberg. Rapid recognition of resistance to the major antituberculosis agents is essential for control of TB. Integration of an MDR increase factor into the TB budget would not dramatically increase the total TB budget. Our data urgently point the the need for drug resistance surveys, followed by continuous drug resistance monitoring in high TB prevalence areas.


Silencing the hydroxyproline-rich glycopeptide systemin precursor in two accessions of Nicotiana attenuata alters flower morphology and rates of self-pollination.
Systemins and their hydroxyproline-rich glycopeptide systemin (ppHS) subfamily members are known to mediate antiherbivore defenses in some solanaceous taxa but not others; functions other than in defense remain largely unexplored. Nicotiana attenuata's ppHS is known not to function in herbivore defense. NappHS transcripts are abundant in flowers, particularly in pistils, and when two N. attenuata accessions from Utah and Arizona were transformed to silence NappHS by RNAi (IRsys), seed capsule production and seed number per capsule were reduced in both accessions. These reductions in reproductive performance could not be attributed to impaired pollen or ovule viability; hand-pollination of all IRsys lines of both accessions restored seed production per capsule to levels found in wild-type plants. Rather, changes in flower morphology that decreased the efficiency of self-pollination are likely responsible: IRsys plants of both accessions have flowers with pistils that protrude beyond their anthers. Because these changes in flower morphology are reminiscent of CORONATINE-INSENSITIVE1-silenced N. attenuata plants, we measured jasmonates (JAs) and their biosynthetic transcripts in different floral developmental stages, and found levels of JA-isoleucine (Ile)/leucine and threonine deaminase transcripts, which are abundant in wild-type pistils, to be significantly reduced in IRsys buds and flowers. Threonine deaminase supplies Ile for JA-Ile biosynthesis, and we propose that ppHS mediates JA signaling during flower development and thereby changes flower morphology. These results suggest that the function of ppHS family members in N. attenuata may have diversified to modulate flower morphology and thereby outcrossing rates in response to biotic or abiotic stresses.


Transdifferentiation and remodeling of post-embryonic C. elegans cells by a single transcription factor.
Terminally differentiated post-mitotic cells are generally considered irreversibly developmentally locked, i.e. incapable of being reprogrammed in vivo into entirely different cell types. We found that brief expression of a single transcription factor, the ELT-7 GATA factor, can convert the identity of fully differentiated, highly specialized non-endodermal cells of the pharynx into fully differentiated intestinal cells in intact larvae and adult Caenorhabditis elegans. Stable expression of intestine-specific molecular markers parallels loss of markers for the original differentiated pharynx state; hence, there is no apparent requirement for a dedifferentiated intermediate during the transdifferentiation process. Based on high-resolution morphological characteristics, the transdifferentiated cells become remodeled to resemble typical intestinal cells at the level of both the cell surface and internal organelles. Thus, post-mitotic cells, though terminally differentiated, remain plastic to transdifferentiation across germ layer lineage boundaries and can be remodeled to adopt the characteristics of a new cell identity without removal of inhibitory factors. Our findings establish a simple model to investigate how cell context influences forced transdifferentiation of mature cells.


Comparative efficacy and tolerability of probiotic, prebiotic, and synbiotic formulations for adult patients with mild-moderate ulcerative colitis in an adjunctive therapy: A network meta-analysis.
Probiotics, prebiotics, and synbiotics (PPS) have been widely used as adjuvant treatments in patients with ulcerative colitis (UC) in recent years. However, the most effective formulations of PPS have yet to be identified. We thus aimed to compare the efficacy and tolerability of different PPS formulations for mild-moderate UC. We searched PubMed, Embase, Web of Science, and Cochrane CENTRAL from inception to June 24, 2023 for double-blind randomized controlled trials. We used a frequentist approach in random-effects models for network meta-analysis and the Grading of Recommendations Assessment, Development, and Evaluation approach to evaluate the certainty of evidence. We analysed data from 20 trials involving 1153 patients. The combinations of specific strains of Lactobacillus and Bifidobacterium (CLB) (odds ratio (OR), 3.85; 95 % confidence interval (CI), 1.40-10.60; low certainty) and combinations of specific strains of Lactobacillus, Bifidobacterium, and Streptococcus (CLBS) (OR, 2.20; 95 % CI, 1.47-3.28; low certainty) significantly increased the clinical remission rate in intention-to-treat analysis (ITT) when compared to placebo. Similarly, compared with placebo, the two combinations significantly reduced clinical activity scores (standardized mean difference (SMD), -1.17 (95 % CI, -1.68 to -0.65), low certainty; and SMD, -1.33 (95 % CI, -1.81 to -0.86), low certainty, respectively). Hierarchical cluster analyses showed the two combinations formed clusters with high efficacy (clinical remission in ITT and clinical activity score) and tolerability (withdrawal due to worsening symptoms) within 12 weeks. In this systematic review, we found CLB and CLBS demonstrated a clinical benefit in adjuvant treatments, with a comparable tolerability and safety profile to placebo. Further trials are needed. CRD42022344905.


The nature of the link between potassium transport and phosphate transport in Escherichia coli.
A series of mutants of Escherichia coli, combining defects in either of the two phosphate transport systems with defects in one or more of the potassium transport systems, was used to study the nature of the previously observed obligatory requirement for each one of these ions in the transport of the other. The results show that no pair of systems is obligatorily linked, and that either ion can be transported by any one of its systems, provided that a means of entry for the other ion is available. Furthermore, in the total absence of Pi, K+ entry accompanies the transport of other anions, such as aspartate, glutamate, sn-glycero-3-phosphate and glucose 6-phosphate. The results indicate that Pi and the other anions enter by symport with protons, and that a simultaneous K+/H+ exchange, which would serve to maintain the intracellular pH, is responsible for the observed K+ 'symport' with these anions.


Unexpected steric effects of "remote" alkyl groups on the rate of conjugate additions to alkyl alpha,beta-ethylenic sulfones, sulfoxides, and esters.
Examination of conjugated ethylenic sulfones, sulfoxides, and esters in Michael-type addition reactions reveals, for the first time, that the size of the heteroatom-attached alkyl group affects the rate of conjugate addition. Molecular modeling strongly suggests that what are generally considered to be "remote" alkyl groups in -CbetaH=CalphaHS(O)n-alkyl systems and -CH2CbetaH=CalphaHCOO-alkyl systems are actually not remote from the beta-carbon atom of the Michael accepting unit. Molecular modeling clearly shows that the alkyl groups in these Michael acceptors shield the beta-carbons in the following order: Et<i-Pr<t-Bu. Competition experiments establish the relative rates of Michael additions to be in the following order: Et>i-Pr>t-Bu.


Intraspinal localization of the somatosensory evoked potential.
Somatosensory evoked potentials (SEPs) are used widely for monitoring neurophysiological function in experimental spinal injury. Yet the spinal pathways for SEP conduction remain unclear. Consequently, we sought to define specific changes in the SEP after interruption of selected spinal pathways. We activated cortical SEPs with sciatic nerve stimulation in 11 anesthetized (25 mg of pentobarbital per kg) cats after a multilevel thoracic laminectomy. The most consistent wave from component was an initial positivity (IP) at a 17- to 19-ms onset latency. We then used a Cavitron ultrasonic surgical aspirator to interrupt specific spinal pathways. A unilateral dorsal column lesion abolished the ipsilateral IP, but did not affect conduction in the contralateral column. Bilateral dorsal column lesions obliterated the IP, but sometimes left some longer latency components. Interruption of all but the ventral columns abolished the SEPs. When we interrupted all spinal pathways but the dorsal columns, an intact IP remained. In fact, a distinct IP was conducted through a single dorsal column after the division of all other spinal cord pathways. We concluded that, in the barbiturate-anesthetized cat: (a) the most consistent SEP wave form is an initial positivity at a 17- to 19-ms onset latency, (b) the integrity of the dorsal columns is both necessary and sufficient to conduct a normal-appearing IP component of the SEP, (c) the lateral columns may carry some longer latency component of the SEP, (d) the ventral columns carry no component of the SEP, and (e) bilateral recording may be useful for detecting asymmetry of injury.


[Satisfactory evolution of a patient diagnosed in childhood with Bruton's disease].
Bruton's agammaglobulinemia is a primary immunodeficiency with a disease onset during the first months of age, when the maternal serum immunoglobulin levels decrease. It is characterized by recurrent infections and agammaglobulinemia. We report the case of a 6-year-old male patient with third-degree consanguinity, product of a third pregnancy and complete immunization scheme. He had a history of oral candidiasis at the age of 3 months, chicken pox at the age of 7 months, and two episodes of complicated bronchopneumonia at the age of 1 year and 6 years. He was admitted to the hospital because of fever and cough. Examination of the chest showed rales and right basilar hypoventilation, and a blood cell count revealed leukocytosis and neutrophilia. The diagnosis of pneumonia was made. He was treated with IV antibiotics. Serum immunoglobulins were reported to be low (IgM 55 mg/dL, IgA 0.9 mg/dL, and IgG 199 mg/dL). With these findings the clinical diagnosis of X-linked agammaglobulinemia (ALX) was concluded. A molecular test was performed fining a BTK gene confirming the diagnosis of Bruton's disease. Therapy with intravenous IgG was started every 21 days. During his evolution, he presented three episodes of rhinosinusitis, one of suppurative otitis media, and four events of pneumonia that required 37 days of hospitalization. After hospital discharge, the patient was free of infections and he returned to his daily activities. In cases of recurrent and severe respiratory infections in children, we must consider primary immunodeficiency disease in the differential diagnosis, mainly antibiotic deficiency. Early diagnosis and treatment improves the survival and quality of life in these patients.


Endothelial nitric oxide synthase mediates the nitric oxide component of reflex cutaneous vasodilatation during dynamic exercise in humans.
Recent data suggests neuronal nitric oxide synthase (nNOS) mediates the NO component of reflex cutaneous vasodilatation with passive heat stress. We tested the hypothesis that nNOS inhibition would attenuate reflex cutaneous vasodilatation during sustained dynamic exercise in young healthy humans. All subjects first performed an incremental V̇O2, peak test to exhaustion on a custom-built supine cycle ergometer. On a separate day, subjects were instrumented with four intradermal microdialysis fibres on the forearm and each randomly assigned as: (1) lactated Ringer's (control); (2) 20 mm Nω-nitro-l-arginine methyl ester hydrochloride (non-selective NOS inhibitor); (3) 5 mm N-propyl-l-arginine (nNOS inhibitor); and (4) 10 mm N(5)-(1-iminoethyl)-l-ornithine dihydrochloride [endothelial NOS (eNOS) inhibitor]. Following microdialysis placement, subjects performed supine cycling with the experimental arm at heart level at 60% V̇O2, peak for a period sufficient to raise core temperature 0.8°C. At the end of cycling, all microdialysis sites were locally heated to 43°C and sodium nitroprusside was perfused to elicit maximal vasodilatation. Mean arterial pressure, skin blood flow via laser-Doppler flowmetry and core temperature via ingestible telemetric pill were measured continuously; cutaneous vascular conductance (CVC) was calculated as laser-Doppler flowmetry/mean arterial pressure and normalized to maximum. There was no significant difference between control (58 ± 2%CVCmax) and nNOS-inhibited (56 ± 3%CVCmax) sites in response to exercise-induced hyperthermia. The increase in CVC at eNOS-inhibited (41 ± 3%CVCmax) and non-selective NOS-inhibited (40 ± 4%CVCmax) sites were significantly attenuated compared to control and nNOS-inhibited (P < 0.001 all conditions) but there was no difference between eNOS-inhibited and non-selective NOS-inhibited sites. These data suggest eNOS, not nNOS, mediate NO synthesis during reflex cutaneous vasodilatation with sustained dynamic exercise.


MicroRNA transcriptome analysis on hypertrophy of ligamentum flavum in patients with lumbar spinal stenosis.
Molecular pathways involved in ligamentum flavum (LF) hypertrophy are still unclarified. The purpose of this study was to characterize LF hypertrophy by microRNA (miRNA) profiling according to the classification of lumbar spinal stenosis (LSS). Classification of patients with LSS into ligamentous and non-ligamentous cases was conducted by clinical observation and the morphometric parameter adopting the LF/spinal canal area ratio (LSAR) from measurements of magnetic resonance imaging (MRI) T2 weighed images. LF from patients with ligamentous stenosis (n=10) were considered as the degenerative hypertrophied samples, and those from patients with non-ligamentous LSS (n=7) and lumbar disc herniation (LDH, n=3) were used as non-hypertrophied controls. Profiling of miRNA from all samples was conducted by Agilent microarray. Microarray data analysis was performed with GeneSpring GX, and pathway analysis was performed using Ingenuity Pathway Analysis. The mean LSAR in the ligamentous group was significantly higher than that in the control group (0.662±0.154 vs 0.301±0.068, p=0.0000171). Ten significantly differentially expressed miRNA were identified and taken as a signature of LF hypertrophy: nine miRNA showed down-regulated expression, and one showed up-regulated expression in the ligamentous LF. Among those, miR-423-5p (rs=-0.473, p<0.05), miR-4306 (rs=-0.628, p<0.01), miR-516b-5p (rs=-0.629, p<0.01), and miR-497-5p (rs=0.461, p<0.05) were correlated to the LSAR. Pathway analysis predicted aryl hydrocarbon receptor signaling (p<0.01), Wnt/β-catenin signaling (p<0.01), and insulin receptor signaling (p<0.05) as canonical pathways associated with the miRNA signature. Classification based on quantification of the MRI axial image is useful for studying hypertrophy of the LF. Aryl hydrocarbon receptor and Wnt/β-catenin signaling may be involved in LF hypertrophy.


Symmetrical peripheral gangrene and Roth spots in a patient with infective endocarditis.
We present a case of a young man with symmetrical peripheral gangrene (SPG) resulting from Streptococcus viridans-related infective endocarditis, an association which has not previously been reported. SPG is associated with up to 40% mortality and may necessitate amputation; early identification and treatment of the precipitating factors is very important.


Establishment of mouse erythroleukemia cell lines expressing complete Influenza C virus CM2 protein or chimeric protein consisting of CM2 and Influenza A virus M2.
The role of the Influenza C virus (ICV) CM2 protein in virus replication as well as its precise function as an ion channel remains to be elucidated. For this purpose, we established a CM2-expressing mouse erythroleukemia (MEL) cell line and determined the biochemical characteristics of the expressed CM2. The features of the expressed CM2 were similar to those of the viral CM2 synthesized in ICV-infected cells. Furthermore, we established MEL cell line expressing a chimeric protein consisting of characteristic regions of CM2 and Influenza A virus (IAV) M2 protein that could be helpful in elucidation of the specific ion conductance properties.


An open label study on the supplementation of Gymnema sylvestre in type 2 diabetics.
Diabetes mellitus is a complex metabolic disorder characterized by chronic hyperglycemia, and associated with long-term damage and dysfunction of various organs. Management of diabetes is therefore vital and involves maintaining euglycemia as much as possible by reducing blood glucose and by increasing insulin sensitivity and peripheral glucose uptake. Ayurveda has promoted the management of diabetes by regulating carbohydrate metabolism using several medicinal herbs, one of which is Gymnema sylvestre (GS). GS has been used in parts of India as a hypoglycemic agent and the results have been encouraging. Accordingly, we planned a quasi-experimental study to investigate the efficacy of the herb among type 2 diabetics. Patients enrolled from free-living population were purposively assigned to experimental or control groups, based on their willingness to participate in the study. The experimental group was supplemented with 500 mg of the herb per day for a period of 3 months, and the efficacy of the herb was assessed through a battery of clinical and biochemical tests. Supplementation of the diet with GS reduced polyphagia, fatigue, blood glucose (fasting and post-prandial), and glycated hemoglobin and there was a favorable shift in lipid profiles and in other clinico-biochemical tests. These findings suggest a beneficial effect of GS in the management of diabetes mellitus.


The neuroimmune guidance cue netrin-1: a new therapeutic target in cardiovascular disease.
Netrins are a family of proteins involved in cell migration and axon guidance during embryogenesis. The different functions and mechanisms of action of this family of proteins have been better characterized with the study of netrin-1. They are chemotropic and act as a bifunctional regulator of neuron migration. Apart from its role in the central nervous system, researchers have proven that netrin-1 plays a role in the development and formation of non-neural tissue; netrin-1 is thereby involved in regulation of cancers, cardiovascular diseases, kidney diseases, and other diseases. Concerning the cardiovascular realm, netrin-1 promotes angiogenesis and accelerates atherosclerosis, protects the heart against ischemia-reperfusion injury, and reduces the infarct size. These findings make the neuroimmune guidance cue netrin-1 an important therapeutic target. This work seeks to review the subject based on studies that have been conducted over the past decade to identify the perspectives and extent of the research on this protein in the field of cardiology.


D-carbon: Ab initio study of a novel carbon allotrope.
By means of ab initio computations and the global minimum structure search method, we have investigated structural, mechanical, and electronic properties of D-carbon, a crystalline orthorhombic sp3 carbon allotrope (space group Pmma [ D2h5 ] with 6 atoms per cell). Total-energy calculations demonstrate that D-carbon is energetically more favorable than the previously proposed T6 structure (with 6 atoms per cell) as well as many others. This novel phase is dynamically, mechanically, and thermally stable at zero pressure and more stable than graphite beyond 63.7 GPa. D-carbon is a semiconductor with a bandgap of 4.33 eV, less than diamond's gap (5.47 eV). The simulated X-ray diffraction pattern is in satisfactory agreement with previous experimental data in chimney or detonation soot, suggesting its possible presence in the specimen.


Hierarchical IL-5 expression defines a subpopulation of highly differentiated human Th2 cells.
Each of the three Th2 cytokine genes, IL-4, IL-5, and IL-13, has different functions. We hypothesized that Th2 heterogeneity could yield Th2 subpopulations with different cytokine expression and effector functions. Using multiple approaches, we demonstrate that human Th2 cells are composed of two major subpopulations: a minority IL-5(+) (IL-5(+), IL-4(+), IL-13(+)) and majority IL-5(-) Th2 (IL-5(-), IL-4(+), IL-13(+)) population. IL-5(+) Th2 cells comprised only 20% of all Th2 cells. Serial rounds of in vitro differentiation initially yielded IL-5(-) Th2, but required multiple rounds of differentiation to generate IL-5(+) Th2 cells. IL-5(+) Th2 cells expressed less CD27 and greater programmed cell death-1 than IL-5(-) Th2 cells, consistent with their being more highly differentiated, Ag-exposed memory cells. IL-5(+) Th2 cells expressed greater IL-4, IL-13, and GATA-3 relative to IL-5(-) Th2 cells. GATA-3 and H3K4me(3) binding to the IL5 promoter (IL5p) was greater in IL-5(+) relative to IL-5(-) Th2 cells, whereas there was no difference in their binding to the IL4p and IL13p. Conversely, H3K27me(3) binding to the IL5p was greater in IL-5(-) Th2 cells. These findings demonstrate Th2 lineage heterogeneity, in which the IL5 gene is regulated in a hierarchical manner relative to other Th2 genes. IL-5(+) Th2 cells are phenotypically distinct and have epigenetic changes consistent with greater IL5p accessibility. Recurrent antigenic exposure preferentially drives the differentiation of IL-5(+) Th2 cells. These results demonstrate that IL-5(+) and IL-5(-) Th2 cells, respectively, represent more and less highly differentiated Th2 cell subpopulations. Such Th2 subpopulations may differentially contribute to Th2-driven pathology.


Spatial and Pregnancy-Related Changes in the Protein, Amino Acid, and Carbohydrate Composition of Bovine Oviduct Fluid.
Knowledge of how the biochemical composition of the bovine oviduct is altered due to the oviduct anatomy or the presence of an embryo is lacking. Thus, the aim of this study was to assess the effect of (І) oviduct anatomy and (ІІ) embryo presence on oviductal fluid (OF) protein, amino acid, and carbohydrate composition. Cross-bred beef heifers (n = 19) were synchronized and those in standing estrus were randomly allocated to a cyclic (non-bred) or pregnant (artificially inseminated) group. All heifers were slaughtered on Day 3 after estrus. The oviducts ipsilateral to the corpus luteum from each animal were isolated, straightened and cut, separating ampulla and isthmus. Each portion was flushed with 500 µl of PBS enabling recovery of the oocyte/embryo. Recovered unfertilized oocytes (cyclic group) and embryos (8-cell embryos; pregnant group) were located in the isthmus of the oviduct. Samples of flushing medium from the isthmus and ampulla were used for proteomic (n = 2 per group), amino acid (n = 5), and carbohydrate (n = 5) analysis. For proteomic analysis, total protein from cyclic and pregnant samples were labelled with different cyanine fluorescent probes and separated according to the isoelectric point using immobilized pH gradient strips (pH 3-10, 17 cm, Protean® IEF cell system, Bio Rad). Second dimension was performed in a polyacrylamide gel (12%) in the presence of SDS using a Protean II XL system (Bio Rad). Images were obtained with a Typhoon 9410 scanner and analyzed with Progenesis SameSpots software v 4.0. Amino acid content in the OF was determined by high performance liquid chromatography (HPLC). Glucose, lactate, and pyruvate were quantified using microfluorometric enzyme-linked assays. For the proteomic assessment, the results of the image analysis were compared by ANOVA. For both amino acid and carbohydrate analyses, statistical analysis was carried out by 2-way ANOVA with the Holm-Sidak nonparametric post hoc analysis. On Day 3 post-estrus, OF composition varied based on (І) anatomical region, where isthmic metabolites were present in lower (i.e., lactate, glycine, and alanine) or higher (i.e., arginine) concentrations compared to the ampulla; and (ІІ) embryo presence, which was correlated with greater, arginine, phosphoglycerate kinase 1, serum albumin, α-1-antiproteinase and IGL@ protein concentrations. In conclusion, data indicate that the composition of bovine OF is anatomically dynamic and influenced by the presence of an early embryo.


Protocols for Investigating the Host-tissue Distribution, Transmission-mode, and Effect on the Host Fitness of a Densovirus in the Cotton Bollworm.
Many novel viruses have been discovered in animal hosts using next-generation sequencing technologies. Previously, we reported a mutualistic virus, Helicoverpa armigera densovirus (HaDV2), in a lepidopteran species, the cotton bollworm, Helicoverpa armigera (Hubner). Here, we describe the protocols that are currently used to study the effect of HaDV2 on its host. First, we establish a HaDV2-free cotton bollworm colony from a single breeding pair. Then, we orally inoculate some neonate larval offspring with HaDV2-containing filtered liquid to produce two colonies with the same genetic background: one HaDV2-infected, the other uninfected. A protocol to compare life table parameters (e.g., larval, pupal, and adult periods and fecundity) between the HaDV2-infected and -uninfected individuals is also presented, as are the protocols for determining the host-tissue distribution and transmission efficiency of HaDV2. These protocols would also be suitable for investigating the effects of other orally transmitted viruses on their insect hosts, lepidopteran hosts in particular.


N-3 PUFA increase bioavailability and function of endothelial progenitor cells.
Recent data suggest that n-3 PUFA exert beneficial effects on endothelial progenitor cell (EPC) biology. We sought to investigate whether these effects might be mediated by enhanced EPC in vitro function and/or in vivo bioavailability. CACs and late-outgrowth EPCs were isolated from peripheral blood mononuclear cells obtained from 12 donor buffy-coats. The effect of n-3 PUFA (EPA:DHA = 0.9:1.5; 9 μM EPA plus 15 μM DHA) was tested on CAC/EPC viability, function (tube-formation) and pro-inflammatory molecule expression. Circulating EPC (cells positive for CD34, CD133 and kinase insert domain receptor - KDR cell-surface antigens by flow cytometry) number was evaluated in 20 healthy subjects (10 F/10 M, 32 ± 5 years), randomized to receive 4 mackerel or sardine portions per week for 6 weeks followed by a 6 week free-diet period. N-3 PUFA improved CAC and late-outgrowth EPC viability (p < 0.05) and the capacity to form tube-like structures in CACs (+38%; p < 0.05) and late-outgrowth EPCs (+15%; p < 0.05). ICAM-1 expression was reduced in both CACs (p < 0.05) and late-outgrowth EPCs (p < 0.05) and VCAM-1 in late-outgrowth EPCs (p < 0.005). N-3 PUFA significantly decreased TNF-α and MCP-1 expression in CACs and IL-8, TNF-α and MCP-1 in late-outgrowth EPCs (p < 0.05). Circulating EPC number significantly improved after 6 weeks of a fish-enriched diet (p < 0.01) and returned to baseline levels after a 6 week free-diet period (p < 0.01). Plasma EPA levels were independently and positively associated with EPC levels (p < 0.005). Our findings support the case of a beneficiary role played by n-3 PUFA in EPC function and bioavailability.


Can chronic irritations of the trigeminal nerve cause musculoskeletal disorders?
In this article, five cases of odontogenous dysfunctions and musculoskeletal complaints are presented. A common finding in all patients of this study was that the presence of joint complaints was related to deficits in the corresponding muscular function. These deficits were determined by manual muscle tests as described by Kendall et al. [Muscles - Testing and Function, ed 4. Baltimore, Williams and Wilkins, 1993] and were eliminated immediately by a neural therapeutic test injection into the disturbed dental region. The therapy provided solely aimed to eliminate the odontogenous dysfunction. No other therapeutic measures were carried out with regard to the patients' respective muscle, tendon, or joint complaints.


A paediatric case of myopericarditis post-COVID-19 mRNA vaccine.
Myopericarditis is a condition, which primarily involves the pericardium, with minimal myocardial involvement. In myopericarditis, chest pain, elevated cardiac enzymes, and electrocardiographic changes occur. Although COVID-19 mRNA vaccines significantly contribute to preventing the COVID-19 disease, rarely myocarditis and/or pericarditis may develop due to these vaccines. We present a previously healthy 14-year-old male patient who developed myopericarditis after receiving the second dose of the COVID-19 mRNA vaccine.


Replisome-Cohesin Interfacing: A Molecular Perspective.
Cohesion is established in S-phase through the action of key replisome factors as replication forks engage cohesin molecules. By holding sister chromatids together, cohesion critically assists both an equal segregation of the duplicated genetic material and an efficient repair of DNA breaks. Nonetheless, the molecular events leading the entrapment of nascent chromatids by cohesin during replication are only beginning to be understood. The authors describe here the essential structural features of the cohesin complex in connection to its ability to associate DNA molecules and review the current knowledge on the architectural-functional organization of the eukaryotic replisome, significantly advanced by recent biochemical and structural studies. In light of this novel insight, the authors discuss the mechanisms proposed to assist interfacing of replisomes with chromatin-bound cohesin complexes and elaborate on models for nascent chromatids entrapment by cohesin in the environment of the replication fork.


Dual consistency loss for contour-aware segmentation in medical images.
Medical image segmentation is a paramount task for several clinical applications, namely for the diagnosis of pathologies, for treatment planning, and for aiding image-guided surgeries. With the development of deep learning, Convolutional Neural Networks (CNN) have become the state-of-the-art for medical image segmentation. However, issues are still raised concerning the precise object boundary delineation, since traditional CNNs can produce non-smooth segmentations with boundary discontinuities. In this work, a U-shaped CNN architecture is proposed to generate both pixel-wise segmentation and probabilistic contour maps of the object to segment, in order to generate reliable segmentations at the object's boundaries. Moreover, since the segmentation and contour maps must be inherently related to each other, a dual consistency loss that relates the two outputs of the network is proposed. Thus, the network is enforced to consistently learn the segmentation and contour delineation tasks during the training. The proposed method was applied and validated on a public dataset of cardiac 3D ultrasound images of the left ventricle. The results obtained showed the good performance of the method and its applicability for the cardiac dataset, showing its potential to be used in clinical practice for medical image segmentation.Clinical Relevance- The proposed network with dual consistency loss scheme can improve the performance of state-of-the-art CNNs for medical image segmentation, proving its value to be applied for computer-aided diagnosis.


Multiple Perforations of the Sinus Floor During Maxillary Sinus Floor Augmentation to Provide Access to the Bone Marrow Space: A Technical Report.
Sinus floor augmentation is a routinely used surgical technique for increasing the bone height/volume of the atrophic posterior maxilla. Optimal integration of the implanted augmentation material within the newly formed bone will-at least partly-depend on adequate vascularization to ensure sufficient recruitment of osteoblast and osteoclast precursor cells. The present technical note describes a modification intended to facilitate increased blood inflow into the augmented space. After preparation of the lateral window and elevation of the Schneiderian membrane, the cortical bone of the sinus floor is perforated several times either by using a piezoelectric device or a microsurgical handpiece with the corresponding tip or bur; these perforations should extend into the trabecular bone. The experiences with this modified technique after 12 patients are presented and discussed. It is expected that by means of this relatively simple technique, increased blood and cell inflow into the augmented space is achieved. This may, in turn, enhance new bone formation and improve the integration of the augmentation material.


Fused RolA protein enhances beta-glucoronidase activity 50-fold: implication for RolA mechanism of action.
We report that the plant oncoprotein RolA from Agrobacterium rhizogenes acts to stabilize beta-glucoronidase (Gus) when the two proteins are expressed as a fusion protein in transformed tobacco. The observed 50-fold increase of Gus activity was shown to be related to protein accumulation, with no significant changes in mRNA abundance, kinetic properties of the enzyme and thermostability. The entire RolA sequence is essential to achieve the full effect since both the N-terminal region, spanning a putative reverse signal-anchor and nuclear targeting sequences, or the contiguous C-terminal portion were shown to increase Gus activity only 10-fold. A possible interference of RolA in the protein degradation pathway regulated by auxin is discussed.


[An echocardiographic study of the aortic root in ankylosing spondylitis].
The inflammatory condition in ankylosing spondylitis (AS) may affect cardiac structures, being the aortic valve mostly studied. Several studies regard the existence of a subaortic ridging as specific for AS. Our objective was to assess abnormalities in the aortic root in AS using echocardiography and its relation to HLA B-27 and clinical parameters of the disease. Thirty patients with no clinical, radiographic or electrocardiographic evidence of cardiovascular disease were studied by monodimensional, bidimensional and Doppler echocardiography. After an initial ultrasound examination to detect subclinical cardiac abnormalities, aortic root dimensions were measured at the aortic annulus, at the tip of the cusps and 1 cm above the cusps. The existence of subaortic ridging was assessed using bidimensional echocardiography. As clinical parameters were estimated duration of AS, sacroiliac joint X-ray involvement and activity of disease. The results are compared with those in a control group of thirty healthy people with same age, sex and corporal surface. No statistical differences were observed in the mean values of aortic root dimensions between the two groups; in patients with AS were not seen significant differences in the echocardiographic measurements related to clinical parameters or HLA B-27. Only in one patient was observed the characteristic subaortic ridging (4%, NS compared to control group). We conclude, in contrast to other authors, that in patients with AS without cardiovascular disease echocardiographic examination of aortic root does not detect significant abnormalities.


Beating to rotational transition of a clamped active ribbon-like filament.
We present a detailed study of a clamped ribbon-like filament under a compressive active force using Brownian dynamics simulations. We show that a clamped ribbon-like filament is able to capture beating as well as rotational motion under the compressive force. The nature of oscillation is governed by the torsional rigidity of the filament. The frequency of oscillation is almost independent of the torsional rigidity. The beating of the filament gives a butterfly-shaped trajectory of the free-end monomer, whereas rotational motion yields a circular trajectory on a plane. The binormal correlation and the principal component analysis reveal the butterfly, elliptical, and circular trajectories of the free end monomer. We present a phase diagram for different kinds of motion in the parameter regime of compressive force and torsional rigidity.


Landfill mining: a critical review of two decades of research.
Landfills have historically been seen as the ultimate solution for storing waste at minimum cost. It is now a well-known fact that such deposits have related implications such as long-term methane emissions, local pollution concerns, settling issues and limitations on urban development. Landfill mining has been suggested as a strategy to address such problems, and in principle means the excavation, processing, treatment and/or recycling of deposited materials. This study involves a literature review on landfill mining covering a meta-analysis of the main trends, objectives, topics and findings in 39 research papers published during the period 1988-2008. The results show that, so far, landfill mining has primarily been seen as a way to solve traditional management issues related to landfills such as lack of landfill space and local pollution concerns. Although most initiatives have involved some recovery of deposited resources, mainly cover soil and in some cases waste fuel, recycling efforts have often been largely secondary. Typically, simple soil excavation and screening equipment have therefore been applied, often demonstrating moderate performance in obtaining marketable recyclables. Several worldwide changes and recent research findings indicate the emergence of a new perspective on landfills as reservoirs for resource extraction. Although the potential of this approach appears significant, it is argued that facilitating implementation involves a number of research challenges in terms of technology innovation, clarifying the conditions for realization and developing standardized frameworks for evaluating economic and environmental performance from a systems perspective. In order to address these challenges, a combination of applied and theoretical research is required.


Identification of Serological Biomarkers for Early Diagnosis of Lung Cancer Using a Protein Array-Based Approach.
Lung cancer (LC) remains the leading cause of mortality from malignant tumors worldwide. Currently, a lack of serological biomarkers for early LC diagnosis is a major roadblock for early intervention and prevention of LC. To undertake this challenge, we employed a two-phase strategy to discover and validate a biomarker panel using a protein array-based approach. In Phase I, we obtained serological autoimmune profiles of 80 LC patients and 20 healthy subjects on HuProt arrays, and identified 170 candidate proteins significantly associated with LC. In Phase II, we constructed a LC focused array with the 170 proteins, and profiled a large cohort, comprised of 352 LC patients, 93 healthy individuals, and 101 patients with lung benign lesions (LBL). The comparison of autoimmune profiles between the early stage LC and the combined group of healthy and LBL allowed us to identify and validate a biomarker panel of p53, HRas, and ETHE1 for diagnosis of early stage LC with 50% sensitivity at >90% specificity. Finally, the performance of this biomarker panel was confirmed in ELISA tests. In summary, this study represents one of the most comprehensive proteome-wide surveys with one of the largest (i.e. 1,101 unique samples) and most diverse (i.e. nine disease groups) cohorts, resulting in a biomarker panel with good performance.


How familiarization and repetition modulate the picture naming network.
A common strategy to reveal the components of the speech production network is to use psycholinguistic manipulations previously tested in behavioral protocols. This often disregards how implementation aspects that are nonessential for interpreting behavior may affect the neural response. We compared the electrophysiological (EEG) signature of two popular picture naming protocols involving either unfamiliar pictures without repetitions or repeated familiar pictures. We observed significant semantic interference effects in behavior but not in the EEG, contrary to some previous findings. Remarkably, the two protocols elicited clearly distinct EEG responses. These were not due to naming latency differences nor did they reflect a homogeneous modulation of amplitude over the trial time-window. The effect of protocol is attributed to the familiarization induced by the first encounter with the materials. Picture naming processes can be substantially modulated by specific protocol requirements controlled by familiarity and, to a much lesser degree, the repetition of materials.


A relative value unit-based cost comparison of treatment modalities for nonmelanoma skin cancer: effect of the loss of the Mohs multiple surgery reduction exemption.
The incidence of skin cancer has increased dramatically, with as many as 2.8 million skin cancers treated in 2005. In an era of decreasing reimbursement, insurer policy changes, and increasing pressure to deliver cost effective care, physicians should understand the total cost of different skin cancer treatment modalities in order to determine which yields the best value for patients. To estimate the costs of treating nonmelanoma skin cancers by multiple modalities based on their assigned relative value unit (RVU) values. The cost analysis was performed for the treatment of two skin cancer examples, a basal cell carcinoma (BCC) on the central cheek and a squamous cell carcinoma (SCC) on the forearm of varying sizes. The estimated costs of treatment of each of the skin cancers was calculated for treatment with electrodessication and curettage (EDC), imiquimod immunotherapy, Mohs micrographic surgery, traditional surgical excision with permanent section margin evaluation in an office setting (with immediate repair or with repair delayed until clear margins are confirmed), surgical excision with frozen section margin control in both an ambulatory surgery center and hospital-based setting, and radiation therapy. The effect of the loss of exemption from multiple surgery reduction on the cost of Mohs surgery is also examined. Our estimation of costs for each of the treatment modalities reveals that EDC is the least expensive option, with average costs of $471 (BCC cheek) and $392 (SCC arm). Imiquimod treatment and office-based excision with immediate repair of the surgical defect have similar total average costs of $959 (BCC cheek) and $931 (SCC arm) and $1006 (BCC cheek) and $907 (SCC arm), respectively. If repair of the defect is delayed until negative surgical margins are confirmed by permanent section, the cost of excision increases to $1170 and $1041. The average cost of Mohs micrographic surgery is $1263 (BCC cheek) and $1131 (SCC arm). Mohs surgery's recent loss of multiple surgery reduction exemption has decreased the cost of Mohs surgery by 9% to 25%. Excision with frozen section margin control in an ambulatory surgery center results in costs of $2334 (BCC cheek) and $2200 (SCC arm). However, if the excision is performed in a hospital operating room, the procedure is substantially more expensive, at $3085 and $2680. The cost of radiation therapy treatment is $2591 to $3460 for the BCC of the cheek and $2559 to $3431 for the SCC of the arm, depending on the fractional dose used. These are cost estimates based on literature examples and 2008 RVU values; variations related to individual practices and procedure valuations by private insurers are expected. Tumor destruction by EDC or imiquimod and office-based procedures, such as traditional surgical excision or Mohs surgery, are the lowest cost options for treatment of nonmelanoma skin cancer.


Hepatitis B and A markers in children with thalassaemia and sickle-cell disease in Riyadh.
Sera from 40 children with beta-thalassaemia major, 54 children with sickle-cell disease and 120 matched controls were examined by radio-immunoassay for hepatitis B and A markers. The overall prevalence of hepatitis B virus markers was highest in children with thalassaemia followed by children with sickle-cell disease and lowest in the control group. The hepatitis A marker showed a reversed pattern, being lowest in the thalassaemic group. The high incidence of hepatitis B infection was presumably due to frequent blood transfusions, frequent exposure to hospital environment and the high carrier rate in the community. Frequent blood transfusions, however, seem to protect against hepatitis A virus infection.


Eye-movement patterns do not mediate size distortion effects in hemispatial neglect: looking without seeing.
Over the last decade a range of studies have shown that some patients with hemispatial neglect subjectively underestimate the size of objects presented in their contralesional hemispace. Recently, it has been suggested that the effect is simply due to either hemianopia [Brain 124 (2001) 527], or the combination of neglect and hemianopia [Neurology 52 (1999) 1845]. In the current study we asked right hemisphere lesioned patients with and without neglect and hemianopia as well as healthy controls to judge either two horizontal or vertical lines presented simultaneously in right and left hemispace and monitored their eye movements. Three out of the six patients showed the predicted size distortion effect for horizontal lines. We found no evidence that the effect was mediated by eye movements. The two neglect patients who showed the strongest left side underestimation showed symmetrical (left, right) scanning of the lines both in terms of number of fixations and fixation time, yet they still failed to judge the relative size veridically. In addition, we did not find strong evidence for a link with hemianopia. We therefore propose that the effect reflects a computational/representational failure of processing for horizontal extent.


Association of gender norms, relationship and intrapersonal variables, and acculturation with sexual communication among young adult Latinos.
Sexual communication is an important strategy in promoting safer sex behavior, but few investigators have explored sexual communication among young adult Latinos. In this cross-sectional study, we examined the role of traditional gender norms, relationship factors (relationship characteristics and relationship power), intrapersonal factors (attitudes and subjective norms), and acculturation as statistical predictors of three different types of sexual communication (sexual health, pleasure discussions, and physical sexual communication) in Latino women and men. The sample was 220 Latinos (111 women and 109 men) ages 18-30 years who were sexually active in heterosexual relationships. In multiple regression, after controlling for relationship power, intrapersonal factors, and acculturation, traditional gender norms did not predict sexual communication for either women or men. For both women and men, pleasure-focused communication (pleasure discussions and physical sexual communication) increased with acculturation. For women, the strongest predictor of all types of sexual communication was their attitudes toward sexual communication. Greater relationship power and lower acculturation were associated with women's sexual health communication. For men, no variables explained sexual health communication or physical sexual communication, and acculturation and attitude toward pleasure discussions predicted pleasure communication. Women who believed they had power in their relationships and had positive attitudes toward pleasure discussions and a high level of acculturation reported more physical sexual communication. Findings suggest the importance of relationship power, attitudes, and acculturation in young adult Latinos' sexual communication. Sexual risk prevention strategies among young adult Latinos should include encouraging sexual communication by supporting positive attitudes toward pleasure-focused communication. © 2015 Wiley Periodicals, Inc.


Epidemiology of multiple sclerosis in US veterans. VIII. Long-term survival after onset of multiple sclerosis.
Survival to 1996 was analysed for nearly 2500 veterans of World War II who were rated as 'service-connected' for multiple sclerosis as of 1956 by the then Veterans Administration. Survival from onset was defined for all white women and black men, and a random sample of white men. Median survival times from onset were 43 years (white females), 30 years (black males) and 34 years (white males). Crude 50-year survival rates were 31.5% (white females), 21.5% (black males) and 16.6% (white males), but only the white females and white males were significantly different. A proportional hazard analysis was used to identify risk factors for mortality from multiple sclerosis onset year. Significant risk factors included male sex (risk ratio: 1.57), older age at onset (risk ratio: 1.05 per year) and high socioeconomic status (risk ratio: 1.05 per socioeconomic status category). There were no statistically significant differences in survival following multiple sclerosis onset by race or latitude of place of entry into military service, both significant risk factors associated with the development of multiple sclerosis. Standardized mortality ratios utilizing national US data (for 1956-96) showed a marked excess for all three race-sex groups of multiple sclerosis cases, with little difference among them, but with a decreasing excess over time. Relative survival rates, used to compare the survival of multiple sclerosis cases with that of other military veterans, did not differ significantly by sex-race group, nor by latitude of place of entry into military service, but did differ significantly by socioeconomic class. The lack of difference in male and female relative survival rates suggests that the significant difference in survival between male and female multiple sclerosis cases is, at least in part, a result of sex per se and not the disease.


Neuronal hypersensitivity to acetylcholinesterase inhibitors induced by a kindling stimulus in the rabbit brain.
Kindling has been studied primarily with electrical stimuli applied to subcortical structures in the limbic system. Several recent pharmacological studies have implicated muscarinic cholinergic synapses in the genesis of limbic epilepsy. The present investigation was designed to test the hypothesis that cholinergic neurons may constitute a critical element in the neuronal circuitry of kindling. Chemitrodes implanted in rabbit brains enabled both electrical and chemical stimulations of limbic structures. The results indicate that only few electrical kindling-stimuli produce prolonged hypersensitivity to intracerebrally injected physostigmine. A subsequent electrical stimulus continues the kindling progression. The cholinergic supersensitivity persisted for a long time and was suppressed only by intramuscular injections of scopolamine.


Clinical evaluation of platelet concentrates stored for one to five days.
There are no large-scale data available describing the increments obtained with platelet concentrates stored for varying durations. Platelet concentrates prepared by standard techniques were stored at 22 degrees C with horizontal agitation in PL-732 bags and administered to clinically stable, nonalloimmunized recipients known to respond well to random donor platelet transfusions. The platelet concentrates were stored in a mean volume of 65.0 mL (range 54-80 mL) with an average yield of .72 X 10(11) platelets per unit of platelet concentrates (N = 100 consecutive units). There was no significant deterioration of pH during storage. Mean corrected count increments ranged between 16,600 (N = 146 transfusions) after 1 day of storage to 13,300 (N = 34 transfusions) after 5 days of storage. Although these differences were statistically significant (P less than .003, analysis of variance), the overall deterioration in increments was quite modest. Platelet concentrates can be stored under standard conditions for 1 to 5 days with acceptable clinical results.