Epidermal lipid metabolism of cultured skin substitutes during healing of full-thickness wounds in athymic mice.
Cultured epidermal keratinocytes provide an abundant supply of biologic material for wound treatment. Because restoration of barrier function is a definitive criterion for efficacy of wound closure and depends on the lipids present in the epidermis, we analyzed lipid composition of the epidermis in cultured skin substitutes in vitro and after grafting to athymic mice. The cultured skin substitutes were prepared from human keratinocytes and fibroblasts attached to collagen-glycosaminoglycan substrates. After 14 days of incubation, cultured skin substitutes were grafted orthotopically onto full-thickness wounds in athymic mice. Samples for lipid analysis were collected after 14 and 34 days of in vitro incubation, and 3 weeks and 4 months after grafting. Both in vitro samples show disproportions in epidermal lipid profile as compared with the native human epidermis, i.e., a low amount of phospholipids (indicating imbalance in proliferation and differentiation); a large excess of triglycerides (storage lipids); and low levels of free fatty acids, gluco-sphingolipids, cholesterol sulfate, and ceramides-suggesting abnormal composition of stratum corneum barrier lipids. Fatty acid analysis of cultured skin substitutes in vitro revealed insufficient uptake of linoleic acid, which resulted in increased synthesis of and substitution with monounsaturated fatty acids, mainly oleic acid. These abnormalities were partially corrected by 3 weeks after grafting; and 4 months after grafting, all epidermal lipids, with some minor exceptions, were synthesized in proportions very similar to human epidermis. Results of this study show that grafting of cultured skin substitutes to a physiologic host permits the recovery of lipid in proportion to that required for barrier formation in normal human epidermis.


Kinetic analysis of high-concentration isopropanol biodegradation by a solvent-tolerant mixed microbial culture.
The ability of a previously enriched microbial population to utilize isopropanol (IPA) as the sole carbon source within a minimal salts medium is studied. The advantage of prior enrichment procedures for the improvement of IPA biodegradation performance is demonstrated for an IPA concentration of up to 24 g L(-1). Results showing the interrelationship between temperature and substrate utilization and inhibition levels at temperatures of between 2 degrees C and 45 degrees C are examined. Models of inhibition based on enzyme kinetics are assessed via nonlinear analysis, in order to accurately represent the growth kinetics of this solvent-tolerant mixed culture. The model that best describes the data is the Levenspiel substrate inhibition model, which can predict the maximum substrate level above which growth is completely limited. This is the first report of IPA treatment of up to 24 g L(-1) by an aerobic solvent-tolerant population.


Case Report: Congenital Methaemoglobinaemia Type 1: Benign Cyanosis?
Type 1 congenital methaemoglobinaemia is a rare cause of cyanosis which may manifest in affected individuals during concomitant illness. Treatment indications and aims differ from that of acquired methaemoglobinaemia. Type 1 methaemoglobinaemia is a distinct condition from the type 2 form which has a high mortality rate in infancy. A 25 year old male with known type 1 congential methaemoglobinaemia presented with cyanosis in the context of Influenza A with raised methaemoglobin levels on arterial blood gas analysis. The patient was assessed based on his level of 'functional haemoglobin' with no acute indication for IV methylene blue or ascorbic acid. Consideration could be given to prescription of these on a cosmetic basis for some patient populations.


Encapsulated Essential Oils as an Alternative to Insecticides in Funnel Traps.
Pheromone-lured funnel traps are widely used for pest monitoring and mass trapping in agricultural fields and stores. DDVP vapona (dichlorvos), the insecticide of choice as a killing agent inside traps, has been banned, and research on new products is being pursued. Essential oils (EO) could be an alternative if properly formulated. To test their potential, beads of encapsulated coriander and basil EO were tested in funnel traps in stores of almonds and pet foods during 2 mo. The number of adult moth (Plodia interpunctella (Hübner) and Ephestia kuehniella Zeller) dead captures was similar with either coriander or basil EO beads and with vapona tablets while there were more insects alive in the control. These preliminary results indicate a good potential for the development of such natural products as an alternative to synthetic insecticides to include them inside funnel traps.


A Ras inhibitor tilts the balance between Rac and Rho and blocks phosphatidylinositol 3-kinase-dependent glioblastoma cell migration.
Glioblastoma multiforme are highly aggressive tumors for which no adequate treatment has yet been developed. Glioblastoma multiforme show large amounts of active Ras, considered an appropriate target for directed therapy. Here, we show that the Ras inhibitor S-trans, trans-farnesyl thiosalicylic acid (FTS) can avert the transformation of human glioblastoma multiforme cells by inhibiting both their migration and their anchorage-independent proliferation. FTS, by down-regulating Ras activity in glioblastoma multiforme cells, inhibited phosphatidylinositol 3-kinase signaling, resulting in decreased activity of Rac-1. At the same time, activation of RhoA was increased. These two small GTPases are known to control the arrangement of the actin cytoskeleton. By tilting the balance between Rac-1 and RhoA activities, FTS caused the glioblastoma multiforme cells to undergo profound changes in morphology, including rearrangement of actin into stress fibers and assembly of focal adhesions, both of which are governed by RhoA signaling. These morphologic changes allowed strong attachment of the cells to the matrix, rendering them immobile. The results show that FTS should be considered as a candidate drug for glioblastoma multiforme therapy because it targets not only cell proliferation but also cell migration and invasion, which together constitute the most problematic aspect of these malignancies.


The Role of O-GlcNAcylation for Protection against Ischemia-Reperfusion Injury.
Ischemia reperfusion injury (IR injury) associated with ischemic heart disease contributes significantly to morbidity and mortality. O-linked β-N-acetylglucosamine (O-GlcNAc) is a dynamic posttranslational modification that plays an important role in numerous biological processes, both in normal cell functions and disease. O-GlcNAc increases in response to stress. This increase mediates stress tolerance and cell survival, and is protective. Increasing O-GlcNAc is protective against IR injury. Experimental cellular and animal models, and also human studies, have demonstrated that protection against IR injury by ischemic preconditioning, and the more clinically applicable remote ischemic preconditioning, is associated with increases in O-GlcNAc levels. In this review we discuss how the principal mechanisms underlying tissue protection against IR injury and the associated immediate elevation of O-GlcNAc may involve attenuation of calcium overload, attenuation of mitochondrial permeability transition pore opening, reduction of endoplasmic reticulum stress, modification of inflammatory and heat shock responses, and interference with established cardioprotective pathways. O-GlcNAcylation seems to be an inherent adaptive cytoprotective response to IR injury that is activated by mechanical conditioning strategies.


Genetic methods in honey bee breeding.
The honey bee Apis mellifera is a rather difficult object for selection due to the peculiarities of its biology. Breeding activities in beekeeping are aimed at obtaining bee colonies with high rates of economically useful traits, such as productivity, resistance to low temperatures and diseases, hygienic behavior, oviposition of the queen, etc. With two apiaries specializing in the breeding of A. m. mellifera and A. m. carnica as examples, the application of genetic methods in the selection of honey bees is considered. The first stage of the work was subspecies identification based on the analysis of the polymorphism of the intergenic mtDNA locus tRNAleu-COII (or COI-COII) and microsatellite nuclear DNA loci Ap243, 4a110, A24, A8, A43, A113, A88, Ap049, A28. This analysis confirmed that the studied colonies correspond to the declared subspecies. In the apiary with A. m. mellifera, hybrid colonies have been identified. A method based on the analysis of polymorphisms of the tRNAleu-COII locus and microsatellite nuclear DNA loci has been developed to identify the dark forest bee A. m. mellifera and does not allow one to differentiate subspecies from C (A. m. carnica and A. m. ligustica) and O (A. m. caucasica) evolutionary lineages from each other. The second stage was the assessment of the allelic diversity of the csd gene. In the apiary containing colonies of A. m. mellifera (N = 15), 20 csd alleles were identified. In the apiary containing colonies of A. m. carnica (N = 44), 41 alleles were identified. Six alleles are shared by both apiaries. DNA diagnostics of bee diseases showed that the studied colonies are healthy. Based on the data obtained, a scheme was developed for obtaining primary material for honey bee breeding, which can subsequently be subjected to selection according to economically useful traits. In addition, the annual assessment of the allelic diversity of the csd gene will shed light on the frequency of formation of new allelic variants and other issues related to the evolution of this gene.


CDK inhibitors for muscle stem cell differentiation and self-renewal.
Regeneration of muscle is undertaken by muscle stem cell populations named satellite cells which are normally quiescent or at the G0 phase of the cell cycle. However, upon signals from damaged muscle, satellite cells lose their quiescence, and enter the G1 cell cycle phase to expand the population of satellite cell progenies termed myogenic precursor cells (MPCs). Eventually, MPCs stop their cell cycle and undergo terminal differentiation to form skeletal muscle fibers. Some MPCs retract to quiescent satellite cells as a self-renewal process. Therefore, cell cycle regulation, consisting of satellite cell activation, proliferation, differentiation and self-renewal, is the key event of muscle regeneration. In this review, we summarize up-to-date progress on research about cell cycle regulation of myogenic progenitor cells and muscle stem cells during embryonic myogenesis and adult muscle regeneration, aging, exercise and muscle diseases including muscular dystrophy and muscle fiber atrophy, especially focusing on cyclin-dependent kinase inhibitors (CDKIs).


Genomic imprinting and environment in hereditary paraganglioma.
Hereditary paraganglioma (PGL) is characterized by the development of slow-growing and vascularized tumors in the paraganglionic system. PGL is caused by germ line heterozygous inactivating mutations in the SDHB (PGL4), SDHC (PGL3), or SDHD (PGL1) genes, which encode three of the four subunits of mitochondrial complex II (succinate dehydrogenase; SDH). Common tumor sites include the carotid body in the neck and paraganglia in the abdomen. The risk of tumor development associated with SDHD mutations is determined by the sex of the transmitting parent, because only a paternal transmission leads to tumorigenesis in the progeny. This transmission pattern suggests operation of genomic imprinting on the SDHD gene. There is also evidence that the risk of tumor development increases at higher altitudes among SDHD mutation carriers. Accordingly, the increased prevalence of SDHD mutations in the Netherlands, attributable to multiple founder mutations, has been explained in part by the low altitudes in this country, which presumably reduce gene penetrance and relax the natural selection. Thus, PGL caused by SDHD mutations represents an unusual example of an inherited monogenic tumor syndrome because the risk of tumorigenesis shows an absolute dependence on the sex of the transmitting parent and may be modified by a ubiquitous environmental factor.


Diffusion-weighted imaging in evaluating the efficacy of concurrent chemoradiotherapy in the treatment of non-small cell lung cancer.
To explore the predictive value of diffusion-weighted imaging (DWI) in evaluating the short-term efficacy of concurrent chemoradiotherapy (CCRT) in the treatment of patients with non-small cell lung cancer (NSCLC). A total of 192 patients with NSCLC were selected and treated with CCRT. Dynamic contrast-enhanced magnetic resonance imaging combined with DWI was performed on all patients before and after CCRT treatment. Correspondingly, apparent diffusion coefficient (ADC) values were recorded before treatment (ADCpre), during treatment (ADCmid), and after treatment (ADCpost). Tumor response was evaluated as complete response (CR), partial response (PR), stable disease (SD), or progressive disease (PD). Receiver operating characteristic (ROC) curves were used to evaluate the diagnostic power of quantitative DWI parameters in predicting the short-term efficacy of CCRT for patients with NSCLC. There were 21 patients with CR, 82 with PR, 77 with SD, and 12 with PD. The ADCpre was negatively correlated with tumor regression rate, whereas ADCmid, ADCpost, and their respective change rates ∆ADCmid and ∆ADCpost were positively related to tumor regression rate. The ROC curve analysis suggested ADCpre = 1.38 × 10-3 mm2/s, ∆ADCmid = 14.14%, and ∆ADCpost = 20.39% as thresholds to predict the short-term efficacy of CCRT, with corresponding areas under the curve of 0.637, 0.743, and 0.752, respectively. These findings indicate that DWI provides promising predictive value in evaluating the short-term efficacy of CCRT in the treatment of patients with NSCLC.


Discovery of three toxin peptides with Kv1.3 channel and IL-2 cytokine-inhibiting activities from Non-Buthidae scorpions, Chaerilus tricostatus and Chaerilus tryznai.
Non-Buthidae venomous scorpions are huge natural sources of toxin peptides; however, only a few studies have been done to understand their toxin peptides. Herein, we describe three new potential immunomodulating toxin peptides, Ctri18, Ctry68 and Ctry2908, from two non-Buthidae scorpions, Chaerilus tricostatus and Chaerilus tryznai. Sequence alignment analyses showed that Ctri18, Ctry68 and Ctry2908 are three new members of the scorpion toxin α-KTx15 subfamily. Electrophysiological experiments showed that Ctri18, Ctry68 and Ctry2908 blocked the Kv1.3 channel at micromole to nanomole levels, but had weak effects on potassium channel KCNQ1 and sodium channel Nav1.4, which indicated that Ctri18, Ctry68 and Ctry2908 might have specific inhibiting effects on the Kv1.3 channel. ELISA experiments showed that Ctri18, Ctry68 and Ctry2908 inhibited IL-2 cytokine secretions of activated T lymphocyte in human PBMCs. Excitingly, consistent with the good Kv1.3 channel inhibitory activity, Ctry2908 inhibited cytokine IL-2 secretion in nanomole level, which indicated that Ctry2908 might be a new lead drug template toward Kv1.3 channels. Together, these studies discovered three new toxin peptides, Ctri18, Ctry68 and Ctry2908, with Kv1.3 channel and IL-2 cytokine-inhibiting activities from two scorpions, C. tricostatus and C. tryznai, and highlighted that non-Buthidae venomous scorpions are new natural toxin peptide sources.


Moving forward with reactive oxygen species involvement in antimicrobial lethality.
Support for the contribution of reactive oxygen species (ROS) to antimicrobial lethality has been refined and strengthened. Killing by diverse antimicrobials is enhanced by defects in genes that protect against ROS, inhibited by compounds that block hydroxyl radical accumulation, and is associated with surges in intracellular ROS. Moreover, support has emerged for a genetic pathway that controls the level of ROS. Since some antimicrobials kill in the absence of ROS, ROS must add to, rather than replace, known killing mechanisms. New work has addressed many of the questions concerning the specificity of dyes used to detect intracellular ROS and the specificity of perturbations that influence ROS surges. However, complexities associated with killing under anaerobic conditions remain to be resolved. Distinctions among primary lesion formation, resistance, direct lesion-mediated killing and a self-destructive stress response are discussed to facilitate efforts to potentiate ROS-mediated bacterial killing and improve antimicrobial efficacy.


A study of the interaction of Cr(III) complexes and their selective binding with B-DNA: a molecular modeling approach.
Molecular modeling and energy minimisation calculations have been used to investigate the interaction of chromium(III) complexes in different ligand environments with various sequences of B-DNA. The complexes are [Cr(salen)(H(2)O)(2)](+); salen denotes 1, 2 bis-salicylideneaminoethane, [Cr(salprn)(H(2)O)(2)](+); salprn denotes 1, 3 bis- salicylideneaminopropane, [Cr(phen)(3)](3+); phen denotes 1, 10 phenanthroline and [Cr(en)(3)](3+); en denotes ethylenediamine. All the chromium(III) complexes are interacted with the minor groove and major groove of d(AT)(12), d(CGCGAATTCGCG)(2) and d(GC)(12) sequences of DNA. The binding energy and hydrogen bond parameters of DNA-Cr complex adduct in both the groove have been determined using molecular mechanics approach. The binding energy and formation of hydrogen bonds between chromium(III) complex and DNA has shown that all complexes of chromium(III) prefer minor groove interaction as the favourable binding mode.


Small extracellular vesicles secreted from senescent cells promote cancer cell proliferation through EphA2.
Cellular senescence prevents the proliferation of cells at risk for neoplastic transformation. However, the altered secretome of senescent cells can promote the growth of the surrounding cancer cells. Although extracellular vesicles (EVs) have emerged as new players in intercellular communication, their role in the function of senescent cell secretome has been largely unexplored. Here, we show that exosome-like small EVs (sEVs) are important mediators of the pro-tumorigenic function of senescent cells. sEV-associated EphA2 secreted from senescent cells binds to ephrin-A1, that is, highly expressed in several types of cancer cells and promotes cell proliferation through EphA2/ephrin-A1 reverse signalling. sEV sorting of EphA2 is increased in senescent cells because of its enhanced phosphorylation resulting from oxidative inactivation of PTP1B phosphatase. Our results demonstrate a novel mechanism of reactive oxygen species (ROS)-regulated cargo sorting into sEVs, which is critical for the potentially deleterious growth-promoting effect of the senescent cell secretome.


Long-chain perfluoroalkyl carboxylates induce cytoskeletal abnormalities and activate epithelial-mesenchymal transition in both renal cell carcinoma 3D cultures and Caki-1 xenografted mouse model.
Exposure to perfluorooctanoate (PFOA; a type of perfluoroalkyl carboxylates [PFACs]) may be correlated with the incidence of kidney cancer in individuals exposed to high levels of PFOA. However, mechanistic studies on the influence of PFACs on renal cell carcinoma (RCC) development are lacking. We explored the effects of five types of PFACs on RCC using in vitro and in vivo models to fill this knowledge gap and provide information for environmental/usage regulations. Using 2D/3D cultures of Caki-1 cells, a human clear cell RCC line, we examined the effects of short-chain (SC) PFACs and long-chain (LC) PFACs on RCC physio/pathological markers, including the cytoskeleton, epithelial-mesenchymal transition (EMT)-related proteins, and Na+/K+-ATPase. We also administered three different PFACs orally to mice harboring Caki-1 xenografts to assess the impact of these compounds on engrafted RCC in vivo. Compared with the effects of SCPFACs, mice with Caki-1 xenografts treated with LCPFACs showed increased EMT-related protein expression and exhibited liver toxicity. Therefore, LCPFACs induced EMT, influencing cancer metastasis activity, and displayed higher toxicity in vivo compared with SCPFACs. These findings improve our understanding of the effects of PFACs on RCC development and their corresponding in vivo toxicity, which is crucial for regulating these substances to protect public health.


Pentraxin 3 in children suffering from allergic rhinitis.
Allergic disorders are typically characterized by an inflammatory response to allergen exposure. PTX3 behaves as an acute-phase response protein as there is a relationship between PTX3 plasma levels and disease severity. Therefore, this study was designed to investigate a possible role of PTX3 in children with allergic rhinitis. One hundred patients (28 females, 72 males, median age 11 years) were enrolled in the study. All patients were monosensitized: 43 (43%) to seasonal allergens (Graminaceae), 57 (57%) to perennial allergens (house dust mites, cat and dog epithelium, alternaria tenuis). Patients' blood samples for assessing serum PTX3 levels were performed during the spring. Children with rhinitis had higher PTX3 levels and there was a significant relationship between symptom severity and serum levels. Therefore, this study shows that PTX3 serum levels could be a reliable marker for symptom severity in children with allergic rhinitis.


Immunologic and clinical effects of injecting mature peptide-loaded dendritic cells by intralymphatic and intranodal routes in metastatic melanoma patients.
A phase I/II trial was conducted to evaluate clinical and immunologic responses after intralymphatic and intranodal injections of mature dendritic cells. Fourteen patients with a metastatic melanoma received matured dendritic cells, loaded with Melan-A/MART-1 and/or NA17-A peptides and keyhole limpet hemocyanin. The cells were matured overnight with Ribomunyl, a toll-like receptor ligand, and IFN-gamma, which ensured the production of high levels of interleukin-12p70. Dendritic cells were injected at monthly intervals, first into an afferent lymphatic and then twice intranodally. Immunologic responses were monitored by tetramer staining of circulating CD8(+) lymphocytes and delayed-type hypersensitivity tests. Dendritic cell vaccination induced delayed-type hypersensitivity reactivity toward NA17-A-pulsed, keyhole limpet hemocyanin-pulsed, and Melan-A-pulsed dendritic cells in 6 of 10, 4 of 11, and 3 of 9 patients, respectively. Four of the 12 patients analyzed by tetramer staining showed a significantly increased frequency of Melan-A-specific T cells, including one patient vaccinated only with NA17-A-pulsed dendritic cells. Furthermore, 2 of the 12 analyzed patients had a significant increase of NA17-A-specific T cells, including one immunized after an optional additional treatment course. No objective clinical response was observed. Two patients were stabilized at 4 and 10 months and three patients are still alive at 30, 39, and 48 months. Injections into the lymphatic system of mature peptide-loaded dendritic cells with potential TH1 polarization capacities did not result in marked clinical results, despite immunologic responses in some patients. This highlights the need to improve our understanding of dendritic cell physiology.


Measuring plasma C4D to monitor immune complexes in lupus nephritis.
Because currently available assays that measure circulating immune complexes (ICx) are suboptimal, a novel assay was recently developed measuring C4d, a stable product of activation of the classical complement pathway. The present study aimed to establish the value of measuring plasma C4d levels in a longitudinal cohort of patients with severe refractory SLE who were treated with a combination therapy of rituximab with belimumab (RTX+BLM). Fifteen patients with SLE who were treated with RTX+BLM in a phase 2A, open label study were included to sequentially measure plasma C4d levels and correlated to well-established markers of ICx-formation, that is, autoantibodies against double-stranded (ds) DNA, autoantibodies against C1q and proteinuria. The performance of plasma C4d measurements, C4 measurements and the ratio of C4d over C4 (C4d:C4) was evaluated. After establishing that on RTX+BLM treatment kinetics of C4d levels was distinct from traditional C3 and C4 levels, we found strong correlation of C4d:C4 with anti-dsDNA (R=0.76, p<0.001) and anti-C1q (R=0.65, p<0.001) autoantibody levels, which outperformed both stand-alone C4 and C4d levels. Additionally, changes in C4d:C4 over time correlated strongly with changes in proteinuria (R=0.59, p<0.001) as well as anti-dsDNA (R=0.46, p=0.003) and anti-C1q (R=0.47, p=0.002). In patients with severe SLE, plasma C4d levels in relation to C4 levels is useful for longitudinal monitoring after RTX+BLM treatment to reflect amelioration of classical complement activation by ICx as well as proteinuria.


Modeling the effect of temperature on survival rate of Listeria monocytogenes in yogurt.
The aim of the study was to (i) evaluate the behavior of Listeria monocytogenes in a commercially produced yogurt, (ii) determine the survival/inactivation rates of L. monocytogenes during cold storage of yogurt and (iii) to generate primary and secondary mathematical models to predict the behavior of these bacteria during storage at different temperatures. The samples of yogurt were inoculated with the mixture of three L. monocytogenes strains and stored at 3, 6, 9, 12 and 15°C for 16 days. The number of listeriae was determined after 0, 1, 2, 3, 5, 7, 9, 12, 14 and 16 days of storage. From each sample a series of decimal dilutions were prepared and plated onto ALOA agar (agar for Listeria according to Ottaviani and Agosti). It was found that applied temperature and storage time significantly influenced the survival rate of listeriae (p<0.01). The number of L. monocytogenes in all the samples decreased linearly with storage time. The slowest decrease in the number of the bacteria was found in the samples stored at 6°C (D-10 value = 243.9 h), whereas the highest reduction in the number of the bacteria was observed in the samples stored at 15°C (D-10 value = 87.0 h). The number of L. monocytogenes was correlated with the pH value of the samples (p<0.01). The natural logarithm of the mean survival/inactivation rates of L. monocytogenes calculated from the primary model was fitted to two secondary models, namely linear and polynomial. Mathematical equations obtained from both secondary models can be applied as a tool for the prediction of the survival/inactivation rate of L. monocytogenes in yogurt stored under temperature range from 3 to 15°C, however, the polynomial model gave a better fit to the experimental data.


Stingless Bee (Heterotrigona Itama) Honey and Its Phenolic-Rich Extract Ameliorate Oxidant-Antioxidant Balance via KEAP1-NRF2 Signalling Pathway.
Diabetes is associated with an imbalance between oxidants and antioxidants, leading to oxidative stress. This imbalance contributes to the development and progression of diabetic complications. Similarly, renal and liver diseases are characterised by oxidative stress, where an excess of oxidants overwhelms the antioxidant defense mechanisms, causing tissue damage and dysfunction. Restoring the oxidant-antioxidant balance is essential for mitigating oxidative stress-related damage under these conditions. In this current study, the efficacy of stingless bee honey (SBH) and its phenolic-rich extract (PRE) in controlling the oxidant-antioxidant balance in high-fat diet- and streptozotocin/nicotinamide-induced diabetic rats was investigated. The administration of SBH and PRE improved systemic antioxidant defense and oxidative stress-related measures without compromising liver and renal functioning. Analyses of the liver, skeletal muscle and adipose tissues revealed differences in their capacities to scavenge free radicals and halt lipid peroxidation. Transcriptional alterations hypothesised tissue-specific control of KEAP1-NRF2 signalling by upregulation of Nrf2, Ho1 and Sod1 in a tissue-specific manner. In addition, hepatic translational studies demonstrated the stimulation of downstream antioxidant-related protein with upregulated expression of SOD-1 and HOD-1 protein. Overall, the results indicated that PRE and SBH can be exploited to restore the oxidant-antioxidant imbalance generated by diabetes via regulating the KEAP1-NRF2 signalling pathway.


Id2 GABAergic interneurons comprise a neglected fourth major group of cortical inhibitory cells.
Cortical GABAergic interneurons (INs) represent a diverse population of mainly locally projecting cells that provide specialized forms of inhibition to pyramidal neurons and other INs. Most recent work on INs has focused on subtypes distinguished by expression of Parvalbumin (PV), Somatostatin (SST), or Vasoactive Intestinal Peptide (VIP). However, a fourth group that includes neurogliaform cells (NGFCs) has been less well characterized due to a lack of genetic tools. Here, we show that these INs can be accessed experimentally using intersectional genetics with the gene Id2. We find that outside of layer 1 (L1), the majority of Id2 INs are NGFCs that express high levels of neuropeptide Y (NPY) and exhibit a late-spiking firing pattern, with extensive local connectivity. While much sparser, non-NGFC Id2 INs had more variable properties, with most cells corresponding to a diverse group of INs that strongly expresses the neuropeptide CCK. In vivo, using silicon probe recordings, we observed several distinguishing aspects of NGFC activity, including a strong rebound in activity immediately following the cortical down state during NREM sleep. Our study provides insights into IN diversity and NGFC distribution and properties, and outlines an intersectional genetics approach for further study of this underappreciated group of INs.


Dupilumab-associated ocular surface disease in atopic dermatitis patients: Clinical characteristics, ophthalmic treatment response and conjunctival goblet cell analysis.
Dupilumab-associated ocular surface disease (DAOSD) is frequently reported as side effect in atopic dermatitis (AD) patients. Therefore, the aim of this study was to investigate the frequency and severity of DAOSD, ophthalmic treatment response and to learn more about the effect of dupilumab on conjunctival goblet cells (GC). This prospective study included dupilumab-treated AD patients between February 2020 and June 2022 from the University Medical Centre Utrecht. Patients were examined by an ophthalmologist and a dermatologist before start (baseline), and after 4 and 28 weeks of dupilumab treatment. Ophthalmological examination was assessed by the Utrecht Ophthalmic Inflammatory and Allergic disease (UTOPIA) score. DAOSD was defined as an increase in UTOPIA score of ≥3 points from baseline. To quantify conjunctival GCs and to investigate the percentage of Cytokeratin 19 (CK19)-CD45-Mucin 5 AC (MUC5AC)+ cells, conjunctival impression cytology samples were analysed. Ocular surface disease (OSD) was present in 91.3% (n = 63/69) patients at baseline. DAOSD was observed in 28.9% (n = 20/69) patients, in whom GC numbers remained stable and the percentage of CK19-CD45-MUC5AC+ cells decreased at onset of DAOSD compared with baseline. After 28 weeks of dupilumab treatment, DAOSD was seen in 14.5% (n = 10/69) patients. Of the 85.5% (n = 59/69) patients without DAOSD or with controlled DAOSD at Week 28, 40.7% (n = 24/59) patients received anti-inflammatory ophthalmic drugs. Ocular surface disease is common in moderate-to-severe AD patients before starting dupilumab. During treatment with dupilumab DAOSD severity improves with early ophthalmic treatment. The decrease in percentage of CK19-CD45-MUC5AC+ cells during dupilumab treatment suggests an impairment of the GC function due to dupilumab treatment.


Anomalous glass transition behavior of SBR-Al₂O₃ nanocomposites at small filler concentrations.
Elastomers filled with hard nanoparticles are of great technical importance for the rubber industry. In general, fillers improve mechanical properties of polymer materials, e.g. elastic moduli, tensile strength etc. The smaller the size of the particles, the larger is the interface where interactions between polymer molecules and fillers can generate new properties. Using temperature-modulated differential scanning calorimetry and dynamic mechanical analysis, we investigated the properties of pure styrene-butadiene rubber (SBR) and SBR/alumina nanoparticles. Beside a reinforcement effect seen in the complex elastic moduli, small amounts of nanoparticles of about 2 wt% interestingly lead to an acceleration of the relaxation modes responsible for the thermal glass transition. This leads to a minimum in the glass transition temperature as a function of nanoparticle content in the vicinity of this critical concentration. The frequency dependent elastic moduli are used to discuss the possible reduction of the entanglement of rubber molecules as one cause for this unexpected behavior.


High-speed feedback control of an oscillating magnetic helicity injector using a graphics processing unit.
A real-time control system has been developed to control the amplitude, phase, and offset of bulk plasma parameters inside an oscillating magnetic helicity injector. Control software running entirely on an Nvidia Tesla P40 graphical processing unit is able to receive digitizer inputs and send response patterns to a Pulse Width Modulation (PWM) controller with a minimum control loop period of 12.8 µs. With an input digitization rate of 10 MS/s, a three-parameter proportional integral differential controller is shown to be sufficient to inform the PWM controller to drive the desired oscillating plasma waveform with a frequency of 16.6 kHz that is located near the resonance of a coupled RLC circuit. In particular, the temporal phase of the injector waveform is held within 10° of the target value. Control is demonstrated over the toroidal modal structure of the imposed magnetic perturbations of the helicity injection system, allowing a new class of discharges to be studied.


The accumulation of the rare earth elements and of scandium in successive needle age classes of Norway spruce.
The endogenous concentrations of Sc, La, Ce, Sm, Eu, Tb, Yb, and Lu were determined by neutron activation analysis in up to five successive needle age classes of Norway spruce (Picea abies). Trees from nine sites over different bedrocks were sampled individually. Concentration values found are generally much lower than those reported in the literature. This is attributed to the careful removal of any aerosols or soil particles from the needle surface prior to analysis. The concentration of each element increases linearly with the needle age class, i.e., the accumulation can be characterized by just one parameter, the yearly increment. This pattern is followed at small as well as at large concentrations. The accumulation behavior of the investigated elements is identical to that of Si. The relative concentrations of the rare earth elements (REE) in the needles are similar to those in the earth crust. There are significant correlations between the individual REE and between Sc and La.


Mannan-binding lectin (MBL)-associated serine protease-1 (MASP-1), a serine protease associated with humoral pattern-recognition molecules: normal and acute-phase levels in serum and stoichiometry of lectin pathway components.
The pattern-recognition molecules mannan-binding lectin (MBL) and the three ficolins circulate in blood in complexes with MBL-associated serine proteases (MASPs). When MBL or ficolin recognizes a microorganism, activation of the MASPs occurs leading to activation of the complement system, an important component of the innate immune system. Three proteins are produced from the MASP1 gene: MASP-1 and MASP-3 and MAp44. We present an assay specific for MASP-1, which is based on inhibition of the binding of anti-MASP-1-specific antibody to MASP-1 domains coated onto microtitre wells. MASP-1 was found in serum in large complexes eluting in a position corresponding to ∼600 kDa after gel permeation chromatography in calcium-containing buffer and as monomers of ∼75 kDa in dissociating buffer. The concentration of MASP-1 in donor sera (n = 105) was distributed log-normally with a median value of 11 µg/ml (range 4-30 µg/ml). Serum and citrate plasma levels were similar, while the values in ethylenediamine tetraacetic acid plasma were slightly lower and in heparin plasma were 1·5 times higher than in serum. MASP-1 was present at adult level at 1 year of age, while it was 60% at birth. In normal healthy individuals the level of MASP-1 was stable throughout a 2-month period. After induction of an acute-phase reaction by operation we found an initial short decrease, concomitant with an increase in C-reactive protein levels, followed by an increase, doubling the MASP-1 concentration after 2 days. The present data prepare the ground for studies on the associations of MASP-1 levels with disease.


Different pattern of gene mutations in Iranian patients with severe congenital neutropenia (including 2 new mutations).
Severe congenital neutropenia (SCN) is a rare primary immunodeficiency disease. Different genes are found to be associated with SCN, including ELA2, HAX1, WAS, GFI1, G-CSFR and G6PC3. The aim of this study was to find different gene mutations responsible for SCN in Iranian patients. Twenty-seven patients with SCN referred to Immunology, Asthma and Allergy Research Institute during a five year priod 5 years (May 2007 and May 2012), were included in this study. Neutropenia related exons and flanking regions of ELA2, HAX1, WAS, GFI1, G-CSFR and G6PC3 were amplified by PCR and the sequences were analyzed. The results showed different mutations including 4 ELANE mutations, 11 HAX1 mutations and 2 G6PC3 mutations. None of the patients had GFI1 mutation and also one mutation was found in G-CSFR in a patient with ELANE mutation. Ten patients had unknown genetic diagnosis which was compatible with other studies. According to these results, most of the patients showed HAX1 mutations and this finding which significantly differed from other reports, might be related to differences in Iranian ethnicity and also in high rate of consanguineous marriages in Iran.


Defining new aims for BME programs in Latin America: the case of UAM-Iztapalapa.
The need for upkeep and management of medical technology has fostered the creation of a large number of under graduate programs in the field of biomedical Engineering. In Latin America alone, there are over 85 programs dedicated to this. This contrasts with programs in other regions where most of the undergraduates continue on to pursue graduate degrees or work as research and development engineers in the biomedical industry. In this work we analyze the situation regarding curricular design in the 48 BME programs in Mexico and compare this to suggestions and classifications of programs according to needs and possibilities. We then focus on a particular institution, Universidad Autónoma Metropolitana and due to its characteristics and performance we propose that it should redefine its aims from the undergraduate program on, in order to not only generate research but also to provide a nurturing environment for a budding biomedical industry in Mexico.


Localization and role of metabotropic glutamate receptors subtype 5 in the gastrointestinal tract.
Metabotropic glutamate receptor subtype 5 (mGluR5) is a Group I mGlu subfamily of receptors coupled to the inositol trisphosphate/diacylglycerol pathway. Like other mGluR subtypes, mGluR5s contain a phylogenetically conserved, extracellular orthosteric binding site and a more variable allosteric binding site, located on the heptahelical transmembrane domain. The mGluR5 receptor has proved to be a key pharmacological target in conditions affecting the central nervous system (CNS) but its presence outside the CNS underscores its potential role in pathologies affecting peripheral organs such as the gastrointestinal (GI) tract and accessory digestive organs such as the tongue, liver and pancreas. Following identification of mGluR5s in the mouth, various studies have subsequently demonstrated its involvement in mechanical allodynia, inflammation, pain and oral cancer. mGluR5 expression has also been identified in gastroesophageal vagal pathways. Indeed, experimental and human studies have demonstrated that mGluR5 blockade reduces transient lower sphincter relaxation and reflux episodes. In the intestine, mGluR5s have been shown to be involved in the control of intestinal inflammation, visceral pain and the epithelial barrier function. In the liver, mGluR5s have a permissive role in the onset of ischemic injury in rat and mice hepatocytes. Conversely, livers from mice treated with selective negative allosteric modulators and mGluR5 knockout mice are protected against ischemic injury. Similar results have been observed in experimental models of free-radical injury and in vivo mouse models of acetaminophen intoxication. Finally, mGluR5s in the pancreas are associated with insulin secretion control. The picture is, however, far from complete as the review attempts to establish in particular as regards identifying specific targets and innovative therapeutic approaches for the treatment of GI disorders.


Multiplex genotyping of CYP3A4, CYP3A5, CYP2C9 and CYP2C19 SNPs using MALDI-TOF mass spectrometry.
Pharmacogenetics is the study of genetic variations that cause alterations in drug level, drug response and adverse drug reactions. SNPs found in CYP450 genes have the greatest genetic influences on interindividual variability in drug bioavailability. The polymorphic nature of these genes may modulate several enzyme levels that affect individual responses to pharmacological treatment. Among them, CYP3A4, CYP3A5, CYP2C9 and CYP2C19 isoforms of CYP450 enzymes are involved in the metabolism of many commonly prescribed drugs. In this study, we would like to develop a CYP450 genotyping platform that could lead a complete definition of a patient's metabolic genotype in order to improve the clinical outcome of some drug treatments. We used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) (Sequenom) to develop a SNP genotyping method. This MALDI-TOF-based multiplexing system allows the simultaneous and efficient genotyping of a set of CYP450 gene polymorphisms. The multiple CYP450 gene testing achieved with this application can be used to develop diagnostic tests to predict drug responses and clinical outcomes.


RNA-processing proteins regulate Mec1/ATR activation by promoting generation of RPA-coated ssDNA.
Eukaryotic cells respond to DNA double-strand breaks (DSBs) by activating a checkpoint that depends on the protein kinases Tel1/ATM and Mec1/ATR. Mec1/ATR is activated by RPA-coated single-stranded DNA (ssDNA), which arises upon nucleolytic degradation (resection) of the DSB. Emerging evidences indicate that RNA-processing factors play critical, yet poorly understood, roles in genomic stability. Here, we provide evidence that the Saccharomyces cerevisiae RNA decay factors Xrn1, Rrp6 and Trf4 regulate Mec1/ATR activation by promoting generation of RPA-coated ssDNA. The lack of Xrn1 inhibits ssDNA generation at the DSB by preventing the loading of the MRX complex. By contrast, DSB resection is not affected in the absence of Rrp6 or Trf4, but their lack impairs the recruitment of RPA, and therefore of Mec1, to the DSB. Rrp6 and Trf4 inactivation affects neither Rad51/Rad52 association nor DSB repair by homologous recombination (HR), suggesting that full Mec1 activation requires higher amount of RPA-coated ssDNA than HR-mediated repair. Noteworthy, deep transcriptome analyses do not identify common misregulated gene expression that could explain the observed phenotypes. Our results provide a novel link between RNA processing and genome stability.


Polyphenols and Intestinal Permeability: Rationale and Future Perspectives.
Increasing evidence links intestinal permeability (IP), a feature of the intestinal barrier, to several pathological or dysfunctional conditions. Several host and environmental factors, including dietary factors, can affect the maintenance of normal IP. In this regard, food bioactives, such as polyphenols, have been proposed as potential IP modulators, even if the mechanisms involved are not yet fully elucidated. The aim of the present paper is to provide a short overview of the main evidence from in vitro and in vivo studies supporting the role of polyphenols in modulating IP and briefly discuss future perspectives in this research area.


[Serum and salivary CEA in benign and malignant lesions of the oral cavity].
Serum carcinoembryonic antigen (CEA) and salivary CEA levels were determined for 18 patients with squamous oral carcinoma, 12 patients with oral leukoplakia, and 14 healthy volunteers. The determination of the CEA-serum levels has no diagnostic significance. Salivary CEA levels are not correlated with CEA-serum values, with cigarette consumption and clinical stage. Salivary CEA determination provides no additional information for the primary diagnosis. Serial determinations may have adjunctive value in monitoring oral cancer.


Loss of functional transforming growth factor (TGF)-beta type II receptor results in insensitivity to TGF-beta1-mediated apoptosis and Epstein-Barr virus reactivation.
Transforming growth factor (TGF)-beta1 induces not only cell growth inhibition or apoptosis but also Epstein-Barr virus (EBV) reactivation in some Burkitt's lymphoma (BL) cell lines. The purpose of this study was to define the role of TGF-beta signaling molecules in response to TGF-beta1-mediated cell growth inhibition, apoptosis, and EBV reactivation in BL cell lines. First, we confirmed the effect of TGF-beta1 on the cell growth and EBV reactivation in six BL cell lines. TGF-beta1 induced cell growth inhibition and EBV reactivation in these cell lines but did not in Akata cells. To elucidate the mechanism of TGF-beta1 unresponsiveness in Akata cells, we studied the expression of TGF-beta receptors and the intracellular signaling molecules Smads. All cell lines expressed TGF-beta type I receptor, Smad2, Smad3, and Smad4. TGF-beta type II receptor (R-II) was expressed in all cell lines except Akata cells. Introduction of the TGF-beta R-II into Akata cells results in sensitivity to TGF-beta1-mediated growth inhibition, apoptosis, and EBV reactivation. In addition, to test a possibility to the transcriptional repression of the TGF-beta R-II gene in Akata cells, the effect of histone deacetylation (HDAC) inhibitor, trichostatin A (TSA) was examined. The expression of TGF-beta R-II in Akata cells was induced by TSA treatment. These results suggest that the lack of functional TGF-beta R-II impedes the progression of signals through TGF-beta1 and becomes a determinant of unresponsiveness to TGF-beta1-mediated growth inhibition and EBV reactivation.


Quality of life after non-nerve-sparing, robot-assisted radical prostatectomy.
To evaluate quality of life (QOL) after non-nerve-sparing, robot-assisted radical prostatectomy (RARP) and factors associated with worsened QOL. Patients who underwent non-nerve-sparing RARP (n = 115) were enrolled. The Expanded Prostate Cancer Index Composite and 8-item Short Form Health Survey were used to evaluate QOL at baseline and postoperative 1, 3, 6, 12, and 24 months. Preoperative factors were evaluated by multivariate analysis for worsened QOL. Postoperative 24-month urinary QOL worsened in 68 of 115 (59%) patients. In multivariate analysis, membranous urethral length ≥10.8 mm [odds ratio (OR): .26, 95% confidence interval (CI): .07-.96] and intravesical prostatic protrusion ≥12 mm (OR: .14, 95% CI .06-.35) were significantly associated with maintaining postoperative 24-month urinary QOL. Significantly more patients with intravesical prostatic protrusion ≥12 mm than < 12 mm had improved urinary QOL. At postoperative 24 months, 43 of 115 (38%) patients had worsened sexual QOL. In multivariate analysis, higher Sexual Health Inventory for Men score (OR: 8.39, 95% CI 2.28-30.9), sexual activity (OR: 6.94, 95% CI: 2.27-21.2), and intercourse activity (OR: 6.81, 95% CI: 1.64-28.3) preoperatively were significantly associated with worsened postoperative 24-month sexual QOL. More patients with high preoperative intercourse activity (12/23,53%) had significantly worsened (p = .009) QOL regarding sexual bother than those who had no preoperative intercourse activity (21/92, 23%). After non-nerve-sparing RARP, patients with long membranous urethral length and/or intravesical prostatic protrusion maintain urinary QOL. Patients with high intercourse activity presented with impairment of sexual QOL after non-nerve-sparing RARP.


Study of the structure and properties of native and hydrothermally processed wild-type, lam and r variant pea starches that affect amylolysis of these starches.
Starches from WT, lam, and r pea mutants differing in amylopectin/amylose contents (70, 90, and 28% amylopectin, respectively) were used in kinetic studies of pancreatic α-amylase action at 37 °C and for investigations of their supramolecular structure and physicochemical properties during heating. For WT and lam starches, amylase accessibility and catalytic efficiencies (CE) increased following hydrothermal processing up to 100 °C. Accessibility changed relatively less in r during heating with increasing K(m) between 60-90 °C. Limiting values of K(m) after gelatinization were very similar for all three mutants, indicating that relative proportions of amylose/amylopectin have little influence on amylase accessibility once ordered structures are lost. For WT and lam, increases in enzyme accessibility and CE paralleled a rise in amorphous content. It is suggested that the complex behavior for r resulted from amylose gel formation between 60-90 °C. Amorphous amylopectin seems a better substrate for amylase than amorphous amylose.


[Reticular erythematous mucinosis-A special subtype of cutaneous lupus erythematosus?]
Reticular erythematous mucinosis (REM syndrome) is a rare skin disease that predominantly affects women. It is clinically characterized by flat, partly reticular, irregularly configurated pale erythema located in the region of the central chest and upper back. The skin alterations are usually asymptomatic or associated with slight pruritus or burning. Extracutaneous diseases and involvement of internal organs do not occur in REM syndrome. Histopathological features include perivascular and periadnexal lymphocytic inflammatory infiltrates as well as increased deposition of mucin in the dermis. Due to the similar histology to lupus erythematosus (LE) tumidus, it is discussed whether REM syndrome can be assigned to the spectrum of cutaneous LE or can be assessed as a lupus-like disease. Moreover, both conditions respond well to treatment with hydroxychloroquine.


Pancreatic ascites in the setting of portal hypertension.
Pancreatic ascites is a rare cause of ascites and develops largely as a complication of chronic pancreatitis or sometimes due to duct injury during surgical procedures. The entity may mimic spontaneous bacterial peritonitis or ascites due to portal hypertension. Here, the authors discuss a case of pancreatic ascites developing in the setting of alcoholic liver disease with portal hypertension. The patient had features of chronic pancreatitis with pancreatic duct fistula and was managed with stenting of the pancreatic duct.


Imbruvica®▾(ibrutinib) patient support programme for chronic lymphocytic leukaemia and mantle cell lymphoma.
Single-agent ibrutinib is an effective therapy for three types of non-Hodgkin lymphoma: chronic lymphocytic leukaemia (CLL) and small lymphocytic lymphoma, both in relapsed and refractory cases and as a frontline treatment; relapsed and refractory mantle cell lymphoma; and Waldenstrom's macroglobulinaemia in patients who have been treated previously with a different medication. This novel agent has changed the landscape for the aforementioned three subtypes of lymphoma therapies as an oral alternative to traditional chemoimmunotherapy. You&i™ is a no-cost support programme, funded by Janssen, that connects patients taking Imbruvica with a nurse who can answer their questions and help address treatment challenges. This programme offers patients information about their disease, their treatment regimen and side effects management by telephone. The You&i programme was tested at an NHS hospital. Case studies of patients and feedback from health professionals who have used this service show its potential benefits to the patient experience and service delivery.


[Characterization of experimental infection with Mycobacterium lepraemurium in 2 strains of mice].
A model of experimental leprosy in two strains of mice, namely CBA/J and CBi, has been developed based on: 1) the histological examination of a granuloma in the hind foot pad 200 days after inoculation of 0.30 microliter of Mycobacterium lepraemurium (6 x 10(8) MLm/ml); 2) the assessment of T lymphocytes in the granuloma identified by the alpha-naphthyl acetate method for esterase, and c) dissemination of the infection. The histological findings in the low resistance CBA/J strain included positive acid fast bacilli vacuolated cells, without lymphocytic infiltration, scarce number of T lymphocytes and a generalized and important dissemination, similarly to the one observed in human lepromatous leprosy. The histological findings in the hind foot pad granuloma of 30-40 per cent of the medium to high resistance CBi strain, consisted of vacuolated cells and lymphocytic infiltration, a large number of T cells and a scarce dissemination, similar to the human borderline leprosy. Both strains present a different susceptibility to a unique challenge with the mycobacterium which could be useful to disentangle the immunogenetic components involved, by means of appropriate selection and crosses. Furthermore, it could be of interest to perform immunoprotection assays in CBi mice, which might have some bearing on the development of a vaccine in human leprosy.


STELLA facilitates differentiation of germ cell and endodermal lineages of human embryonic stem cells.
Stella is a developmentally regulated gene highly expressed in mouse embryonic stem (ES) cells and in primordial germ cells (PGCs). In human, the gene encoding the STELLA homologue lies on chromosome 12p, which is frequently amplified in long-term cultured human ES cells. However, the role played by STELLA in human ES cells has not been reported. In the present study, we show that during retinoic acid (RA)-induced differentiation of human ES cells, expression of STELLA follows that of VASA, a marker of germline differentiation. By contrast, human embryonal carcinoma cells express STELLA at a higher level compared with both karyotypically normal and abnormal human ES cell lines. We found that over-expression of STELLA does not interfere with maintenance of the stem cell state of human ES cells, but following retinoic acid induction it leads to up-regulation of germline- and endodermal-associated genes, whereas neural markers PAX6 and NEUROD1 are down-regulated. Further, STELLA over-expression facilitates the differentiation of human ES cells into BE12-positive cells, in which the expression of germline- and endodermal-associated genes is enriched, and suppresses differentiation of the neural lineage. Taken together, this finding suggests a role for STELLA in facilitating germline and endodermal differentiation of human ES cells.


Community mental health centres: a qualitative study of professionals' experiences.
This article presents findings from a qualitative study of mental health work in community mental health centres with a multiprofessional workforce and aims of active user participation in the service. User participation implies collaborative relationships and different roles than that of expert professionals and dependent patients. How do professionals working in these services experience their work and professional role? This question was explored in order to highlight important aspects of community mental health work. A group of six experienced professionals from three community mental health centres in a Norwegian city were interviewed twice. The informants highlighted the complexity of community mental health work and the need to be flexible when working to support people with mental health problems in their everyday life situation. To see the service users as people and to facilitate social interaction was important. Their work was described as 'liberating' compared to working in institutions. However, although in their experience they found that professional knowledge and skills were important in their work situation, all had experienced that this was not always acknowledged by professionals in other services.


T cell cytokine patterns in active tuberculosis patients in a northeastern area of Romania.
Developing countries like Romania have a high incidence of tuberculosis. Literature data suggest that people in these countries have an important Th2-type immune background which prevents a protective Th1 response of the host against Mycobacterium tuberculosis. Our study is the first attempt in Romania to identify cytokine patterns in active tuberculosis. The study included 15 patients with active pulmonary tuberculosis and 11 healthy volunteers. Peripheral blood mononuclear cells (PBMC), stained with carboxyfluoresceine-diacetate-succinimidylester (CFSE), were incubated for 7 days with purified protein derivate (PPD) or with medium alone. Intracellular synthesis of IFNgamma and IL-4 in proliferated (CFSE(low)) T cells was detected by flowcytometry. The results showed that both Th1 (IFNgamma) and Th2 (IL-4) cytokines are produced in response to in vitro PPD-stimulation of PBMC from pulmonary tuberculosis patients and healthy controls. Moreover, the proportion between IFNgamma and IL-4 is tilted in favour of IL-4 in PPD-activated (CD3+ CFSE(low)) cells from healthy persons, who did not develop active tuberculosis during the two-year study time interval. This predominance of Th2 effectors points to the need to further investigate the role of IL-4 in the M. tuberculosis infection.


Relation between respiratory neural output and tidal volume.
We recently described a model for the relation between respiratory neural and mechanical outputs (J. Appl. Physiol.: Respirat. Environ. Exercise Physiol. 51: 963-1001, 1981). In this communication we utilize the model to address the following questions. 1) How sensitive is tidal volume (VT) to shape and timing of respiratory neural output (N)? 2) How effective are shape and timing characteristics of N in load compensation? 3) For a given VT, what is the most economical shape and timing of N? Using different values of passive respiratory mechanics, we generated the VT profiles associated with theoretical N waveforms having different shape and timing parameters. We found that 1) with normal mechanics VT is moderately sensitive to inspiratory time (TI) but not to shape of N, whereas with high resistance and short TI, VT is very sensitive to shape and timing; 2) changes in shape, within the physiological range, can serve as potent load-compensatory mechanisms; and 3) for a given VT, the most economical (lowest mean pressure) N pattern is one with a very short TI and a rising phase that is convex to time axis. This holds true even with high resistance.


Evaluation of complementary and alternative medicine trials registered in clinicaltrials.gov database.
Complementary and alternative medicine (CAM) products are increasingly used because they are perceived as natural, relatively low-cost and probably effective therapies for various diseases including cancer. We aimed to determine the quantity and major characteristics of recent herbal/alternative medicine trials registered in clinicaltrials. gov in patients with cancer. "Cancer AND (herbal OR complementary OR alternative)" key words were used to query clinicaltrials. gov (access date 17 April 2015). From the results, 163 trials which have been conducted in patients with the diagnosis of cancer were identified and included in this analysis. At the date of access, 72 trials were completed, 37 trials were still recruiting patients and 10 trials had been withdrawn. Most common cancer type was breast cancer. Eighty-eight percent of trials were interventional and 60% of trials were randomized. The rate of new trial submission were similar for 5-year periods after 2000. The majority of the trials were conducted in United States of America (55%) and People's Republic of China (11%). Nine and 4 of 37 recruiting trials were recorded as phase II and phase III, respectively. When browsing was restricted to "recruiting" and "interventional" studies, the ratio of herbal/complementary treatment trials to all chemotherapy trials was 1.8 %. CAM research in patients with cancer is currently limited, both in terms of quantity and quality. Until high quality scientific and clinical research establishes safety and efficacy of CAM practices, physicians should rigorously inform patients and the public on potential risks and caveats associated with CAM practices.


Arterioarterial Prosthetic Loop as an Alternative Approach for Hemodialysis Access.
In the present study, we performed an arterioarterial prosthetic loop (AAPL) between the femoral artery and deep femoral artery as a new access in patients who did not have adequate vascular conditions for creating an arteriovenous fistula or graft.Between April 2005 and June 2014, 18 patients received AAPL as a vascular access. During the procedure, a polytetrafluoroethylene graft was anastomosed to the femoral artery and deep femoral artery and looped on the thigh. We assessed the reliability and safety of AAPLs by analyzing complication, primary and secondary patency rates, and postoperative blood flow.Eighteen patients (median age, 66 years; range, 43-96 years) underwent AAPL access placement under the general or local anesthesia. All patients were followed up for 3 to 38 months (mean, 24 months). Primary and secondary patency rates at 6 months were 94.5% and 88.8%, respectively, and at 3 years were 61% and 72%, respectively. After operation, one patient had infection, and another one had fat necrosis at the surgical incision site. To maintain the AAPL function, 5 surgical procedures in 4 grafts, including revision, thrombectomy, excision, and repair for bleeding were performed. More than 5000 hemodialyses were performed efficiently in our center.Our study shows that AAPL loop is an unusual but effective and safe procedure that may be a good alternative for the patients who do not allow the conventional hemodialysis access.


Organ-sparing treatment in muscle-invasive bladder cancer.
Organ-sparing treatment of bladder cancer by a trimodality approach is feasible and effective. In this study, the results of a series of patients are reported, who were, in the majority, not suitable for major surgery. In the period from June 1995 through December 2003, 68 patients (64 males, four females) with urothelial bladder cancer were treated with curative intent. The median age was 68 years (range 42-82 years). Clinical T-category was 32x T2, 20x T3, and 16x T4. Transurethral resection was performed in all cases, and a complete TUR-BT (transurethral resection of bladder tumor) was attempted, if possible. Radiotherapy was administered in conventional fractionation (five fractions of 1.8 Gy per week) up to 50.4 Gy to bladder, and regional nodes and the whole bladder received a boost up to 54-59.4 Gy. 34 patients received concurrent cisplatin-based chemotherapy (25 mg/m(2) on days 1-5 and 29-33), and patients with impaired renal function were either treated with irradiation alone (n = 7) or received paclitaxel as alternative to cisplatin in a phase II protocol or on an individual decision (n = 27). The median follow-up was 34 months (range 2-104 months). A histologically confirmed complete remission (CR) on restaging cystoscopy was observed in 40/46 patients (87%) who underwent restaging cystoscopy. CR rates were not significantly correlated to T-category (CR: 24/32 T2, 9/19 T3, and 9/16 T4 tumors) or clinical nodal status. Patients with non-radical resection and macroscopic residual tumor (R2 resection) achieved a CR in only 39% (12/31); this figure was significantly lower as compared to patients with radical R0 TUR-BT (CR: 15/16, 94%, p = 0.013) Furthermore, age and preexisting anemia had no impact on response. The overall survival of the whole group was 45% after 5 years, and survival according to clinical T-category was 62% for T2, 43% for T3, and 19% for T4 (p = 0.015). In eleven patients, local disease progression or relapse was observed. So far, only one salvage cystectomy has been performed, due to contraindications to surgery in the majority of patients. The data obtained in this study confirm the high efficacy of TUR and radiochemotherapy for locally advanced bladder cancer.


Metformin inhibits the growth of human pancreatic cancer xenografts.
Pancreatic ductal adenocarcinoma is a devastating disease, with an overall 5-year survival rate of only 3% to 5%. As the current therapies offer very limited survival benefits, novel therapeutic strategies are urgently required to treat this disease. Here, we determined whether metformin administration inhibits the growth of PANC-1 and MiaPaCa-2 tumor xenografts in vivo. Different xenograft models, including orthotopic implantation, were used to determine whether intraperitoneal or oral administration of metformin inhibits the growth of pancreatic cancer in vivo. We demonstrate that metformin given once daily intraperitoneally at various doses (50-250 mg/kg) to nude mice inhibited the growth of PANC-1 xenografts in a dose-dependent manner. A significant effect of metformin was obtained at 50 mg/kg and maximal effect at 200 mg/kg. Metformin administration also caused a significant reduction in the phosphorylation of ribosomal S6 protein and ERK in these xenografts. Metformin also inhibited the growth of pancreatic cancer xenografts when administered orally (2.5 mg/mL) either before or after tumor implantation. Importantly, oral administration of metformin also inhibited the growth of MiaPaCa-2 tumors xenografted orthotopically. The studies presented here provide further evidence indicating that metformin offers a potential novel approach for pancreatic ductal adenocarcinoma prevention and therapy.


Erythrocyte transfusions are associated with retinopathy of prematurity in extremely low gestational age newborns.
Retinopathy of prematurity (ROP) is a major morbidity in preterm infants causing visual impairment including blindness. Prevention and timely treatment are critical. We investigated the potential role of red blood cell (RBC) transfusions as risk factor for ROP development. Retrospective cohort study of data from 68 tertiary level neonatal intensive care units in Germany. Preterm infants born at 22 + 0 to 28 + 6 weeks of gestation between January 2009 and December 2021 were enrolled. We included n = 12 565 infants. Prevalence of any ROP was 49.2% with most infants being diagnosed with stage 1 (21.5%) and 2 disease (17.2%). ROP stage 3 was present in 10.2%, stage 4 in 0.3%, and ROP requiring treatment in 6.6%. Infants with ROP had significantly more frequently a history of RBC transfusions. Adjusting for confounders, RBC transfusions were associated with increased odds of ROP (OR 1.4, p < 0.001), ROP progression (OR 2.1, p < 0.01) and ROP requiring treatment (OR 3.6, p < 0.001). Restrictive transfusion approaches correlated with decreased (OR 0.7, p < 0.001), liberal regimes with increased odds (OR 1.2, p = 0.001). The present study confirmed an association of RBC transfusions and ROP. Our findings emphasise the need for anaemia prevention and critical re-evaluation of transfusion practices in preterm infants.


Single Nucleotide Polymorphisms Associated with Reading Ability Show Connection to Socio-Economic Outcomes.
Impairments in reading and in language have negative consequences on life outcomes, but it is not known to what extent genetic effects influence this association. We constructed polygenic scores for difficulties with language and learning to read from genome-wide data in ~6,600 children, adolescents and young adults, and tested their association with health, socioeconomic outcomes and brain structure measures collected in adults (maximal N = 111,749). Polygenic risk of reading difficulties was associated with reduced income, educational attainment, self-rated health and verbal-numerical reasoning (p < 0.00055). Polygenic risk of language difficulties predicted income (p = 0.0005). The small effect sizes ranged 0.01-0.03 of a standard deviation, but these will increase as genetic studies for reading ability get larger. Polygenic scores for childhood cognitive ability and educational attainment were correlated with polygenic scores of reading and language (up to 0.09 and 0.05, respectively). But when they were included in the prediction models, the observed associations between polygenic reading and adult outcomes mostly remained. This suggests that the pathway from reading ability to social outcomes is not only via associated polygenic loads for general cognitive function and educational attainment. The presence of non-overlapping genetic effect is indicated by the genetic correlations of around 0.40 (childhood intelligence) and 0.70 (educational attainment) with reading ability. Mendelian randomization approaches will be important to dissociate any causal and moderating effects of reading and related traits on social outcomes.


[Ala499Val (C > T) and Lys939Gln (A > C) polymorphisms of the XPC gene: their correlation with male infertility].
The Ala499Val (C > T) and Lys939Gln (A > C) of the XPC gene are two potentially functional nonsynonymous polymorphisms, which affect the rate of DNA repair and might change XPC production and activity. This study aimed to explore the distribution of these two polymorphisms in the Chinese Han population and their relationship with male infertility. We genotyped the two polymorphisms of the XPC gene by the PCR-restriction fragment length polymorphism (PCR-RFLP) method in 318 infertile patients and 228 fertile male controls, detected the frequency of the alleles, and analyzed both the individual and the joint contribution of the two polymorphisms to male infertility. For the Ala499Val (C > T) polymorphism, the frequencies of the CC, CT, and TT genotypes were significantly different in distribution between the patients and the controls (P = 0.020). Males with the TT genotype had a lower risk of male infertility than those with the CC genotype (adjusted OR = 0.49, 95% CI: 0.23-0.88), and even lower than those with both CC and CT genotypes (adjusted OR = 0.39, 95% CI: 0.22-0.71). The Lys939Gln (A > C) polymorphism was not related with male infertility. The combined genotype analysis showed that the individuals with 1-4 risk alleles had a significantly higher risk of male infertility (adjusted OR = 2.75, 95% CI = 1.50-5.04) than those with 0 risk allele. The Ala499Val (C > T) polymorphism of the XPC gene is correlated with male infertility and may be a potential genetic risk factor for male infertility in the Chinese Han population.


An Extended Kalman Filter to Estimate Human Gait Parameters and Walking Distance.
In this work, we present a novel method to estimate joint angles and distance traveled by a human while walking. We model the human leg as a two-link revolute robot. Inertial measurement sensors placed on the thigh and shin provide the required measurement inputs. The model and inputs are then used to estimate the desired state parameters associated with forward motion using an extended Kalman filter (EKF). Experimental results with subjects walking in a straight line show that distance walked can be measured with accuracy comparable to a state of the art motion tracking systems. The EKF had an average RMSE of 7 cm over the trials with an average accuracy of greater than 97% for linear displacement.


[SOD1 gene mutations in patients with amyotrophic lateral sclerosis: potential for the method of molecular].
The molecular modeling method is promising for the assessment of protein structure, being able to present an energetically beneficial protein conformation with atomic precision. This method is of great importance for studying molecular interactions and confirming pathogenic significance of the changes in the protein structure caused by particular mutations. In the present study we used molecular modeling for the assessment of mutations in the SOD1 gene in patients with amyotrophic lateral sclerosis (ALS), a severe neurodegenerative disorder characterized by the loss of the spinal and cerebral motor neurons. The product of SOD1 is a cytosolic dimeric enzyme Cu/Zn superoxide dismutase (SOD1) responsible for detoxification of the cellular superoxide radicals. We showed that all 8 revealed coding point mutations of the gene led to moderate or significant changes of the SOD1 protein energy. Mutation His49Arg increased the protein energy, and reconstruction of the respective model pointed out to spatial destabilization of the molecule and abnormal interaction with the metal ion inside the active center. The other 7 mutations (Gly17Ala, Leu85Mal, Asn87Ser, Asp91Ala, Serl06Leu, Glu134Gly, and Leul45Phe), on the contrary, led to decrease of the protein energy and increase of the spatial stability of SOD1, which is usually accompanied by increased propensity of the 'inert' mutant molecule to misfolding and cellular aggregation. Thereby, the results of in silico analysis of the SOD1 gene mutations confirm staying of ALS within the class of the so-called conformational diseases of the central nervous system, a characteristic feature of which is forming of cytotoxic insoluble protein inclusions in neurons.


Advanced MRI Techniques for the Hip Joint: Focus on the Postoperative Hip.
Imaging the hip joint with optimized MRI protocol parameters provides the radiologist with the ability to reliably diagnose complex hip abnormalities. The institution of appropriate metal artifact reduction techniques and dedicated sequences can enhance visualization of the periprosthetic bone and soft tissues and allow improved detection of the more frequently encountered total hip arthroplasty-related conditions, such as mechanical loosening, polyethylene wear, and adverse local tissue reactions. Finally, through the use of dynamic contrast-enhanced MRI, abnormal femoral head perfusion, one of the most common complications associated with femoral neck fracture internal fixation, can be detected before the development of femoral head osteonecrosis, subchondral collapse, and secondary osteoarthritis. Thus, through the institution of the techniques described in this review article, the challenges associated with MRI of hip arthroplasty and instrumentation can be overcome, allowing for the diagnosis of common associated complications.


Genome-Wide Single-Nucleotide Polymorphism Data Unveil Admixture of Chinese Indigenous Chicken Breeds with Commercial Breeds.
The conservation and development of chicken have received considerable attention, but the admixture history of chicken breeds, especially Chinese indigenous breeds, has been poorly demonstrated. In this study, we aimed to evaluate the genetic diversity and population structure of eight chicken breeds (including conserved chicken breeds) from different geographic origin and to identify admixture within these breeds using a 600-K single-nucleotide polymorphism panel for genotyping. Using the genotype of 580,961 single-nucleotide polymorphism markers scored in 1,200 animals, we evaluated the genetic diversity (heterozygosity and proportion of polymorphic markers), linkage disequilibrium decay, population structure (principal component analysis and neighbor-joining tree), genetic differentiation (FST and genetic distance), and migration events (TreeMix and f-statistics) of the eight domesticated chicken breeds. The results of population analytical methods revealed patterns of hybridization that occurred after divergence in Tibetan chicken. We argue that chicken migration and admixture, followed by trade, have been important forces in shaping the genomic variation in modern Chinese chicken. Moreover, isolation by distance might play a critical role in shaping the genomic variation within Eurasia continent chicken breeds. Moreover, genetic information provided in this study is valuable resources for production applications (genomic prediction, and breeding strategy) and scientific research (genetic basis detection, studying evolution, or domestication).


Spontaneous abortion as a screening device. The effect of fetal survival on the incidence of birth defects.
To make sound inferences about the distribution, causes and risks of congenital anomalies requires an understanding of selective survival among fetuses. Studies of spontaneous abortions are therefore advocated. A simple model is proposed of the distribution, among spontaneous abortions and births, of conceptuses with and without anomalies. The resulting equation accomodates both maternal and fetal factors. Interaction between maternal and fetal factors is hypothesized, in the form of a maternal device for screening out anomalous conceptuses. A number of the known facts about congenital anomalies are shown to be plausibly interpreted in terms of the proposed model. Finally, some clinical and epidemiologic implications of the model for the study of spontaneous abortions are pointed out.


Hydrogen peroxide contrast echocardiography: quantification in vivo of myocardial risk area during coronary occlusion and of the necrotic area remaining after myocardial reperfusion.
During sustained coronary occlusion in canine preparations, the extent of regions that fail to show contrast enhancement when imaged by supra-aortic hydrogen peroxide contrast echocardiography (SHPCE) has been shown to correlate well for single cross sections with the extent of malperfused myocardium "at risk" of infarction. In the present study, SHPCE was investigated as a means of determining the fraction of total left ventricular mass at risk during occlusion. Since necrotic tissue has low blood flow even when reperfused, we also investigated the potential of quantitating the extent of infarcted myocardium by measuring the extent of contrast defects seen with SHPCE performed during reperfusion. In 20 dogs the fraction of the left ventricle showing a contrast defect during coronary occlusion correlated well with the fraction of the left ventricular mass "at risk" by an autoradiographic technique (autoradiography = 0.83 echocardiography + 8.6%; r = .89, SEE = 4.5%). SHPCE was also performed after 3 hr of reperfusion following occlusions varying in duration from 60 to 150 min. The fraction of the ventricle showing a contrast defect during reperfusion predicted the infarcted portion of the left ventricle as shown by triphenyl tetrazolium chloride staining (% left ventricle infarcted = 0.81 echocardiography + 3.3%; r = .84, SEE = 5.3%). Observer variability for the fraction of the ventricle showing a contrast defect was excellent during both occlusion and reperfusion. The ratio of the left ventricular extent of contrast-negative regions during reperfusion and occlusion was used to calculate a necrosis-to-risk index in vivo that correlated relatively well with the myocardial necrosis-to-risk ratio determined morphologically (r = .77, SEE = 16%).(ABSTRACT TRUNCATED AT 250 WORDS)


Observations of formic and acetic acids at three sites of Mexico City.
Levels of atmospheric carboxylic acids in gas and particulate matter were measured at three sites in Mexico City within the month of March 2000. An annular denuder system was used for sampling and the analytical method was HPLC with UV detection. Formic and acetic acids were present in the PM2.5 fraction and in the gas phase. Total concentration of formic acid was between 0 and 7 ppbV and total concentration of acetic acid was between 1 and 17 ppbV. On average 53% of the formic acid and 67% of the acetic acid were present in particulate matter.


CT mimics of peritoneal carcinomatosis.
Peritoneal carcinomatosis is a term used to describe widespread metastases of cancerous tumors in the peritoneal cavity. It is most common in carcinomas of the gastrointestinal tract (GIT) and ovaries, and must be considered to be the main diagnosis even when the primary is not known. A wide variety of disease processes mimic peritoneal carcinomatosis. Precise diagnosis based on imaging alone is often difficult and very often the final diagnosis is only obtained after appropriate histopathology or microbiology.


Effect of population density on the early post-implantation mouse embryo growth in vitro.
To investigate the relationship between embryo population density and embryo development in vitro. Swiss Webster albino mice were used and blastocysts were flushed from the uterine horns. Various numbers of blastocysts were randomly assigned to separate 35 mm dishes and cultured for 9 days. The effect of duration of culture to the mean success rates was different as the number of embryos in the dish was changed. In early egg cylinder (EEC) stage, the mean success rate was a flat function of the number of embryos. But the mean success rate was a decreasing function in late egg cylinder (LEC) stage and the decreasing rate was faster in early somite (ES) stage. Interestingly, the mean success rate to ES stage was nearly constant when more than 10 embryos were cultured in one dish. The success rate to each developmental stage in vitro decreased as the number of embryos in the dish and the duration of cultured increased.


Electro-reduction of nitrogen on molybdenum nitride: structure, energetics, and vibrational spectra from DFT.
We used density functional theory to study the electrochemical conversion of nitrogen to ammonia on the (001), (100/010), (101), and (111) surfaces of γ-Mo2N. Based on the calculated free energy profiles for the reduction of nitrogen by the associative and dissociative mechanisms, reactivity was found to decrease in the order (111) > (101) > (100/010) ≈ (001). Namely, the cell potentials needed to drive the reduction to ammonia increase in the following order: -0.7 V on (111), -1.2 V on (101), and -1.4 V on (100/010) and (001) surfaces. The (111) surface was found to be the most reactive for nitrogen due to (i) its ability to adsorb the N2 in the side-on position which activates N-N bonding and (ii) its high affinity for N-adatoms which also prevents accumulation of H-adatoms on the catalytic surface at low cell potentials. We have also calculated vibrational frequencies of different NxHy species adsorbed on various γ-Mo2N surfaces. The frequencies are found to depend strongly on the type of the binding sites available on the crystal facets. A comparison of the calculated frequencies with the frequencies of the corresponding species in transition metal complexes and other metal surfaces shows that the frequencies of several signature modes fall in a similar region and might be used to assign the spectra of hydrogen and nitrogen containing surface species on different metal surfaces.


Dynamic properties of excitation and inhibition in the cochlear nucleus.
The dynamic properties of inhibition and excitation of single units in the cochlear nucleus were studied using tones that were amplitude modulated either sinusoidally or with pseudorandom noise. The cross-correlation analysis of the unit discharge rate and the pseudorandom noise modulation of the stimuli showed that the dynamic properties determined from the responses to a single modulated excitatory tone (at CF), to a modulated excitatory tone together with an unmodulated inhibitory tone (above CF), and to a modulated inhibitory tone together with an an unmodulated excitatory tone were almost identical with regard to latency as well as to delay of the peak of the cross-covariance function. On the basis hereof it is inferred that the inhibition is either a cochlear phenomenon or that it is transmitted over pathways with identical temporal properties as those of the excitation. When 2 tones were presented simultaneously the modulation of the excitatory tone usually gave a higher degree of modulation of the discharge rate than did modulation of an inhibitory tone. Addition of an unmodulated inhibitory tone to a modulated tone at CF resulted in an extension of the range of intensities over which the maximal gain was relatively constant. In many units this range extended from about 10dB above the unit's threshold to 60 dB or more above.


A new promoter-probe vector for Saccharomyces cerevisiae using fungal glucoamylase cDNA as the reporter gene.
A system is described for the selection of DNA sequences showing promoter activity in the yeast Saccharomyces cerevisiae using a heterologous reporter enzyme which is efficiently secreted by the yeast host. A multicopy shuttle plasmid of the YEp-type was constructed so as to carry multiple unique cloning sites at the 5' end of the Aspergillus awamori glucoamylase cDNA. Glucoamylase can only be expressed upon insertion at one of these unique cloning sites of a DNA fragment from any source, provided it is endowed with promoter function in S. cerevisiae. As the glucoamylase signal-peptide is functional in S. cerevisiae, the enzyme is efficiently secreted by the yeast transformants. This phenotype can be very easily detected on plate assays and accurately quantified by spectrophotometric analysis of the culture supernatant. Since S. cerevisiae naturally lacks amylolytic activity, any wild-type strain can be used as a host in this system. To evaluate the system, a DNA pool of random fusions was created by ligating sau 3A digested S. cerevisiae genomic DNA to the BglII-linearized vector. The resulting hybrid plasmids were transformed into S. cerevisiae and several transformants secreting glucoamylase to varying degrees were obtained.


Physicochemical properties and phenolic content of honey from different floral origins and from rural versus urban landscapes.
The composition of honey influences how beneficial it is to human health. This study evaluated the physiochemical properties and total phenolic content (TPC) of single vs. multi-floral Irish and selected international honeys, and whether properties varied according to hive location. Oilseed rape honey had the lowest TPC of Irish unifloral honeys. Heather honey had the highest TPC, similar to Manuka honey (Mean ± SD = 68.16 ± 2.73 and 62.43 ± 10.03 respectively), and the TPC of ivy honey was approximately half that of heather. Urban multi-floral honeys contained higher TPC (28.26 ± 13.63) than rural honeys (20.32 ± 11.54). Physiochemical properties varied according to floral origin, and whether hives were in urban or rural sites. Irish heather honey had similar physiochemical characteristics to Manuka honey. This first examination of Irish honey confirms that TPC and physiochemical properties vary with honey type and hive location, and suggests that Irish heather honey should be examined for potential health benefits.


[Perception of the community health agent about the Healthy Family Program].
This study aims at investigating the perceptions of the Community Health Agent regarding the Healthy Family Program in order to identify strengths and weaknesses, for changing the health care attention model. This is a qualitative study, carried out in the Healthy Family Unit of São Sebastião, Federal District. Data were collected by observing the process of work and from interviews with four Community Health Agents, analyzed through the technique of the Collective Subject Speech. The results demonstrated that the object of the Healthy Family Program are the community health problems; basic actions are developed, whose objective is to reduce congestion in health services. It was concluded that the Healthy Family Program is moving toward changes, above all because it promotes the creation of liaisons within the community.


Isolation and characterization of the sheep erythrocyte receptor in man.
Properties of the sheep erythrocyte receptor on human lymphocytes were examined from two different aspects: inhibition of E-rosette formation by antisera, and extraction of T and non-T cells with 3 M KCl. Antisera to several T cell populations produced significant inhibition of E-rosette formation by human peripheral blood lymphocytes, whereas anti-B cell, anti-beta2-microglobulin, anti-I, and anit-i were ineffective. Sera with activity to HL-A subgroups on the HPBL tested produced complete rosette inhibition, although nonspecific anti-HL-A produced no inhibition. T cell preparations extracted by a 3M KCl technique contained activity that could be bound to SRBC and produce inhibition of rosette formation by HPBL. This inhibitory component could be quickly eluted from the SRBC and retain its inhibitory activity. No inhibitory activity could be demonstrated in extracts of B cells and non-lymphoid cells. These results suggest that 1) T cell-specific and HL-A specific antisera inhibit E-rosette formation; 2) beta2-M is not involved in the SRBC receptor as an active component; and 3) the receptor may be isolated by 3 M KCl extraction, and partially purified by the use of SRBC as a specific immunoadsorbent, without loss of biologic activity.


Human male genital tract secretions: both mucosal and systemic immune compartments contribute to the humoral immunity.
In contrast to numerous studies of female genital tract secretions, the molecular properties of Abs and the magnitude of humoral responses in human male genital tract secretions to naturally occurring Ags and to mucosal and systemic immunizations have not been extensively investigated. Therefore, seminal plasma (SP) collected from healthy individuals was analyzed with respect to Ig levels, their isotypes, molecular forms of IgA, and for the presence of Abs to naturally occurring Ags, or induced by systemic or mucosal immunizations with viral and bacterial vaccines. The results indicated that in SP, IgG and not IgA, is the dominant Ig isotype, and that IgM is present at low levels. IgA is represented by secretory IgA, polymeric IgA, and monomeric IgA. In contrast to the female genital tract secretions in which IgA2 occurs in slight excess, the distribution of IgA subclasses in SP resembles that in plasma with a pronounced preponderance of IgA1. The IgG subclass profiles in SP are also similar to those in serum. Thus, SP is an external secretion that shares common features with both typical external secretions and plasma. Specifically, SP contains naturally occurring secretory IgA Abs to environmental Ags of microbial origin and to an orally administered bacterial vaccine, and plasma-derived IgG Abs to systemically injected vaccines. Therefore, both mucosal and systemic immunization with various types of Ags can induce humoral responses in SP. These findings should be considered in immunization strategies to induce humoral responses against sexually transmitted infections, including HIV-1.


Pretreatment of chemically-synthesized Aβ42 affects its biological activity in yeast.
The tendency of amyloid β (Aβ42) peptide to misfold and aggregate into insoluble amyloid fibrils in Alzheimer's disease (AD) has been well documented. Accumulation of Aβ42 fibrils has been correlated with abnormal apoptosis and unscheduled cell division which can also trigger the death of neuronal cells, while oligomers can also exhibit similar activities. While investigations using chemically-synthesized Aβ42 peptide have become common practice, there appear to be differences in outcomes from different preparations. In order to resolve this inconsistency, we report 2 separate methods of preparing chemically-synthesized Aβ42 and we examined their effects in yeast. Hexafluoroisopropanol pretreatment caused toxicity while, ammonium hydroxide treated Aβ42 induced cell proliferation in both C. glabrata and S. cerevisiae. The hexafluoroisopropanol prepared Aβ42 had greater tendency to form amyloid on yeast cells as determined by thioflavin T staining followed by flow cytometry and microscopy. Both quiescent and non-quiescent cells were analyzed by these methods of peptide preparation. Non-quiescent cells were susceptible to the toxicity of Aβ42 compared with quiescent cells (p < 0.005). These data explain the discrepancy in the previous publications about the effects of chemically-synthesized Aβ42 on yeast cells. The effect of Aβ42 on yeast cells was independent of the size of the peptide aggregates. However, the Aβ42 pretreatment determined whether the molecular conformation of peptide resulted in proliferation or toxicity in yeast based assays.


Diffusion tensor tractography of the limbic system.
The limbic system, relevant to memory and emotion, is an interesting subject of study in healthy and diseased individuals. It consists of a network of gray matter structures interconnected by white matter fibers. Although gray matter components of this system have been studied by using MR imaging, the connecting fibers have not been analyzed to the same degree. Cerebrospinal fluid (CSF) signal intensity contamination of the fornix and cingulum, the 2 major white matter tracts of the limbic system, can alter diffusion-tensor imaging (DTI) measurements and affect tractography. We investigated the effect of CSF signal intensity suppression on fiber tracking of the limbic connections and characterized the diffusion properties of these structures in healthy volunteers. Nine healthy individuals were scanned with standard and CSF-suppressed DTI. Tractography of the fornix and cingulum was performed for both acquisition methods. We report mean diffusivity and fractional anisotropy measurements of the crus, body, and columns of the fornix, and descending, superior, and anterior portions of the cingulum. Diffusion measurements were improved and tractography was facilitated by using CSF-suppressed DTI. In particular, tract volume increased, whereas decreases of the mean diffusivity and increases of diffusion anisotropy more accurately represented the underlying tissue by minimizing deleterious partial volume averaging from CSF. This was particularly true for the fornix because it is in closest contact to CSF. Diffusion measurements throughout the limbic connections were consistent in healthy volunteers. We recommend the use of CSF suppression when performing diffusion-tensor tractography of the limbic system.


Collagens, the basic proteins of the human body.
Collagens are structural elements of many tissues in the human body. The family of collagens can be divided into fibrillar and non-fibrillar collagens. The criterion of the classification is the structure of these proteins. Mutations in the genes encoding collagens cause a variety of human diseases that include osteogenesis imperfecta, some forms of osteoporosis, chondrodysplasias, some types of Ehlers-Danlos syndrome, arterial and intracranial aneurysms, epidermolysis bullosa and the renal disease known as Alport syndrome. The detection of mutations is important both scientifically and clinically. Defining the molecular defects underlying a disorder helps in the understanding of not only the properties of the mutated protein but also the function of the normal protein. Even though many mutations in the genes encoding collagens have been described, the pathogenic consequences of some of the mutations are not fully understood. The important rationale for mutation detection is the clinical use of molecular diagnostics in genetic counselling and differential diagnosis.


Association between parental satisfaction and antibiotic prescription for children with cough and cold symptoms.
Providers' interest in satisfying parents may provide an impetus for unnecessary antibiotic use in children. To determine (1) whether receipt of antibiotics at a visit for cough and cold symptoms was associated with increased satisfaction and (2) whether nonreceipt of antibiotics at an initial visit but subsequent receipt of antibiotics in the course of the same illness episode was associated with decreased satisfaction. Prospective cohort study of patients 2-10 years of age presenting to a university-affiliated pediatric clinic with cough and cold symptoms. Parents were enrolled at the index visit and then followed up by phone at least 7 days later (mean time to follow-up, 14.9 days). Satisfaction with the index visit on a 10 point scale was the primary outcome. The primary predictors were whether antibiotics were prescribed at the index visit and, if not, whether they were prescribed since that visit. Linear and median regression were used to adjust for income, child age, parental race and individual provider. A total of 539 parents were enrolled in the study, and 378 (70%) completed follow-up interviews. The mean age of participating children was 4.67 years (SD 2.16). Overall 47% of patients received antibiotics at the index visit, and 8% of those that did not reported receiving them between the index visit and the follow-up assessment. In the regression model, receiving antibiotics at the index visit trended toward being associated with higher satisfactions scores (0.28; P = 0.08). Among those who did not receive antibiotics initially, receiving them subsequently was associated with significantly lower median satisfaction score for the index visit (-3.0; P < 0.01). Receiving antibiotics after an initial visit for cough and cold symptoms at which antibiotics were not prescribed is associated with decreased satisfaction. Use of contingency prescriptions may be an important intervention.