Detection of human herpesviruses HHV-6, HHV-7 and HHV-8 in whole blood by real-time PCR using the new CMV, HHV-6, 7, 8 R-gene kit.
Human herpesviruses 6 (HHV-6), 7 (HHV-7) and 8 (HHV-8) are lymphotropic herpesviruses that may cause opportunistic diseases in immunosuppressed patients such as transplant or AIDS patients. The new commercial CMV HHV-6, 7, 8 R-gene kit (Argene, Varilhes, France) for the simultaneous quantitation of HHV-6 and qualitative detection of HHV-7 and HHV-8 was evaluated using whole blood samples (respectively, n=175, 100 and 161) and using different extraction and real-time PCR platforms in two Centers A and B. In comparison with HHV-6 in-house real-time PCR the commercial kit showed agreements of 96% (n=75) and 85% (n=100) in A and B, respectively, with significant Spearman's correlation between both techniques (in A: r=0.97 [p<0.001]; in B: r=0.70 [p<0.001]). The Bland-Altman test results and prospective monitoring of patients confirmed the accuracy of these HHV-6 real-time PCR techniques. The agreement between the in-house HHV-7 PCR and commercial kit was of 86% (n=100). In comparison with in-house HHV-8 real-time PCRs, the commercial kit showed agreements of 100% (n=61) and 93.7% (n=96) in A and B, respectively. These results demonstrate that the new commercial CMV HHV-6, 7, 8 R-gene kit was an efficient and reliable tool for the diagnosis of herpesvirus 6, 7, 8 infections.


Ecological risk assessment of microplastics in coastal, shelf, and deep sea waters with a consideration of environmentally relevant size and shape.
This study assessed the ecological risk posed by microplastics in surface and subsurface seawaters in coastal, continental shelf, and deep-sea areas of South Korea. The target microplastics for risk assessment were specified as only non-spherical type microplastics in the size range 20-300 μm, because this type was predominantly observed in our study areas, and adverse biological effects have previously been reported. Exposure data for non-spherical microplastics were obtained from a previous study or were measured for microplastics of sizes down to 20 μm. A predicted no-effect concentration (PNEC) of 12 particles/L was derived by employing a species sensitivity distribution approach. Then the results were compared to the in situ observed concentrations at each site. The detected microplastic concentrations did not exceed the derived PNEC, i.e., the current pollution levels of fragment and fiber microplastics in the size range 20-300 μm would not pose a significant threat to the marine ecosystem in South Korea. However, predictions are that microplastic pollution will increase to 50-fold by 2100 at the current rates, and in this scenario, the microplastic concentration is expected to far exceed the derived PNEC values for marine ecosystems. It is therefore urgent to take precautionary actions to prevent a further increase in microplastic concentrations in these environments.


The relationship between osteoclastogenic and anti-osteoclastogenic pro-inflammatory cytokines differs in human osteoporotic and osteoarthritic bone tissues.
Pro-inflammatory cytokines possess osteoclastogenic or anti-osteoclastogenic activities. They influence osteoclasts directly or via the receptor activator of nuclear factor κB (RANK), RANK ligand (RANKL) and osteoprotegerin (OPG) system. Recent evidence suggests that inflammation may play a role in osteoporosis (OP) and osteoarthritis (OA). We aimed therefore to determine whether there is a difference between both groups: first, in the expression of the osteoclastogenic and anti-osteoclastogenic cytokines, second, in correlation of these cytokines with bone mineral density (BMD) and levels of bone turnover markers (BTM) and third, in correlation between the expression of these cytokines and osteoclast specific genes and RANK/RANKL/OPG genes. Human bone samples from 54 age and sex matched patients with OP or OA were collected during hip arthroplasty surgery. The expression of 25 genes encoding pro-inflammatory cytokines, their receptors, osteoclast specific genes and RANK/RANKL/OPG genes was measured using quantitative real-time PCR. Total hip, femoral neck and lumbar spine BMD and BTM in blood samples were measured. The comparison between OP and OA was assessed using Student's t-test or Mann-Whitney U test and correlations between gene expression, BMD and BTM were determined using nonparametric correlation. The results demonstrated a higher expression of interleukin (IL)-6 and IL-1α in OP, and interferon (IFN)-γ in OA (p < 0.0005). Negative correlations of total hip BMD with tumor necrosis factor-α (TNF-α) in OA and with RANKL/RANK in OP were found (p < 0.05). Significant correlations with BTM were shown for IL-1α and IFN-γ in OP (rho = 0.608 and -0.634) and for TNF-α, IL-6 and transforming growth factor-β1 (TGF-β1) in OA (rho = 0.591, -0.521 and 0.636). Results showed OP specific negative correlations (IFN-γ with ITGB3, IFN-β1 with CTSK, tartrate resistant acid phosphatase (TRAP), CALCR, RANK, RANKL, IL-1α with CTSK, OPG, IL-17A with CALCR) and positive (TGF-β1 with CTSK, TRAP, RANK), and OA specific negative (IL-1α with osteoclast associated immunoglobulin-like receptor (OSCAR), TNF-α with RANK, RANKL, OPG) and positive (IL-6 with RANK, RANKL, OPG) correlations. Our results demonstrate that the relationship between osteoclastogenic and anti-osteoclastogenic pro-inflammatory cytokines differs in human OP and OA bone and could present an important factor for characteristics of OP and OA bone phenotypes.


[Evaluation of phase transition models in lipid-aqueous systems by the optical analogy method].
There are two models of phase transition in mesomorphic lipid-water systems. The first one describes the transition in terms of trans-gauche isomerisation, the second-by the appearance of gauche-trans-gauche isomers (kinks). Both were argumented theoretically, but experiments cannot as yet offer an unambiguous decision. We have tried to evaluate each of the assumptions based on the qualitative analysis of X-ray and optical diffraction pictures from models and real objects. We have had in mind the early proved reason in the application of optical modeling of liquid crystal structures in a range of which our dipalmitoyllecithin-water system could be included. Results of comparing optical diffraction of laser beam (gamma = 630 nm) on two-dimensional models with our X-ray pictures of the system mentioned in L alpha and L beta modifications have shown that high-temperature L alpha phase consists of molecules with gauche-conformation of aliphatic fragments. In view of these data the kink model seems less realistic.


Determining flexor-tendon repair techniques via soft computing.
An SC-based multi-objective decision-making method for determining the optimal flexor-tendon repair technique from experimental and clinical survey data, and with variable circumstances, was presented. Results were compared with those from the Taguchi method. Using the Taguchi method results in the need to perform ad-hoc decisions when the outcomes for individual objectives are contradictory to a particular preference or circumstance, whereas the SC-based multi-objective technique provides a rigorous straightforward computational process in which changing preferences and importance of differing objectives are easily accommodated. Also, adding more objectives is straightforward and easily accomplished. The use of fuzzy-set representations of information categories provides insight into their performance throughout the range of their universe of discourse. The ability of the technique to provide a "best" medical decision given a particular physician, hospital, patient, situation, and other criteria was also demonstrated.


Solitary extramedullary plasmacytoma (a case report).
Solitary plasmacytoma of the upper air and food passages is a rare tumour. This paper deals with such a case along with a brief account of the etiology, clinical features and therapeutic measures.


Genome-wide expression profiles of Pyropia haitanensis in response to osmotic stress by using deep sequencing technology.
Pyropia haitanensis is an economically important marine crop grown in harsh intertidal habitats of southern China; it is also an excellent model system for studying mechanisms of stress tolerance. To understand the molecular mechanisms underlying osmotic tolerance and adaptation to intertidal environments, a comprehensive analysis of genome-wide gene expression profiles in response to dehydration and rehydration in Py. haitanensis was undertaken using digital gene expression profile (DGE) approaches combined with de novo transcriptome sequencing. RNA-sequencing of the pooled RNA samples from different developmental phases and stress treatments was performed, which generated a total of 47.7 million clean reads. These reads were de novo assembled into 28,536 unigenes (≥ 200 bp), of which 18,217 unigenes (63.83 %) were annotated in at least one reference database. DGE analysis was performed on four treatments (two biological replicates per treatment), which included moderate dehydration, severe dehydration, rehydration, and normal conditions. The number of raw reads per sample ranged from 12.47 to 15.79 million, with an average of 14.69 million reads per sample. After quality filtering, the number of clean reads per sample ranged from 11.83 to 15.04 million. All distinct sequencing reads were annotated using the transcriptome of Py. haitanensis as reference. A total of 1,681 unigenes showed significant differential expression between moderate dehydration and normal conditions, in which 977 genes were upregulated, and 704 genes were downregulated. Between severe dehydration and normal conditions, 1,993 unigenes showed significantly altered expression, which included both upregulated (1,219) and downregulated genes (774). In addition, 1,086 differentially expressed genes were detected between rehydration and normal conditions, of which 720 genes were upregulated and 366 unigenes were downregulated. Most gene expression patterns in response to dehydration differed from that of rehydration, except for the synthesis of unsaturated fatty acids, several transcription factor families, and molecular chaperones that have been collectively implicated in the processes of dehydration and rehydration in Py. haitanensis. Taken together, these data provide a global high-resolution analysis of gene expression changes during osmotic stress that could potentially serve as a key resource for understanding the biology of osmotic acclimation in intertidal red seaweed.


Evaluation of Pregnancy Malaria Vaccine Candidates: The Binding Inhibition Assay.
The parasite-binding inhibition assay is designed to evaluate the acquisition of naturally acquired functional antibodies that block Plasmodium falciparum binding to endothelial or placental receptors. The assay is also used to assess functional activity by antibodies induced by immunization, for example antibodies raised against pregnancy malaria vaccine candidates like VAR2CSA. Here we describe a plate-based assay to measure the levels of adhesion-blocking antibodies. This assay format can be adapted to any lab that is minimally equipped for short-term parasite culture.


Regulation of macrophage cholesterol efflux and liver X receptor α activation by nicotine.
This study aims to investigate the characteristics of liver X receptor α (LXRα) and its target gene expression, as well as cholesterol efflux in human macrophages treated by nicotine. Human monocyte-derived macrophages were collected. Before apoA-I-mediated human monocyte-derived macrophage cholesterol efflux, and mRNA expression of LXRα, and some of its target genes being detected, the macrophages were induced with or without nicotine. Pre-incubation of Human monocyte-derived macrophages with nicotine, cholesterol efflux was suppressed to apolipoprotein AI. Nicotine also inhibited LXRα and some of its target genes mRNA expression involved cholesterol metabolism, and facilitated some inflammatory genes expression. The changed function of cholesterol efflux and some genes expression may be the pathogenetic cause, and LXR activity of macrophage may offer potential therapeutic benefit in the treatment of atherosclerosis. Thus nicotine can regulate foam cell formation by inhibiting LXR pathway.


The structure and stability of small mammal faunas.
I examined questions about the local and global stability and resilience characteristics of six small mammal faunas and their relationship to connectance, average interaction strength, community covariance, and indirect competitive pathways. Community matrices estimated elsewhere were used in the analyses. The faunas had from three to nine species and represented several habitat types. The following properties were observed: (1) all community matrices were globally stable, (2) return rates to equilibrium were generally maximized, and (3) competitive interactions were strictly hierarchical. Global stability resulted from a reduction in connectance with increasing species and not from structural characteristics (i.e., community covariance). Average interaction strength did not change with the number of species in these faunas. Increased resilience was due largely to reduced community covariance. Two of the six faunas showed some evidence for indirect pathways, but this appeared to be artifactual. Randomizations of the original community matrices showed that indirect pathways have a high probability of occurrence. Monte Carlo simulations indicated that the probability of indirect pathways increases with increasing number of species or magnitude of competition. These results bring into question previous observational studies invoking indirect competitive pathways. Communities without indirect pathways have a special linear hierarchical structure. This structure has greater resilience than when indirect pathways are present. The observed patterns in community structure are discussed with regard to current theories of habitat selection.


Emerging nanotherapeutics alleviating rheumatoid arthritis by readjusting the seeds and soils.
Rheumatoid arthritis (RA) is a complicated autoimmune disease that results in severe joint inflammation, synovial hyperplasia, pannus formation, cartilage and bone destruction, and other RA-associated complications. Although the pathogenesis of RA remains unclear, increasing reports have shown that inflammation-relevant cells and the microenvironment of inflamed joints play a critical role in the formation and aggravation of RA. Recently, numerous nanotherapeutics have been engineered to overcome these intractable challenges by readjusting inflammation-related seeds (endothelial cells, macrophages, neutrophils, antigen-presenting cells, fibroblasts, osteoclasts, T cells, B cells, and chondrocytes) and inflamed soils (NO, cell-free DNA, hypoxia, ROS, and pro-inflammatory cytokines). In this review, we first present a detailed pathogenesis of RA, with an emphasis on the emerging advances in regulating seeds or remodeling soils for RA treatment. We then outline these intelligent therapeutics via synergistic seed-soil adjustment, particularly for spatiotemporally cascade-responsive or all-in-one integrational nanosystems. Finally, we briefly discuss the ongoing challenges and prospects for the clinical development and translation of seed soil-based therapies.


Human papillomavirus type 18 is associated with less apoptosis in fibroblast tumours than human papillomavirus type 16.
In human cervical neoplasia human papillomavirus (HPV) type 18 has a higher cancer/cervical intraepithelial neoplasia (CIN) prevalence ratio than HPV 16. Fibrosarcomas derived from rat fibroblasts transfected with HPV 16 or 18 genomes showed increased apoptosis compared with controls. However, HPV 18 was associated with significantly less apoptosis than HPV 16, affording one possible explanation for the more rapidly progressive cervical neoplasia associated with HPV 18.


Tracing sulfate origin and transformation in an area with multiple sources of pollution in northern China by using environmental isotopes and Bayesian isotope mixing model.
Sulfate (SO42-) contamination in groundwater and surface water is an environmental problem of widespread concern. In this study, we combined stable isotope analyses of SO42- (δ34S and δ18O) and water (δ2H and δ18O) with a Bayesian mixing model (SIAR), for the first time, to identify sources and transformation of SO42- in an area of northern China with multiple potential sources of pollution. The overall values of δ34S and δ18O-SO42- ranged from 1.3‰ to 16.3‰ and -3.8‰-8.8‰ in groundwater, and from -1.1‰ to 9.3‰ and 2.7‰-9.2‰ in surface waters, respectively. Analyses of SO42- isotopes and water chemistry indicated that SO42- in groundwater and surface water mainly originated from mixing of oxidation of sulfate, sewage, chemical fertilizers, dissolution of evaporite and precipitation. There was no significant correlation between δ34S and δ18O and SO42- concentration in groundwater, indicating that bacterial sulfate reduction did not affect the SO42- isotopic composition. SIAR model showed the main sources of SO42- in groundwater and surface water comprised oxidation of sulfide minerals and sewage. In groundwater, oxidation of sulfide minerals and sewage accounted for 37.5-44.5% and 35.5-42.7% of SO42-, respectively. In regard to surface waters, the contribution of oxidation of sulfide minerals to SO42- was higher in the wet season (31.8 ± 9.9%) than in the intermediate (22.4 ± 7.8%) and dry (20.9 ± 8.2%) seasons, but the contribution proportion of sewage was slightly lower in the wet season (19.9 ± 8.5%) than in the intermediate (23.8 ± 8.7%) and dry (24.2 ± 8.5%) seasons. This study indicates that it is necessary for local government to improve the treatment infrastructure for domestic sewage and optimize methods of agricultural fertilization and irrigation to prevent SO42- contamination of groundwater and surface water.


An age-related decline in interleukin-10 may contribute to the increased expression of interleukin-6 in brain of aged mice.
The DNA-binding activity of nuclear factor kappaB (NFkappaB) is elevated in brain of aged mice, resulting in elevated levels of the inflammatory cytokine interleukin (IL)-6. The purpose of this study was to determine if in the brain of aged mice a decrease in the anti-inflammatory cytokine IL-10 contributes to the increase in IL-6. In initial studies coronal brain sections and glial cells from adult (6-months-old) and aged (24-months-old) mice were incubated in the presence or absence of lipopolysaccharide (LPS) and the concentrations of IL-6 and IL-10 in supernatants were determined. In subsequent studies, the effect of recombinant murine IL-10 on constitutive and inducible NFkappaB activity, IL-6 mRNA expression and IL-6 protein secretion by glia cultured from brains of adult and aged mice was determined. Coronal brain sections and glia from aged mice secreted more IL-6 and less IL-10 than brain sections and glia from adults. This effect of age was evident in the absence and presence of LPS and suggested that a decrease in IL-10 production permitted increased IL-6 production. Consistent with this idea, treatment of glia from aged mice with recombinant IL-10 decreased both constitutive and inducible binding of NFkappaB to the IL-6 gene promoter. The decrease in NFkappaB activity was associated with a reduction of IL-6 mRNA and protein. Exogenous IL-10, however, had no effect on NFkappaB activity, which was undetectable in unstimulated glia from adult mice, and did not decrease steady-state levels of IL-6 mRNA or IL-6 protein secretion. Collectively, these studies suggest that IL-10 constrained IL-6 gene expression in the adult brain, but in the aged brain it decreased and thus enabled a cascade of intracellular events that increased expression of the IL-6 gene.


Neural Mechanisms of Material Perception: Quest on Shitsukan.
In recent years, a growing body of research has addressed the nature and mechanism of material perception. Material perception entails perceiving and recognizing a material, surface quality or internal state of an object based on sensory stimuli such as visual, tactile, and/or auditory sensations. This process is ongoing in every aspect of daily life. We can, for example, easily distinguish whether an object is made of wood or metal, or whether a surface is rough or smooth. Judging whether the ground is wet or dry or whether a fish is fresh also involves material perception. Information obtained through material perception can be used to govern actions toward objects and to make decisions about whether to approach an object or avoid it. Because the physical processes leading to sensory signals related to material perception is complicated, it has been difficult to manipulate experimental stimuli in a rigorous manner. However, that situation is now changing thanks to advances in technology and knowledge in related fields. In this article, we will review what is currently known about the neural mechanisms responsible for material perception. We will show that cortical areas in the ventral visual pathway are strongly involved in material perception. Our main focus is on vision, but every sensory modality is involved in material perception. Information obtained through different sensory modalities is closely linked in material perception. Such cross-modal processing is another important feature of material perception, and will also be covered in this review.


A novel nudivirus infecting the invasive demon shrimp Dikerogammarus haemobaphes (Amphipoda).
The Nudiviridae are a family of large double-stranded DNA viruses that infects the cells of the gut in invertebrates, including insects and crustaceans. The phylogenetic range of the family has recently been enhanced via the description of viruses infecting penaeid shrimp, crangonid shrimp, homarid lobsters and portunid crabs. Here we extend this by presenting the genome of another nudivirus infecting the amphipod Dikerogammarus haemobaphes. The virus, which infects cells of the host hepatopancreas, has a circular genome of 119,754 bp in length, and encodes a predicted 106 open reading frames. This novel virus encodes all the conserved nudiviral genes (sharing 57 gene homologues with other crustacean-infecting nudiviruses) but appears to lack the p6.9 gene. Phylogenetic analysis revealed that this virus branches before the other crustacean-infecting nudiviruses and shares low levels of gene/protein similarity to the Gammanudivirus genus. Comparison of gene synteny from known crustacean-infecting nudiviruses reveals conservation between Homarus gammarus nudivirus and Penaeus monodon nudivirus; however, three genomic rearrangements in this novel amphipod virus appear to break the gene synteny between this and the ones infecting lobsters and penaeid shrimp. We explore the evolutionary history and systematics of this novel virus, suggesting that it be included in the novel Epsilonnudivirus genus (Nudiviridae).


SDF Factor-1α Promotes the Migration, Proliferation, and Osteogenic Differentiation of Mouse Bone Marrow Mesenchymal Stem Cells Through the Wnt/β-Catenin Pathway.
Bone marrow mesenchymal stem cells (BMSCs) are thought to have great potential in the treatment of many diseases and may serve as a cell source for tissue engineering. These cells may be regulated by stromal cell-derived factor-1α (SDF-1α), which has been shown to promote the migration, proliferation, and osteogenic differentiation of BMSCs in inflammation-associated diseases. However, the specific mechanism underlying this process remains unclear. We herein transduced lentivirus carrying SDF-1α, empty vector, or siRNA-SDF-1α into mouse BMSCs and then performed transwell, CCK-8, cell cycle, alkaline phosphatase activity, and Alizarin Red staining experiments on the three groups of samples. Overexpression of SDF-1α promoted the migration, proliferation, and osteogenic differentiation of BMSCs, and SDF-1α upregulated the expression of Wnt pathway-related factors and downstream target genes as determined by western blot, real-time polymerase chain reaction, and immunofluorescence. The effect of low SDF-1α expression on BMSCs was significantly weakened. In addition, we transduced lentivirus carrying siRNA-Wnt3a into BMSCs and treated them with SDF-1 drugs. After inhibiting the Wnt pathway, SDF-1 significantly weakened the migration, proliferation, and osteogenic differentiation of BMSCs. From this, we concluded that high SDF-1 expression can promote the migration, proliferation, and osteogenic differentiation of BMSCs, at least in part by activating the Wnt pathway.


Overexpression of the Arabidopsis and winter squash superoxide dismutase genes enhances chilling tolerance via ABA-sensitive transcriptional regulation in transgenic Arabidopsis.
The winter squash (Cucurbita moschata, Cm) superoxide dismutase (SOD) CmSOD gene and Arabidopsis thaliana (At)SOD gene were transferred under a ubiquitin promoter into Arabidopsis via Agrobacterium tumefaciens. The expression and amount of SOD and the SOD activities in the AtSOD and CmSOD transgenic lines were significantly higher than those of non-transgenic (NT) plants exposed to 23 or 4°C treatment for 6 ~ 192-h periods. Furthermore, expressions of the cold-inducible gene (AtCBF2) and desiccation-responsible transcription factors (AtRD29A/B) were also activated in all transgenic lines compared to NT plants after chilling treatments. Compared to NT plants under chilling stress, superoxide (•O2-) accumulation was significantly lower, and chlorophyll (Chl) contents were significantly higher in all transgenic lines with higher SOD activity. Moreover, Arabidopsis seedlings overexpressing AtSOD and CmSOD also displayed greater resistance to chilling and less oxidative injury than NT plants under chilled conditions, indicating that the overexpression of AtSOD and CmSOD in Arabidopsis enhanced chilling tolerance by eliminating •O2-. The expression of AtRD29A was strongly up-regulated only in AtSOD transgenic plants treated with abscisic acid (ABA), while it was repressed in other transgenic plants, indicating ABA-sensitive AtCBF2 and AtRD29A/B transcriptional regulation signaling pathways in transgenic Arabidopsis under chilling conditions.


Identification of a new class of nitrogen fixation genes in Rhodobacter capsulatus: a putative membrane complex involved in electron transport to nitrogenase.
DNA sequence analysis of a 12236 bp fragment, which is located upstream of nifE in Rhodobacter capsulatus nif region A, revealed the presence of ten open reading frames. With the exception of fdxC and fdxN, which encode a plant-type and a bacterial-type ferredoxin, the deduced products of these coding regions exhibited no significant homology to known proteins. Analysis of defined insertion and deletion mutants demonstrated that six of these genes were required for nitrogen fixation. Therefore, we propose to call these genes rnfA, rnfB, rnfC, rnfD, rnfE and rnfF (for Rhodobacter nitrogen fixation). Secondary structure predictions suggested that the rnf genes encode four potential membrane proteins and two putative iron-sulphur proteins, which contain cysteine motifs (C-X2-C-X2-C-X3-C-P) typical for [4Fe--4S] proteins. Comparison of the in vivo and in vitro nitrogenase activities of fdxN and rnf mutants suggested that the products encoded by these genes are involved in electron transport to nitrogenase. In addition, these mutants were shown to contain significantly reduced amounts of nitrogenase. The hypothesis that this new class of nitrogen fixation genes encodes components of an electron transfer system to nitrogenase was corroborated by analysing the effect of metronidazole. Both the fdxN and rnf mutants had higher growth yields in the presence of metronidazole than the wild type, suggesting that these mutants contained lower amounts of reduced ferredoxins.


[Differentially expressed genes in immunosuppressive and healthy mice after intranasal inoculation of Aspergillus fumigatus].
To screen out the differential expression genes between the immune suppressive host and healthy host after intranasal inoculation of Aspergillus fumigatus (A fumigatus). Six immune suppressive mice and 6 healthy mice were inoculated with A, fumigatus intranasally. Three days after the inoculation the mice were killed and autopsied. The total RNA of the lungs was isolated. Gene chip technology was used to examine the differentially expressed genes. 66 differentially expressed genes were screened out of the 4098 genes. In the experimental group the genes encoding CD22, CD53, and 24p3 and the genes related to engulfment and digestion of macrophage or monocytes were up-regulated. The genes encoding surfactant associated protein C, immunoglobulin lambda chain and the genes related to pulmonary metabolism were down-regulated. The genes encoding IL18, CD8, and CD28 were not differentially expressed. The up-regulation of CD22 gene may be one of the ways by which the host defense is inhibited. CD53 may take part in the regulation of engulfment of phagocytes. 24p3 protein, an acute stress protein, protects the host from excessive inflammatory response.


Circulating immune complexes in patients with cystic fibrosis in relation to clinical features.
Immune complexes have been previously reported in the serum of patients with cystic fibrosis. This study was undertaken to relate the finding of complexes with the clinical features of the disease. Immune complexes detected by the 125I-C1q binding assay were found in the sera of 17/60 (28%) of patients with cystic fibrosis (CF). There was no association between the finding of raised levels of immune complexes and duration of chest symptoms, duration of daily sputum production, age, sex, weight, atopy, the presence or absence of malabsorption, pneumothorax, diabetes, Aspergillus precipitins or specific bacterial pathogens in the sputum. There was however a correlation between the finding of increasing circulating immune complexes and decrease in the respiratory function; forced expiratory volume in 1 sec (P less than 0.001) and forced vital capacity (P less than 0.005); also with weight (P less than 0.02). It is possible that the finding of immune complexes, at low levels, in cystic fibrosis are the result of tissue damage rather than its cause.


Dermatomycosis in dogs.
During the routine examination of dogs for cutaneous lesions, 205 dogs were screened for fungi other than dermatophytes. Twenty-two dogs (10.8%) revealed the presence of non-dermatophytic fungi suspicious for representing the etiologic agents of the skin lesions. The fungi isolated were Alternaria sp. (2.9%), Penicillium sp. (2.4%), Aspergillus fumigatus (2.0%), Mucor sp. (1.5%), Cladosporium sp. (1.5%) and Fusarium sp. (0.5%). No dermatophyte was isolated in association with these fungi. The incidence of these infections was found to be greater in warm and humid climate.


A pharmacologic approach to the infusion of ritodrine.
The recommended regimen for the infusion of ritodrine leads to unnecessarily high plasma concentrations. A review of basic pharmacologic principles provides a basis for changing the infusion regimen. A regimen is proposed that results in lower average and peak concentrations and reduces the total amount of drug given. These changes may decrease the risk of developing side effects without adversely affecting the ability to inhibit preterm labor.


Use of protein A-treated sera in unmasking herpes simplex virus type 1 (HSV-1) immunoglobulin A and identifying HSV-1 immunoglobulin G as the predominant neutralizing antibody.
Treatment of human sera with protein A reduced the amounts of immunoglobulin G (IgG), IgA, and IgM detected by radial immunodiffusion. This treatment also decreased the amount of herpes-specific IgG and IgM detected by radioimmunoassay, whereas it increased and even unmasked the amount of herpes-specific IgA detected. Comparison of protein A-treated sera with untreated sera indicated that herpes simplex virus type 1 IgG was responsible for more than 92 to 99% of the serum neutralizing activity.


Carbachol, an acetylcholine receptor agonist, enhances production in rat aorta of 2-arachidonoyl glycerol, a hypotensive endocannabinoid.
The production of 2-arachidonoyl glycerol, an endogenous cannabinoid, is enhanced in normal, but not in endothelium-denuded rat aorta on stimulation with carbachol, an acetylcholine receptor agonist. 2-Arachidonoyl glycerol potently reduces blood pressure in rats and may represent an endothelium-derived hypotensive factor.


Serpin-9 and -13 regulate hemolymph proteases during immune responses of Manduca sexta.
Serpins are a superfamily of proteins, most of which inhibit cognate serine proteases by forming inactive acyl-enzyme complexes. In the tobacco hornworm Manduca sexta, serpin-1, -3 through -7 negatively regulate a hemolymph serine protease system that activates precursors of the serine protease homologs (SPHs), phenoloxidases (POs), Spätzles, and other cytokines. Here we report the cloning and characterization of M. sexta serpin-9 and -13. Serpin-9, a 402-residue protein most similar to Drosophila Spn77Ba, has R366 at the P1 position right before the cleavage site; Serpin-13, a 444-residue ortholog of Drosophila Spn28Dc, is longer than the other seven serpins and has R410 as the P1 residue. Both serpins are mainly produced in fat body and secreted into plasma to function. While their mRNA and protein levels were not up-regulated upon immune challenge, they blocked protease activities and affected proPO activation in hemolymph. Serpin-9 inhibited human neutrophil elastase, cathepsin G, trypsin, and chymotrypsin to different extents; serpin-13 reduced trypsin activity to approximately 10% at a molar ratio of 4:1 (serpin: enzyme). Serpin-9 was cleaved at Arg366 by the enzymes with different specificity, but serpin-13 had four P1 sites (Arg410 for trypsin-like proteases, Gly406 and Ala409 for the elastase and Thr404 for cathepsin G). Supplementation of induced cell-free hemolymph (IP, P for plasma) with recombinant serpin-9 did not noticeably affect proPO activation, but slightly reduced the PO activity increase after 0-50% ammonium sulfate fraction of the IP had been elicited by bacteria. In comparison, addition of recombinant serpin-13 significantly inhibited proPO activation in IP and the suppression was stronger in the fraction of IP. Serpin-9- and -13-containing protein complexes were isolated from IP using their antibodies. Hemolymph protease-1 precursor (proHP1), HP6 and HP8 were found to be associated with serpin-9, whereas proHP1, HP2 and HP6 were pulled downed with serpin-13. These results indicate that both serpins regulate immune proteases in hemolymph of M. sexta larvae.


Remnant-Tensioning Single-Bundle Anterior Cruciate Ligament Reconstruction Provides Comparable Stability to and Better Graft Vascularity Than Double-Bundle Anterior Cruciate Ligament Reconstruction in Acute or Subacute Injury: A Prospective Randomized Controlled Study Using Dynamic Contrast-Enhanced Magnetic Resonance Imaging.
To compare the clinical, second-look arthroscopic, magnetic resonance imaging (MRI), and dynamic-contrast-enhanced MRI (DCE-MRI) findings between remnant-tensioning single-bundle (RT-SB) and double-bundle (DB) anterior cruciate ligament reconstruction (ACLR). Sixty-seven patients with acute or subacute anterior cruciate ligament (ACL) injury were randomized to undergo RT-SB or DB ACLR. Twenty-six patients in the RT-SB group and 28 in the DB group were evaluated using stability tests (Lachman test, pivot-shift test, and KT-2000 arthrometer) and multiple clinical scores. One year postoperatively, all 54 patients underwent MRI for evaluation of graft continuity and graft signal/noise quotient and DCE-MRI for the calculation of normalized area under the curve (nAUC) as a marker of graft vascularity. Among them, 41 patients underwent second-look arthroscopy for the evaluation of graft continuity, graft tension, and synovialization. The results were compared between the 2 groups. At the minimum 2-year follow-up (28.7 ± 6.4 months), the stability tests, clinical scores, second-look arthroscopic findings, and MRI findings were not significantly different between the groups. However, the mean nAUC values on DCE-MRI for the ACL graft were significantly higher in the RT-SB group than those in the DB group in all 3 zones (nAUCproximal, P = .005; nAUCmiddle, P = .021; nAUCdistal, P = .027; and nAUCaverage, P = .008). For acute or subacute ACL injury, the RT-SB ACLR showed an outcome comparable to that of DB ACLR in terms of knee stability, clinical scores, MRI findings, and second-look arthroscopic findings. Moreover, RT-SB ACLR showed better graft vascularity 1 year postoperatively than DB ACLR using DCE-MRI. II, prospective randomized controlled trial.


[Epidural anaesthesia associated with induction of labour. A clinical study. Electronic monitoring of the physical and biological parameters (author's transl)].
73 case histories of induction of labour under epidural anaesthesia have been studied very thoroughly from the different parameters of the uterine contractions as well as the fetal heart rate and the acid base balance of the fetus. From this study it is clear that the basal tone, the intensity and the frequency of contractions and the activity of the uterus are increased in comparison with different studies of normal labour. This increase however is within physiological limits and does not change in any marked way the cardiac rhythm nor the acid base balance of the fetus. The Apgar score at birth is equal to or above 8 in 89 p. 100 of cases at 1 minute and in 100 p. 100 of cases at 5 minutes. Finally, in the last phases of the labour dilatation is quicker than in normal delivery. It follows therefore that maternal comfort has not been bought at the expense of the fetus.


Effects of testosterone in the treatment of immune-mediated sensorineural hearing loss.
Testosterones have beneficial effects not only on the modulation of expression of autoimmunity of B/W mice, but on other unrelated models such as autoimmune thyroiditis as well. This study was performed to evaluate the effect of testosterone in the treatment of immune-mediated sensorineural hearing loss (IMSNHL). To induce IMSNHL, 29 female Wistar rats were sensitized with bovine inner ear antigen three times weekly. Fourteen of 29 rats were injected subcutaneously three times a week with 250 microg of testosterone propionate from 1 week prior to the first sensitization to 1 week following the second booster sensitization. An auditory brain-stem response (ABR) test was performed prior to each antigen challenge and at 1, 2, 3, 4, 6 and 8 weeks following sensitization. On ABR testing, the testosterone-treated animals showed better results than the control animals with IMSNHL in the degree of hearing loss (10 to 20 dB vs. 10 to 40 dB) and incidence of hearing loss over 20 dB (25.0% vs. 53.3%). None of testosterone-treated animals showed a hearing loss over 20 dB, whereas 40% of the control animals with IMSNHL revealed hearing loss over 20 dB. These results suggest that testosterone propionate may be effective in the prevention of and early recovery from IMSNHL and may be used in the future as a one treatment modality for IMSNHL.


Stereoselective effects of the enantiomers, quinidine and quinine, on depolarization- and agonist-mediated responses in rat isolated aorta.
1. The effects of the two enantiomers, quinidine and quinine, were studied on depolarization- and agonist-induced isometric contractions in rat isolated thoracic aortic rings. 2. Quinidine or quinine (10(-6)M-3 x 10(-4)M) produced a concentration-dependent relaxation of 80 mM KCl-contracted rings, the pD2 values being 4.89 and 4.23, respectively. Thus, quinidine was about 4-5 times more potent than quinine. 3. The voltage-dependence of quinidine- and quinine-induced inhibition was studied in rings that had been incubated in 5 or 40 mM KCl Ca(2+)-free solution and then contracted by changing the bath solution to 100 mM KCl and 2 mM Ca2+. The inhibitory effects of quinidine were significantly enhanced when the rings were preincubated in 40 mM KCl (depolarizing conditions), when compared to normally polarized rings. In contrast, the effects of quinine were similar in 5 or 40 mM KCl solution. 4. The antagonism of noradrenaline (NA)-induced contractions by low concentrations of quinidine (< 10(-4)M) and quinine (< 3 x 10(-4)M) was competitive, as demonstrated by the concentration-dependent parallel rightward shift of the NA concentration-response curves (pA2 values 6.20 and 5.68, respectively, P < 0.05). 5. At low concentrations (< or = 3 x 10(-5)M), quinidine and quinine did not shift the concentration-response curve to 5-hydroxytryptamine (5-HT) or endothelin-1, whereas at higher concentrations they produced a downward shift of these curves. Quinidine and quinine (> 10(-4)M) inhibited to a similar extent both the phasic (induced in Ca(2+)-free media) and tonic responses (after restoring extracellular Ca2+) induced by 5-HT. 6. In conclusion, quinidine and quinine produced a stereoselective inhibition of depolarization and NA-induced contractions, quinidine being more potent than quinine. The inhibition of KCl-induced contractions could be attributed to inhibition of Ca2+ entry. Both drugs also behaved as competitive antagonists of alpha 1D-adrenoceptors. At high concentrations, quinidine and quinine also decreased the contractions induced by endothelin-1 and 5-HT in a non-stereoselective manner.


Parsing within & between-person dynamics of therapy homework completion and clinical symptoms in two cognitive behavioral treatments for adults with anhedonia.
Homework is a key theoretical component of cognitive-behavioral therapies, however, the effects of homework on clinical outcomes have largely been evaluated between-persons rather than within-persons. The effects of homework completion on treatment response were examined in a randomized trial comparing Behavioral Activation Treatment for Anhedonia (BATA, n = 38), a novel psychotherapy, to Mindfulness-Based Cognitive Therapy (MBCT, n=35). The primary endpoint was consummatory reward sensitivity, measured weekly by the Snaith Hamilton Pleasure Scale (SHAPS), up to 15 weeks. Multilevel models evaluated change in SHAPS scores over time and the effects of clinician-reported and participant-reported homework. BATA and MBCT resulted in significant, equivalent reductions in SHAPS scores. Unexpectedly, participants who completed greater mean total amounts of homework did not improve at a faster rate (i.e., no between-person effect). However, sessions with greater than average participant-reported homework completion were associated with greater than average reductions in SHAPS scores (i.e., a within-person effect). For clinician-reported homework, this effect was only evident within the BATA condition. This study shows psychotherapy homework completion relates to symptomatic improvement in cognitive-behavioral treatments for anhedonia when session-to-session changes are examined within-person. On the contrary, we found no evidence that total homework completion predicted greater improvements between-person. When possible, psychotherapy researchers should evaluate their constructs of interest across multiple sessions (not just pre/post) to allow more direct tests of hypotheses predicted by theoretical models of individual change processes.


Liver-derived FGF21 is essential for full adaptation to ketogenic diet but does not regulate glucose homeostasis.
Fibroblast growth factor 21 (FGF21) is expressed in several metabolically active tissues, including liver, fat, and acinar pancreas, and has pleiotropic effects on metabolic homeostasis. The dominant source of FGF21 in the circulation is the liver. To analyze the physiological functions of hepatic FGF21, we generated a hepatocyte-specific knockout model (LKO) by mating albumin-Cre mice with FGF21 flox/flox (fl/fl) mice and challenged it with different nutritional models. Mice fed a ketogenic diet typically show increased energy expenditure; this effect was attenuated in LKO mice. LKO on KD also developed hepatic pathology and altered hepatic lipid homeostasis. When evaluated using hyperinsulinemic-euglycemic clamps, glucose infusion rates, hepatic glucose production, and glucose uptake were similar between fl/fl and LKO DIO mice. We conclude that liver-derived FGF21 is important for complete adaptation to ketosis but has a more limited role in the regulation of glycemic homeostasis.


Transoral robotic surgery versus upper airway stimulation in select obstructive sleep apnea patients.
Transoral robotic surgery (TORS) has been used to treat obstructive sleep apnea (OSA) since 2009, with recent meta-analysis showing an average reduction of apnea-hypopnea index (AHI) from 44.3 to 17.0. In 2014, upper airway stimulation surgery (UAS) was approved for OSA treatment, with results showing an average AHI reduction from 32.0 to 15.3. Given there was a period when TORS was available and UAS was not, we looked at a subset of patients treated with TORS but who could have qualified for UAS and compared their outcomes to patients who received UAS. This is a retrospective chart review comparing TORS to UAS in treatment of OSA performed by a single surgeon between 2011 and 2016. Inclusion criteria were a body mass index less than 35 and AHI between 20 and 65 consistent with criteria for UAS implantation. Patients who received TORS and met the inclusion criteria had their preoperative drug-induced sleep endoscopy recordings re-evaluated. Patients with anteroposterior retropalatal collapse that would have qualified them for UAS had their outcomes compared to patients who received UAS. Results between TORS and UAS showed an average AHI reduction of 12.7 and 33.3, respectively. Overall cure rate, defined as AHI < 5, was 10.0% and 70.3%, respectively. Results of this study indicate that, when met with criteria for both TORS and UAS, patients receiving UAS had greater improvement in several objective measures of OSA. Studies like this may help direct future treatment algorithms for surgical management of OSA. 4 Laryngoscope, 129:256-258, 2019.


Identification of Cyclocybe erebia metabolites that affect the circadian rhythm of Eluc expression under control of Bmal1 promoter in mouse fibroblast cells.
Pharmacological intervention of circadian rhythms is a potentially useful approach for ameliorating various health problems caused by disturbed circadian rhythms including sleep disorder and metabolic diseases. To find compounds that affect circadian rhythms, we screened mushroom extracts using mouse cells expressing the luciferase gene under the control of the mouse Bmal1 promoter. The culture filtrate extract from the basidiomycete Cyclocybe erebia enhanced the oscillation of bioluminescence caused by the expression of the luciferase gene and prolonged the period of bioluminescence. Bioassay-guided fractionation of the extract resulted in purification of compounds 1 and 2. Spectroscopic analyses along with single-crystal X-ray diffraction analysis, revealed that these compounds were diterpenoids with a unique skeleton and a fused ring system comprising 3-, 7-, and 5-membered rings. Compounds 1 and 2 were named cyclocircadins A and B, respectively. These findings suggested that natural diterpenoids could be a source of compounds with the activity affecting circadian rhythms.


Disinhibition in the rat septum mediated by M1 muscarinic receptors.
Acetylcholine (ACh) has been documented as an important central neurotransmitter. We have investigated the actions of ACh within the dorsolateral septal nucleus of the rat to examine its actions within this nucleus, specifically how it may interact to modulate the inhibitory action of gamma-aminobutyric acid (GABA), the known inhibitory transmitter in this area. Our results demonstrate that ACh, acting on M1 muscarinic receptors leads to disinhibition by decreasing GABA release.


Angiogenic response of endothelial cells seeded dispersed versus on beads in fibrin gels.
Induction of an inter-connected microvessel network in a tissue-engineered construct prior to implantation may be an alternative to improve the success rate of cell/tissue survival and wound integration. Conditions of endothelial cell-seeding density and distribution were investigated in two 3-D angiogenesis culture systems. Endothelial cells were either seeded dispersed in a fibrin gel, or subconfluent on micro-beads (Cytodex) prior to being embedded in fibrin. Human fibroblasts and growth factors were introduced to optimize angiogenesis. A density higher than 4 x 10(4) cells/ml of fibrin was necessary to induce angiogenic-like structures (i.e., sprouting, cord-, lumen-like structures) by 14 days in the dispersed cell model. Endothelial cells on micro-beads also exhibited angiogenic-like structures that were inter-connected to those on neighboring beads. The sizes of the angiogenic-like structures were larger on beads compared to those found in the dispersed cell model. High cell density was needed in angiogenesis when cells were seeded separately, whereas the association of endothelial cells on bead surfaces significantly reduced the cell density used. Moreover, increasing bead density was not necessary to facilitate further angiogenic formation. Micro-spheres may represent a potential support for endothelial cells in microvessel networking, with subsequent applications in the pre-vascularization of bio-implants.


Lower arterial catheter-related infection in brachial than in femoral access.
Recent guidelines do not establish a recommendation about the arterial catheter site to minimize the arterial catheter-related infection risk. In this prospective and observational study, we found a higher arterial catheter-related infection in 1085 arterial femoral sites than in 141 arterial brachial sites (5.08 vs 0 per 1000 catheter-days, respectively; odds ratio, 6.18; 95% confidence interval: 1.11-infinite; P = .02). Thus, arterial brachial access should be used in preference to femoral access.


Sentinel lymph node biopsy with a gamma probe in patients with high-risk cutaneous squamous cell carcinoma: follow-up results of sentinel lymph node-negative patients.
Lymphoscintigraphy (LS) and sentinel lymph node biopsy have become the standard of care for melanoma and breast cancer. However, the data on patients with cutaneous squamous cell carcinoma (SCC) are limited. We aimed to evaluate and identify the role of LS and sentinel lymph node biopsy in patients with high-risk cutaneous SCC. Nineteen patients (13 men, six women; 47-87 years of age, mean age 67.5 ± 12.3) with SCC were included in the study. LS was performed on all patients after intracutaneous injection of Tc-99m nanocolloid. Primary lesions and sentinel lymph nodes (SLNs) were excised with the help of a gamma probe. A total of 26 SLNs and 32 secondary lymph nodes were imaged on LS and were marked. During surgery, 29 SLNs, 21 secondary lymph nodes and three nonactive lymph nodes were excised. In total, 53 lymph nodes were removed surgically. A histopathological study revealed that all lymph nodes were negative for metastasis. Patients were followed up for an average of 41.1 ± 22.2 months (7-80 months). Until the time of data collection, 14 patients were alive and had no regional lymph node or distant metastasis. Local recurrence was seen in only one patient. He was reoperated upon 38 months ago. The feasibility of determining SLNs using LS and an intraoperative gamma probe in patients with cutaneous SCC was shown. Unnecessary elective lymph node dissection and possible complications could be avoided in 19 patients.


Verapamil induces glucose intolerance in conscious, nondiabetic rats.
Intravenous verapamil administration in normal rats significantly impaired the rats' ability to respond to a glucose tolerance test, and blocked the glucose-stimulated release of insulin into the blood. The results suggest that verapamil can impair pancreatic endocrine function in vivo.


HIV antibody testing and its correlates among heterosexual attendees of sexually transmitted disease clinics in China.
This study was conducted to determine the prevalence of HIV antibody testing and associated factors among heterosexual sexually transmitted disease (STD) clinic attendees in China. A self-administered questionnaire was administered among 823 attendees of 4 STD clinics of Zhejiang Province, China in October to December 2007. Psychosocial and behavioural factors associated with HIV antibody testing were identified in both genders using univariate and multivariate analyses. Of all 823 STD clinic attendees, 9.3% of male and 18.0% of female attendees underwent HIV antibody testing in the most recent 6 months, and 60% of the participants had gotten no educational/behavioral intervention related to HIV prevention. The correlates for HIV antibody testing in the most recent 6 months as identified by multivariate analysis were ever condom use [odds ratio (OR), 10.37; 95% confidence interval (CI), 1.32-81.22]; ever anal/oral sex (OR, 3.13; 95% CI, 1.03-9.50) during their lifetime; having ever received three to seven types of behavioural interventions in the most recent 6 months (OR, 3.70; 95% CI, 1.32-10.36) among male subjects; and ever condom use (OR, 12.50; 95% CI, 2.20-71.01), STD history (OR, 3.86; 95% CI, 1.26-11.86) over their lifetime, or having ever received three to seven types of behavioural interventions in the most recent 6 months (OR, 8.68; 95% CI, 2.39-31.46) in female subjects. A lifetime experience of casual/commercial sex partners was strongly negatively associated with HIV testing in female subjects (OR, 0.08; 95% CI, 0.01-0.83). The low prevalence of HIV antibody testing and behavioural intervention among STD clinic attendees indicates a need for more targeted, intensive behavioural interventions to promote HIV antibody testing in this population.


Comparing long-term educational effects of two early childhood health interventions.
We compare the educational effects of two medical protocols that mitigate long-term consequences of prematurity or low birth weight. The two protocols are Traditional Care (TC), which uses incubators, and Kangaroo Mother Care (KMC) which replaces incubators for 24-hour skin-to-skin contact between newborns and caregivers. We concentrate on educational outcomes addressing contradictory results in previous contributions. We use a randomized controlled trial implemented in 1993 that randomly assigned children to either TC or KMC. OLS results suggest that KMC children spent more time in preschool, had fewer temporary school absences, and showed lower math test scores. Both groups observed similar effects on high-school graduation and language test scores. We correct for attrition, small sample, and multiple outcomes. Effects on preschool attendance and school absenteeism are robust, particularly for more vulnerable infants (birth weight ≤ 1,800 g). The other effects lose statistical significance due to multiple outcome testing or attrition corrections.


Transforming growth factor-β receptors.
Transforming growth factor beta (TGF-β) binds specifically and with high affinity to several different cell surface proteins. Low M(r) proteins of 50,000 and 80,000 have been termed type I and type II receptors. Intermediate sized binding components of 115,000-140,000 M(r) and a high binding components of approximately 250,000 M(r) in subunit size have been termed type III receptors. The high M(r) component is a proteoglycan containing the glycosaminoglycan chains of heparan sulfate and chondroitin sulfate and the intermediate sized components are its core proteins. Although almost all cells have TGF-β receptors, binding of TGF-β to the type III binding components is restricted to cells of fibroblastic, osteoblastic and chondroblastic origin. The physiological relevance of each individual binding class is unclear. However, recent data indicate that the type III protein does not transmit signals to inhibit cell proliferation, induce protein synthesis, or promote cytomorphological change and that these activities may be mediated through the type I receptor. The mechanism of signal transduction remains unknown, but it does not appear to be associated with tyrosine phosphorylation or phosphorylation of the 40s ribosomal protein S6.


Intestinal calcium-binding protein (CaBP) and bone calcium mobilization in response to 25R,26 and 25S,26-dihydroxycholecalciferol in intact and nephrectomized rats.
Since intestinal calcium-binding protein (CaBP) can be regarded as an expression of the hormone-like action of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) on the duodenal enterocyte we have investigated the potential biological activity of 25R and 25S,26-(OH)2D3 (two recently synthesized epimers of vitamin D3 metabolite) to promote intestinal CaBP production as compared to bone calcium mobilization in vitamin D and calcium-deficient rats. In our assay steroids exhibited a 72 hour calcemic response. Our results show a linear relationship between CaBP synthesis and the logarithm of the dose (130-2080 pmol dose range) of either 25R or 25S epimer. The CaBP response was comparable for both epimers. Similarly bone calcium mobilization response was dose related as a linear function of the logarithm of the administered dose. Again, calcemic response was comparable for both epimers. In our model these two epimers were about as active on intestine to increase CaBP amount as on bone to elevate serum calcium level. Bilateral nephrectomy abolished CaBP response to a large dose (1040 pmol) of either 25R or 25S epimer but did not abolish it to a 130 pmol dose of 1alpha,25-(OH)2D3.


Human cardiac gap junctions: isolation, ultrastructure, and protein composition.
Recent experiments from our laboratory have shown that the ultrastructure and protein composition of gap junctions isolated from rat ventricles are tissue specific, i.e., markedly different from gap junctions of liver and lens. The differences include a cytoplasmic surface component characteristic for cardiac gap junctions; this component can be visualized by two ultrastructural techniques: as a fuzzy layer in electron micrographs of thin-sectioned junctional pellets and as cytoplasmic surface particles in deep-etched freeze-fractured junctions. The component corresponds to a Mr 17,500 cytoplasmic surface domain of each of the six (Mr 47,000) rat heart gap junctional channel protein subunits that make up the gap junctional channel hexamer known as a connexon. The cytoplasmic surface component is localized at the carboxy-terminal of the subunit. Within the cytoplasmic surface component, rat cardiac gap junctions are cross-linked by disulfide linkages between subunits of the same connexon and between subunits of adjacent connexons. By contrast, the Mr 28,000 liver gap junctional subunit lacks a comparably large cytoplasmic surface component, cytoplasmic surface fuzz, cytoplasmic surface particles, and intra- and interconnexon disulfide linkages. Most of these unique characteristics of cardiac gap junctions were discovered in junctions isolated from rat ventricles. Unlike liver and lens gap junctions, cardiac gap junctions from humans, non-human primates, or other large mammals have not previously been isolated and characterized. Here we report the isolation of unproteolyzed gap junctions from the ventricle of a 24 year-old man with advanced cardiomyopathy whose heart was removed for replacement by a transplanted heart.(ABSTRACT TRUNCATED AT 250 WORDS)


[Antigen enzyme coupling: a few results (author's transl)].
Various technics of coupling (carbodiimide, glutaraldehyde, periodate) have been used for the grafting of human casein on glucose oxidase. The various types of product obtained were identified by gel filtration and used for drawing up titration curves. The authors compared radioimmunoassay and enzymoimmunologic estimation showing the importance of the molecular size of the conjugates obtained.


HER2-specific affibody-conjugated thermosensitive liposomes (Affisomes) for improved delivery of anticancer agents.
Thermosensitive liposomes are attractive vehicles for the delivery and release of drugs to tumors. To improvethe targeting efficacy for breast cancer treatment, an 8.3-kDa HER2-specific Affibody molecule (Z(HER2:342)-Cys) was conjugated to the surface of liposomes. The effects of this modification on physical characteristics and stability of the resulting nanoparticles denoted as "Affisomes" were investigated. Thermosensitive small unilamellar vesicle (SUV) liposomes of (80-100 nm) a diameter consisting of dipalmitoyl phosphatidylcholine (DPPC, Tm 41 degrees C) as the matrix lipid and a maleimide-conjugated pegylated phospholipid (DSPE-MaL-PEG2000) were prepared by probe sonication. Fluorescent probes were incorporated into liposomes for biophysical and/or biochemical analysis and/or triggered-release assays. Affibody was conjugated to these liposomes via its C-terminal cysteine by incubation in the presence of a reducing agent (e.g., tributylphosphine) for 16-20 hours under an argon atmosphere. Lipid-conjugated affibody molecule was visible as an 11.3-kDa band on a 4-12% Bis/Tris gel under reducing conditions. Affibody conjugation yields were approximately 70% at a protein-lipid ratio of 20 microg/mg, with an average number of 200 affibody molecules per Affisome. Affibody conjugation to thermosensitive liposomes did not have any significant effect on the hydrodynamic size distribution of the liposomes. Thermosensitivity of Affisomes was determined by monitoring the release of entrapped calcein (a water-soluble fluorescent probe, lambdaex/em 490/515 nm) as a function of temperature. Calcein was released from Affisomes (thermosensitive liposomes with affibody-Targeted SUV) as well as nontargeted SUV (thermosensitive liposomes without affibody) in a temperature-dependent manner, with optimal leakage (90-100%) at 41 degrees C. In contrast, liposomes prepared from Egg phosphatidyl choline (Egg PC, Tm approximately 0 degrees C) under similar conditions released only 5-10% calcein at 41 degrees C. Affisomes, when stored at room temperature, retained > 90% entrapped calcein up to 7 days. Moreover, incubation of liposomes in phosphate-buffered saline, supplemented with 10% heat-inactivated serum (fetal bovine serum) did not result in a destabilization of liposomes. Therefore, Affisomes present promising, novel drug-delivery candidates for breast cancer targeting.


Methods to Identify Rat and Mouse Retinal Ganglion Cells in Retinal Flat-Mounts.
The identification of distinct retinal ganglion cell (RGC) populations in flat-mounted retinas is key to investigating pathological or pharmacological effects in these cells. In this chapter, we review the main techniques for detecting the total population of RGCs and various of their subtypes in whole-mounted retinas of pigmented and albino rats and mice, four of the animal strains most studied by the scientific community in the retina field. These methods are based on the studies published by the Vidal-Sanz's laboratory.


Postextubation atelectasis: a retrospective review and a prospective controlled study.
To determine the role of chest physiotherapy in the prevention of postextubation atelectasis in neonates intubated for greater than 24 hours, a retrospective survey compared the incidence of this complication in a newborn intensive care unit prior to and following the institution of a routine of chest physiotherapy. Eight of 23 infants extubated developed atelectasis in the "pre-physio" period, whereas only one collapse occurred in 20 infants treated with a routine of physiotherapy at extubation (P less than 0.025). Subsequently a prospective controlled trial compared the use of a routine of physiotherapy at extubation with no physiotherapy. Eight of 21 infants not receiving physiotherapy developed postextubation atelectasis and none of 21 infants receiving physiotherapy developed atelectasis (P less than 0.01). Seventy-six percent of the collapses involved the right upper lobe. A vigorous program of chest physiotherapy, including postural drainage emphasizing the positions of the right upper lobe and chest vibrations, will significantly reduce the incidence of postextubation atelectasis.


Radioimmunologic localization of vasoactive intestinal polypeptide in hypothalamic and extrahypothalamic sites in the rat brain.
Using a specific radioimmunoassay (RIA), we have quantitated the hypothalamic and extrahypothalamic distribution of vasoactive intestinal polypeptide (VIP) in the rat brain. Highest concentrations of VIP were found in cerebral cortex, suprachiasmatic nucleus, and medial amygdaloid nucleus. Lower concentrations were found in the anterior hypothalamic area, medial preoptic area, peri- and paraventricular nuclei, and hippocampus. Significant levels of VIP were also found in the anterior pituitary. Very little VIP was detected in median eminence and cerebellar cortex. This distribution of VIP in areas of the rat brain known to be important to neuroendocrine function suggests a role for VIP in the control of anterior pituitary events.


Neuroprotective effects of caffeine in MPTP model of Parkinson's disease: A (13)C NMR study.
Parkinson's disease (PD) is a neurodegenerative disorder characterized by degeneration of nigrostriatal dopaminergic neurons with an accompanying neuroinflammation leading to loss of dopamine in the basal ganglia. Caffeine, a well-known A2A receptor antagonist is reported to slow down the neuroinflammation caused by activated microglia and reduce the extracellular glutamate in the brain. In this study, we have evaluated the neuroprotective effect of caffeine in the MPTP model of PD by monitoring the region specific cerebral energy metabolism. Adult C57BL6 mice were treated with caffeine (30 mg/kg, i.p.) 30 min prior to MPTP (25 mg/kg, i.p.) administration for 8 days. The paw grip strength of mice was assessed in order to evaluate the motor function after various treatments. For metabolic studies, mice were infused with [1,6-(13)C2]glucose, and (13)C labeling of amino acids was monitored using ex vivo(1)H-[(13)C]-NMR spectroscopy. The paw grip strength was found to be reduced following the MPTP treatment. The caffeine pretreatment showed significant protection against the reduction of paw grip strength in MPTP treated mice. The levels of GABA and myo-inositol were found to be elevated in the striatum of MPTP treated mice. The (13)C labeling of GluC4, GABAC2 and GlnC4 from [1,6-(13)C2]glucose was decreased in the cerebral cortex, striatum, olfactory bulb, thalamus and cerebellum suggesting impaired glutamatergic and GABAergic neuronal activity and neurotransmission of the MPTP treated mice. Most interestingly, the pretreatment of caffeine maintained the (13)C labeling of amino acids to the control values in cortical, olfactory bulb and cerebellum regions while it partially retained in striatal and thalamic regions in MPTP treated mice. The pretreatment of caffeine provides a partial neuro-protection against severe striatal degeneration in the MPTP model of PD.


Seroprevalence of SARS-CoV2 in IBD Patients Treated with Biologic Therapy.
A similar course of COVID-19 in patients with inflammatory bowel diseases [IBD] and in the general population has been reported. However, disease prevalence in IBD patients is presently unknown. In this prospective observational study, we aimed at determining SARS-CoV2 infection prevalence in IBD patients treated with biologic therapy. From IBD patients under biologic therapy and recruited from three different locations in Italy and Germany, 354 sera were evaluated for antibody presence by RBD ELISA. Control groups were: i] age-matched healthy subjects tested in the same time period in Milan, Italy; ii] healthy subjects collected in the pre-COVID era; iii] IBD patients under biologic therapy collected in the pre-COVID era. Eight out of 354 patients tested positive for the anti-RBD-SARS-CoV2 IgG antibody [prevalence 2.3%]. The percentage of IgG-positive patients among those recruited from Milan was significantly higher than among those recruited from other locations [prevalence 5.4% vs 0.4%, p <0.005]. IgG-positive patients reported a significantly higher incidence of fever, anosmia, and ageusia, and were more likely to have entered into close contact with COVID-19-positive subjects before the study enrolment. Seroprevalence of SARS-CoV2 in IBD patients treated with biologic therapy reflects values measured in the local general population. Specific symptoms and contact history with SARS-CoV2-infected individuals strongly increase the likelihood of SARS-CoV2 seropositivity.


Injury to the anteroinferior glenohumeral capsule during anterior dislocation.
Glenohumeral dislocation commonly results in permanent deformation of the glenohumeral capsule. Knowing the location and extent of tissue damage may aid in improving diagnostic and repair procedures for shoulder dislocations. Therefore, the objectives of this study were to determine: (1) the strain in the anteroinferior capsule at dislocation and (2) the location and extent of injury to the anteroinferior capsule due to dislocation by quantifying the resulting non-recoverable strain. A robotic/universal force-moment sensor testing system was used to anteriorly dislocate six cadaveric shoulders. The magnitude of the maximum principle strain at dislocation and the resulting non-recoverable strain due to dislocation in the anteroinferior capsule were measured by tracking the change in the location of a grid of strain markers from a reference position. The glenoid side of the capsule experienced higher strains at dislocation than the humeral side. The greatest strains at dislocation were found on the glenoid side of the anterior band (strain ratio of 0.60), but the greatest non-recoverable strains were found in the posterior axillary pouch (strain ratio of 0.34 on the glenoid side and 0.31 on the humeral side). These findings suggest that even though the glenoid side of the anterior band undergoes more deformation during anterior dislocation, the most permanent deformation occurs in the posterior axillary pouch, and surgeons should consider also plicating the posterior axillary pouch when performing repair procedures following anterior dislocation. In the future, the mechanical properties of the normal and injured glenohumeral capsules will be compared.


Experimental yield and evaluation of proton induced reactions for neutron production and synthesis of beryllium-7 using lithium compounds as target material.
We had investigated the nuclear processes occur during nuclear reactions of protons with lithium containing targets. Experimental measurements were undertaken to obtain the thick-target yield of 7Be and infer upon the neutron production. Three candidate target materials having the chemical compositions LiF, LiBO2, and Li2B4O7 were used. The 7Be yield was probed using its 477 keV day γ-line while the neutron yield was probed using 63Cu(n,γ)64Cu. The 7Be and neutron yields were found different among the different compositions. Discussion on the details of Li (p,n) reactions and decay of exit channels are given and evaluated in order to explain the difference in yield among different target compositions and evaluate the nominal value of yield. Based on experimental results and evaluation, accumulation of 7Be in the target material limits its operation time for neutron production. The LiF targets gave the highest 7Be yield compared to water soluble LiBO2 and Li2B4O7. Neutron production may be undertaken with LiF target as well as lithium metaborate target with special attention to the target operation time.


[Peptides--inhibitors of carboxycathepsin (peptidyl-dipeptidase A) and their role in clinical medicine].
Properties of the carboxycathepsin inhibitors teprotide, captopryl, enalacryl are considered. Presence of low molecular thermo- and acid stable carboxycathepsin inhibitors of peptide nature in blood serum and lymphocytes is discussed.


Genetic screening of lactic acid bacteria of oenological origin for bacteriocin-encoding genes.
A total of 330 lactic acid bacteria isolated from South African red wines during alcoholic and malolactic fermentations and 9 commercial malolactic bacteria starter cultures were screened for antimicrobial activity. Of the entire screened isolates, 26 strains, belonging to the species Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus hilgardii and Oenococcus oeni, showed activity towards various wine-related and non-wine-related indicator strains. A PCR-based screening revealed the presence of the plantaricin encoding genes plnA, plnEF, plnJ and plnK in five selected Lb. plantarum strains. Furthermore, a co-culture experiment with Lb. plantarum and Enterococcus faecalis was performed. A complete inhibition of cell growth of Ent. faecalis was observed within 72 h. Four putative bacteriocin-encoding genes in the genome of O. oeni were identified and sequenced.


Subjective knee apprehension is not associated to physical parameters 6-12 months after anterior cruciate ligament reconstruction.
Anterior cruciate ligament (ACL) rupture is a common injury and psychological parameters measured at 6-8 months are said to be almost more predictive for return to sport (RTS) than physiological. Purpose was 1) to evaluate the correlation between knee apprehension using ACL-RSI and physical factors after ACL reconstruction (ACLR), 2) to assess the correlation between ACL-RSI and patient parameters (age, pivot-sport, BMI), and 3) to evaluate ACL-RSI over time. Patients with ACLR with or without meniscal repair between 2013 and 2020 were retrospectively analyzed. Including criteria were RTS testing battery, assessed at least 6 months after surgery, including physical parameters (strength, triple hop test, side hop test, and bilateral knee stability) and psychological parameters (ACL-RSI). 5 subgroups were analyzed to assessed factors such as age, BMI, pivot sport, time interval between two RTS testing battery. Three hundred three patients (212 male, 91 female) presenting ACLR were included. Mean age at surgery was 27 (± 8) years. 258 patients practiced pivot-sport activity and 45 non-pivot-sport activity. The mean interval between ACL rupture and surgery was 6.5 (± 4.5) months. RTS testing battery were performed at 8 (± 7) months after ACLR. Mean ACL-RSI was 58 (± 28). 1) ACL-RSI was not influenced by muscle strength, coordination and stability of the knee. 2) ACL-RSI was significantly better in lower BMI and non-pivot-sport activities. No correlation was found between graft type, age, sex, and ACL-RSI assessment. 3) For patients who performed two RTS testing battery at 8 and 12 months, ACL-RSI did not significantly increase over time (56 to 64 points, p = 0.22) in spite of significant increased quadriceps (127 to 151 Nm/kg, p = 0.005) and hamstring (93 to 105 Nm/kg, p = 0.05) strength. Psychological readiness before RTS, measured upon ACL-RSI does not correlate with any physical parameter at 8-12 months postoperatively. Although quadriceps and hamstring strength increased significantly over time, ACL-RSI does not and must therefore be routinely assessed.


Statistical model of pesticide penetration through woven work clothing fabrics.
Statistical models estimating the level of protection and thermal comfort performance of woven fabrics were developed using simple fabric and liquid parameters. Eighteen woven fabrics were evaluated against three pesticide mixtures of atrazine and pendimethalin at different concentrations. Using three mixtures that represent a range of both surface tension and viscosity, percentages of pesticide penetration are measured, along with fabric thickness, fabric cover factor, yarn twist factor, yarn packing factor, solid volume fraction, wicking height, and air permeability. Statistical analyses are performed to examine the relationship between liquid/fabric parameters and pesticide penetration. Statistical analyses show that fabric cover factor, yarn twist factor, viscosity of pesticide mixture, critical surface tension of solid, and wicking height are significant parameters affecting pesticide penetration. For this purpose, cover factor and twist factor are better parameters in describing the geometry of woven fabrics than solid volume fraction. Modeling of comfort performance of woven fabric based on simple textile parameters shows that the combination of fabric thickness, cover factor, yarn twist factor and yarn packing factor can be used to estimate air permeability of woven fabric. These findings could be used for developing selection charts or tools as guidelines for the selection of personal protective equipment for use in hot, humid environments.


Inheritance of the acoustic signal parameters in interspecific hybrids of the bank (Myodes glareolus) and the Tien Shan (M. centralis) voles.
The continuity of behavioral responses in the traits of offspring can be interpreted ambiguously because animal behavior can be transmitted from generation to generation genetically or can be trained. Inheritance of the sound signals characteristics in the absence of directional selection is of particular interest, and that was the purpose of the present work. Comparisons of distress signals of hybrids of the Tien Shan vole females Myodes centralis (Kyrgyzstan) and the bank vole males M. glareolus suecicus (the Tver region), and the parent species were made. Acoustic signals of the hybrids and parent species were compared using the variance and discriminant analysis. The distress signals of the Tien Shan voles were shorter, had lower peak frequency than the signals of the bank voles. The peak frequency of hybrids signals was closer to that of the bank voles. However, the duration of the hybrids sounds, on the contrary, was closer to that of the Tien Shan voles. The expression of the noise component in the sounds of the hybrids occupied an intermediate position. The discriminant analysis of distress signals in the Tien Shan and bank voles showed 94% of correct attributions. Our results confirm that the acoustic signals parameters were inheritance intermediate. The paternal genome (the bank voles) had greater influence on parameters of the sound signals in hybrids.


Hospital admissions due to vasomotor and allergic rhinitis in England and Wales between 1999 and 2019: an ecological study.
Allergic rhinitis is among the most prevalent chronic disorders in high-income countries. Its estimated cost is €30-€50 billion annually in the European Union and in the UK, it is approximately €1.8 billion per year. To study hospital admissions for vasomotor and allergic rhinitis in England and Wales during the past 20 years. This was an ecological study using publicly available data extracted from the Hospital Episode Statistics (HES) database in England and the Patient Episode Database for Wales (PEDW). The trend in hospital admissions was assessed using a Poisson model. Admission rate increased by 2.14-fold [from 2.59 (95% CI 2.46-2.73) in 1999 to 8.16 (95% CI 7.93-8.39) in 2020 per 100,000 persons, trend test, p < 001]. The most prevalent hospital admission reasons were allergic rhinitis due to pollen, other allergic rhinitis, and unspecified allergic rhinitis, which accounted for 56.4%, 22.3%, and 15.7%, respectively. The age group 15-59 years accounted for 69.5% of the total number of admissions. Admission rate between males increased by 2.25-fold. Admission rate between females increased by 2.02-fold. Vasomotor and allergic rhinitis are common cause of hospital admissions in England and Wales that showed a clear increase in the rate of their admissions in the past 20 years. Allergic rhinitis due to pollen was the most dominant cause of admission, which warrants further investigation to identify its preventable risk factors and decrease the probability of the exacerbation of patients' cases and the need for hospitalization.


Optimal waist circumference cut-off points and ability of different metabolic syndrome criteria for predicting diabetes in Japanese men and women: Japan Epidemiology Collaboration on Occupational Health Study.
We sought to establish the optimal waist circumference (WC) cut-off point for predicting diabetes mellitus (DM) and to compare the predictive ability of the metabolic syndrome (MetS) criteria of the Joint Interim Statement (JIS) and the Japanese Committee of the Criteria for MetS (JCCMS) for DM in Japanese. Participants of the Japan Epidemiology Collaboration on Occupational Health Study, who were aged 20-69 years and free of DM at baseline (n = 54,980), were followed-up for a maximum of 6 years. Time-dependent receiver operating characteristic analysis was used to determine the optimal cut-off points of WC for predicting DM. Time-dependent sensitivity, specificity, and positive and negative predictive values for the prediction of DM were compared between the JIS and JCCMS MetS criteria. During 234,926 person-years of follow-up, 3180 individuals developed DM. Receiver operating characteristic analysis suggested that the most suitable cut-off point of WC for predicting incident DM was 85 cm for men and 80 cm for women. MetS was associated with 3-4 times increased hazard for developing DM in men and 7-9 times in women. Of the MetS criteria tested, the JIS criteria using our proposed WC cut-off points (85 cm for men and 80 cm for women) had the highest sensitivity (54.5 % for men and 43.5 % for women) for predicting DM. The sensitivity and specificity of the JCCMS MetS criteria were ~37.7 and 98.9 %, respectively. Data from the present large cohort of workers suggest that WC cut-offs of 85 cm for men and 80 cm for women may be appropriate for predicting DM for Japanese. The JIS criteria can detect more people who later develop DM than does the JCCMS criteria.


Comparison of sea turtle thrombocyte aggregation to human platelet aggregation in whole blood.
The endangered sea turtles are living "fossils" that afford us an opportunity to study the hemostatic process as it likely existed millions of years ago. There are essentially no data about turtle thrombocyte aggregation prior to our studies. Thrombocytes are nucleated cells that serve the same hemostatic functions as the anucleated mammalian platelet. Sea turtle thrombocytes aggregate in response to collagen and beta-thrombin. Ristocetin induces an agglutination/aggregation response indicating the presence of a von Willebrand-like receptor, GPIb, found in all mammalian platelets. Samples treated with alpha-thrombin plus gamma-thrombin followed by ristocetin results in a rapid, stronger response than ristocetin alone. These responses are inhibited by the RGDS peptide that blocks fibrinogen cross-linking of mammalian platelets via the fibrinogen receptor, GPIIb/IIIa. Three platelet-like proteins, GPIb, GPIIb/IIIa and P-selection are detected in sea turtle thrombocytes by fluorescence activated cell sorting. Turtle thrombocytes do not respond to ADP, epinephrine, serotonin, thromboxane A2 mimetic, U46619, trypsin, or alpha-thrombin and gamma-thrombin added alone. Comparison of hemostasis in sea turtles to other vertebrates could provide a framework for understanding the structure/function and evolution of these pathways and their individual components.


In vitro binding and in vivo clearance of human alpha 2-macroglobulin after reaction with endoproteases from four different classes.
The binding of human alpha 2-macroglobulin complexed with trypsin, papain, thermolysin and cathepsin-D to murine macrophages was studied at 4 degrees C. Similar dissociation constants (0.4 nM) were determined for all of the complexes except alpha 2-macroglobulin-cathepsin-D (0.7 nM). Radioiodinated alpha 2-macroglobulin-protease complexes were injected into mice, and the clearance studied. Native alpha 2-macroglobulin cleared slowly, as previously reported, while greater than 50% of the complexes formed with trypsin, papain and thermolysin cleared in less than 5 min. The clearance of alpha 2-macroglobulin-cathepsin-D was biphasic, suggesting that only about half the alpha 2-macroglobulin was present in a reacted complex.


Influence of ceramic firing on marginal gap accuracy and metal-ceramic bond strength of 3D-printed Co-Cr frameworks.
The marginal gap and ceramic bond strength of metal-ceramic restorations are important for success. However, studies evaluating the marginal gap and ceramic bond strength of fixed partial dentures (FPDs) produced with 3D printing technologies such as selective laser melting (SLM) are scarce. The purpose of this in vitro study was to investigate the marginal gap of cobalt-chromium (Co-Cr) alloy frameworks produced by SLM technology before and after ceramic firing. Additionally, the metal-ceramic bond strength was evaluated with the Schwickerath crack-initiation test according to the International Standards Organization (ISO) 9693-1:2012. Conventional impressions were made, and the definitive cast of a patient requiring a 4-unit FPD was scanned. After designing the FPD, the files were sent to a service center for the fabrication of a metal master model, 80 Co-Cr frameworks, and 80 flat specimens (25×3×0.5 mm) with SLM technology. The marginal gap between frameworks and the abutment tooth of the metal master model was nondestructively measured by using an optical coordinate-measuring machine. A total of 80 sets, consisting of 1 framework and 1 flat specimen, were sent to 80 dental laboratory technicians for ceramic firing. Detailed instructions for correct manipulation of the framework and flat specimen were provided. The marginal gap was remeasured, and the 3-point bend test was used to evaluate metal-ceramic bond strength. Only 28 of the 80 dental technicians returned the specimens within a prespecified time and/or in adequate condition. The mean ±standard deviation marginal gap of the framework before ceramic firing was 25 ±9 μm and 34 ±12 μm after firing. The difference was statistically significant (P=.001). The mean ±standard deviation 3-point bend strength was 33 ±9 MPa. Ceramic firing affected the marginal gap; however, all Co-Cr frameworks had a marginal gap lower than 120 μm, which is reported to be a clinically acceptable limit. Most of the specimens (80%) had a metal-ceramic bond strength value higher than the 25-MPa ISO 9693 requirement. Five of 28 dental laboratory technicians were not able to comply with ceramic firing instructions.


Frontiers in lymphatic microsurgery.
The Authors report an overview on the modern surgical treatment of peripheral lymphoedema. The aim of lymphatic microsurgical operations is to drain the lymph either toward the venous circulation (lympho-venous shunts) or the lymphatic collectors above the obstacle to the lymph flow, with the interposition of lymphatic or venous grafts (lymphatic-venous-lymphatic plasty). Selection of candidate patients for lymphatic microsurgery is based on an adequate diagnostic investigation, which includes above all lymphoscintigraphy, conventional oil contrast lymphangiography, Doppler venous flowmetry and manometry, and, if necessary (angiodysplasias), an accurate study also of the artery circulation. The clinical outcome of lymphatic microsurgery, assessed by water volumetry and lymphangioscintigraphy, performed at variable distance of time from operations till over 5 years after surgery, shows a significant reduction of edema volume and improvement of lymph flow in all patients and that the more precocious the microsurgical treatment the better the results.


Rapid on-site evaluation improves needle aspiration sensitivity in the diagnosis of central lung cancers: a randomized trial.
Few prospective studies have evaluated the role of endobronchial needle aspiration in diagnosing central airways neoplasms. Rapid on-site evaluation has long been used in transbronchial needle aspiration of adenopathies and peripheral lesions, but its role in sampling central malignancies has not been substantiated yet. In this study we evaluated if endobronchial needle aspiration may increase the sensitivity of bronchoscopy for diagnosing central airways neoplasms when added to conventional diagnostic methods (forceps biopsy, brushing and bronchial washing), and if rapid on-site evaluation may be beneficial in patients undergoing endobronchial needle aspiration. 125 patients (77% males, aged 70 ± 7 years; mean ± SD) with central lung cancers were randomized to undergo bronchoscopy including conventional diagnostic methods and needle aspiration, with or without rapid on-site evaluation, stratifying the patients on the basis of the neoplasm growth pattern (exophytic and submucosal/peribronchial disease). Needle aspiration significantly increased the sensitivity of bronchoscopy when added to conventional methods (from 76 to 91%; p < 0.001), primarily resulting from differences in submucosal/peribronchial diseases (68 vs. 90%; p < 0.001) and independently from the presence of rapid on-site evaluation; needle aspiration guided by rapid on-site evaluation guaranteed a higher improvement in bronchoscopy sensitivity than conventional needle aspiration (98 vs. 84%, respectively; p = 0.004). Needle aspiration guided by rapid on-site evaluation showed a significantly higher sensitivity than the conventional method (97 vs. 76%, respectively; p = 0.001). Needle aspiration increases the sensitivity of bronchoscopy in diagnosing central airways malignancies when added to conventional diagnostic methods, with a further significant improvement when guided by rapid on-site evaluation.


The use of simulation technology in the education of nursing students.
This article provides a broad overview of simulation use in nursing education. An example of a simulation experience focusing on the development and debriefing of a medication safety exercise is presented based on a theoretic approach. Questions for future consideration by nursing educators and researchers are also presented with the ultimate goal of producing safer patient care and more deliberate reflecting practitioners.


Influence of Mycoplasma arginini infection on the induction of aryl hydrocarbon hydroxylase by TCDD in rat hepatoma cell cultures.
Mycoplasma arginini was eliminated from a rat hepatoma cell line (H-4-II-E) by plating at low cell density and treatment with chlortetracycline (250 micrograms/ml), kanamycin (250 micrograms/ml), tylosin (100 micrograms/ml), 3% M. arginini antiserum and 5% fresh guinea-pig serum. The induction of AHH activity in the cell culture was measured in response to increasing concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The ED50 values (estimated doses that produce 50% maximum enzyme induction) were calculated to be 0.256, 0.452 and 0.344 pmol TCDD/plate for original, mycoplasma-free and reinfected cells, respectively. Although the absence of M. arginini in the rat hepatoma cell line makes the cells slightly less responsive to AHH induction by TCDD, this decrease does not detract from the use of the method to screen food extracts and environmental samples for the presence of certain toxic planar organic compounds.


Breast Radiation Therapy Using a Breast Cup: Moving the Target, Not the Patient.
To compare whole-breast radiation therapy dose distributions with and without the use of breast cups to reposition the breast itself. A fitted plastic breast cup repositions the breast on the anterior chest wall, reducing the lateral fall and ptosis of the breast. Eighteen patients requiring breast cups were identified on an initial simulation computed tomography scan (without breast cup); subsequently, a second scan with the breast cup fitted was performed. An optimal treatment plan was then created on both scans with the differences in radiation dose to underlying organs compared. In vivo measurements were performed to assess any possible bolusing effect on skin dose owing to the breast cup. Breast cups significantly reduce dose to both lung and heart for all left-sided cases. All lung dose metrics decreased for all right-sided cases. Right-sided heart dose metrics did not significantly decrease with the use of breast cups; however, heart mean dose for these cases was all under 100 cGy. A 16% increase in skin surface dose was observed in an anthropomorphic phantom when using a breast cup. Use of the breast cup in breast radiation therapy decreases dose to underlying organs at risk and is cost-effective. It can be easily integrated with deep inspiration breath hold, intensity modulated radiation therapy, and volumetric-modulated arc therapy techniques. With use of the device, a supine patient position can be maintained, meaning nodal regions can be treated mono-isocentrically.


A Chemical Time Machine for Mucosal Healing.
Mucosal healing regenerates the mucosal complex upon injury. Mucosal repair requires sequential biologic steps paradoxically enabled or blocked by prostaglandin E2. In this issue of Cell Host & Microbe, Jain et al. (2018) find that the microbial metabolite, deoxycholate, regulates prostaglandin E2 and thus coordinates the steps of mucosal healing.


Inclusions in topaz from miarolitic pegmatites of the Volodarsk-Volynski Massif (Ukraine)--a Raman spectroscopic study.
The differently coloured (colourless, brown-pinkish and blue-pinkish) crystals of topaz from granitic pegmatites of Volodarsk-Volynsky Massif (VVM) have been investigated by scanning electron microscopy (SEM) and Raman microspectroscopy (RS) methods. Topaz (287, 522, 855, 929 cm(-1)), goethite (390 cm(-1)), pyrite (377-379 cm(-1)), marcasite (397 and 331 cm(-1)) and monazite (460 and 970-1070 cm(-1)) were identified as mineral inclusions in analysed crystals. On the basis of RS spectra some of this inclusions contain also organic matter, represented by carbonaceous matter (D-band at ca. 1320-1340 cm(-1) and G-band at ca. 1590-1600 cm(-1)) and liquid simple hydrocarbons consisting of aliphatic and aromatic groups (1240, 1325 and 1420 cm(-1)). Other solid phases found the host topaz, i.e. quartz, orthoclase, very rare minerals (micas) as lepidolite, zinnwaldite and also beryl and rutile, were identified with SEM-EDS analyses. All these mineral inclusions have been formed by post-magmatic, fluid-induced processes, extended from pegmatite to hydrothermal stages of magma crystallization.


Referrals to a pediatric emergency department of a tertiary care teaching hospital before and after introduction of a referral education module - a quality improvement study.
Provision of timely care to critically ill children is essential for good outcome. Referral from smaller peripheral hospitals to higher centers for intensive care is common. However, lack of an organized referral and feedback system compromises optimal care. We studied the quality of referral letters coming to our Emergency Department (ED) with respect to their demography, association with severity of illness and mortality before and after referral education. Our study was completed in three phases in the Pediatric ED; Pre-intervention, Intervention and Post intervention phases. Quality of referral letter was matched with a quality checklist proforma and graded as 'good', 'fair' and 'poor' if it scored > 7, 5-7 and < 5 points respectively. A peer reviewed referral education module was prepared using case studies, expert opinions, and lacunae observed in the first phase and administered to health care providers (HCP's) of referring hospitals. Quality of referral letter was compared between pre and post intervention phases. Most referrals belonged to the neighboring states of Punjab (48.2%) and Haryana (22.4%). Major referring hospitals were from public sector (80.9%), of which the teaching hospitals topped the list (53.6%). Government run ambulance services (85.5%) was commonest mode of transport used and need for a PICU bed and/or mechanical ventilation (50.4%) was the commonest reason for referral. The post intervention phase saw a significant decline in the proportion of poor (93.2 vs.78.2%; p = 0.001) and a significant increase in the proportion of fair (6.1 vs 18%; p = 0.001) and good referral letters (0.7 vs 18%; p = 0.001). The proportion of children with physiological decompensation at triage had reduced significantly in the post intervention phase [513 out of 1403 (36.5%) vs. 310 out of 957 (32.3%); p = 0.001]. Referral education had significantly improved the quality of referral letters. Proportion of children with physiological decompensation at triage had decreased significantly after referral module. This change suggests sensitization of the peripheral hospitals towards a better referral process. Continued multifaceted approach will be required for sustained and increased benefits.


Lumbar and thoracic spine pain in the athlete: radiographic evaluation.
A comprehensive but basic and updated systematic radiographic approach to various pain-producing thoracolumbar diseases is presented. The advantages and limitations of available diagnostic modalities are discussed in relation to the clinically considered etiologies.


Synthesis of some newer derivatives of substituted quinazolinonyl-2-oxo/thiobarbituric acid as potent anticonvulsant agents.
5-[1'-[3"-Aminoacetyl-2"-methyl-6",8"-dihalosubstitutedquinazolin-4"(3"H)-onyl]-thiosemicarbazido]-2-oxo/thiobarbituric acids 3a-3h and 5-[2'-amino-5'-[3"-aminomethylene-2"-methyl-6",8"-dihalosubstitutedquinazolin-4"(3"H)-onyl]-1',3',4'-thiadiazol-2'-yl]-2-oxo/thiobarbituric acid 5a-5h were prepared by incorporating 1-[3'-aminoacetyl-2'-methyl-6",8"-dihalosubstituted-quinazolin-4'(3'H)-onyl]-thiosemicarbazides 2a-2d and 2-amino-5-[3'-aminomethylene-2'-methyl-6',8'-dihalosubstituted-quinazolin-4'(3'H)-onyl]-1,3,4-thiadiazoles 4a-4 h respectively at 5(th) position of 2-oxo/thiobarbituric acids (via Mannich reaction). All the newly synthesized compounds were screened for their anti-convulsant activity in MES and PTZ models and were compared with standard drugs phenytoin sodium and sodium valproate. Interestingly, these compounds were found to be devoid of sedative and hypnotic activities when tested. Out of the compounds studied, the most active compound 5h, that is 5-[2'-amino-5'-[3"-aminomethylene-2"-methyl-6",8"-dibromoquinazolin-4"(3"H)-onyl]-1',3',4'-thiadiazol-2'-yl]-2-thiobarbituric acid showed activity (90%) more potent than the standard drug.


[Postcoital contraception in primates. I. Action mechanism of a potential postovulatory fertility-inhibiting substance STS 456 in the baboon (Papio hamadryas)].
With the substance STS 456, an estrogenic active steroid, a high fertility inhibition could be obtained in the pavian, when administered p. o. over a 5 day period postcoital. The effectiveness and also the side effects were dose dependent. The antifertility mechanism is based on an luteolytic effect, demonstrated by analytical hormonal investigations. The inhibition of the synthesis of steroids in the ovary affected not only progesterone but also the estrogens.