Relationship between tumor cell infiltration and 5-aminolevulinic acid fluorescence signals after resection of MR-enhancing lesions and its prognostic significance in glioblastoma.

PURPOSE: This study investigated the degree of tumor cell infiltration in the tumor cavity and ventricle wall based on fluorescent signals of 5-aminolevulinic acid (5-ALA) after removal of the magnetic resonance (MR)-enhancing area and analyzed its prognostic significance in glioblastoma. METHODS: Twenty-five newly developed isocitrate dehydrogenase (IDH)-wildtype glioblastomas with complete resection both of MR-enhancing lesions and strong purple fluorescence on resection cavity were retrospectively analyzed. The fluorescent signals of 5-ALA were divided into strong purple, vague pink, and blue colors. The pathologic findings were classified into massively infiltrating tumor cells, infiltrating tumor cells, suspicious single-cell infiltration, and normal-appearing cells. The pathological findings were analyzed according to the fluorescent signals in the resection cavity and ventricle wall. RESULTS: There was no correlation between fluorescent signals and infiltrating tumor cells in the resection cavity (p = 0.199) and ventricle wall (p = 0.704) after resection of the MR-enhancing lesion. The median progression-free survival (PFS) and median overall survival (OS) were 12.5 (± 2.1) and 21.1 (± 3.5) months, respectively. In univariate analysis, the presence of definitive infiltrating tumor cells in the resection cavity and ventricle wall was significantly related to the PFS (p = 0.002) and OS (p = 0.027). In multivariate analysis, the absence of definitive infiltrating tumor cells improved PFS (hazard ratio: 0.184; 95% CI: 0.049-0.690, p = 0.012) and OS (hazard ratio: 0.124; 95% CI: 0.015-0.998, p = 0.050). CONCLUSIONS: After resection both of the MR-enhancing lesions and strong purple fluorescence on resection cavity, there was no correlation between remnant fluorescent signals and infiltrating tumor cells. The remnant definitive infiltrating tumor cells in the resection cavity and ventricle wall significantly influenced the prognosis of patients with glioblastoma. Aggressive surgical removal of infiltrating tumor cells may improve their prognosis.

Influence of Gingival Contour on Marginal Fit of CAD-CAM Zirconia Copings on Implant Stock Abutments.

PURPOSE: Intraoral scanning subgingival finish lines has been described as challenging. The purpose of this study was to assess the impact of gingival contour around margins of implant stock abutments on marginal fit of Computer-aided Design - Computer-aided Manufacturing (CAD-CAM) zirconia copings. This in-vitro study was conducted on 40 analogues of implant stock abutments that were embedded into individual phantoms composed by a resin block. All 40 phantoms underwent two intraoral scans - one with (test group) and one without artificial gingiva (control group) - using a closed system intraoral scanner. Zirconia copings were then digitally designed and milled, followed by high-speed sintering, before being analyzed for marginal adaptation and internal surface roughness with scanning electron microscopy. Statistically significant differences between groups were assessed with the Mann-Whitney test. Median marginal gap values were 149.78 µm (95% CI: 112.39-216.66) for the test group and 94.90 µm (95% CI: 83.89-107.74) for the control group. A statistically significant difference was found between groups (p=0.0001). However, there were no subjective differences between groups for internal surface roughness. Within the limitations of this study, the present findings suggest that a gingival contour one millimeter higher than the finishing line affects marginal adaptation of CAD-CAM zirconia copings.

Simulating the behavior of patients who leave a public hospital emergency department without being seen by a physician: a cellular automaton and agent-based framework.

The objective of this study was to develop an agent based modeling (ABM) framework to simulate the behavior of patients who leave a public hospital emergency department (ED) without being seen (LWBS). In doing so, the study complements computer modeling and cellular automata (CA) techniques to simulate the behavior of patients in an ED. After verifying and validating the model by comparing it with data from a real case study, the significance of four preventive policies including increasing number of triage nurses, fast-track treatment, increasing the waiting room capacity and reducing treatment time were investigated by utilizing ordinary least squares regression. After applying the preventing policies in ED, an average of 42.14% reduction in the number of patients who leave without being seen and 6.05% reduction in the average length of stay (LOS) of patients was reported. This study is the first to apply CA in an ED simulation. Comparing the average LOS before and after applying CA with actual times from emergency department information system showed an 11% improvement. The simulation results indicated that the most effective approach to reduce the rate of LWBS is applying fast-track treatment. The ABM approach represents a flexible tool that can be constructed to reflect any given environment. It is also a support system for decision-makers to assess the relative impact of control strategies.

Interregional correlations of glucose metabolism between the basal ganglia and different cortical areas: an ultra-high resolution PET/MRI fusion study using 18F-FDG

Basal ganglia have complex functional connections with the cerebral cortex and are involved in motor control, executive functions of the forebrain, such as the planning of movement, and cognitive behaviors based on their connections. The aim of this study was to provide detailed functional correlation patterns between the basal ganglia and cerebral cortex by conducting an interregional correlation analysis of the 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) data based on precise structural information. Fifteen participants were scanned with 7-Tesla magnetic resonance imaging (MRI) and high resolution research tomography (HRRT)-PET fusion system using 18F-FDG. For detailed interregional correlation analysis, 24 subregions of the basal ganglia including pre-commissural dorsal caudate, post-commissural caudate, pre-commissural dorsal putamen, post-commissural putamen, internal globus pallidus, and external globus pallidus and 80 cerebral regions were selected as regions of interest on the MRI image and their glucose metabolism were calculated from the PET images. Pearson's product-moment correlation analysis was conducted for the interregional correlation analysis of the basal ganglia. Functional correlation patterns between the basal ganglia and cerebral cortex were not only consistent with the findings of previous studies, but also showed new functional correlation between the dorsal striatum (i.e., caudate nucleus and putamen) and insula. In this study, we established the detailed basal ganglia subregional functional correlation patterns using 18F-FDG PET/MRI fusion imaging. Our methods and results could potentially be an important resource for investigating basal ganglia dysfunction as well as for conducting functional studies in the context of movement and psychiatric disorders.

Interregional correlations of glucose metabolism between the basal ganglia and different cortical areas: an ultra-high resolution PET/MRI fusion study using 18F-FDG.

Basal ganglia have complex functional connections with the cerebral cortex and are involved in motor control, executive functions of the forebrain, such as the planning of movement, and cognitive behaviors based on their connections. The aim of this study was to provide detailed functional correlation patterns between the basal ganglia and cerebral cortex by conducting an interregional correlation analysis of the 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) data based on precise structural information. Fifteen participants were scanned with 7-Tesla magnetic resonance imaging (MRI) and high resolution research tomography (HRRT)-PET fusion system using 18F-FDG. For detailed interregional correlation analysis, 24 subregions of the basal ganglia including pre-commissural dorsal caudate, post-commissural caudate, pre-commissural dorsal putamen, post-commissural putamen, internal globus pallidus, and external globus pallidus and 80 cerebral regions were selected as regions of interest on the MRI image and their glucose metabolism were calculated from the PET images. Pearson's product-moment correlation analysis was conducted for the interregional correlation analysis of the basal ganglia. Functional correlation patterns between the basal ganglia and cerebral cortex were not only consistent with the findings of previous studies, but also showed new functional correlation between the dorsal striatum (i.e., caudate nucleus and putamen) and insula. In this study, we established the detailed basal ganglia subregional functional correlation patterns using 18F-FDG PET/MRI fusion imaging. Our methods and results could potentially be an important resource for investigating basal ganglia dysfunction as well as for conducting functional studies in the context of movement and psychiatric disorders.

Emergence of CTNNB1 mutation at acquired resistance to KIT inhibitor in metastatic melanoma.

PURPOSE: The KIT inhibitor, imatinib, has shown promising efficacy in patients with KIT-mutated melanoma; however, acquisition of resistance to imatinib occurs rapidly in the majority of patients. The mechanisms of acquired resistance to imatinib in melanoma remain unclear. METHODS: We analyzed biopsy samples from paired baseline and post-treatment tumor lesions in one patient with KIT-mutated melanoma who had had an initial objective tumor regression in response to imatinib treatment followed by disease progression 8 months later. RESULTS: Targeted deep sequencing from post-treatment biopsy samples detected an additional mutation in CTNNB1 (S33C) with original KIT L576P mutation. We examined the functional role of the additional CTNNB1 S33C mutation in resistance to imatinib indirectly using the Ba/F3 cell model. Ba/F3 cell lines transfected with both the L576P KIT mutation and the CTNNB1 S33C mutation demonstrated no growth inhibition despite imatinib treatment, whereas growth inhibition was observed in the Ba/F3 cell line transfected with the L576 KIT mutation alone. CONCLUSIONS: We report the first identification of the emergence of a CTNNB1 mutation that can confer acquired resistance to imatinib. Further investigation into the causes of acquired resistance to imatinib will be essential to improve the prognosis for patients with KIT-mutated melanoma.

Effects of dietary fermented garlic on the growth performance, relative organ weights, intestinal morphology, cecal microflora and serum characteristics of broiler chickens

The present study was conducted to evaluate the effects of feeding broilers with garlic fermented by Leuconostoc citreum SK2556. A total of 250 male broiler chicks was randomly housed into 25 floor pens. Five dietary treatments with five replicates of 10 chicks each (n=50 chicks/treatment). A corn and soybean meal based diet was used as the control diet (NC). The experimental diets were formulated by mixing the basal diet either with antibiotics (10 ppm; PC) or fermented garlic (FG) at the concentrations of 0.1% (FG1), 0.3% (FG3) or 0.5% (FG5) in diets. Daily weight gain, feed intake, and feed:gain ratio were not affected by any of the dietary treatments. Average daily gain on day 21 linearly increased (p= 0.024) with increasing FG levels. The relative weight of the bursa of Fabricius showed a progressive decline with increasing the FG levels. Jejunal villus height was not influenced by dietary treatments. Villus width linearly decreased as FG levels increased (p= 0.17). Jejunal crypt depth was significantly lower (p < 0.05) in the FG1 and FG3 groups compared with the NC group. Villus height: crypt depth ratio linearly increased (p= 0.018) with increasing FG levels. The population of cecal microflora was not altered by dietary treatments. Broiler chickens fed the FG5 diet exhibited (p < 0.05) higher blood levels of total protein and cholesterol compared with those fed the NC diet. Collectively, the results show that dietary FG marginally affected growth performance, especially during the first days rearing, improved intestinal morphology, and altered blood characteristics of broiler chickens.(AU)

Effects of dietary fermented garlic on the growth performance, relative organ weights, intestinal morphology, cecal microflora and serum characteristics of broiler chickens

The present study was conducted to evaluate the effects of feeding broilers with garlic fermented by Leuconostoc citreum SK2556. A total of 250 male broiler chicks was randomly housed into 25 floor pens. Five dietary treatments with five replicates of 10 chicks each (n=50 chicks/treatment). A corn and soybean meal based diet was used as the control diet (NC). The experimental diets were formulated by mixing the basal diet either with antibiotics (10 ppm; PC) or fermented garlic (FG) at the concentrations of 0.1% (FG1), 0.3% (FG3) or 0.5% (FG5) in diets. Daily weight gain, feed intake, and feed:gain ratio were not affected by any of the dietary treatments. Average daily gain on day 21 linearly increased (p= 0.024) with increasing FG levels. The relative weight of the bursa of Fabricius showed a progressive decline with increasing the FG levels. Jejunal villus height was not influenced by dietary treatments. Villus width linearly decreased as FG levels increased (p= 0.17). Jejunal crypt depth was significantly lower (p < 0.05) in the FG1 and FG3 groups compared with the NC group. Villus height: crypt depth ratio linearly increased (p= 0.018) with increasing FG levels. The population of cecal microflora was not altered by dietary treatments. Broiler chickens fed the FG5 diet exhibited (p < 0.05) higher blood levels of total protein and cholesterol compared with those fed the NC diet. Collectively, the results show that dietary FG marginally affected growth performance, especially during the first days rearing, improved intestinal morphology, and altered blood characteristics of broiler chickens.

Characterization of microRNAs and their target genes associated with transcriptomic changes in gamma-irradiated Arabidopsis.

MicroRNAs (miRNAs) regulate gene expression in response to biotic and abiotic stress in plants. We investigated gamma-ray-responsive miRNAs in Arabidopsis wild-type and cmt3-11t mutant plants using miRNA microarray analysis. miRNA expression was differentiated between the wild-type and cmt3-11t mutants. miR164a, miR169d, miR169h, miR172b*, and miR403 were identified as repressible in the wild-type and/or cmt3-11t mutant in response to gamma irradiation, while miR827, miR840, and miR850 were strongly inducible. These eight miRNA genes contain UV-B-responsive cis-elements, including G-box, I-box core, ARE, and/or MBS in the putative promoter regions. Moreover, Box 4, MBS, TCA-element, and Unnamed_4, as well as CAAT- and TATA-box, were identified in these eight miRNA genes. However, a positive correlation between the transcriptions of miRNAs and their putative target genes was only observed between miR169d and At1g30560 in the wild-type, and between miR827 and At1g70700 in the cmt3-11t mutant. Quantitative RT-PCR analysis confirmed that the transcription of miR164a, miR169d, miR169h, miR172b*, miR403, and miR827 differed after gamma irradiation depending on the genotype (wild-type, cmt3-11t, drm2, drd1-6, and ddm1-2) and developmental stage (14 or 28 days after sowing). In contrast, high transcriptional induction of miR840 and miR850 was observed in these six genotypes regardless of the developmental stage. Although the actual target genes and functions of miR840 and miR850 remain to be determined, our results indicate that these two miRNAs may be strongly induced and reproducible genetic markers in Arabidopsis plants exposed to gamma rays.

Analysis of 'Fuji' apple somatic variants from next-generation sequencing.

The domesticated apple (Malus x domestica Borkh.) is a major fruit crop of temperate regions of the world. 'Fuji' apple (Ralls Genet x Delicious), a famous apple cultivar in Korea, has been very popular since its promotion in Japan in 1958. 'Fuji' and its bud mutant cultivars possess variable levels of genetic diversity. Nonetheless, the phenotypes of each group, which are classified into the bud mutation groups: early season, fruiting spur, and coloring, are similar. Despite attempts to identify these bud mutation cultivars, molecular markers, which were developed before the emergence of next-generation sequencing technology, have not been able to distinguish each cultivar easily. In this study, we adopted the resequencing technique using the 'Golden Delicious' (Grimes Golden x Unknown) apple genome as a reference. SNPs (single nucleotide polymorphisms) and InDels (insertions or deletions) of 'Fuji' apple and its bud mutant cultivar were detected and SNPs and unique InDels distinct to each cultivar were identified. Data from this study may be used to identify bud mutant cultivars of 'Fuji' apples and be useful for further breeding of apples.

Assessment of apple core collections constructed using phenotypic and genotypic data.

Several types of information can be used to select core collections, including passport data, agronomic data, and molecular data. However, little is known about the ability of core collections to retain the genetic diversity and structure of the whole collection for characters that were not considered during the selection, particularly when molecular markers are used. In this study, two core subsets were established for the apple (Malus spp) germplasm bank curated at the Apple Research Station, National Institute of Horticultural and Herbal Science, Korea, based upon genetic diversity estimated with 14 simple sequence repeat markers, and phenotypic diversity based on 23 traits. Comparisons between these two subsets and with the whole collection were used to determine the effect of the data used in the selection on phenotypic and genetic diversity, and population structure. The two subsets had a similar diversity and did not differ from the original collection, according to the Nei and Shannon diversity indices. Allele and class frequencies were also maintained in the two subsets. Overall, the type of data used to construct the core collection had little influence on the phenotypic and genetic diversity retained. Therefore, in the case of apple collections, the use of molecular markers is preferable, because they allow rapid and reliable characterization.

Gene expression profiles of primary retinal pigment epithelial cells from apolipoprotein E knockout and human apolipoprotein E2 transgenic mice.

Age-related macular degeneration (AMD) causes visual impairment in the elderly. In non-neovascular AMD, studies involving human subjects have suggested potential involvement of aberrant lipid metabolism. However, there have been no reports on gene expression patterns in animal models of non-neovascular AMD with abnormal lipid metabolism such as apolipoprotein E knockout and human apolipoprotein E2 transgenic mice. Transcriptome analysis was performed using retinal pigment epithelium cells of apoE knockout and apolipoprotein E2 mice using microarray analysis. C57BL/6, Rxrb, Pparbp, Vldlr, and Edf1, which are primarily related to lipid metabolism, were upregulated, while Tgfbr1 and Pdgfb, which are related to pathologic angiogenesis in AMD, were downregulated in both types of mice. Apolipoprotein E knockout and apolipoprotein E2 mice showed characteristic gene expression patterns in the transcriptome analysis of primary retinal pigment epithelium cells. These results suggest that specific genes associated with lipid metabolism and angiogenesis are involved in the pathogenesis and progression of AMD.

Laparoscopic excision of a renal subcapsular abscess presenting as a subcapsular haematoma.

Renal subcapsular abscess is a very rare entity that is defined by a suppurative process localized to a space between the renal capsule and the renal parenchyma. The pathogenesis and aetiology of this entity remain speculative. To our knowledge, only five cases have been reported in the English literature. We describe a 74-year old woman with renal subcapsular abscess treated with laparoscopic removal and do a review of the literature.

Ependymoma in children: molecular considerations and therapeutic insights.

A multi-modality approach that encompasses maximal surgical resection in combination with adjuvant therapy is critical for achieving optimal disease control in children with ependymoma. In view of its complex biology and variable response to therapy, ependymoma remains a challenge for clinicians involved in the care of these patients. Meanwhile, translation of molecular findings can characterize unique features of childhood ependymoma and their natural history. Furthermore, understanding the biology of pediatric ependymoma serves as a platform for development of future targeted therapies. In line with these goals, we review the molecular basis of pediatric ependymoma and its prognostic implications, as well as novel therapeutic advances in the management of ependymoma in children.

Phylogeography, population history and conservation genetics of jaguars (Panthera onca, Mammalia, Felidae).

The jaguar (Panthera onca), the largest felid in the American Continent, is currently threatened by habitat loss, fragmentation and human persecution. We have investigated the genetic diversity, population structure and demographic history of jaguars across their geographical range by analysing 715 base pairs of the mitochondrial DNA (mtDNA) control region and 29 microsatellite loci in approximately 40 individuals sampled from Mexico to southern Brazil. Jaguars display low to moderate levels of mtDNA diversity and medium to high levels of microsatellite size variation, and show evidence of a recent demographic expansion. We estimate that extant jaguar mtDNA lineages arose 280 000-510 000 years ago (95% CI 137 000-830 000 years ago), a younger date than suggested by available fossil data. No strong geographical structure was observed, in contrast to previously proposed subspecific partitions. However, major geographical barriers such as the Amazon river and the Darien straits between northern South America and Central America appear to have restricted historical gene flow in this species, producing measurable genetic differentiation. Jaguars could be divided into four incompletely isolated phylogeographic groups, and further sampling may reveal a finer pattern of subdivision or isolation by distance on a regional level. Operational conservation units for this species can be defined on a biome or ecosystem scale, but should take into account the historical barriers to dispersal identified here. Conservation strategies for jaguars should aim to maintain high levels of gene flow over broad geographical areas, possibly through active management of disconnected populations on a regional scale.

Partial cross-resistance between Strongyloides venezuelensis and Nippostrongylus brasiliensis in rats.

Rats were immunized through an initial infection with 1,000 filariform larvae (L3) of Nippostrongylus brasiliensis and after complete expulsion of worms they were challenged with 1,000 L3 of Strongyloides venezuelensis to investigate whether cross-resistance developed against a heterologous parasite. Nippostrongylus brasiliensis-immunized rats developed a partial cross-resistance against S. venezuelensis migrating larvae (MSL3) in the lungs and adult worms in the small intestine. The population of MSL3 in the lungs were significantly lower (P < 0.05) in immunized rats (22.0 +/- 7.4) compared with controls (105.0 +/- 27.6). The populations of adult worms, egg output and fecundity were initially decreased but from day 14 post-challenge they did not show any significant difference between immunized and control rats. However, the length of worm in immunized rat was revealed as retardation. Peripheral blood eosinophilia was significantly decreased (P < 0.05) on day 7 post-challenge and then gradually increased, which peaked on day 42 post-challenge when most of the worms were expelled. These results suggest that peripheral blood eosinophilia is strongly involved in the worm establishment and expulsion mechanisms.

Disparate phylogeographic patterns of molecular genetic variation in four closely related South American small cat species.

Tissue specimens from four species of Neotropical small cats (Oncifelis geoffroyi, N = 38; O. guigna, N = 6; Leopardus tigrinus, N = 32; Lynchailurus colocolo, N = 22) collected from throughout their distribution were examined for patterns of DNA sequence variation using three mitochondrial genes, 16S rRNA, ATP8, and NADH-5. Patterns between and among O. guigna and O. geoffroyi individuals were assessed further from size variation at 20 microsatellite loci. Phylogenetic analyses using mitochondrial DNA sequences revealed monophyletic clustering of the four species, plus evidence of natural hybridization between L. tigrinus and L. colocolo in areas of range overlap and discrete population subdivisions reflecting geographical isolation. Several commonly accepted subspecies partitions were affirmed for L. colocolo, but not for O. geoffroyi. The lack of geographical substructure in O. geoffroyi was recapitulated with the microsatellite data, as was the monophyletic clustering of O. guigna and O. geoffroyi individuals. L. tigrinus forms two phylogeographic clusters which correspond to L.t. oncilla (from Costa Rica) and L.t. guttula (from Brazil) and which have mitochondrial DNA (mtDNA) genetic distance estimates comparable to interspecific values between other ocelot lineage species. Using feline-specific calibration rates for mitochondrial DNA mutation rates, we estimated that extant lineages of O. guigna diverged 0.4 million years ago (Ma), compared with 1.7 Ma for L. colocolo, 2.0 Ma for O. geoffroyi, and 3.7 Ma for L. tigrinus.

Brazilin modulates immune function mainly by augmenting T cell activity in halothane administered mice.

Previously we reported that brazilin, the main principle of Caesalpinia sappan, was able to improve the altered immune functions caused by halothane administration in mice. To elucidate the mechanisms of its immunomodulating activities, the effects of brazilin on the functions of T cells and splenic cellularity were investigated. Brazilin decreased splenic cellularity and IL-2 production which had been augmented in mice treated with halothane (21.5% in olive oil, 10 mmol/kg) for 4 consecutive days whereas the reduced expression of IL-2 receptors by ConA or standard IL-2 was increased by brazilin treatment. These data indicate that halothane induced a dysfunction of T cells resulting in abnormal immune responses and these altered immune functions might be improved mainly by affecting the function of T cells.

Myopathy in tuberculosis: two presumptive cases and a review of the literature.

Mycobacterium tuberculosis infrequently infects muscle. Two cases of muscle involvement complicating tuberculosis (TB), one as a result of infection and a second resulting from the treatment of TB, are reported. Thirteen previous cases are summarized. The pathology, pathogenesis, clinical features, differential diagnosis, and treatment of TB myopathy are discussed.

Status of intestinal parasite infections in inhabitants of Palmar, Guayas Province, Ecuador.

Status of intestinal protozoan and helminthic infections was surveyed in mestizo population living in rural parish of Palmar and its nearby recintos(villages). Three of the surveyed villages were at the Pacific coast and 5 villages were in inner pasture land, located about 100 km west of Guayaquil, the second largest city in Ecuador. One stool sample was examined by one cellophane thick smear for helminth ova and one direct smear stained with Lugol's iodine solution for protozoan cysts. Of 325 persons examined, 66.1% were positive for any ova or cyst. The positive rates were: 18.1% for Ascaris lumbricoides, 19.4% for Trichuris trichiura, 0.6% for hookworm, 3.7% for Hymenolepsis nana, 1.8% for Taenia sp., 19.4% for Entamoeba histolytica, 28.6% for Entamoeba coli, 5.5% for Endolimax nana, 1.5% for Iodamoeba buetschlii, 11.1% for Giardia lamblia and 0.6% for Chilomastix mesnili. Poor supply of potable water was considered the main cause of high prevalence of intestinal protozoan infections.