ABSTRACT
After discharge from neonatal intensive care units (NICUs), the parents of pre-term newborns have to provide developmentally supportive care (DSC) to their children; thus, educational support for parents is essential. This study aimed to explore the lived experiences of parents providing DSC to their children born as pre-term newborns at home and to investigate their parenting-related needs. This study included 10 mothers who were identified through theoretical sampling. In-depth interviews were conducted for data collection. For data analysis, grounded theory was used according to Corbin and Strauss's process. The mother's perception and educational needs were characterized by the phenomena "Coexistence of familiarity and unfamiliarity" and "Desire for expert support". Causal conditions include the "Incomplete education system" and "Gap between expectations and reality". Contextual conditions include the "Fear of developmental disability" and "Lack of good evaluation criteria". Intervening conditions include the "Difficulty in obtaining useful information". Action/interaction strategies include the "Active information seeking" and "Continuing to provide DSC". The consequences were the "Needs for professional educational support". The core category was the "Parenting routine that continues without awareness" and "Hope to establish parenting system supported by multidisciplinary experts". These results may provide the preliminary evidence base for suitable educational programs and for developing a social support system for parents.
ABSTRACT
This study aimed to identify the level of mothers' smartphone dependency and determine its correlation with preschoolers' problem behavior and emotional intelligence. From 1 November to 30 December 2020, 141 mothers of preschool children (aged three to six years) were recruited to complete questionnaires that assessed their smartphone dependency and their child's problem behavior and emotional intelligence. The result revealed that the younger the mother and the higher the perception of boredom in daily living, the higher was the level of her smartphone dependency. Maternal smartphone dependency was also significantly correlated with the aggression, oppositional, and emotional instability subscales of the tool assessing children's problem behavior. To prevent problem behaviors among preschoolers, strategies to reduce mothers' smartphone dependency are needed.
ABSTRACT
The government ordered various restrictions to limit the spread of coronavirus disease 2019 (COVID-19), thus, affecting the mental health status and lifestyle of people with diabetes. This study identifies COVID-19 effects on mental health problems and unhealthy behavioral changes among patients with diabetes. The subjects of this cross-sectional study were adults aged 19 years or older who participated in the 2020 Korean Community Health Survey. Stress, depression, and changes in unhealthy behavior in diabetic patients (N = 26,839) because of COVID-19 were compared with controls (N = 26,834). The association between stress and depression and unhealthy behaviors among patients with diabetes was investigated. During the COVID-19 pandemic, 20.3% and 4.2% of diabetic patients reported higher levels of stress and depression, respectively, than controls. Diabetic patients showed decreased physical activity and sleep time, and increased smoking. Among diabetic patients, stress and depression are associated with unhealthy behavior changes during COVID-19. Measures to promote healthy lifestyles along with stress and depression management strategies must be implemented for the health care of diabetic patients during the pandemic.
ABSTRACT
BACKGROUND: Recently, monoclonal-antibody-conjugated immunomagnetic separation (IMS) procedure combined with quantitative reverse transcription-polymerase chain reaction (qRT-PCR) has been used for quantifying non-cultivated human noroviruses (HuNoVs). METHODS: We examined the efficacy of 27 commercially available disinfectants and a prototype against GII.4 strain HuNoV through the IMS/qRT-PCR assay. RESULTS: The average log reduction in viral titer in vitro varied among the disinfectants. The prototype was the most effective with an average log reduction of 6.86 log. CONCLUSIONS: The IMS/RT-qPCR assay is an effective method to evaluate the activities of disinfectants against GII.4 HuNoV in vitro. Further work is needed to enhance the virucidal activity of the prototype disinfectant against more resistant HuNoV strains.
Subject(s)
Disinfectants/pharmacology , Immunomagnetic Separation/methods , Norovirus/drug effects , Real-Time Polymerase Chain Reaction/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , Humans , Norovirus/genetics , Norovirus/isolation & purification , Viral Load , Virus InactivationABSTRACT
Xanthomonas citri subsp. citri is the causal agent of citrus canker, which has a significant impact on citrus production. In this study, we characterized the galU gene of X. citri subsp. citri. Two galU mutants (F6 and D12) were identified in an X. citri subsp. citri EZ-Tn5
Subject(s)
Bacterial Proteins/metabolism , Citrus/microbiology , Plant Diseases/microbiology , Polysaccharides, Bacterial/biosynthesis , UTP-Glucose-1-Phosphate Uridylyltransferase/metabolism , Xanthomonas/enzymology , Xanthomonas/pathogenicity , Bacterial Proteins/genetics , Biofilms/growth & development , Citrus paradisi/microbiology , DNA Transposable Elements , Gene Deletion , Gene Expression Profiling , Genetic Complementation Test , Mutagenesis, Insertional , Plant Leaves/microbiology , UTP-Glucose-1-Phosphate Uridylyltransferase/genetics , Virulence , Virulence Factors/biosynthesis , Xanthomonas/growth & development , Xanthomonas/metabolismABSTRACT
Citrus greening or huanglongbing (HLB) is a devastating disease of citrus. HLB is associated with the phloem-limited fastidious prokaryotic alpha-proteobacterium 'Candidatus Liberibacter spp.' In this report, we used sweet orange (Citrus sinensis) leaf tissue infected with 'Ca. Liberibacter asiaticus' and compared this with healthy controls. Investigation of the host response was examined with citrus microarray hybridization based on 33,879 expressed sequence tag sequences from several citrus species and hybrids. The microarray analysis indicated that HLB infection significantly affected expression of 624 genes whose encoded proteins were categorized according to function. The categories included genes associated with sugar metabolism, plant defense, phytohormone, and cell wall metabolism, as well as 14 other gene categories. The anatomical analyses indicated that HLB bacterium infection caused phloem disruption, sucrose accumulation, and plugged sieve pores. The up-regulation of three key starch biosynthetic genes including ADP-glucose pyrophosphorylase, starch synthase, granule-bound starch synthase and starch debranching enzyme likely contributed to accumulation of starch in HLB-affected leaves. The HLB-associated phloem blockage resulted from the plugged sieve pores rather than the HLB bacterial aggregates since 'Ca. Liberibacter asiaticus' does not form aggregate in citrus. The up-regulation of pp2 gene is related to callose deposition to plug the sieve pores in HLB-affected plants.
Subject(s)
Citrus/microbiology , Plant Diseases/microbiology , Protein Array Analysis , Rhizobiaceae/physiology , Expressed Sequence Tags , Gene Expression Regulation, Plant/immunology , Plant Diseases/immunology , Plant Leaves/microbiology , Plant Proteins/metabolismABSTRACT
Severe fever with thrombocytopenia syndrome (SFTS) is a tick-borne viral disease with a high mortality rate. Infection can also occur through close contact with an infected patient. Scrub typhus is an acute febrile illness caused by Orientia tsutsugamushi, a bacterium transmitted to humans through chigger mite bites. South Korea is an endemic region of SFTS and scrub typhus. In this study, we confirmed that a patient was coinfected with SFTS virus and two (Boryong and Taguchi) genotypes of O. tsutsugamushi.
Subject(s)
Bunyaviridae Infections/diagnosis , Coinfection/microbiology , Coinfection/virology , Scrub Typhus/diagnosis , Aged , Anti-Bacterial Agents/therapeutic use , Azithromycin/therapeutic use , Bites and Stings , Ceftriaxone/therapeutic use , Coinfection/diagnosis , Female , Genotype , Humans , Leukopenia , Molecular Diagnostic Techniques , Orientia tsutsugamushi/drug effects , Orientia tsutsugamushi/genetics , Phlebovirus/genetics , Phlebovirus/isolation & purification , Phylogeny , Republic of Korea , Thrombocytopenia , Tick-Borne Diseases/diagnosis , Tick-Borne Diseases/microbiology , Treatment OutcomeABSTRACT
Rapid and sensitive detection techniques for foodborne pathogens are important to the food industry. However, traditional detection methods rely on bacterial culture in combination with biochemical tests, a process that typically takes 4 to 7 days to complete. Thus, this study was conducted to address the issue of time lag inherent in traditional methods by developing a novel PCR assay for each of five foodborne pathogenic bacteria. This new system consists of a simultaneous screening method using multiplex PCR in a single reaction tube for the rapid and sensitive detection of each of the five bacteria. Specific primers for multiplex PCR amplification of the Shiga-like toxin (verotoxin type II), femA (cytoplasmic protein), toxR (transmembrane DNA binding protein), iap (invasive associative protein), and invA (invasion protein A) genes were designed to allow simultaneous detection of Escherichia coli O157:H7, Staphylococcus aureus, Vibrio parahaemolyticus, Listeria monocytogenes, and Salmonella, respectively. To confirm the specificity of each primer pair for the respective target gene, three types of experiments were carried out using boiled cell lysates and their DNAs. In the multiplex PCR with mixed DNA samples, specific bands for corresponding genes were simultaneously detected from a single reaction. The detection of all five foodborne pathogenic bacteria could be completed in less than 24 h with this novel PCR method.
Subject(s)
Food Contamination/analysis , Polymerase Chain Reaction/methods , DNA Primers , Escherichia coli O157/genetics , Escherichia coli O157/isolation & purification , Food Microbiology , Gene Amplification , Listeria monocytogenes/genetics , Listeria monocytogenes/isolation & purification , Reproducibility of Results , Salmonella/genetics , Salmonella/isolation & purification , Sensitivity and Specificity , Species Specificity , Staphylococcus aureus/genetics , Staphylococcus aureus/isolation & purification , Time Factors , Vibrio parahaemolyticus/genetics , Vibrio parahaemolyticus/isolation & purificationABSTRACT
Linkage group identities and homologies were determined for metaphase chromosomes of Sorghum bicolor (2n = 20) by FISH of landed BACs. Relative lengths of chromosomes in FISH-karyotyped metaphase spreads of the elite inbred BTx623 were used to estimate the molecular size of each chromosome and to establish a size-based nomenclature for sorghum chromosomes (SBI-01-SBI-10) and linkage groups (LG-01 to LG-10). Lengths of arms were determined to orient linkage groups relative to a standard karyotypic layout (short arms at top). The size-based nomenclature for BTx623 represents a reasonable choice as the standard for a unified chromosome nomenclature for use by the sorghum research community.
Subject(s)
Chromosomes, Plant , Sorghum/genetics , Terminology as Topic , Centromere/genetics , Chromosome Mapping , Genetic Linkage , Genetic Markers , In Situ Hybridization, Fluorescence , Karyotyping , Metaphase , Nucleolus Organizer Region/genetics , Sorghum/cytologyABSTRACT
To integrate genetic, physical, and cytological perspectives of the Sorghum bicolor genome, we selected 40 landed bacterial artificial chromosome (BAC) clones that contain different linkage map markers, 21 from linkage group 2 (LG-02) and 19 from linkage group 8 (LG-08). Multi-BAC probe cocktails were constructed for each chromosome from the landed BACs, which were also preevaluated for FISH signal quality, relative position, and collective chromosome coverage. Comparison to the corresponding linkage map revealed full concordance of locus order between cytological and prior segregation analyses. The pericentromeric heterochromatin constituted a large quasi-uniform block in each bivalent and was especially large in the bivalent corresponding to LG-08. Centromere positions in LG-02 and LG-08 were progressively delimited using FISH to identify landed BACs for which the FISH signals visibly flanked the centromere. Alignment of linkage and cytological maps revealed that pericentromeric heterochromatin of these sorghum chromosomes is largely devoid of recombination, which is mostly relegated to the more distal regions, which are largely euchromatic. This suggests that the sorghum genome is thus even more amenable to physical mapping of genes and positional cloning than the C-value alone might suggest. As a prelude to positional cloning of the fertility restorer, Rf1, FISH of BAC clones flanking the Rf1 locus was used to delimit the chromosomal position of the gene. FISH of BACs that contain the most proximal linkage markers enabled localization of Rf1 to a approximately 0.4-Mbp euchromatic region of LG-08. Cytogenetic analyses of Rf1 and other trait loci will aid in assessing the feasibility of positional cloning and help formulate strategies required for cloning this and other agriculturally critical genes.
Subject(s)
Chromosomes, Plant , Genes, Plant , Genetic Linkage , Physical Chromosome Mapping , Sorghum/genetics , Centromere , Chromosomes, Artificial, Bacterial , DNA, Plant , Genetic Markers , Genome, Plant , Heterochromatin/genetics , In Situ Hybridization, FluorescenceABSTRACT
PURPOSE: Developmental care has been recognized as a very important component for the development and health promotion of preterm infants. However, research on how to assess developmental nursing competency has not been studied as expected. This study was done to develop and evaluate a new scale to measure nursing competency for developmental support of preterm infants. METHODS: Concept analysis was done with using the Hybrid model of Schwartz-Barcott and Kim (2000), from which a preliminary new scale (30 items) was developed. To test the validity and reliability of the new scale being developed, data were collected from 122 NICU nurses at 4 hospitals in 3 cities in the Republic of Korea, from December, 2014 to March, 2015. RESULTS: The final version of the Developmental Support Competency Scale for Nurses (DSCS-N) caring for premature infants was a 4-point Likert type scale, consisting of 19 items, and categorized as 6 factors, explaining 62.5% of the total variance. Each of the factors were named as follows; 'environmental support' (4 items), 'parental support' (3 items), 'interaction' (3 items), 'critical thinking' (3 items), 'professional development' (3 items), and 'partnership' (3 items). The Cronbach's α coefficient for the scale was .83 and the reliability of the subscales ranged from .60~.76. CONCLUSION: The psychometric evaluation of the new scale demonstrated an acceptable validity and reliability. Findings indicate that the DSCS-N can be used as the tool to test the effect of educational programs for nurses and contribute to advance developmental care for preterm infants.
Subject(s)
Infant Care , Nursing Staff, Hospital/psychology , Adult , Clinical Competence , Female , Hospitals , Humans , Infant , Infant, Premature , Intensive Care Units, Neonatal , Interviews as Topic , Job Satisfaction , Program Development , Social Support , Surveys and QuestionnairesABSTRACT
To assess the role of 8-oxoguanine glycosylase (OGG1) in the cell defense against radiation injury, the radiation-induced cytotoxicities were compared between the mutant type KG-1 featuring a loss of OGG1 activity due to a homozygous mutation of Arg 229 Gln, and the wild type U937. While the following three obvious toxicities were displayed in KG-1, they were observed only minimally in U937. These were: a dramatic arrest at the G2/M phase indicated by a marked increase in both the number of G2/M cells and the expression of cyclin B1, cdc2, and mitotic phosphoprotein monoclonal-2 (MPM-2)-reactive proteins; a severe apoptosis shown by a marked increase in the number of cells with hypo-diploid DNA and DNA fragmentation; and as a result, a severe inhibition of cell growth and proliferation measured by the MTT test and [(3)H]-thymidine uptake assay. As expected, KG-1 exhibited a significant increase in the 8-hydroxyguanine level in DNA whereas U937 did not. However, the level of irradiation-induced lipid peroxidation was almost the same in both cell lines. All of these symptoms shown by KG-1 were observed in Molt-4 and CEM-CM3, which were also found to feature low OGG1 activity. These findings suggest that OGG1 plays an important role in cell survival from radiation-induced damage and are also indicative of the capability of 8-hydroxyguanine in DNA to induce cellular toxicities.
Subject(s)
Leukemia/enzymology , N-Glycosyl Hydrolases/metabolism , Radiation Tolerance , Apoptosis/radiation effects , Blotting, Western , CDC2 Protein Kinase/metabolism , Cell Cycle/radiation effects , Cell Division/radiation effects , Cell Line , Cell Survival/radiation effects , Cyclin B/metabolism , Cyclin B1 , DNA Damage/radiation effects , DNA-Formamidopyrimidine Glycosylase , Dose-Response Relationship, Radiation , Flow Cytometry , Gamma Rays , Humans , Jurkat Cells , Leukemia/genetics , Lipid Peroxidation/radiation effects , Protein Serine-Threonine Kinases/metabolism , Time Factors , Tumor Cells, CulturedABSTRACT
The surface characteristics of ethylene-vinyl acetate (EVA) were modified by argon, air, and oxygen plasma at atmospheric pressure. The surface energies of the EVA were evaluated by contact angles according to a sessile-drop method and adhesion energy (G(IC)) was estimated by a 180 degrees peel test with polyurethane (PU). After the plasma treatments, the surface free energies (or specific polar component) of the EVA increased about five times compared to that of virgin EVA. The adhesion between the EVA and the PU is significantly improved by the plasma treatment. Especially, Ar/air/O(2) plasma treatment increases G(IC) of EVA/PU up to about 600% compared to that of the sample using virgin EVA.
ABSTRACT
We previously reported that 8-oxo-2'-deoxyguanosine (8-oxo-dG) suppressed airway hyperresponsiveness and allergy-associated immune responses in ovalbumin-induced allergic mice by inactivating Rac. In the present study, 8-oxo-dG was investigated for its suppression of inflammation and remodeling in lung tissues induced by allergic reaction in mice. Mice were sensitized and challenged with ovalbumin without or with oral administration of 8-oxo-dG. The mice without 8-oxo-dG administration showed the following inflammatory and airway remodeling signs: infiltration of inflammatory cells into peribronchial area, hyperplasia of mucus-secreting goblet cells in bronchial walls, increase of expressions of Muc5ac and vascular cell adhesion molecule (VCAM)-1, collagen deposition and protein expression, and matrix metalloproteinase (MMP)-2/-9 expressions. We also observed an increase of various inflammation-mediating proteins, namely IL-4, IL-5, IL-8, IL-13, TNF-α and IFN-γ, and activation of STAT1 and NF-κB. Production of reactive oxygen species and nitric oxide (NO(.)) was increased as indicated by a dramatic increase in formation of nitro-tyrosine. Importantly, Rac1 and 2 were also markedly activated. However, 8-oxo-dG suppressed all these inflammatory and tissue remodeling signs as well as activation of Rac1 and 2. These results indicate that 8-oxo-dG can inhibit allergy-induced inflammation and remodeling in airway and lung tissues through Rac inactivation.
Subject(s)
Airway Remodeling/drug effects , Deoxyguanosine/analogs & derivatives , Hypersensitivity/pathology , Immunosuppressive Agents/pharmacology , Lung/drug effects , 8-Hydroxy-2'-Deoxyguanosine , Airway Remodeling/genetics , Airway Remodeling/immunology , Animals , Collagen/metabolism , Cytokines/metabolism , Deoxyguanosine/pharmacology , Enzyme Activation/drug effects , Enzyme Activation/immunology , Female , Gene Expression Regulation/drug effects , Gene Expression Regulation/immunology , Goblet Cells/drug effects , Goblet Cells/immunology , Goblet Cells/pathology , Hyperplasia/drug therapy , Hypersensitivity/genetics , Hypersensitivity/immunology , Hypersensitivity/metabolism , Inflammation/genetics , Inflammation/immunology , Inflammation/metabolism , Inflammation/pathology , Lung/immunology , Lung/metabolism , Lung/pathology , Matrix Metalloproteinase 2/genetics , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/genetics , Matrix Metalloproteinase 9/metabolism , Mice , Mice, Inbred BALB C , Mucin 5AC/genetics , Mucin 5AC/metabolism , NF-kappa B/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , STAT1 Transcription Factor/metabolism , Tyrosine/analogs & derivatives , Tyrosine/biosynthesis , Vascular Cell Adhesion Molecule-1/genetics , Vascular Cell Adhesion Molecule-1/metabolism , rac GTP-Binding Proteins/metabolismABSTRACT
BACKGROUND: Citrus Huanglongbing (HLB) is one of the most devastating diseases on citrus and is associated with Candidatus Liberibacter spp.. The pathogens are phloem limited and have not been cultured in vitro. The current management strategy of HLB is to remove infected citrus trees and reduce psyllid populations with insecticides to prevent the spreading. This strategy requires sensitive and reliable diagnostic methods for early detection. RESULTS: We investigated the copy numbers of the 16S rDNA and 16S rRNA of the HLB pathogen and the implication of improving the diagnosis of HLB for early detection using Quantitative PCR. We compared the detection of HLB with different Quantitative PCR based methods with primers/probe targeting either 16S rDNA, beta-operon DNA, 16S rRNA, or beta-operon RNA. The 16S rDNA copy number of Ca. Liberibacter asiaticus was estimated to be three times of that of the beta-operon region, thus allowing detection of lower titer of Ca. L. asiaticus. Quantitative reverse transcriptional PCR (QRT-PCR) indicated that the 16S rRNA averaged 7.83 times more than that of 16S rDNA for the same samples. Dilution analysis also indicates that QRT-PCR targeting 16S rRNA is 10 time more sensitive than QPCR targeting 16S rDNA. Thus QRT-PCR was able to increase the sensitivity of detection by targeting 16S rRNA. CONCLUSION: Our result indicates that Candidatus Liberibacter asiaticus contains three copies of 16S rDNA. The copy number of 16S rRNA of Ca. L. asiaticus in planta averaged about 7.8 times of 16S rDNA for the same set of samples tested in this study. Detection sensitivity of HLB could be improved through the following approaches: using 16S rDNA based primers/probe in the QPCR assays; and using QRT-PCR assays targeting 16S rRNA.
ABSTRACT
To study genome evolution and diversity in barley (Hordeum vulgare), we have sequenced and compared more than 300 kb of sequence spanning the Rph7 leaf rust disease resistance gene in two barley cultivars. Colinearity was restricted to five genic and two intergenic regions representing <35% of the two sequences. In each interval separating the seven conserved regions, the number and type of repetitive elements were completely different between the two homologous sequences, and a single gene was absent in one cultivar. In both cultivars, the nonconserved regions consisted of approximately 53% repetitive sequences mainly represented by long-terminal repeat retrotransposons that have inserted <1 million years ago. PCR-based analysis of intergenic regions at the Rph7 locus and at three other independent loci in 41 H. vulgare lines indicated large haplotype variability in the cultivated barley gene pool. Together, our data indicate rapid and recent divergence at homologous loci in the genome of H. vulgare, possibly providing the molecular mechanism for the generation of high diversity in the barley gene pool. Finally, comparative analysis of the gene composition in barley, wheat (Triticum aestivum), rice (Oryza sativa), and sorghum (Sorghum bicolor) suggested massive gene movements at the Rph7 locus in the Triticeae lineage.
Subject(s)
Genetic Variation , Genome, Plant , Haplotypes/genetics , Hordeum/genetics , Conserved Sequence/genetics , Contig Mapping , DNA, Plant/chemistry , DNA, Plant/genetics , Evolution, Molecular , Genes, Plant/genetics , Molecular Sequence Data , Oryza/genetics , Physical Chromosome Mapping , Sequence Analysis, DNA , Sorghum/genetics , Triticum/geneticsABSTRACT
In this work, the effect of anodic surface treatment on activated carbons and its Cr(VI) adsorption properties were investigated under reaction-treatment time conditions with 35 wt% HCl solution. The acid-base surface values were determined by Boehm's titration technique. The pore and surface characteristics were studied in terms of BET volumetric measurement with N(2) adsorption. As an experimental result, the acidic surface functional groups of activated carbons increased with increasing the HCl reaction-treatment time. It was found that the surface characteristics, including specific surface area, total pore volume, net heat of adsorption, and BET's C, slightly decrease in anodic surface treatments on activated carbons. In addition, an increase in reaction-treatment time led to increases of the first rate (K(1)) of Cr(VI) adsorption and diffusion coefficient (D). These values are evidence that adsorption is controlled more by the acid-base interactions between electron-donor substances and acidized activated carbons as electron acceptor than by the pore and surface structures of activated carbons. Copyright 2001 Academic Press.
ABSTRACT
Aloe vera continues to be used for wound healing as a folk medicine. We previously reported that A. vera gel has angiogenic activity. In this study, we report upon the isolation of an angiogenic component beta-sitosterol from A. vera and examination of its effect upon damaged blood vessels of the Mongolian gerbil. In a chick embryo chorioallantoic membrane assay, beta-sitosterol was found to have an angiogenic effect. It enhanced new vessel formation in gerbil brains damaged by ischaemia/reperfusion, especially in the cingulated cortex and septal regions, in a dose-dependent fashion (up to 500 microg/kg, p < 0.05, n = 34 - 40). beta-Sitosterol also enhanced the expressions of proteins related to angiogenesis, namely von Willebrand factors, vascular endothelial growth factor (VEGF), VEGF receptor Flk-1, and blood vessel matrix laminin (p < 0.05, n = 6). In addition, the intraperitoneal administration of beta-sitosterol at 500 microg/kg/day for a period of 19 days significantly improved the motion recovery of ischaemia/reperfusion-damaged gerbils as assessed by rota-rod testing (p < 0.001, n = 10). Our results suggest that beta-sitosterol has therapeutic angiogenic effects on damaged blood vessels.
Subject(s)
Aloe , Brain/drug effects , Plant Extracts/pharmacology , Sitosterols/pharmacology , Angiogenesis Inducing Agents/metabolism , Animals , Behavior, Animal/drug effects , Blood Vessels/drug effects , Blotting, Western , Brain/blood supply , Brain/metabolism , Chick Embryo , Dose-Response Relationship, Drug , Endothelial Growth Factors/metabolism , Gerbillinae , Immunohistochemistry , Laminin/drug effects , Laminin/metabolism , Lymphokines/drug effects , Lymphokines/metabolism , Plant Leaves/chemistry , Receptor Protein-Tyrosine Kinases/drug effects , Receptor Protein-Tyrosine Kinases/metabolism , Receptors, Growth Factor/drug effects , Receptors, Growth Factor/metabolism , Receptors, Vascular Endothelial Growth Factor , Reperfusion Injury/physiopathology , Sitosterols/isolation & purification , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth Factors , von Willebrand Factor/drug effects , von Willebrand Factor/metabolismABSTRACT
The reliability of genome analysis and proficiency of genetic manipulation are increased by assignment of linkage groups to specific chromosomes, placement of centromeres, and orientation with respect to telomeres. We have endeavored to establish means to enable these steps in sorghum (Sorghum bicolor (L.) Moench), the genome of which contains ca. 780 Mbp spread across n = 10 chromosomes. Our approach relies on fluorescence in situ hybridization (FISH) and integrated structural genomic resources, including large-insert genomic clones in bacterial artificial chromosome (BAC) libraries. To develop robust FISH probes, we selected sorghum BACs by association with molecular markers that map near the ends of linkage groups, in regions inferred to be high in recombination. Overall, we selected 22 BACs that encompass the 10 linkage groups. As a prelude to development of a multiprobe FISH cocktail, we evaluated BAC-derived probes individually and in small groups. Biotin- and digoxygenin-labeled probes were made directly from the BAC clones and hybridized in situ to chromosomes without using suppressive unlabelled C0t-1 DNA. Based on FISH-signal strength and the relative degree of background signal, we judged 19 BAC-derived probes to be satisfactory. Based on their relative position, and collective association with all 10 linkage groups, we chose 17 of the 19 BACs to develop a 17-locus probe cocktail for dual-color detection. FISH of the cocktail allowed simultaneous identification of all 10 chromosomes. The results indicate that linkage and physical maps of sorghum allow facile selection of BAC clones according to position and FISH-signal quality. This capability will enable development of a high-quality molecular cytogenetic map and an integrated genomics system for sorghum, without need of chromosome flow sorting or microdissection. Moreover, transgeneric FISH experiments suggest that the sorghum system might be applicable to other Gramineae.