How Prime Numbers Are Interconnected and Built with Two Equations: Addition and Subtraction Rules the Function (6µ)

Are all prime numbers linked by four simple functions? Can we predict when a prime will appear in a sequence of primes? If we classify primes into two groups, Group 1 for all primes that appear before ζ (such that , for instance 5, ), an even number divisible by 3 and 2, and Group 2 for all primes that are after ζ (such that , for instance 7), then we find a simple function: for each prime in each group, , where n is any natural number. If we start a sequence of primes with 5 for Group 1 and 7 for Group 2, we can attribute a μ value for each prime. The μ value can be attributed to every prime greater than 7. Thus for Group 1, and . Using this formula, all the primes appear for , where μ is any natural number.

Left atrial appendage occluder detachment treated with transthoracic ultrasound combined with digital subtraction angiography guided catcher:A case report

BACKGROUND There are very few cases of cardiac occluder detachment,and it is rare to completely remove the occluder using interventional methods without undergoing thoracotomy surgery after detachment.This case innovatively used ultrasound guidance combined with digital subtraction angiography(DSA)to completely remove the occluder,accumulating some experience.CASE SUMMARY The patient underwent left atrial appendage occlusion surgery in our hospital due to atrial fibrillation.After the surgery,the occluder fell off and became free in the left ventricle,which is very dangerous.We innovatively used ultrasound guidance,combined with DSA,and interventional surgery to successfully capture the free occluder using a catcher,completely remove it,and then re implant a new left atrial appendage occluder.After the surgery,the patient recovered very well.CONCLUSION The size selection of the occluder is slightly conservative,and the shape of the left atrial appendage opening is irregular.

Isolating Soil Drought-Induced Genes from Maize Seedling Leaves Through Suppression Subtractive Hybridization

In this study, a forward cDNA library was constructed by suppression subtractive hybridization using seedling leaves of CN165, a drought-tolerant maize inbred line. In the suppression subtractive hybridization （SSH） library, 672 positive clones were picked up randomly. After polymerase chain reaction （PCR） of each clone, all the single clones were sequenced. Totally 598 available sequences were obtained. After cluster analysis of the EST sequences, 80 uniESTs were obtained, among which 57 uniESTs were contigs and 23 uniESTs were singlets. The results of BLASTN showed that all the uniESTs had homologous sequences in the nr database. The BLASTX results indicated that 68 uniESTs had significant protein homology, 8 uniESTs with homology of unknown proteins and putative proteins, and 4 uniESTs without protein homology. Those drought stress-induced genes were involved in many metabolism pathways to regulate plant growth and development under drought stress.

Identifcation of up-regulated genes in human uterine leiomyoma by sup-pression subtractive hybridization

In searching for differentially expressed genes in human uterine leiomyomas (ULs), suppression sub-tractive hybridization was used to construct an UL up-regulated library, which turned out to represent 88genes. After two rounds of screening by reverse Northern analysis, twenty genes were proved to be up-regulated, including seventeen known genes and three genes with unknown function. All these genes werefirstly associated with UL. Three genes with notable difference were selected for Northern confirmationOur results proved the authenticity of the twenty genes. One gene named Phospholipase A2 (PLA2) showedup-regulation in 4/6 of the patients and investigation of tissue distribution indicated that it had obviousexpression in prostate, testis, liver, heart and skeletal muscle.

Identification of differentially expressed genes associated with bud dormancy release in tree peony(Paeonia suffruticosa) by suppression subtractive hybridization

A subtractive cDNA library was developed to study genes associated with bud dormancy release in tree peonies. In order to identify genes that are highly expressed in buds released from dormancy, 588 clones were examined by differential screening. Of these, 185 clones were selected to be sequenced. A total of 37 unique sequences were obtained of which only 31 sequences have matches in the NCBI database or the Arabidopsis thaliana protein database. Semi-quantitative RT-PCR was used to confirm further the expression profiles for 12 transcripts identified within the subtractive cDNA library. Gene ontology analyses indicated that many of the different genes identified have unknown or hypothetical functions while it is speculated that other genes play different mo- lecular roles. In our study, genes involved in bud dormancy release were growth-related or stress-responsive, while low-temperature-induced ribosomal proteins may also play a role in bud dormancy release. Our results provide interesting information for further understanding of the molecular mechanism of bud dormancy release in tree peonies.

Construction of Two Suppression Subtractive Hybridization Libraries and Identification of Salt-Induced Genes in Soybean

Soybean is planted worldwide and its productivity is significantly hampered by salinity. Development of salt tolerant cultivars is necessary for promoting soybean production. Despite wealth of information generated on salt tolerance mechanism, its basics still remain elusive. A continued effort is needed to understand the salt tolerance mechanism in soybean using suitable molecular tools. To better understand the molecular basis of the responses of soybean to salt stress and to get an enrichment of critical salt stress responsive genes in soybean, suppression subtractive hybridization libraries （SSH） are constructed for the root tissue of two cultivated soybean genotypes, one was tolerant and the other was sensitive to salt stress. To compare the responses of plants in salt treatment and non-treatment, SSH1 was constructed for the salt-tolerant cultivar Wenfeng 7 and SSH2 was constructed for the salt-sensitive cultivar Union. From the two SSH cDNA libraries, a total of 379 high quality ESTs were obtained. These ESTs were then annotated by performing sequence similarity searches against the NCBI nr （National Center for Biotechnology Information protein non-redundant） database using the BLASTX program. Sixty-three genes from SSH1 and 49 genes from SSH2 could be assigned putative function. On the other hand, 25 ESTs of SSH1 which may be not the salt tolerance-related genes were removed by comparing and analyzing the ESTs from the two S SH libraries, which increased the proportion of the genes related to salt tolerance in S SH 1. These results suggested that the novel way could realize low background of SSH and high level enrichment of target cDNAs to some extent.

Identification of Differentially Expressed Genes in the Salivary Gand of Rhipicephalus haemaphysaloides by the Suppression Subtractive Hybridization Approach

For the purpose of screening and analyzing the differentially expressed genes from the salivary gland of Rhipicephalus haemaphysaloides, two salivary gland-subtracted cDNA libraries of partially fed female ticks and fed male ticks were constructed using suppression subtractive hybridization （SSH）. A total of 247 female expression sequence tags （ESTs） and 168 male ESTs were obtained from the two SSH cDNA libraries. It is predicted that 25 female ESTs and 44 female ESTs contain the 5＂ and 3＂ ends, respectively, and that 53 male ESTs and 74 male ESTs contain the 5＂ and 3＂ ends, respectively. To identify the subtraction rate of the two SSH cDNA libraries, the RT-PCR method was used to test 24 female ESTs and 21 male ESTs selected randomly but not repeatedly. The results showed that there were 13 upregulated or differentially expressed genes in the partially fed salivary gland of the female R. haemaphysaloides and that the differentially expressed rate was 54%. In addition, they indicated that there were 9 upregulated or differently expressed genes in the fed salivary gland of the male R. haemaphysaloides and that the differentially expressed rate was 43%. Putative translations of 141 （57%） female ESTs and 125 （74%） male ESTs had similarity to GenBank sequences, and 32 （23%） female ESTs and 29 （23%） male ESTs exhibited similarity to tick proteins, which showed that most of the proteins in the libraries were mainly related to the feeding blood physiology of the ticks.

Subsurface Defect Evaluation of Selective-Laser-Melted Inconel 738LC Alloy Using Eddy Current Testing for Additive/Subtractive Hybrid Manufacturing

New materials and manufacturing technologies require applicable non-destructive techniques for quality assurance so as to achieve better performance.This study comprehensively investigated the effect of influencing factors includ-ing excitation frequency,lift-off distance,defect depth and size,residual heat,and surface roughness on the defect EC signals of an Inconel 738LC alloy produced by selective laser melting(SLM).The experimental investigations recorded the impedance amplitude and phase angle of EC signals for each defect to explore the feasibility of detecting sub-surface defects by merely analyzing these two key indicators.Overall,this study revealed preliminary qualitative and roughly quantitative relationships between influencing factors and corresponding EC signals,which provided a prac-tical reference on how to quantitively inspect subsurface defects using eddy current testing(ECT)on SLMed parts,and also made solid progress toward on-line ECT in additive/subtractive hybrid manufacturing(ASHM)for fabricating SLMed parts with enhanced quality and better performance.

Soil Microbial Dynamics Modeling in Fluctuating Ecological Situations by Using Subtractive Clustering and Fuzzy Rule-Based Inference Systems

Microbial population and enzyme activities are the significant indicators of soil strength.Soil microbial dynamics characterize microbial population and enzyme activities.The present study explores the development of efficient predictive modeling systems for the estimation of specific soil microbial dynamics,like rock phosphate solubilization,bacterial population,and ACC-deaminase activity.More specifically,optimized subtractive clustering(SC)and Wang and Mendel's(WM)fuzzy inference systems(FIS)have been implemented with the objective to achieve the best estimation accuracy of microbial dynamics.Experimental measurements were performed using controlled pot experiment using minimal salt media with rock phosphate as sole carbon source inoculated with phosphate solubilizing microorganism in order to estimate rock phosphate solubilization potential of selected strains.Three experimental parameters,including temperature,pH,and incubation period have been used as inputs SC-FIS and WM-FIS.The better performance of the SC-FIS has been observed as compared to the WM-FIS in the estimation of phosphate solubilization and bacterial population with the maximum value of the coefficient of determination(0.9988)2 R=in the estimation of previous microbial dynamics.

Multi-face detection based on downsampling and modified subtractive clustering for color images

This paper presents a multi-face detection method for color images. The method is based on the assumption that faces are well separated from the background by skin color detection. These faces can be located by the proposed method which modifies the subtractive clustering. The modified clustering algorithm proposes a new definition of distance for multi-face detection, and its key parameters can be predetermined adaptively by statistical information of face objects in the image. Downsampling is employed to reduce the computation of clustering and speed up the process of the proposed method. The effectiveness of the proposed method is illustrated by three experiments.

Screening for metronidazole-resistance associated gene fragments of H pylori by suppression subtractive hybridization

AIM: To screen for metronidazole (MTZ)-resistance associated gene fragments of H pylori by suppression subtractive hybridization (SSH). METHODS: Five MTZ-resistant (tester, T) and 1 MTZ-susceptible (driver, D) clinical H pylori isolates were selected. Genomic DNAs were prepared and submitted to RsaⅠdigestion. Then two different adaptors were ligated respectively to the 5'-end of two aliquots of the tester DNA fragments and SSH was made between the tester and driver DNAs. The specific inserts of tester strains were screened and MTZ-resistance related gene fragments were identified by dot blotting. RESULTS: Among the randomly selected 120 subtractive colonies, 37 DNA fragments had a different number of DNA copies (≥ 2 times) in resistant and susceptible strains and 17 of them had a significantly different number of DNA copies (≥ 3 times). Among the sequences obtained from the 17 DNA fragments, new sequences were found in 10 DNA fragments and duplicated sequences in 7 DNA fragments, representing respectively the sequences of depeptide ABC transporter periplasmic dipeptide-binding protein (dppA), permease protein (dppB), ribosomal protein S4 (rps4), ribonuclease Ⅲ (rnc), protease (pqqE), diaminopimelate epimerase (dapF), acetatekinase (ackA), H pylori plasmid pHP51 and H pylori gene 1334. CONCLUSION: Gene fragments specific to MTZ-resistant H pylori strains can be screened by SSH and may be associated with MTZ-resistant H pylori.

Identification of genes upregulated by recombinant interferon-alpha in HepG2 cells by suppressive subtractive hybridization analysis

BACKGROUND: Interferon-alpha (IFN-alpha) is an important cytokine with multiple functions, but the target genes transactivated by IFN-alpha remain largely unknown. A study of such genes will help to understand the mechanism of function of IFN-alpha. To isolate the gene transcripts specifically upregulated by IFN-alpha in HepG2 cells, we conducted suppressive subtractive hybridization (SSH) analysis. METHODS: SSH was used to analyze the target genes transactivated by recombinant IFN-alpha protein, and a subtractive cDNA library was constructed from HepG2 cells treated with recombinant IFN-alpha (rIFN-alpha, 2000 IU/ml) for 16 hours as tester, and cells not treated with rIFN-alpha as driver. The SSH PCR products from the library were cloned into pGEM-T easy vector and with BLASTX, the positive clones were randomly selected, sequenced and compared to the database in GenBank of the 35 differentially expressed gene fragments from the library, 6 clones showed significant homology to other known proteins. RESULTS: The subtractive cDNA library of genes upregulated by IFN-alpha was constructed successfully, rIFN-alpha upregulated the expression of the RAN binding protein 5 (RANBP5), NADH dehydrogenase, exosome component 3 (EXOSC3), zinc finger RNA binding protein, Dickkopf homolog 1 (DKK1) and acetyl-coenzyme A acetyltransferase 2 (ACAT2). CONCLUSIONS: These results suggest that rIFN-alpha can upregulate the expression of important genes to exert its functions, and provide new clues for discovering the molecular mechanisms of action of IFN-alpha.

Construction of Suppression Subtractive Hybridization Library for Testis of Male Tilapia Under the Stress of Methomyl

[Objectives]This study was conducted to construct forward and reserve libraries of suppression subtractive hybridization (SSH) in the testis of male tilapia under the stress of methomyl by using SSH technology.[Methods]Using male tilapia as the test animal,the forward and reserve libraries of SSH in the testis of tilapia under the stress of methomyl were constructed by using the SSH technology.[Results]45 expressed sequence tags (ESTs) were obtained,and 25 expressed sequence tags were successfully noted,including 13 forward libraries and 12 reverse libraries.The genes with confirmed functions were classified into five types.Genes related to catalytic activity and cell characteristics were up-regulated,while genes related to structural molecule's activity and biological process were down-regulated.The expression amount of integrin β1 was up-regulated,while serine/threonine protein kinase pim-3,Ca^2+-ATPase,Na^+-K^+-ATPase and ribosomal protein L22 were down-regulated.[Conclusions]The research results could lay a foundation for revealing the molecular mechanism of methomyl's reproductive toxicity to tilapia.

Suppression Subtractive Hybridization Reveals Different Responses of Two Varieties of Gossypium arboreum L. Under Apolygus lucorum Stress

Plants reshape their transcriptomes, proteomes and metabolomes in response to insect damage. In this study, we used suppression subtractive hybridization to investigate the transcriptomes of two cotton varieties （CCRI41 and CCRI23） under Apolygus lucorum damage. From the CCRI23 libraries we obtained 92 transcripts and from the CCRI41 libraries we obtained 96 transcripts. 26 and 63 of the transcripts from CCRI23 and CCRI41, respectively, had known functions. Using reverse transcription PCR, we detected expression proifle of genes with known functions. Ultimately, we identiifed eight signiifcantly regulated genes, including one downregulated and four upregulated genes from the CCRI41 libraries, and one downregulated and two upregulated genes from the CCRI23 libraries. Only the gene encoding the polyphenol oxidase （PPO） is involved in plant defense against insect herbivores, and the others are related to improving tolerance to insect damage. Quantitative real-time PCR was used to study changes in expression levels during A. lucorum damage in CCRI23 and CCRI41. Signiifcantly regulated genes from CCRI23 showed a response in CCRI23 but not response in CCRI41. Similarly, signiifcantly regulated genes from CCRI41 showed a response in CCRI41 but not response in CCRI23. The results showed that, among transcriptomes of cotton varieties, there are different responses to A. lucorum damage.

Using a Suppression Subtractive Library-Based Approach to Identify Non-Heading Chinese Cabbage Genes Up-Regulated in Early Response to Elicitor PB90

Monitoring expression at the transcriptional level is the first essential step for the functional analysis of plant genes. Genes-encoding proteins directly involved in early response to elicitor constitute only a small fraction of all the genes affected by elicitor. Transcriptional responses to various elicitors have been extensively studied in different plants including Nicotiana and Arabidopsis thaliana; however, corresponding data aren＇t available for non-heading Chinese cabbage. To address this problem, we describe a suppression subtractive library-based approach to isolate the plant＇s ESTs up-regulated in the early induction/execution of the HR induced by elicitor PB90 from Phytophthora boehmeriae. According to their putative identification in BLAST searches against the three genome databases, 70 up-regulated genes were classified into 9 parts： some aspect of primary ＇metabolism＇ or ＇energy＇ production; ＇protein synthesis＇ or ＇protein fate＇; cellular communication/signal transduction mechanism; cell fates including Beclin, SPT1, and SPT2; HLA-B and AGO1 which participate in transcription; cellular transport and hypothetical proteins or proteins for which a function has yet to be determined. Seven selected genes such as Beclin, thioredoxin, HLA-B, MAP3K, SPT1, SPT2, and AGO1 were up-regulated induced by PB90, suggesting that the genes may play an important role in PB90-triggered HR.

Construction and Analysis of Suppression Subtractive cDNA Library from Fertile Disk Floret Buds of Zinnia elegans

In order to isolate genes related to anther development and understand molecular basis of male sterility,cDNA library of Zinnia elegans was constructed using suppression subtractive hybridization（SSH） approach.672 different expressed clones were selected from the fertile disk floret buds.PCR results showed that cDNA inserts were ranged from 100 to 750 bp.303 positive clones screened by dot-blot hybridization were sequenced.273 out of 303 sequenced clones produced readable sequences;these sequences represent 87 non-repetitive sequences.The homology alignment showed that 76 expressed sequence tags（ESTs） had functional annotations in GenBank,the other 11 ESTs without any homology to the known gene.In addition,87 ESTs were divided into 17 groups according to MIPS of Arabidopsis thaliana database.Sequence data from the cDNA library have been deposited with the GenBank under the accession numbers GT067016-GT067085.As an important result in this study,7 genes related to anther development were isolated.Results from semi-quantitative RT-PCR showed 6 genes were expressed only in disk florets of fertile plants compared with that of male sterile plants.These ESTs obtained provide important clues for further isolation and identification of fertility-related genes in Z.elegans.

Identification of differentially expressed genes after partial rat liver ischemia/reperfusion by suppression subtractive hybridization

AIM: To identify potential diagnostic target genes in early reperfusion periods following warm liver ischemia before irreversible liver damage occurs.METHODS: We used two strategies (SSH suppression subtractive hybridization and hybridization of cDNA arrays)to determine early changes in gene expression profiles in a rat model of partial WI/R, comparing postischemic and adjacent nonischemic liver lobes. Differential gene expression was verified (WT/R; 1 h/2 h) and analyzed in more detail after warm ischemia (1 h) in a reperfusion time kinetics (0, 1, 2 and 6 h) and compared to untreated livers by Northern blot hybridizations. Protein expression was examined on Western blots and by immunohistochemistry for four differentially expressed target genes (Hsp70,Hsp27, Gadd45a and IL-1rl).RESULTS: Thirty-two individual WI/R target genes showing altered RNA levels after confirmation by Northern blot analyzes were identified. Among them, six functionally uncharacteristic expressed sequences and 26 known genes (12 induced in postischemic liver lobes, 14 with higher transcriptional expression in adjacent nonischemic liver lobes). Functional categories of the verified marker genes indicate on the one hand cellular stress and tissue damage but otherwise activation of protective cellular reactions (AP-1 transcription factors, apoptosis related genes, heat shock genes). In order to assign the transcriptional status to the biological relevant protein level we demonstrated that Hsp70, Hsp27, Gadd45a and IL-1rI were clearly up-regulated comparing postischemic and untreated rat livers, suggesting their involvement in the WI/R context.CONCLUSION: This study unveils a WI/R response gene set that will help to explore molecular pathways involved in the tissue damage after WI/R. In addition, these genes especially Hsp70and Gadd45a might represent promising new candidates indicating WI/R liver damage.

Differential Regulation of Proteins and a Possible Role for Manganese Superoxide Dismutase in Bioluminescence of Panellus stipticus Revealed by Suppression Subtractive Hybridization

Suppression subtractive hybridization (SSH) was employed to investigate bioluminescence in Panellus stipticus (Bull.) P. Karst. by detecting proteins differentially expressed in bioluminescent and luminescent strains. Comparisons of luminescent and non-luminescent monokaryon cultures of North American strains revealed differences in transcript levels of proteins responsible for post-translational modification (PTM) of enzymes. A similar comparison of a luminescent strain of P. stipticus from North America with a non-luminescent European strain revealed the presence of extracellular manganese superoxide dismutase (MnSOD) in the luminescent form, in addition to proteins involved in PTM. The application of MnSOD-specific inhibitors to luminescent mycelium resulted in the rapid loss of luminescence. The relevance to luminescence of proteins involved in PTM is discussed, together with a possible role for MnSOD that considers the potential for SODs to form stable complexes with catechols revealed in previously published research. In light of the recent discovery that hispidine may be the precursor of fungal luciferin, we consider a hypothetical mechanism for fungal luminescence in which the ο-hydroquinone moiety of a hispidine derivative ligates with the extracellular form of MnSOD producing a semiquinone-radical complex, with the resultant semiquinonato complex potentially reacting with molecular oxygen or other reactive oxygen species to produce sufficiently excited intermediates to emit light on relaxation.

Identifcation of up-regulated genes in human uterine leiomyoma by sup-pression subtractive hybridization

In searching for differentially expressed genes in human uterine leiomyomas (ULs), suppression sub-tractive hybridization was used to construct an UL up-regulated library, which turned out to represent 88genes. After two rounds of screening by reverse Northern analysis, twenty genes were proved to be up-regulated, including seventeen known genes and three genes with unknown function. All these genes werefirstly associated with UL. Three genes with notable difference were selected for Northern confirmationOur results proved the authenticity of the twenty genes. One gene named Phospholipase A2 (PLA2) showedup-regulation in 4/6 of the patients and investigation of tissue distribution indicated that it had obviousexpression in prostate, testis, liver, heart and skeletal muscle.

Novel robust approach for constructing Mamdani-type fuzzy system based on PRM and subtractive clustering algorithm

A novel approach for constructing robust Mamdani fuzzy system was proposed, which consisted of an efficiency robust estimator(partial robust M-regression, PRM) in the parameter learning phase of the initial fuzzy system, and an improved subtractive clustering algorithm in the fuzzy-rule-selecting phase. The weights obtained in PRM, which gives protection against noise and outliers, were incorporated into the potential measure of the subtractive cluster algorithm to enhance the robustness of the fuzzy rule cluster process, and a compact Mamdani-type fuzzy system was established after the parameters in the consequent parts of rules were re-estimated by partial least squares(PLS). The main characteristics of the new approach were its simplicity and ability to construct fuzzy system fast and robustly. Simulation and experiment results show that the proposed approach can achieve satisfactory results in various kinds of data domains with noise and outliers. Compared with D-SVD and ARRBFN, the proposed approach yields much fewer rules and less RMSE values.