Clinical Description

Four main symptoms are observed in CADASIL migraines with aura, ischemic attacks, mood disturbances, and cognitive decline or dementia. The clinical presentation is highly variable between and within families. The age of onset varies greatly, depending on the criterion used for the onset of the disease. 1-3 Migraine attacks may begin even before age 10 years, but more commonly during the second and third decades. The most constant finding is a history of ischemic events, transient or completed,...

Introduction

Prader-Willi syndrome (PWS), a condition initially described by Prader, Labhard, and Willi in 1956, 1 is a neurodevelopmental disorder with a clinical presentation that includes neonatal hypotonia, obesity, mental retardation, and behavioral abnormalities. After introduction of high-resolution chromosome analysis, PWS was the first abnormal phenotype found to be associated with a submicroscopic chromosomal deletion. 2 The PWS-asso-ciated microdeletion spans about 4 Mbp in the chromosomal region...

Clinical Features In Hereditary Breast And Ovarian Cancer Syndrome

BRCA mutations occur in approximately 20 of families with inherited susceptibility to breast cancer. Personal and family characteristics associated with an increased likelihood that a BRCA mutation will be identified are listed in Table 3. Breast cancers in BRCA1 carriers often have medullary features, are more likely to be poorly differentiated with high mitotic rates and S-phase fraction, lack an in situ component, have low estrogen and progesterone receptors score, and node-positive. BRCA2...

Screening In Hereditary Breast And Ovarian Cancer Syndrome

Although overall population screening reduces breast cancer mortality by 25-40 in women between the ages of 50 and 70 years, no data are available on the outcomes of interventions to reduce risk in HBOC syndrome. No data are available to demonstrate that surveillance for ovarian cancer in high-risk women reduces mortality 3 because transvaginal ultrasound and CA-125 lack sensitivity and specificity. However, recommended screening in BRCA gene mutation carriers includes monthly breast Table 4...

Substrate Reduction Therapy

Oral substrate reduction therapy (SRT) involves attenuation of the rate of synthesis to achieve a balance with reduced endogenous glucocerebrosidase. A pivotal trial of SRT was carried out in 28 adult patients (from four centers Cambridge, UK Amsterdam Prague and Jerusalem) naive to enzyme therapy, with mild to moderate type I Gaucher disease, who were unable or unwilling to receive enzyme treatment. There was significant reduction in spleen and liver, although amelioration of anemia and...

Bladder Cancer Studies Using In Vitro Models

Expression profiling using bladder cancer cell lines has been used to gain insight into the molecular events associated with clinical disease states, assigning potential functional roles to novel genes in both tumorigenic and tumor progression processes. Certain studies have focused on gene and pathway discovery associated to genistein 10 or 5-aza-2'-deoxycytidine 11 exposure. Other reports describe the functional classification of genes comparing the expression patterns of p53-mediated...

Microarrays As Target Identification Tools In Cancer

Microarrays constitute a group of technologies characterized by the common availability of measuring hundreds or thousands of items, including DNA sequences, RNA transcripts, or proteins, within a single experiment using miniaturized devices. Hybridization-based methods and the microarray format constitute together an extremely versatile platform provided for both static and dynamic views of DNA structure, as well as RNA and protein expression patterns in cultured cancer cells and tumor

Prevalence And Clinical Diversity

OI is an autosomal genetic disorder characterized by brittleness of bones and susceptibility to bone fractures from very mild trauma. Fractures may be rare or frequent (more than 200 prior to puberty), and bone fragility may be accompanied by reduced life span and various connective tissue abnormalities short stature, blue sclerae, joint laxity, dentinogenesis imperfecta, easy bruising, and hearing loss. OI represents extreme variation in severity from lethal perinatal to barely detectable....

Udpglucuronosyltranferase 1a1

UDP-glucoronosyltransferases are enzymes responsible for detoxification and elimination of various metabolites and drugs. These enzymes also catalyze the inactivation of irinotecan (CPT-11), an analog of the alkaloid campto-thecin. CPT-11 is clinically used for the treatment of refractory and advanced colorectal cancers. Dose-limiting toxicities are diarrhea and pancytopenia. CPT-11 is metabolized to its active product, SN-38, by the enzyme carboxylesterase 2. As depicted in Fig. 3, UGT1A1 is...

The Need For Biomarkers In Bladder Cancer

Bladder tumors are pathologically stratified based on stage, grade, tumor size, presence of concomitant carcinoma in situ, and multicentricity.1-2-1 The chances of tumor progression are augmented with the increase of these pathological variables. Pathologically, most bladder tumors are transitional cell carcinomas. There is, however, increasing recognition of the prognostic importance associated with the metaplastic variants displaying squamous and glandular differentiation as part of their...

Chemoprevention In Hereditary Breast And Ovarian Cancer Syndrome

Women who desire risk reduction intervention for breast cancer have the option of chemoprevention with tamoxifen therapy (20 mg day for 5 years) or prophylactic surgery. The National Surgical Adjuvant Breast and Bowel Project prevention trial in healthy women 35 years and older with a 1.7 or greater cumulative 5-year risk for developing breast cancer demonstrated short-term reduction in risk of developing estrogen-receptor-positive breast cancer by 49 . The utility of tamoxifen for breast...

Genotypephenotype Correlation

The lower limit of the normal range for factor XI is commonly given at about 70 . Individuals with levels up to 15-20 are regarded as partially deficient, while individuals with lower levels are severely deficient and represent homozygotes or compound heterozygotes. 6,7 In general, bleeding is negatively correlated with factor XI level. 7 However, only at a very low factor XI level is the risk for excessive bleeding clearly increased, probably because the quantity of factor XI necessary for...

Pharmacogenetics Of Drugmetabolizing Enzymes

Polymorphism of genes encoding drug-metabolizing enzymes is the major known genetic cause for the interindividual variability in drug response. 1-6 Based on the occurrence of mutations in these genes, gene deletions, and gene duplications, the populations can be divided into poor (PM), intermediate (IM), efficient (EM), or ultrarapid (UM) drug metabolizers. The PMs lack the enzyme in question, the IMs are heterozygous for a defect gene, the EMs have two functional alleles, whereas the UMs have...

Bladder Cancer Studies Using Clinical Specimens

Microarray analyses have been used to correlate changes in the expression of specific genes and groups of genes within distinct bladder subclasses. Such signature genes would ideally provide a molecular basis for classification, yielding insight into the molecular events underlying different clinical bladder cancer phenotypes. The first report monitored the expression patterns of superficial and invasive tumor cell suspensions prepared from individuals and pools of normal and bladder tumors of...

The Challenge Of Integrating Genomic Innovation

It is commonly believed that innovations succeed on their own merit. Those without real merit fail in the market place those with superior merit succeed simply on the basis of their superiority over existing alternatives. However, experience shows otherwise. Technological capability and superior engineering do not, in fact, guarantee acceptance (Table 1). Great innovations themselves are often insufficient to ensure usability, effectiveness, or user satisfaction. In other words, good technology...

Famccggcccagccaccatggtamra

Table 3 Sequences of the probes used for real-time PCR of CYP1A2 Detected allele Sequence of probe 5'-3' Reference Phenotyping using caffeine showed no difference in CYP1A2*1F metabolic activity between different genotypes in nonsmokers but a significant difference in smokers homozygous for the A allele compared with the other genotypes in terms of a 1.6-fold increased activity.1-17-1 Unexpectedly, another study demonstrated a lower in vivo activity in patients with colorectal cancer than in...

Molecular Characterization Of Pathogen And Molecular Testing

E. histolytica and E. dispar genome shotgun sequencings are underway as a joint effort from TIGR 11 and the Sanger Centre, 12 funded by the National Institute for Allergy and Infectious Diseases and the Wellcome Trust, respectively. The genome is around 18-20 Mb in size in 14 chromosomes. 11-13 However, partial genomic data from these organisms are available since the end of the 1980s. 14,15 These data allowed designing molecular assays that are sensitive to detection and, more interestingly,...

Optimization Of Liposomal Delivery

Efficient in vivo nucleic acid-liposome complexes have unique features, including their morphology, mechanisms for cell and nuclear entry, targeted delivery, and ability to penetrate across tight barriers and throughout target tissues. Liposomes have different morphologies based on their composition and the formulation method that contribute to their ability to deliver nucleic acids in vivo. Formulations frequently used for the delivery of nucleic acids are lamellar structures, including small...

Metabolism of Chemotherapeutic Agents

Tenable, although the failure to define precise roles for individual GST has hampered assessment of the implications of polymorphism as selection of diseases whose pathogenesis may be affected is subjective. Thus while associations between GST genotypes and clinical pheno-type have been reported they cannot be predicted, and it is unclear why some, but not other, links are significant. Further, study reproducibility is a problem, probably because of variable definition of phenotypes and...

Type Of Pathogen

Candida species are primarily saprophytic eukaryotic organisms and only occasional opportunistic pathogens. Many species exhibit a bitrophic existence, spending much of their life as common commensals of the gastrointestinal (GI) tract and mucous membranes of humans only to become pathogenic because of debilitation of the host. Indeed, the most significant source of Candida causing candidosis is endogenous. 2 Despite the main cause of invasive candidosis (IC Candida albicans) being considered...

Comparative Genomic Hybridization In Cancer Cytogenetics

Genetic alterations associated with neoplasia have been well defined in hematological malignancies by both classical and molecular cytogenetics.1-5'6-1 In contrast, there is significantly less information known about the cytogenetics and molecular cytogenetics of solid tumors. This is because of technical difficulties in the production of metaphase spreads from these tumor cells. Karyotype analysis requires viable, proliferating cells that can be arrested in the metaphase stage of the cell...

Microsatellite Instability In Hnpcc And In Sporadic

The MSI-H phenotype is a defining feature of HNPCC patients and is a result of predisposing constitutional mutations in the MMR genes MSH2, MLH1, MSH6, hPMS2, and hMSH3. 8 Most mutations were found in MSH2 and MLH1 and less frequently in MSH6, hPMS2, and hMSH3. Germline mutations in MMR genes predispose not only to CRC but also to endometrial, upper urinary tract, and ovarian malignancies. Small intestinal and gastric cancers as well as tumors of the biliary tract are also seen in HNPCC, but...

Estimation Of Infectivity

Another important reason for HBV viral load determination is the assessment of the infectivity of hepatitis B carriers. Without intervention, more than 90 of HBeAg-positive female chronic HBV carriers transmit the virus to their infants 17 of these, 85-90 develop chronic HBV infection in most cases asymptomatic themselves, thus perpetuating the infection in high-endemicity settings. Immediate postpartum immunization of the infant can efficiently prevent transmission. It has to be considered...

Assay Formats Direct Hybridization

Several formats of wet-ware were adopted to bead-based nucleic acid testing. The simplest method is the direct hybridization of labeled nucleic acids to beads carrying a sequence-specific oligonucleotide. An assay for gene expression analysis using color-coded microspheres was developed using this technique. In Ref. 4 , labeled cRNA was generated which was hybridized to a set of 20 bead populations carrying specific capture sequences of 25 bp for 20 Arabidopsis genes. The obtained expression...

Bio Robot Workstations

The BioRobot EZ1 workstation (Qiagen, Hilden, Germany) purifies nucleic acids from many clinically relevant specimens one to six samples can be processed within 15 min of time. This rapid purification is feasible using a magnetic particle technology nucleic acids in sample lysates are isolated in one step through their binding to the silica surface of magnetic particles. The automated protocols for nucleic acid purification are provided on special preprogrammed cards. The BioRobot M48 M96...

Impact Of Fish Analysis In Different Research Areas

More than 5 million sperm cells from more than 500 normal men have been analyzed by a number of laboratories from around the world. 7 These studies have included the frequency and distribution of aneuploidy in sperm cells, and the effect of donor age, lifestyle factors, and geographical and ethnic factors. Frequency and Distribution of Aneuploidy The frequency and distribution of disomy in human sperm cells is of interest to determine the normal baseline values and to determine if all...

Clinical Diagnosis

Becker muscular dystrophy patients show normal growth and development in their early childhood. In BMD, affected men start to show disturbance of walking due to muscle weakness at 20s or over. He maintains to walk but his muscle strength gradually decreases. Dilated cardio-myopathy is sometimes an initial clinical sign for the diagnosis of BMD. The muscle weakness involved in BMD follows a mild downward course with patients living near normal lives. 1 Serum creatine kinase (CK) is markedly...

Clinical Description of Infection

HHV-6B is the principle cause of the exanthem roseola infantum (exanthem subitum), an illness characterized by high fever and development of a rash after fever resolves. 8 HHV-7 can also cause roseola. However, the majority of children with HHV-6B or HHV-7 infection develop an undifferentiated fever, but this may be complicated in some by febrile convulsions, encephalopathy, and hepatitis. Primary infection is rare in adults but can occur including an infectious mononucleosis-type illness....

Oxidative Stress Related Disorders

Oxidative stress is implicated in the inflammatory demy-elination that characterizes multiple sclerosis suggesting GST polymorphisms may be associated with disability. In 177 patients with disease duration over 10 years, GSTM3 AA (OR 2.4) and homozygosity for both GSTM1*0 and GSTP1*Ile105-encoding allele (OR 5.0) were linked with severe disability suggesting that long-term prognosis in MS is influenced by GST-mediated ability to remove toxic products of oxidative stress. 1 Exposure to...

Epidemiology And Clinical Features

415 N-terminal amino acids but shows variability thereafter. The polypeptide is cleaved proteolytically to 20-40-kDa structural proteins, and, for example, the 26-kDa structural protein expresses a neutralization epitope. Currently, eight serotypes of human astrovirus are recognized (HuAst 1-8) based upon epitopes on ORF2. It is also possible to genotype HuAst by examination of sequence data of ORF1a, ORF1b, and the 5' end of ORF2, 12 and genotypes equivalent to the eight serotypes have been...

Hcv Rna Quantitation

Quantitation of HCV RNA by bDNA technology is possible with the commercial VERSANT HCV RNA 3.0 assay. The performance characteristics of this test were established in our laboratory during the so-called ''beta trials'' before licensing and are given in Table 1. 4 Our findings were essentially confirmed by other studies, 11,12 and showed that the assay, from an analytical point of view, can be used as a routine tool for HCV RNA quantitation in clinical laboratory settings. Table 1 Analytical...

Dm2

Although less severe, DM2 closely resembles adult-onset DM1 with common features, including progressive weakness, myotonia, cardiac arrhythmias, cataract, male hypogonadism, insulin insensitivity, and hypogammaglo-bulinemia. 2 The most frequent manifestations are muscle pain, muscle stiffness, myotonia, and weakness. 2 Muscle pain is frequently fluctuating or episodic. Weakness characteristically affects the neck flexors, elbow extensors, deep finger flexors, hip flexors, and hip extensors....

Infectious Disease Applications

The development and ongoing improvement of bDNA technology during the past decade was mainly driven by the increasing demand for quantitation of nucleic acids in clinical virology. Understanding the natural history and pathogenesis of chronic viral infections caused, for instance, by hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) has been greatly supported by accurate determinations of viral load. Consequently, measurements of HBV DNA, HCV RNA, and HIV...

Molecular Genetics

The NF1 gene is located on chromosome 17 and encompasses more than 300 kb of genomic DNA with 60 exons. The protein product is referred to as ''neuro-fibromin,'' a 2818 amino acid protein. Neurofibromin includes a GTPase-activating protein (GAP) domain, which regulates the conversion of Ras-GTP to Ras-GDP. Neurofibromin therefore appears to be a regulator of Rasmediated intracellular signaling. 12 The NF1 gene behaves as a classic tumor suppressor. Affected individuals are heterozygous for NF1...

Replication And Infectivity

Orthopoxvirus genomes encode about 176 to 266 proteins, including a number of enzymes and factors that are necessary for self-replication and maturation. The central region of the genome contains highly conserved genes that are essential for viral replication, and the terminal regions contain less conserved genes that are important for virus-host interactions. Although it is well known that replication occurs in the cytoplasm, it is not clear how or if host cell nuclear factors are involved in...

Studies In Renal Disease

The individual response to treatment with ACE inhibitors is highly variable. 20 Therefore, several studies have been conducted in which the efficacy of ACE inhibitor therapy was studied in relation to ACE genotype in patients with kidney diseases (Table 2). Yoshida et al. 21 studied the response to ACE inhibitor therapy in 21 Japanese patients with IgA-nephropathy, who were treated with lisinopril (10 mg day). After 4 years of therapy, only patients with DD genotype showed a significant...

Biology Of The Organisms

Chlamydiae are obligate intracellular bacteria of limited metabolic capability, characterized by a dimorphic growth cycle. The infectious form is called the elementary body (EB) and is a condensed sporelike spheroid. The EB is metabolically inactive, consisting of a tightly compacted chromosome or nucleoid, and an outer membrane of covalently linked lipopolysaccharides. When it encounters host cells, the EB is taken in by receptor-mediated endocytosis. It has been suggested that a trimolecular...

Early Diagnosis Of Hemochromatosis Phenotyping Vs Genotyping

Early diagnosis of hemochromatosis permits treatment of iron overload and thereby prevents premature death due to hepatic cirrhosis (and primary liver cancer) and diabetes mellitus complications.1-1,2-1 Preventing iron overload may also reduce the frequency or severity of arthropathy, hypogonadotrophic hypogonadism and other endocrinopathic disorders, and cardiac abnormalities. 1,2 The cornerstone of case detection in medical care and in population screening has been the measurement of serum...

Clinical Presentation

Wilson disease is a protean disorder with a broad clinical spectrum owing to the toxic effect of copper on the various organ systems. The disorder occurs worldwide with an estimated prevalence in Caucasians of approximately 1 per 30,000 to 1 per 100,000 live births and a carrier frequency of about 1 per 90 to 1 per 150. Most patients with Wilson disease present between 5 and 35 years of age. However, Wilson disease should be included in the differential diagnosis in individuals with liver...

Other Keratin Disorders

The spectrum of keratin disorders also encompasses a number of extracutaneous diseases. Meesmann corneal dystrophy (MSD) is inherited in an autosomal-dominant fashion with incomplete penetrance. Slit lamp identification of typical fine, round cysts in the corneal epithelium aids in differentiating this disorder from other inherited corneal dystrophies. 33 Although vision is only rarely impaired to a serious degree in MSD, cyst rupture can cause corneal erosions and intermittent decrease in...

Clinical Findings

Classical branching enzyme deficiency most frequently presents during the first few months of life, with hepatosplenomegaly and failure to thrive. Progressive liver cirrhosis, with portal hypertension, ascites, esopha-geal varices, and death, usually occurs before 5 years of age. However, there are patients who have survived without apparent progressive liver disease. The neuromuscular system may also be involved. Severe cardiomyopathy as the predominant symptom has also been reported. 20 The...

Potential Future Applications

Because FISH analysis is simple and provides data on large sample sizes of sperm cells, it has been suggested as a technique for assessing the effects of potential mutagens on human sperm. We and others have studied cancer patients treated by chemotherapeutic agents and determined that these agents can cause a significant increase in the frequency of sperm chromosome abnormalities, particularly during chemotherapy and up to 2 years following treatment. 27-29 The effects of pesticides have also...

Ocular Manifestations

The most common finding in AGS patients is posterior embryotoxon, which is the more minor form of eye anterior chamber abnormality. Posterior embryotoxon has no consequence on visual acuity and is present in 10-15 of normal individuals. All the spectrum of eye anterior chamber abnormalities may be observed including glaucoma. Retinal changes such as pigmentary retinitis were formerly ascribed to vitamin deficiency, but progressive blindness may occur even with normal vitamin level. 6

Genitourinary Chlamydial Infection

Chlamydia trachomatis infection of the genitourinary tract is the most common bacterial STD in both the United States (> 4 million new cases per year) and worldwide (> 50 million new cases per year). Many infected individuals of either sex are asymptomatic but symptomatic urethritis is common in men, whereas women may present with a mucopurulent cervicitis and acute salpin-gitis. The most serious complication of infection in men is epididymitis. Without treatment, more than 40 of infected...

Clinical Features

Patients with PWS have central muscle hypotonia that is manifested prenatally as decreased fetal movements, ab normal fetal position, and difficult delivery. The hypotonia is evident in the newborn period, accompanied by poor sucking and failure to thrive. Hypogonadism resulting in cryptorchidism and scrotal hypoplasia in males and labial hypoplasia in females are also seen in the newborn period. Hyperphagia and food-seeking behavior start at about 1 year of age and lead to obesity by 1-6...

Management Of Hbvinfected Patients

No molecular biology-based assays are necessary for the diagnosis of acute hepatitis B, which is based on serological testing. Chronic hepatitis B is defined by HBsAg persistence in serum for more than 6 months. In this setting, HBV DNA detection-quantification is necessary to determine whether or not HBV is replicating. Table 1 Commercially available HBV DNA quantification assays HBV Hybrid Capture II (Digene, Gaithersburg, MD) Versant HBV DNA (Bayer, Ludwigshafen, Germany) Cobas Amplicor HBV...

Clinical Description Dm1

Ninety percent of DM1 patients present at adulthood with delayed muscle maturation, distal muscle weakness, wasting, myotonia, cataracts, cardiac abnormalities, smooth muscle dysfunction, insulin resistance, daytime sleepiness, testicular atrophy (low reproductive fitness), ''difficult'' personality, neuropsychiatric disturbances, and frontal balding. 1 Ten percent of the patients present at infancy with hypotonia (floppy infant), oromotor dysfunction, tent-shaped mouth, feeding and respiratory...

Visceral Signs And Symptoms

Splenomegaly exists in all patients, and is typically associated with hypersplenism, early satiety, and, in children, growth (height) retardation. Anemia and throm-bocytopenia, resulting in fatigue and bleeding tendency, respectively, are among the earliest presenting signs and most prominent features of Gaucher disease. In almost all symptomatic patients, there is some degree of hepatomegaly but only rarely are liver function tests abnormal. Bone involvement is among the more variable and...

Conclusion

The discovery that subtelomeric rearrangements are a significant cause of unexplained MR has led to a vast number of studies aimed at providing diagnoses in this group. Results from over 3000 patients show that rearrangements involving the ends of chromosomes can be detected in about 5 of unselected cases, that the prevalence of subtelomeric anomalies is higher in patients with moderate to severe MR (about 7 ), and that in about half of those with subtelomeric anomalies, the rearrangements...

Method 1 Nonorganic Solvents Method Puregenetm Dna Isolation

The PUREGENE DNA Isolation kit (Gentra Systems Minneapolis, MN) is used to extract DNA from mummified tissues. 1,10 In this kit, purification of genomic DNA is carried out using ammonium acetate as a substitute for toxic organic solvents in the protein precipitation step. The pulverized tissue is mixed with cell lysis solution containing Tris-HCl, ethylenediaminetetraacetic acid (EDTA), and sodium dodecyl sulfate (SDS), and incubated at 65 C for 60 min. After proteinase K solution (20 mg mL)...

Mugpcr

Abbreviations ADPL allele discrimination by primer length PAMSA PCR amplification of multiple specific alleles CB competitive blocker CTPP confronting two-pair primer MUG multiplex universal genotyping. Abbreviations ADPL allele discrimination by primer length PAMSA PCR amplification of multiple specific alleles CB competitive blocker CTPP confronting two-pair primer MUG multiplex universal genotyping. allele-specific reaction internal control reaction Fig. 1 Principle of ARMS-PCR (see text for...

Clinical Significance Of Legionella

Legionella causes two different manifestations of pulmonary disease a mild, flulike illness, called Pontiac fever, and pneumonia. Pontiac fever has an incubation period of 1-2 days and is characterized by malaise, myalgias, fever, headache, and, sometimes, nonproductive cough. Only symptomatic therapy is required, and complete recovery within 1 week can be expected. Pneumonia, on the other hand, is the predominant manifestation of the more severe form of legionellosis, Legionnaires' disease....

The Virus

The family Astroviridae encompasses small nonenveloped viruses that infect a variety of animal species. They were originally classified among the small round structured viruses (SRSV) causing diarrhea in humans. They are round with icosahedral symmetry and 27-30 nm in diameter. They were first described by Madeley and Cosgrove 1 as a potential cause of diarrheal disease in human infants and named for the characteristic five- or six-pointed star (astron is Greek for a star), visible on the...

Multiplex Nasba And

Greijer et al. 5 designed a multiplex real-time NASBA using molecular beacon probes labeled with three different fluorophores to quantify human cytomegalovirus (CMV) IE1 mRNA by competitive coamplification of wild-type and calibrator RNA. A simultaneous detection of late pp67mRNA in whole-blood samples of CMV-infected lung transplant patients was also obtained. Despite the somewhat lower sensitivity of the real-time NASBA compared with the conventional NASBA, the simultaneous quantification of...

Retargeting Of Adenoviral Vectors

The adenovirus contains a protein capsid with protruding fibers. These fiber proteins form a trimer and the C-terminal region of these fibers is called the fiber knob. The fiber knob recognizes and binds to the host cell via the coxsackievirus-adenovirus receptor (CAR), which is present on many cell types. Upon binding to CAR, secondary interactions between the adenovirus and cell surface receptors cause the internalization of the adeno-virus. By shielding or modifying the fiber of the...

Overview Technical Description

Ramification amplification (RAM), 1,2 also termed hyperbranched rolling circle amplification1-3-1 or cascade rolling circle amplification,1-4-1 utilizes a closed C-probe or padlock probe and a DNA polymerase under isothermal conditions. The choice of DNA polymerase is based on the following requirements a good strand displacement activity in order to separate downstream DNA from the template strand and a high processivity (i.e., incorporating nucleotides continuously on a given primer without...

Tobacco Related Diseases

GSTM1 and GSTT1 have been particularly studied as risk candidates for tobacco-related cancers. 14,15 While some studies show significantly increased risk for GSTM1*0 and GSTT1*0 homozygotes, others have not replicated these findings prompting the use of meta-analysis. For example, analysis using 43 studies indicates that GSTM1 null is not associated with increased lung cancer risk and that there is no evidence for an interactive effect between the genotype and tobacco consumption.1-15-1...

Nat And Disease

The role of NATs in human disease has recently been extensively reviewed. 3 The following is a summation of findings to date. The association between acetylator status and the risk of various diseases has been extensively reported. Altered risk with either the slow or rapid phenotype has been observed for bladder, colon and breast cancer, systemic lupus erythematosis, diabetes, age-related cataracts, Gilbert's disease, Parkinson's disease, and Alzheimer's disease. These associations imply a...

Practical RCA formats

Generally, the RCA-based diagnostics can be classified into two groups some of them operate with the preformed circular probes Fig. 1D and E for the peptide nucleic acid (PNA)-assisted, nick-induced RCA and immuno-RCA, Fig. 1 Schematics of different RCA processes the arrowhead symbolizes the DNA polymerase. Given a small, < 100-nt size of DNA minicircles used in RCA and the strong rigidity of dsDNA fragments with these lengths, only part of the circular probe can be base-paired at any given...

Pathophysiologic And Genetic Features

The first step in the telangiectasia formation consists a focal dilatation of postcapillary venules with predominant stress fibers in pericytes along the luminal border. As the venules enlarge, they become convoluted and form excessive layers of smooth muscle cells without elastic fibers and connect directly with the dilated arterioles.1-1-1 The molecular basis consists of a defect of the TGF-p signaling pathway which, in normal conditions, plays an important role in vascular remodeling and...

Conclusion Genomics And Vaccination

Despite the advances associated with molecular techniques, meningococcal disease remains a globally significant health problem, and the prospect of comprehensive vaccination remains elusive. Whereas vaccines based on the capsular polysaccharide are either licensed or under development against meningococci associated with serogroups A, C, and Y, as well as W-135, attempts to develop a polysaccharide-based serogroup B vaccine have proven unsuccessful. This fact is especially relevant as much of...

Principles Of Rna Storage

Basically, RNA can be stored in purified form after its extraction or synthesis, or unprocessed samples containing the RNA of interest are stored. However, even in the latter case, proper care must be taken to avoid (re) contamination of RNA with RNases once it is extracted from the stored sample. Little information is available on the long-term stability of isolated and purified RNA during storage. Most studies have been carried out focusing on various storage conditions of unprocessed samples...

Principle Of The Taqman Assay

Like several eubacterial DNA polymerases, Thermus aquaticus (Taq) polymerase is a bifunctional enzyme consisting of a polymerization domain and a nuclease domain, and it is the latter that is crucial for the TaqMan assay. 1 In essence, the TaqMan assay is identical to a standard PCR reaction powered by Taq polymerase, except for the addition of the TaqMan probe, a nonextendable fluorescently labeled oligonucleotide hy bridizing on the target DNA between the forward and reverse primers 2-3 (Fig....

Sample Preparation for PFGE

Standard sample preparation procedures for DNA isolation involving mechanical forces are inappropriate for the analysis of fragile, high-molecular weight DNA molecules that easily break into small pieces. To prevent DNA damage, intact cells mixed with warmed, liquid agarose are pipetted into plug molds, 10 x 5 x 1.5 mm in size. Cells embedded in the agarose plugs are lysed in situ by detergents and enzymes. The agarose matrix keeps the large DNA molecules intact, while permitting diffusion of...

Kras

Associated malignancies and their penetrance Associated malignancies and their penetrance Gastric cancer (70 ) Lobular breast cancer in females (39 ) Colorectal cancer (70-85 ) Endometrial cancer (40-60 ) Ovarian cancer (12 ) Stomach, uroepithelial, hepatobiliary, brain, small bowel cancers Colorectal cancer Ovarian stromal tumors Uterine and cervical cancer Gastrointestinal tract polyposis and cancers, breast and testis, cancers Thyroid cancer Breast cancer Gastric cancer Leukemia Breast...

Principles Of Qfish

As stated above, Q-FISH requires the use of fluorescently labeled telomeric PNA probes. The most commonly used probe is Cy3-labeled (CCCTAA)3. Following hybridization of this probe with the metaphase chromosome preparations, high stringency washes are performed to eliminate background fluorescence that may interfere with the quantitation of telomere fluorescence. Separate images of metaphase chromosomes and telomeres are then acquired using appropriate image-acquisition software packages and...

Dominantly Inherited PD

Golbe et al. 7 described the first large kindred of autosomal dominant PD with autopsy verification in 1990 and later reported a clinical genetic analysis of 60 individuals in five generations. The kindred originated from Contursi, a hill town in the Salerno province of the southcentral region of Italy. There was severe neuronal loss and Lewy bodies in the substantia nigra, typical of PD. Mean age at onset was 46.5 years 15 years younger than that of PD in the community and the disease had a...

The Fish Technique

Fish Labelled Probes

The FISH technique is based on the ability of single-stranded DNA to anneal to complementary DNA. In the case of FISH, the target DNA is the nuclear DNA of interphase cells or the DNA of metaphase chromosomes that are affixed to a glass microscope slide. FISH can be accomplished with bone marrow or peripheral blood smears, or other fixed tissue, and modifications of FISH methodology have also permitted the analysis of nuclei extracted from paraffin-embedded tissues or frozen sections from solid...

Ar

Partial loss or complete skipping of exon 6 Procollagen-N-proteinase ADAMTS-2 gene Molecular screening for mutations in COL5A1 or COL5A2 Not available Biochemical analysis of type III collagen by SDS-PAGE Molecular screening for mutations in COL3A1 Increased ratio deoxypyridoline to pyridoline cross-links in urine by HPLC Measurement of lysyl hydroxylase-1 activity Molecular screening of PLOD1 gene Biochemical analysis of type I collagen by SDS-PAGE Detection of complete or partial skipping of...

Genotype To Phenotype Correlations

Atrium Proper

In 1930, Wolff, Parkinson, and White 1 described patients with electrocardiograms showing a short PR interval, a delta wave, and a wide QRS complex. It was postulated an accessory atrial ventricular pathway would explain the particular ECG and the recurrent tachycardias. Electro-physiological investigations, cardiac electrical mapping, surgical findings, anatomic studies, and results of ablation treatment have confirmed the presence of accessory conduction pathways (Fig. 1). The normal cardiac...

Snp Analysis In Coding And Noncoding Dna Regions

Within the chromosomal DNA, SNPs are highly abundant occurring on average 1 base in every 1000. 11,12 As the human genome is now known to be around 3 billion bases Fig. 2 Analysis of two SNP loci on the human mitochondrial genome. (Taken from research work by Dr. Barbara Llewellyn at the University of Strathclyde.) The top panel shows the results of hybridization of a segment of DNA mitochondrial DNA to oligonucleotide sequences on a silicone chip. The position on the chip where the DNA binds...

Consequences of Immune System Cells Stimulation by Superantigens

The excessive activation of an unusual high proportion of T lymphocytes and APCs in vivo and in vitro upon binding the SAg molecules triggers massive production of a wide array of inflammatory and other Th1 Th2 and APC-derived cytokines, by the stimulated cells. This process takes place through a complex of upregulated and downregulated immunological networks, activation signals, and the cooperation of adhesion molecules on target cells leading to a cascade of events including further release...

Herpes Simplex Resistance History

Acyclovir (ACV) is the gold standard for the treatment of HSV infections. This drug is a nucleoside analogue of guanosine that has to be phosphorylated three times. The first phosphorylation is achieved by the thymidine kinase (TK) encoded by the virus this step is important as it allows ACV to become active only in infected cells. The second and third phosphorylations are carried out by cellular thymidylate kinases. Acyclovir triphosphate is a competitive inhibitor of viral DNA polymerase and...

FISH for Aneuploidy Screening

Cytogenetic studies have shown that up to 60 of the early spontaneous abortions in humans are caused by numerical chromosomal abnormalities and that as many as 70 of all human preimplantation embryos may contain aneuploid cells. 5 This prompted the development of PGD for aneuploidy screening of oocytes and preimplantation embryos (PGD-AS). Fluorescent in situ hybridization is Fig. 2 Polar body biopsy. Removal of the first and second polar body. Fig. 2 Polar body biopsy. Removal of the first and...

Info

The TMA has been divided into four subsections to facilitate navigation during microscopy. 2. Taking a cylindrical sample from the tissue sample (donor) paraffin block. 3. Placing the cylindrical tissue sample in the premade hole in the recipient block. Commercially available or various kinds of homemade tissue-arraying systems are used for TMA manufacturing. Exact positioning of the tip of the tissue cylinder at the level of the recipient block surface is crucial...

Familial Adenomatous Polyposis Syndrome

Familial adenomatous polyposis syndrome is characterized by accelerated tumorigenesis from normal mucosa to polyps. It occurs in 1 8000 to 1 13,000 live births and shows no racial, gender, or geographical predilection. It has autosomal dominant inheritance with 100 pene-trance, and if left untreated, patients affected with FAP will inevitably develop colorectal cancer, typically by the age of 35-40 (mean age 39 years). 1 It accounts for about 1 of all colorectal cancers. Approximately 15 , 75 ,...

Heteroduplex Analysis Matrices

At its earliest inception, the sensitivity of HA was limited by the availability of suitable gel matrices. Slab-gel systems are limited to agarose and polyacrylamide. The improved MDE gel solution (Cambrex) is a polyacryl-amide-like matrix with a high sensitivity for DNA conformational changes. The gel's unique structure allows DNA separation on the basis of both size and conformation, thus increasing the probability of detecting sequence differences from as low as 15 achieved in standard...

X

Fig. 3 Microarray-CGH profile of BT474 cell line (test), reference normal kidney DNA (A), oligonucleotides as probe DNAs spotted on the glass slide moving average applied to the log2 ratio (B), BAC PAC PCR representations as probe DNAs spotted on the glass slide. Vertical bars indicate spacing between chromosomes. (View this art in color at www.dekker.com.) oncogene mapping to the amplified region at 20q13.2 in breast tumors. 26 This amplicon was also analyzed in stomach tumors by array-CGH, as...

Singlecell Fish Analysis

Preimplantation genetic diagnosis for the age-related aneuploidies is currently performed by FISH analysis using commercially available chromosome-specific probes (Abbott, Downers Groves, IL). It was first applied in 1991 for gender determination using DNA probes specific either for the X or Y chromosome. 9 Because testing for only one of the sex chromosomes could lead to misdiagnosis of gender because of a possible failure of hybridization, a dual FISH was introduced, involving the...

Procedures For Msms Screening

Specimen preparation for MS-MS screening requires an extraction and derivatization process, 3 which is performed using 96-well polypropylene microplates in routine analysis. 4-6 One 3.2-mm disk of dried blood spot is punched out into the individual wells of a microplate. To the disk in each well, a methanol solution containing known concentrations of stable isotope-labeled standards is added. The microplate is covered and the samples are mixed. The extract is transferred to a second microplate...

Outline Of The Method

The comet assay can be conducted on virtually any nucleated cell that can be brought into suspension. Thus, both cultured or freshly isolated cells from any organ can be used. Isolating cells from solid tissues usually involves time-consuming procedures, often implying the use of extracellular matrix-digesting enzymes, which can damage DNA. Instead, for the study of endogenous levels of DNA damage, it is necessary to use rapid isolation techniques to reduce the onset of artefacts, and, for some...

Myc Gene Amplification In Medulloblastoma

MYC has been the subject of several published studies attempting to identify biological prognostic markers in human MB. Most show that MYC gene amplification is uncommon in MB, with an incidence of 8 in primary tumors. 5,6 The incidence of MYC gene amplification in MB cell lines and xenografts may be as high as 67 , 7 suggesting that MYC gene amplification correlates with cell line establishment and tumorigenicity. MYC gene amplification has been suggested as an indicator of poor prognosis in...

Molecular Genetic Pathogenesis

For many years, RS was thought to be a result of a defect in the Muller cell that acted as a cellular scaffold within the retinal architecture. Recent studies on gene expression and immunolocalization of retinoschisin indicate that it is expressed in all major classes of adult retinal neural cells, with the possible exception of horizontal cells, and that there is no retinoschisin detected in Muller cells or processes, 22 although this is reported by others. 33,34 RS is abundantly expressed in...

Mitochondrial Dna And Forensic Science

Mitochondrial Gene Panel

The current situation in forensic science is that there are a number of autosomal STR loci that can be analyzed leading to possible links between a crime scene sample and that taken from a suspect. Mitochondrial DNA has been used in forensic science for human identification and Single Nucleotide Polymorphisms SNPS Single Nucleotide Polymorphisms SNPS Fig. 1 Example of minisequencing of 12 SNP sites on the human mitochondrial genome. Panel 1 shows a DNA sequence trace from four samples of...

Immunotherapy

Immune responses to infection are vital as evidenced by the fact that healthy, immunocompetent individuals resolve infection. An interesting correlation between administration of highly active antiretroviral therapy (HAART) and a decrease in cryptosporidiosis in AIDS patients has been observed. HAART does not directly affect the parasite but exerts its effects through enhancing the immune system of these patients. A picture is slowly emerging of the immune responses during infection. CD4+ T...

Future Of Personalized Medicine

Future medicine will be practiced in an environment in which molecular classification of diseases based on genomic analysis will replace the one based on symptoms. Molecular diagnostics to predict the results of therapy will be based on patient's genomic profile. There is still a considerable amount of work to be done before personalized medicine is established in clinical practice. Enables the monitoring of viral load and detection of resistance to treatment in personalized protocols for...

Extracellular Fetal Dna Clinical Applications

The discovery of cell-free fetal DNA quickly led to a clinical application, namely the determination of the fetal RhD genotype in pregnancies at risk for hemolytic disease of the fetus and newborn (HDN). 10 This is largely because of the fortunate constellation existing in Caucasians where the RhD gene is generally absent in RhD individuals. Therefore the analysis of the fetal RhD genotype in pregnancies with an RhD constellation (father RhD, mother RhD) is similar to the facile detection of Y...

Evidence Examination

Irrespective of possible chain of custody rules, examination of evidence starts with photographic and or drawn descriptions of the items received by the forensic biologist. In every photograph, an absolute scale must be visible (millimeters centimeters no pennies, no matches). Resolution should be > 2816 x 2112 pixels 4 MPixel to allow blowing up of the pictures. Flash light should be avoided because brighter parts of the objects will flash out (become white a digital artefact). Biological...

Duplications Of Proximal 15q

The region of chromosome 15q11-13 has long been known to be chromosomally unstable, being the site of deletions, duplications, triplications, translocations, inversions, and the breakpoints for supernumerary chromosomes. The resulting phenotype of such rearrangements is further complicated by the abundance of imprinted genes in the region. Thus, paternal deletions of the region result in Prader-Willi syndrome (PWS), whereas maternal duplications of the exact same region cause the...

Discussion

Fetal DNA has been known to be present in maternal blood since the work of Lo et al. 9 in 1997. They first showed that Y-chromosomal sequences could be amplified from DNA isolated from the plasma of pregnant women carrying a male fetus. Fetal DNA is present in maternal serum at 5 weeks' gestation 10 and the concentration of fetal DNA increases with gesta-tional age. Fetal DNA represents 3.4 (0.4-12 ) and 6.2 (2.3-11.5 ) of total plasma DNA in the first and third trimesters of pregnancy,...

Detection Of Homoplasmic And Heteroplasmic Mutations In Mitochondrial Genome

Energy metabolism disorders caused by mutations in mtDNA represents a group of heterogeneous diseases. The most frequently found mtDNA point mutations are A3243G for mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS), A8344G for myoclonus epilepsy, ragged red fibers (MERRF), T8993G C for neuropathy, ataxia, retinitis pigmentosa (NARP), and Leigh syndrome, and G11778A for Leber's hereditary optic neuropathy (LHON). 16,22 Molecular analysis of the known...

MagNA Pure Lc Instrument

Magnetism is the underlying principle of the automated nucleic acid isolation performed by the MagNA Pure system (Roche Diagnostics, Mannheim, Germany). The instrument has a completely closed housing as well as an automatic clot and tip loss detection. No filtration, centrifugation, or vacuum pumps are necessary, which minimizes the risk of cross-contamination. Up to 32 nucleic acid isolations (the eight-nozzle pipette head allows a variable number of samples from 1 to 32 per run) can be...

Genetic Testing And Counseling In Hereditary Breast And Ovarian Cancer Syndrome

Genetic testing for HBOC has become generally accepted as part of standard clinical practice and should be offered to any patient that has personal or family history features suggestive of HBOC (Table 3) and when genetic testing results could influence the medical management of that patient or the patient's family members. Clinical laboratories employ a variety of molecular techniques to detect germline mutations in BRCA1 and BRCA2 in a peripheral blood sample (Table 4). The sensitivity of...

Identification of E faecalis in GPCPCPositive Blood Culture Bottles

VRE have become a serious nosocomial pathogen because of the widespread use of vancomycin and require aggressive therapy with novel antibiotics, such as line-zolid and quinupristin dalfopristin. Resistance to vanco-mycin and also ampicillin among enterococci is primarily associated with Enterococcus faecium and a few other rare-occurring Enterococcus species, whereas E. faecalis, the most frequent species accounting for approximately two thirds of enterococcal isolates, is rarely resistant to...

Features Of Fraxa And Fraxe

Fmr1 Southern Blot

Clinical Phenotype, Genetics, and Biochemistry The fragile X syndrome affects males to a greater extent than females in males, learning and social deficits are moderate to severe, with social impairments usually preventing them from fathering offspring although sperm production is thought to be unaffected. In adult males, visible signs of the condition often include a large head, long face, large ears, and macroorchidism however, in children, the phenotype is likely to be restricted to...

Staphylococcal Enterotoxins

In addition to their nature as superantigens, SEs function as potent gastrointestinal toxins causing SFP, which has a major public health impact. 1,3-5 Superantigenicity and emesis activities of SEs have been shown to result from distinct regions of the toxin protein. SEs comprise a family of homologous but antigenically different exotox-ins, which are heat stable and resistant to inactivation by gastrointestinal proteases. SEs have also been implicated in several allergic and autoimmune...

Molecular Testing

With an increasing number of cases of IC but lack of a sensitive detection method, it has become essential to develop techniques that overcome traditional limitations. Molecular methods detecting nonculturable nonviable cells or circulating free fungal DNA have been developed, which may provide an alternative to culture-based techniques. Owing to these culture limitations, most diagnostic molecular mycology techniques have focused on detecting IC, with little work performed on detecting...

In Vivo Applications Liver Transduction

The liver is an attractive gene therapy target because the fenestrated endothelium permits exposure to intravenously delivered vector, hepatocytes are well suited for secretion of therapeutic proteins into the circulation for systemic delivery, and it is the affected organ in many genetic disorders. Helper-dependent adenoviral vectors are particularly attractive vectors for liver-directed gene therapy because of their ability to efficiency transduce hepatocytes following intravenous injection....

In Vitro Diagnostic Applications

The generic PNA-FISH assay procedure allows PNA-FISH to be applied for various blood pathogens without procedural changes, such that separate tests can be performed in parallel. Using the results from Gram staining, appropriate PNA-FISH tests can be selected to identify or rule out relevant pathogens (Fig. 2). Fig. 2 Application of PNA-FISH tests for direct identification of positive blood culture bottles. The result of the Gram stain provides a basis for the selection of appropriate PNA-FISH...

References

Human Leprosy. In Granulomatous Infections and Inflammations Cellular and Molecular Mechanisms Boros, D.L., Ed. ASM Press Washington, DC, 2003 207-676. 2. World Health Organization Leprosy Elimination Project Status Report 2003. http www.who.int lep Reports s20042.pdf (accessed March 2004). 3. Kerrie, T. Meisner, S. Siddiqui, M.R. Balakrishnan, K. Ghei, S. Golding, M. Sengupta, U. Pitchappan, R.M. Hill, A.V. Region of chromosome 20 is linked to leprosy susceptibility...