Leprosy

Leprosy (Hansen's Disease) is a chronic infectious disease caused by infection with Mycobacterium leprae affecting the skin, mucous membranes, and peripheral nerves. 1 The predilection of M. leprae for peripheral nerves can result in nerve damage leading to sensory and motor paralysis which in turn can cause deformities of the hands and feet and blindness. It is estimated that 2-3 million patients suffer from irreversible nerve damage as a long-term consequence of M. leprae infection and that...

Metabonomics Data Acquisition Methods

The two principal methods used comprise 1H nuclear magnetic resonance (NMR) spectroscopy and liquid-chromatography mass spectrometry (LC-MS). Nuclear magnetic resonance spectroscopy is nondestructive and provides detailed information on molecular structure, molecular dynamics, and mobility, especially in complex mixture analysis. Mass spectrometry is more sensitive than NMR spectroscopy and is used extensively for molecule identification, but in complex mixtures of very variable composition,...

The Neurological Continuum

Although type I is, by definition, nonneuronopathic, and the presence of the N370S allele obviates neurological involvement, nonetheless, recent data are suggestive of a phenotypical continuum that includes some neurological signs even in patients with type I. Among the earliest anecdotal reports were those of parkinsonism that is marked by early onset, aggressive progression, and refractoriness to conventional anti-Parkinson therapy in patients with very mild type I Gaucher disease. 6...

Dnabinding Cyanine Dyes

In recent years, unsymmetrical cyanine dyes have received considerable interest because of their excellent nucleic acid staining properties. In 1986, Lee et al. 3 showed that the unsymmetrical cyanine dye Thiazole Orange (TO Fig. 2) exhibits 3000-fold enhancement in fluorescence intensity upon binding to RNA. Later, Rye et al. 4 showed that TO, and the similar dye, Oxazole Yellow (YO) with a benzoxazolium moiety instead of benzothiazolium, have a dramatic increase in fluorescence also upon...

Therapeutical Approaches for HCM Are Symptom Oriented

P-Blockers act as negative inotropic drugs and are used as first-line drugs in patients with or without LV obstruction, and are the preferred treatment in symptomatic HCM patients with an LV outflow tract gradient only by exertion. 5 Verapamil, a calcium antagonist with negative inotropic properties, is used in both obstructive and nonobstructive forms in patients without severely disabling symptoms. Disopyramide, a class IA antiarrhyth-mic drug with negative inotropic effects, is reserved for...

Symptomatic Management

Whereas in the past, splenectomy was frequently performed for the management of severe thrombocytopenia and or for relief of mechanical compression by the greatly enlarged spleen, by removing the main reservoir for Gaucher cells, splenectomy may induce or aggravate liver and bone involvement. Today, splenectomy is rarely indicated. Total joint replacement is the most important orthopedic intervention, both in the past and in the present, for patients with avascular necrosis of the joints who...

Genetic Variation In Echinococcus

E. granulosus comprises a number of intraspecific variants or strains that exhibit considerable variation at the genetic level. 2-4 Conversely, there is very limited genetic variation within E. multilocularis 2,3 although recent microsatellite analysis has shown such markers can be used to examine E. multilocularis population structure.1-5-1 There are no available data to indicate that either Echinococcus vogeli or Echinococcus oligarthrus, the other recognized Echinococcus species, is...

Info

Fig. 3 Analysis of SYBR Green 1 dissociation curve. The first derivative view with respect to temperature provides a clear view of the rate of SYBR Green 1 loss and the temperature range over which this occurs. The example shows a view of data between 72 C and 90oC. The small peak at 74.5 C is probably due to primer dimer product formation as this is the only peak to occur in the NTC sample. The main peaks occur around 85.55 C although there are some with a distinctly different profile and a...

Differential Diagnosis

The diagnosis of Alagille syndrome is primarily based on the presence of at least three out of the five main features. The differential diagnosis of each feature of the syndrome will result in a very long and tedious list. Association of butterfly vertebra and cardiac defects has been reported in patients with deletions of 22q11. Atrial septal defect associated with eye anterior chamber anomaly may be observed in Turner syndrome. The main issue of differential diagnosis concerns diagnosis of...

Design and Production of AAV Vectors

Adeno-associated virus vector production was enabled by the molecular cloning of double-strand AAV DNA into bacterial plasmids followed by transfection into helper virus-infected mammalian cells. 1,6 This results in replication of the AAV genome free of any plasmid sequence to yield AAV particles. For AAV vector construction, the cis-acting ITR sequences must be retained and the unique sequence is replaced with foreign DNA. 1,6 There is a limit of about 5 kb of DNA that can be packaged in an...

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...

Diagnosis

The diagnosis of human brucellosis is conventionally performed using serological and bacteriological tests. The Rose Bengal test, 15 the serum agglutination test, and the complement fixation test are the most accepted serolog-ical tests used worldwide. These tests are based on a reaction between Brucella cell antigen and antibodies produced in response to the infection. The accuracy of the Rose Bengal test is over 99 , but it can give false positive results with sera from patients infected with...

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

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...

Molecular Characterization of C botulinum

Molecular characterization of C. botulinum may be used to study the genetic diversity of the organism, 4,5 and in tracing the causative agents of botulism outbreaks. 9,25 Pulsed field gel electrophoresis (PFGE) provides excellent discriminatory power and reproducibility, whereas the PCR-based method (randomly amplified polymorphic DNA assay, or RAPD) is less reproducible but can be quickly performed. The application of rRNA gene restriction pattern analysis (ribotyping) has been used to...

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...

Cm

0 2 4 6 8 10 12 14 16 18 log2 Signal Cy3 microarray experiments, thus omitting the need for RNA amplification. Currently, amplification is necessary for RNA samples smaller than 4 pg. Another way of dealing with a limited size of samples is downscaling the size of the microarrays. Subsequent to the initial diagnostic research that has been performed on large, whole genome arrays, often a ''signature'' set of a limited number of genes that is specific for predicting a disease condition can be...

Biological Relevance

Biopsies contain a range of different cell types a problem exacerbated in heterogeneous tumor samples that include normal and inflammatory cells as well as diversely evolved cell populations. In addition, normal cells adjacent to a tumor may be phenotypically normal but genotypically abnormal, or exhibit altered gene expression profiles because of their proximity to the tumor. 37 Hence expression profiling of such biopsies provides a composite of the whole population, and this may result in the...

Biology

H. ducreyi preferentially infects keratinized stratified squamous epithelium. Epidermal microabrasions formed during sexual intercourse or otherwise damaged skin is the portal of entry for infection. After entry into the skin, the bacteria stimulate target cells such as keratinocytes, fibroblasts, and endothelial cells to secrete IL-6 and IL-8. 1 Polymorphonuclear cells are recruited within 24 hr to the epidermis and dermis and form small micropustules. A dermal infiltrate of T cells,...

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....

Enteritis Necroticans Disease Summary

Enteritis necroticans (also known as necrotizing enteritis, Darmbrand, or Pigbel) caused by C. perfringens type C isolates is a rare but potentially lethal enteric disease of humans. This illness was first recognized after World War II in Darmbrand, Germany and later in Papua New Guinea. 20 The p -toxin produced by type C isolates is considered as the primary virulence factor based on the relative efficacy of a p-toxoid vaccine. 20 Although the incidence of enteritis necroticans is low, risk...

Fluoroquinolone Cellular Targets

Fluoroquinolones function by inhibiting two type 2 topoisomerases DNA gyrase and topoisomerase IV. 1-3 Type 2 topoisomerases cause staggered single-stranded breaks in both strands of the bacterial chromosome, pass an intact segment of the DNA strand through the break, and reseal the DNA. 1,3-5 Fluoroquinolones bind to the DNA-enzyme complex containing the broken DNA strands forming what is termed the cleavable complex. 3,5-7 DNA repair and replication are inhibited and the resulting release of...

Advantages

Colony PCR is a whole-cell-based assay, and thus will save a considerable amount of time, labor, and chemical reagents. The possibility of experimental errors or contamination is also reduced because DNA purification is not required. 3 A comparison study with two DNA extraction methods using mycobacterial isolates indicated that the heating method is more sensitive and no less specific than a conventional chemical method (phenolchloroform). The loss of DNA during purification and the presence...

Background

PCR is an enzyme-based in vitro method used to perform the following 1) to amplify specific DNA fragments for molecular diagnosis of heritable disorders 2) to screen for susceptibility to disease 3) to identify bacterial and viral pathogens or specific target genes 4) to biomonitor environmental changes and gene expression 5) to study the evolutionary process at the nucleotide level. Furthermore, this technique can be used to analyze recombinant vectors to determine both the presence and...

Epidemiology

The epidemiology of N. meningitidis has been reviewed by Cartwright. 1 Humans are the sole natural host for the meningococcus, which is carried asymptomatically as part of the commensal nasopharyngeal microflora in approximately 10 of the general population. The meningococ-cus may occasionally cause invasive disease by traversing the nasopharyngeal mucosa into the bloodstream, where it multiplies rapidly. Although invasive behavior is poorly understood and unpredictable in the individual, it...

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....

Molecular Characterization Of B Pertussis And B Parapertussis

Molecular characterization and typing of B. pertussis and B. parapertussis are used to obtain information on the changes in the phenotype and genotype of circulating isolates and in epidemiological studies. When polymorphisms in a number of B. pertussis genes encoding for virulence factors and surface-associated proteins as well as in housekeeping genes were studied, B. pertussis proved to be a rather conserved bacterium. Polymorphisms in recent isolates are restricted to genes encoding...

Viral Hemorrhagic Fevers

The term ''viral hemorrhagic fever'' (VHF) describes a variety of viral diseases which are characterized by fever and bleeding in humans. This syndrome is caused by RNA viruses belonging to the families Filoviridae (Ebola virus and Marburg virus), Arenaviridae (Lassa virus, Junin virus, Machupo virus, Guanarito virus, and Sabia virus), Bunyaviridae CCHF virus, Rift Valley fever (RVF) virus, and Hantaviruses , and Flaviviridae (yellow fever virus and dengue virus). After transmission from their...

Molecular Characterization

Methicillin resistance in S. aureus and other staphylococci is caused by the production of penicillin-binding protein 2a (PBP2a, also known as PBP2'), a novel 78-kilo Dalton penicillin-binding protein encoded by the mecA gene. 10 PBP2a, along with PBPs normally produced by S. aureus, are enzymes that catalyze peptidoglycan cross-linking reactions in the bacterial cell wall, resulting in a rigid structure that protects the bacterium against osmotic cell lysis and death. 10 However, PBP2a is...

Main Destructive Influences On

Under the influence of UV light (including sunlight) and acids, DNA contained in biological stains as well as extracted DNA breaks into pieces (degrades). Depending on the intensity of fragmentation, PCR might still be possible. Humidity does not directly affect DNA but will allow mold and bacteria to destroy the sample including the DNA within days. Frequent freezing and unfreezing of stains or extracted DNA will also lead to degradation. Household use of detergents and cleaners does not...

Lamivudineassociated Mutations In The Hbv S Gene

Four open reading frames, the S, C, X, and P genes, have been identified within the HBV genome. Because of the overlap of the P and S genes, lamivudine-associated mutations produce changes in the S gene, too. Consequently, lamivudine-resistant HBV isolates show alterations in the ''a'' determinant of the HBsAg protein. 16 Lamivudine-selected HBsAg protein changes may have important virological and clinical implications They may escape neutralization by vaccine-induced anti-HBs antibodies and...

Molecular Detection of C botulinum

PCR and DNA probes provide a sensitive tool for the detection of C. botulinum. The greatest sensitivities of PCR protocols for C. botulinum in various sample materials vary from 2.5 pg of DNA 20 to 0.1 cfu (or spore) g food 21 and 10 cfu g feces. 21,22 A multiplex PCR assay that enables the simultaneous detection of all human pathogenic serotypes A, B, E, and F has provided a marked improvement in the PCR detection of C. botulinum. 21 Nested PCR involves several subsequent amplifications and...

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...

Introduction

Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant, inherited disorder characterized by progressive muscle weakness, including atrophy of the facial muscles, upper arm, and shoulder girdle. Disease progression may eventually involve weakness and atrophy of the pelvic girdle and the foot extensor. FSHD is the third most common form of inherited neuromuscular disorder, after Duchenne and myotonic muscular dystrophies.1-1-1 It is unusual for the disease to become clinically...

References

Sillekens, P. Characteristics and applications of nucleic acid sequence-based amplification (NASBA). Mol. Biotechnol. 2002, 20, 163-179. 2. Loens, K. Ursi, D. Ieven, M. van Aarle, P. Sillekens, P. Oudshoorn, P. Goossens, H. Detection of Mycoplasma pneumoniae in spiked clinical samples by Nucleic Acid Sequence-Based Amplification. J. Clin. Microbiol. 2002, 40, 1339-1345. 3. van Gemen, B. Van Beuningen, R. Nabbe, A. van Strijp, D. Jurriaans, S. Lens, P. Kievits, T. A one...

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...

Conclusions

The conundrum facing research into the potential use of circulatory fetal cells for prenatal diagnostic purposes is whether it will be possible to develop a test that is rapid, reliable, and cheap enough to compete with the current high levels of sensitivity (> 90 ) and very low false-positive rate (< 1 ) for the detection of trisomy 21 fetuses, obtained by a combination of different ultrasound markers (nuchal translucency and nasal bone) and serum analytes. 23 However, it is likely that...

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....

Fetal Cells Diagnosis Or Screening

The scarcity of fetal cells in the maternal circulation and the failure to reliably retrieve these cells from maternal blood samples are amply underscored by the sobering results of the NIFTY Study. As even more disappointing results were obtained in the study initiated by Applied Imaging, which used state-of-the-art technology for the automated detection of putative fetal erythroblasts, 5 questions have been raised as to whether the pursuit of fetal cells can have any future diagnostic value....

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...

Genotypephenotype Correlation

No correlations are found between a mutation and a clinical phenotype because a family unique presentation such as migraine or stroke is also found with several other mutations. 2,3 Additionally, we did not notice neither different clinical presentations nor different skin vessel wall lesions in CADASIL patients with mutations located either in the first or very last exons. Interestingly, a homozygous patient followed in Finland was indistinguishable when compared with an age-matched...

Classificationtype Of Pathogen

The Epstein-Barr virus (EBV) belongs to the family of herpesviruses.1-1-1 With respect to morphology, EBV is hardly distinguishable from other family members. Based on pathogenesis, characteristics of replication, and host cell tropism, herpesviruses can be classified into three subfamilies alpha-, beta-, and gamma-herpesvirus. EBV belongs to the genus Lymphocrypto-virus of gamma-herpesviruses, which are characterized by a narrow host (cell) range and a slow rate of replication in cell culture....

Ethical And Social Aspects Of Genetic Profiling

Genetic profiling for pharmacogenetic purposes is less likely to raise objections than genetic testing for primary disease risk assessment. Nevertheless, ethical issues concerning patient confidentiality, possible misuse of geno-typing data, and possible harm to patient's social and insurability status need to be resolved before the implementation of pharmacogenetics in healthcare. Holding sensitive information on someone's genetic makeup raises questions of privacy and security and ethical...

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...

Molecular Genetics

Studies focusing on large families with the rare, early-onset, autosomal dominant form of the disease led to the discovery that mutations in the amyloid precursor protein (APP) 9 Presenilin 1 (PSEN1), 10,11 and Presenilin 2 (PSEN2) 12 genes cause AD. Although these autosomal dominant families represent only a small percentage of the total AD cases, functional studies of the effects of the identified mutations have become an important way to dissect the causes and underlying disease mechanisms...

Template DNA Preparation

Isolates should be pure and phenotypically identified as S. aureus prior to testing. Control strains of MRSA (ATCC 33591), MSSA (ATCC 25923), and mecA-negative Staphylococcus epidermidis (ATCC 14990), in addition to a no-template (water) control, should be included with each run. Prepare a 1.0 McFarland standard suspension of the organism in 0.85 NaCl. Extract DNA using either commercial nucleic acid extraction kits or Chelex-100 resin. For the latter, Chelex-100 is added to 800 pL of bacterial...

Clinical Significance Natural Habitat and Reservoir

S. aureus is widely distributed in the environment, although it may also be found as part of the normal bacterial flora of humans or other mammals. Strains may colonize various anatomic sites including the skin, mucous membranes, and the upper respiratory, gastrointestinal, and genitourinary tracts. 24 Nevertheless, S. aureus has a particular predilection for colonizing the anterior nares of humans. 15,24 A constellation of host factors is believed to play a role in the ability of an individual...

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...

Polymerase Chain Reaction Primer Design

The task of designing PCR primers 2 starts from the identification of the target sequence (amplicon) of interest. It is assumed here that the target sequence is known from previous work. The region near each end of the target is then analyzed for promising primer sequences. Variables at this stage include the starting location and primer length. The base composition and particular sequence of each potential primer impact the ability of the primer to perform well through their influence on...

Resistance Induced By Ribosomal Mutations

Macrolide antibiotics inhibit protein synthesis through their interactions with the 50S subunit of the bacterial ribosome by binding near the entrance of the peptide passage channel. The interactions have mainly been mapped by chemical footprinting experiments and X-ray crystallography1-1-1 to the peptidyl transferase region within domain V of 23S rRNA. Positions A2058 and A2059 (Escherichia coli numbering) appeared to be essential for macrolide binding. Interaction with domain II of 23S rRNA...

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...

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...

Diagnostics Of Botulism And C Botulinum

The diagnosis of botulism is primarily based on the history of eating suspected foods, as well as detecting BoNT in patients and in suspected food samples. 10 The detection of C. botulinum cells in clinical and food specimens strongly supports the diagnosis. Electroneur-omyography may be used to distinguish botulism from similar neurological diseases. 10 The culture method for the detection and isolation of C. botulinum includes broth culturing in anaerobic media, such as...

Description Of The Infection Spectrum

Whooping cough is acquired by inhalation of aerosols containing B. pertussis. The infection is characterized by bacterial colonization of ciliated respiratory epithelia in the trachea and bronchi. The incubation period is 6-20 days. Classical pertussis is an illness of three stages. It starts with a catarrhal stage, with nonspecific symptoms similar to those of the common cold (duration 1 to 2 weeks). At the paroxysmal stage (duration 2 to 6 weeks), the cough becomes more prominent with...

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...

Spectrum Somatic Mutations in Retinoblastomas and Other Tumors

It is possible that all retinoblastomas have mutations in both alleles of the RB1 gene. RB1 gene mutations have also been identified in several other tumor entities including osteogenic sarcoma and small cell lung cancer. The spectrum of somatic mutations comprises that of germ-line mutations with two important additions More than 60 of retinoblastoma show loss of constitutional heterozygosity (LOH) at polymorphic loci located on chromosome 13. In these tumors, one RB1 allele is lost in...

Pulmonary Disease

Predisposing factors 6 for the development of pulmonary disease include alcoholism, bronchiectasis, cyanotic heart disease, cystic fibrosis, prior mycobacterial disease, pulmonary fibrosis, smoking, and chronic obstructive lung disease. Radiological features include thin-walled cavities, parenchymal infiltrates, and pleural involvement. High-resolution computed tomography of the chest shows a combination of multifocal bronchiectasis and nodular parenchymal disease. 7

Management

So far, there is no causal treatment. Physiotherapy is recommended as soon as weakness or myotonia occurs. Orthoses may be helpful to stabilize knees or ankles. Treatment with nonsteroidal antiinflammatory drugs, carbamazepine, or corticosteroids ameliorates muscle pain in single DM1 patients. If myotonia predominates in DM1, mexiletine, phenytoin, or carbamazepine may be beneficial, but no data supporting its effectiveness in DM2 are available. For muscle cramps, quinine sulfate may be tried....

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...

Reproducibility Precision

Zhang and colleagues, 3 working at Cetus, reasoned that probes bound by UV or heat, were attached in random fashion to the filter and sometimes to each other, making it difficult to determine consistent or optimal hybridization conditions. Until then, Saiki and his collaborators1-4-9-1 had been using nucleotidyl transferase-tailed polyT oligonucleotides under UV radiation to conjugate amino-derivatized membranes (Biodyne B) but Zhang et al. showed that oligonucleotides could be...

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...

Botulinum

Classification and the Microbial Ecology of C. botulinum C. botulinum is an anaerobic gram-positive rod-shaped bacterium that forms heat-resistant spores. 1 The taxo-nomic denominator is the production of botulinum neurotoxin (BoNT). Based on the serological properties of the BoNT they produce, C. botulinum strains are divided into types A-F, with types A, B, E, and F being human pathogens, and types C and D being animal pathogens. Based on their metabolic properties, C. botulinum strains are...

Adenoassociated Virus

Adeno-associated virus is a small, DNA-containing parvovirus that has been isolated from humans and other animal species. 1-4 Several serotypes of AAV have been isolated from humans and nonhuman primates. 1,4 Adeno-associated virus is defective and replicates only in host cell nuclei when certain functions are provided by a coinfecting helper adenovirus (Ad) or herpesvirus.1-1,2-1 The mechanism of the helper function is not clearly defined but only a limited set of adenovirus genes, E1, E2A,...

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...

Clinical Description

In symptomatic individuals, certain clinical manifestations are common. In women, menorrhagia is frequently reported and there is an increased risk of bleeding after childbirth. 9 Epistaxis may occur, whereas petechiae and bruising are rather unusual. 9 Bleeding can be excessive and frequently manifests postoperatively, in particular after surgical procedures involving tissues with high fibrinolytic activity such as dental extractions, tonsillec-tomies, prostatectomies, or urinary tract...

Comparison Of Viral Load Assays

A number of studies have compared the performance of these assays in clinical situations. Schuurman et al. 4 showed that interlaboratory reproducibility of the Roche, Organon, and Versant assays was very good. Others have shown that each of the assays has relatively low intra- and interassay variability.1-5-1 The biological variation of HIV-1 RNA in plasma in clinically stable individuals is approximately threefold (0.5 log) and this variation was not associated with diurnal fluctuations.1-6-1...

Complications of Primary Infection

In rare cases, mostly in children, EBV may cause fatal IM or infection takes a severe chronic active course (SCAEBV). 4- Affected individuals develop life-threatening complications suffering from extensive lymphadenop-athy, hepatosplenomegaly, pancytopenia, polyclonal gammopathy, as well as B cell or T cell lymphoprolifer-ative diseases. EBV-specific antibody titers (anti-VCA and anti-EA) are extremely high, indicating uncontrolled systemic viral replication. Another milder form of chronic...

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...

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...

Genomics And Proteomics

The amount of DNA sequence data available and the methods used to exploit it have grown exponentially in recent years. The aim of this work is to understand how the various components of an organism interact to function properly, not just to catalogue a list of genes and how they function. Genomics describes the study of the genetic makeup of an organism, whereas proteomics is the study of the protein composition of the cell. There are more proteins in an organism than there are genes because...

Pathophysiology

Mutations in the ABCC6 gene have recently been found to underlie PXE in nearly all affected individuals.1-2-4,7-9-1 ABCC6 is located on chromosome 16p13.1, and its 31 exons are dispersed over 73 kb of genomic sequence. 2 Its translated region spans 4.5 kb and encodes multidrug resistance-associated protein 6, MRP6, 10,11 an ATP-dependent efflux pump that is expressed at the basolateral membranes of hepatocytes and the proximal tubules in the kidneys. 12 The protein is composed of three...

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...

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...

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...

Clinical Manifestations

Trophozoites of these parasites are located in the duodenum, jejunum, and upper ileum. When symptoms occur, they vary from mild to severe abdominal discomfort, diarrhea, cramping, and bloating. 1,2 Infants may have anorexia, weight loss, or a malabsorption syndrome that resembles sprue. 3,4 When a child is evaluated because of failure to thrive or is immunocompromised, the presence of Giardia should be considered. 5 Lactose intolerance may develop in these children and persist after elimination...

Genes And Parkinsonism

Parkinsonism is a syndrome of rest tremor, rigidity, bradykinesia, loss of postural reflexes, and the freezing phenomenon.1-1-1 Most often, it is a manifestation of Parkinson's disease (PD), which is characterized neuro-pathologically by brainstem Lewy bodies. Recent epide-miological and molecular data suggest that this is not one disorder, but is comprised of at least several disorders with genetic, environmental, or mixed etiologies. Genes play a complex role in PD. Most twin studies, which...

What Do Genes In Autosomal Dominant Pd Tell Us About Pathogenesis

The cause and pathogenesis of PD are incompletely understood, but rare mutations in familial cases (Table 1) are beginning to unravel the variety of cellular processes involved in the accumulation of Lewy bodies and dopaminergic cell death. It is not possible to discuss this fully without reference to the genetic discoveries in autosomal recessive parkinsonism, which have occurred in parallel with the finding of dominant genes and are outside the scope of this article. The subject has been...

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,...

Psychiatric Disorders

Upon treatment with antipsychotic drugs, adverse reactions are more likely to be seen in PMs for CYP2D6. In addition, the costs for treatment of patients are higher and estimates have been made that costs 4000-6000 more per year to treat patients of the variant UM and PM phenotypes. 9 Parkinsonism-like side effects are Relative importance of polymorphisms in human P450s involved in drug metabolism

Substrate Specificity

NAT1 and NAT2 have different but overlapping substrate specificities, although no substrate is exclusively acetylat-ed by one isozyme or the other, and no clear structural motif that determines substrate specificity for the different isoforms has been identified. Substrates preferentially N-acetylated by human NAT1 are p-aminobenzoic acid, p-aminosalicylic acid, sulfamethoxazole, and the folate catabolite p-aminobenzoyl glutamate. Substrates primarily N-acetylated by human NAT2 include a number...

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....

Immunotherapy of EBVAssociated Malignancies

EBV-associated monoclonal tumors in otherwise immu-nocompetent individuals are known to be mostly invisible from an immunological point of view. In comparison, lymphoproliferative diseases after hematopoetic stem cell transplantation or solid-organ transplantation usually arise as polyclonal proliferations of EBV-infected immortalized cells that are subject to immune control. Immediate partial reconstitution of immune surveillance is the first action to take. Without treatment, benign...

Localized Skin and Soft Tissue Infection

Most species of NTM have been described causing cutaneous disease 9,10 (Table 2) or nosocomial skin and soft tissue infection, postinjection abscess, surgical wound infections, and infections because of long-term intravenous or peritoneal catheters. M. chelonae causes disseminated nodular skin disease in immunocompromised patients. M. marinum causing ''swimming pool granuloma'' or ''fish tank granuloma'' gets implanted while cleaning fish tanks or handling fish. Solitary papules get Table 1...

Management of Thalassemia Major

Patients affected by thalassemia major are treated with regular blood transfusions and iron chelation therapy with desferrioxamine B (DFO). Life expectancy with this treatment extends to the third decade. The alternative oral iron chelator deferiprone (L1) is indicated only in patients with proven allergy or toxicity from DFO. Alternative chelation strategies and drugs, including the combination of deferiprone and DFO or ICL670 alone, are under investigation. Bone marrow transplantation (BMT)...

Treatment

Treatment of BCCs includes surgical excision, curettage and electrodesiccation, cryosurgery, Mohs' micrographic surgery, and ultrapulsed CO2 laser treatment. These methods, although effective, have economic, physical, and psychological implications, primarily because of the risk of scarring and recurrence. New, more inclusive selection criteria for surgery, combined with more effective neo adjuvant chemotherapy based on 5-fluoruracil and topically applied 5 imiquimod, are proving useful in...

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....

Solid Phase Hybridization Techniques

Peptide nucleic acids can be used in many of the same hybridization applications as natural or synthetic DNA probes but with the added advantages of tighter binding and higher specificity. This leads to faster and easier procedures in most standard hybridization techniques, such as Southern and Northern blotting. An alternative to Southern analysis is also the PNA pregel hybridization process, which significantly simplifies the procedure of Southern hybridization. 15 Labeled PNAs are then used...

Restriction Fragment Length Polymorphism

Fig. 1 Schematic representation of HLA class I and class II loci in the MHC that comprises > 200 genes on the short arm of chromosome 6. The corresponding number of antigens (as defined by serology) and alleles (as defined by nucleotide sequence) is indicated for each locus. (From Ref. 1 .) About 10 of the alleles assigned by the Nomenclature Committee are characterized by silent substitutions. Three HLA class I genes (A, B, and C) located at the telomeric end of the MHC encode, respectively,...

Genotype To Phenotype Correlations

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...

Applications in Serotyping Genotyping and Subgrouping Viruses

Multiplex PCR has been shown to be a powerful and cost-effective tool for typing and subtyping virus strains in different epidemiological studies. Fig. 2 gB Genotyping of clinical strains of CMV. From left to right lanes gB2 (613 bp), gB 1 (420 bp), gB3 (190 bp), gB4 (465 bp), gB1 and gB3, gB1, molecular weights in base pairs. (View this art in color at www.dekker.com.) Fig. 2 gB Genotyping of clinical strains of CMV. From left to right lanes gB2 (613 bp), gB 1 (420 bp), gB3 (190 bp), gB4 (465...

Melanoma Pancreatic Cancer Syndrome

Melanoma susceptibility has also been reported in conjunction with gastrointestinal cancer and breast cancer, and an excess of pancreatic carcinoma is frequently being observed. Comparison of CDKN2A mutation-positive and -negative melanoma families showed an increased risk of pancreatic cancer in the mutation-positive group. 11 Interestingly, the increased risk for pancreatic cancer seems to be restricted to only some of the CDKN2A mutation-positive families, implying other genes and or...

Handheld Nucleic Acid Analyzers For Clinical

Simultaneously with DoD-sponsored research, a number of private companies have pursued the development and fabrication of handheld nucleic acid analyzers with an eye toward capturing what is expected to be a very lucrative and burgeoning market in ''point-of-care'' (POC) diagnostics. Convincing physicians and clinical laboratory directors that POC nucleic acid analyzers, much less molecular methods per se, are worth embracing has traditionally been difficult. 12 Sample processing, particularly...

Sequence And Positioning Of Taq Man PROBE

Many of the rules that apply to PCR primer design also apply to TaqMan probe design (see Primer Probe Design chapter in EDGP). The sequence of the TaqMan probe should have GC contents within 40-60 preferably. Long sequences of identical nucleotides should be avoided in the probe. The sequence should be devoid of self-complementarity, or complementarity with the forward and reverse primers. The TaqMan probe should anneal as close as possible to the forward primer without overlapping. The size of...

Threat Agent Detection Impetus For Handheld Nucleic Acid Analyzer Development

Successful performance of PCR obviously depends on the ability of the thermal cycling platform to maintain accurate gradations of temperature in the manifold or block containing the reaction tubes. The heating and cooling apparatus necessary to provide these temperature changes places constraints on the size and weight of the thermal cycler. ''First-generation'' thermal cyclers, such as the Perkin Elmer 480 model instrument, while reasonably light in weight and small in footprint, were not...

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 ,...

G390v F418s

Fig. 1 Mutations found in the serum cholinesterase gene. (Mutations F28I, N96Y, 317-318insA, R386C, R424X, and E460K come from Ref. 7 . All of the others come from Ref. 2 .) serum cholinesterase is 2.2 and low FN or DN FN is 1.1 in these peoples. 1 These values show that polymorphism is very common in normal populations. Distinct ethnic differences in genetic mutations of the human BChE gene have been reported. The P100S, L330I, G365R, and R515C were only found in Japanese, whereas A (G70D),...

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...

Other Mutations In The 12S rRna Gene

Deletion of a T at position 961 and insertion of varying numbers of G were observed in a patient during a search in the mtDNA in 35 sporadic Chinese cases with AID. 30 The same mutation was not detected in 799 control samples. 30 This mutation was later found in an Italian family, in which five maternal relatives became deaf after aminoglycoside exposure. 31 However, in a recent study, 961delTinsC(n) was detected in 7 of 1173 anonymized blood samples in the United States. 24 This result...

Clinical Diagnostic Criteria for Cowden Syndrome

The International CS consortium has developed diagnostic criteria for this condition. 3,11 The criterion is less sensitive in children and young adults as findings appear generally in the third decade. Studies of patients who only partially meet these criteria, 'CS-like', or who have multiple primary cancers, have yielded very few germline PTEN mutations, thus providing indirect evidence of the usefulness of these criteria. 12,13 Clinical Management of Cowden Syndrome Clinical management of a...

Gsd Type Xi Hepatic Glycogenoses With Renal Fanconibickel Syndrome

This rare GSD is caused by defects in the facilitative glucose transporter 2 (GLUT-2), which transports glucose in and out of hepatocytes, pancreatic cells, and the basolateral membranes of intestinal and renal epithelial cells. 30 The disease is characterized by proximal renal tubular dysfunction, impaired glucose and galactose utilization, and accumulation of glycogen in liver and kidney. The affected child presents in the first year of life with failure to thrive, rickets, hepato- and...