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Co-Director, Eastern Virginia Medical School

Of those 305 infants, 287 had no information reported, 17 were reported as missed or not completed, and one provided unclear information. Review of demographic information (such as birth hospital, home town, and ethnicity) did not reveal commonality between the 5 cases, except that all R75X Inconclusive cases were female. Due to the sheer number of result codes, a pre-test phase was initiated to include test messages for a subset of the result codes to test configuration. Typically this meant that interworking and interconnection of equipment from multiple vendors was met with challenges of compatibility and often required some form of protocol conversion or separate data manipulation process. Advances in computer networking, cloud based software services and distributed computing allows a new regime. The software architecture opens up choices for a wider range of vendor equipment with a flexible and modular approach, and provides numerous benefits including better management of software updates and system expansion. Improved efficiencies also result because vendors need only support a library interface rather than complete software functionality. A system is demonstrated using existing laboratory punchers whereby the distributed software system allows independent configuration and operation of punching from separate workstations and provides a common punching platform for all punchers in the laboratory. The demonstrated system is based on storing the configuration and assay parameters for all punchers on a server. The Punching module then uses the server settings for that Puncher and Assay to correctly drive the puncher and punching. The settings also define the number of dots to be punched based on ample Type, Test and Specimen type and the size of punch to be used. It also has the advantage of storing the Plate details and information in the database as opposed to a series of text based files. In addition, challenge the newborn screening program community to consider our enhanced obligation and commitment to meeting the specific needs of the sickle cell disease community. Background: Wide variation exists in communicating trait results found through newborn screening. Studies have demonstrated significant issues, including pain management in emergency departments and risk for maternal death during pregnancy. We continue to measure the four defined outcomes for a complete notification: 1) infant is seen in clinic, 2) newborn screening report is provided to the clinic directly, 3) a discussion about the result occurs between the provider and the parents, and 4) the recommended follow-up testing is completed. In addition to measuring these outcomes, initiatives to address further needs for the sickle cell community were observed and identified. We changed the punch and assay plates to a Corning Polypropylene to reduce static during the punching step. Using the Zephyr, the extract is quenched and transferred to a deep well plate before performing a liquid-liquid extraction to remove proteins, salts, and uncleaved substrates. Numerous interventions were required to identify and repair intermittent poor performance. Routine screening and outcome data will be analyzed on an ongoing basis in order to optimize the cut-offs. Results: Sensitivity, Specificity, Precision, Accuracy, Reference Range and Reportable Range were validated. The first two months of screening (n=7951) found 5 borderline samples, 4 of which resolved with a normal repeat specimen, and 2 presumptive positive samples, of which both have been clinically confirmed. We intend to eliminate the borderline cut-off pending additional screening and outcome data. Background: Upon the notification of an abnormal newborn screening result, providers were faxed both a provider and family fact sheet about the condition. Additionally, we were making assumptions regarding what information they would find useful. We sought to better understand provider usage of the fact sheets and the content they found important, with the goal of revising fact sheets to better serve the needs of our providers. Providers who had a patient with a positive newborn screening result in the past six months (based on newborn screening follow-up data) were invited to participate in the survey. Provider emails were obtained from the Minnesota Board of Medical Practice and Board of Nursing. Results and Conclusion: the survey was emailed to 103 providers and 20 completed the survey. The majority recalled receiving both the provider (15) and family (19) fact sheet.

Hence, it is essential that users immerse themselves in basic knowledgeaboutstainingmaterialsandmethodssotheycancontrol thequalityofresults. Component Mix in order at room temperature 1 2 3 4 5 6 Distilled water Ethylene glycol Hematoxylin, anhydrous Sodium iodate Aluminum sulfate Glacial acetic acid No. H&e is used universally on sectioned materials, most often in histopathologybutalsoincytopathology. While the theoretical resolution fortransmittedlightaspredictedbyabbehasbeenachievedfor over100years,recentdevelopmentshavepermittedresolutionof structureswellbelowthesetransmittedlightresolutions. For optimum results when examining stained specimens, the microscopemustbeusedinapropermanner. When assessing H&e-stained sections from differing organs, Hematoxylinandeosin(H&e)isthe"breadandbutter"staininmost histopathologylaboratories. Pituitary the pituitary contains three principle cell types that should be apparentwithagoodH&e:acidophils(40%)withpinkcytoplasm, basophils(10%)withapalebluecytoplasmandchromophobes(50%) withclearcytoplasm. Heart theheartisrichinbloodvesselsaswellascardiacmuscleand collagen enabling good eosin differentiation. Bone Bone, apart from the rigors of fixation and processing, is also subjectedtoacidicdecalcifyingsolutions. Prostate the stroma of the prostate contains both collagen and smooth muscleandtheseshouldbeeasilydifferentiatedonanH&e. Many of the above recommendations are based on personal experienceandassuchshouldbeopenfordebate. Unfortunately,despiterapidpenetrationoftissues,thechemical reactions of formaldehyde with tissue proteins, especially the formationofmethylenebridges,occurslowly. Fixationprofoundlyaffectshistologicalandimmunohistochemical staining, technicians, pathologists and research workers must thereforedecideonthemostappropriatemethod. Formalin also contains about 10% methanol, which is added bythemanufacturertoretardtheformationofhigherpolymers, which eventually fall out of solution as paraformaldehyde. A paraffin section of Mercuric Chloride eachfixativepreservesmorphologydifferently,thustherearemultiple options. Zincsulfatehasbeen usedformorethanacenturyinastringentlotionsandeyedrops, which reduce the swelling of inflamed surfaces by coagulating extravasatedplasmaproteins. Smears Histotechnicans sometimes perform special stains on cytology smears,bloodfilmsandcytoprepsfromotherdepartmentswithinthe laboratory. One critical aspect of using microwave techniques is ensuring implementation of microwave processing technology, partly for staffingreasons. Specimen Processing staining quality can be depreciated by inadequate fixation and similarlybypoortissueprocessing. Process Solution Time Retort a) Conventional, room temperature processing schedule, 3mm biopsy. Fixation Fixation Fixation Formalin, 10% Alcoholic formalin Alcoholic formalin Alcohol, 95% Alcohol, 95% Alcohol, absolute Alcohol, absolute Xylene Xylene Paraffin Paraffin Paraffin 120 min 60 min 60 min 60 min 45 min 45 min 60 min 60 min 60 min 30 min 60 min 90 min -vacuum -vacuum -vacuum -vacuum Tissue Processing inordertoprepareatissueforembedding,itmustbeinfiltrated with paraffin. Because water and paraffin are not miscible, the specimensmustbegraduallydehydratedtoachievereplacement ofwaterwithalcoholbeforetheclearingagentisintroduced. Dehydrate Dehydrate Dehydrate Dehydrate Clearing agent Clearing agent Infiltrate Infiltrate Infiltrate 148 pecialstainsandH&e s specialstainsandH&e 149 Fixation and Tissue Processing Fixation and Tissue Processing Table 2. Embedding and Microtomy Oncethetissuehasbeenprocesseditisreadytobeorientated Solution Time Retort after sectioning, the tissue slide is drained and may be gently heatedtoevaporatethelayerofwaterbetweenthesectionsandthe glass. Dehydrate Dehydrate Dehydrate Dehydrate Dehydrate Clearing agent Clearing agent Infiltrate Infiltrate Infiltrate Alcohol, 65% Alcohol, 95% Alcohol, 95% Alcohol, absolute Alcohol, absolute Xylene Xylene Paraffin Paraffin Paraffin 15 min 15 min 15 min 15 min 15 min 15 min 15 min 15 min 15 min 15 min c) Microwave processing schedule, 1mm biopsy. Horobin, PhD Technical Considerations there are some technical tips that can apply to all preparative proceduresinmicrotechnique. Update on the immunocytochemical identification of lymphocytes in tissue sections.

Drosophila has proved to be a useful model in drug-feeding experiments to evaluate new antiepileptic drugs (Ackermann et al. In spite of the drug delivery challenge, a number of studies have used Drosophila in toxicological evaluations (Ackermann et al. Endocrine-active chemicals are present in the environment and some therapeutic agents have endocrine effects; in both cases, these compounds disrupt normal endocrine function in mammals. Changes in sperm counts, breast cancer, congenital abnormalities of the genitalia, and other pathologic conditions in humans are linked to these chemicals and therapeutic agents. Invertebrates can be used to study endocrine effects of drugs and environmental contaminants (Avanesian et al. For example, researchers have observed adverse reproductive effects in Drosophila adults and cell lines exposed to a variety of insecticides (Gupta et al. Similarly, Avanesian and colleagues (2009) studied methotrexate toxicity in flies and found ovarian impairment comparable to that observed in mammalian models. The freshwater mud snail (Potamopyrgus antipodarum) and Daphnia have also served as models for the assessment of environmental chemicals (Duft et al. Many gender-related differences in toxicant 138 effects have been noted in humans, and these effects can be modeled in many different invertebrates including insects, nematodes, crustaceans, molluscs, corals, and echinoderms (McClellan-Green et al. Environmental impacts on the lysosomal-autophagic system have been studied in bivalve molluscs (Moore et al. Additionally, many aquatic species serve as biomarkers for organic xenobiotic and metal contamination in both fresh and saltwater (Rainbow 2007; Raisuddin et al. Similarities in the genes involved in development of the hearing structures of flies and mammals have led to speculation that study of fly hearing may provide insights into deafness in humans (Boekhoff-Falk 2005). The creation of mechanical models has enabled modeling of auditory transducer dynamics, which will be used to test auditory performance in both vertebrates and Drosophila (Nadrowski and Gopfert 2009). Olfaction There are many similarities-structural, functional, and physiologic-between the olfactory systems of vertebrates and insects (Kay and Stopfer 2006), making insects excellent models for the study of olfaction. Investigators have studied insect olfactory systems to understand interactions between animals and their environment (de Bruyne and Baker 2008). Other studies have focused on odor processing and the capacity to detect odor blends using moths and honeybees (Lei and Vickers 2008), and C. In many decapod crustaceans, neurogenesis occurs in the olfactory system throughout life, so these animals are being used to study lifelong neurogenesis, with the aim of understanding how it occurs (Sandeman and Sandeman 2003; Schmidt 2007). This research may elucidate methods to stimulate neurogenesis in humans to treat neurological diseases or injuries. All organisms are exposed to an enormous diversity of chemicals in the environment, and understanding the way the nervous system recognizes and responds to these chemical signals is challenging. The chemosensory system of insects has become an important area of research, and Drosophila is the primary model under study (Benton 2008). Crustaceans are also effective models for the study of chemoreception, which is based in their olfactory system (Derby and Sorensen 2008). Taste and Satiation Researchers study Drosophila to understand taste perception and the neural circuits (in particular one called hugin) that affect feeding behavior. Touch and Temperature Sensing Responses to mechanical forces, including touch, are poorly understood at the molecular level. All animals have the ability to detect changes in environmental temperature based on the presence of neuronal and molecular substrates that affect thermosensation. Graham as their model for studying the optic nerve, and in 1967 Hartline, Ragnar Granit, and George Wald were awarded the Nobel Prize in Medicine for their research into visual processes of the eye. Limuli have large, easily accessible retinal neurons, allowing for electrophysiological study. Study of the horseshoe crab has provided insight into the operation of human vision, particularly adaptation to light and lateral inhibition (Liu and Passaglia 2009). Their sucker chemotactile systems have been compared to the mammalian eye (Packard 1972; Sanders et al. Other Models Space Biology Invertebrates are useful in studying the effects of ionizing radiation, both on earth and in space. It has also been used to study natural space radiation exposure, and the resulting mutants have been genetically evaluated (Nelson et al. Tardigrades (water bears, moss piglets) have also served as models for open space research, because they have the ability to survive desiccation, extreme cold, and radiation, all of which occur in space. Additional invertebrate species showing potential for use in outer space biological experiments are the sleeping chironomid (Polypedilum vanderplanki), the brine shrimp (Artemia salina and A.

Negative results do not rule out the presence of acute or ongoing Mycoplasma pneumoniae-associated disease. If testing is performed too early following primary infection, IgM may not be detectable. Negative Clinical References: Smith T: Mycoplasma pneumoniae infections: diagnosis based on immunofluorescence titer of IgG and IgM antibodies. Useful For: Rapid, sensitive, and specific identification of Mycoplasmoides genitalium from genitourinary and reproductive sources this test is not intended for medicolegal use. It is a cause of upper respiratory infection, pharyngitis, and tracheobronchitis, particularly in children, and has been associated with approximately 20% of cases of community acquired pneumonia. The disease is usually self-limited although severe disease has been reported in immunocompromised patients. The development of IgM antibodies takes approximately 1 week and the IgM response in adults may be variable or it may be decreased in immunosuppressed individuals. Useful For: Diagnosis of infections due to Mycoplasmoides pneumoniae Interpretation: A positive result indicates the presence of Mycoplasmoides pneumoniae. A negative result does not rule out the presence of M pneumoniae and may be due to the presence of inhibitors within the specimen matrix, or the presence of organisms at numbers below the limits of detection of the assay. Daxboeck F, Krause R, Wenisch C: Laboratory diagnosis of Mycoplasma pneumoniae infection. It is a B-cell neoplasm that can exhibit excess production of serum IgM symptoms related to hyper viscosity, tissue filtration, and autoimmune-related pathology. Useful For: the prognosis and clinical management of lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia Interpretation: Mutations detected or not detected. Useful For: Establishing the diagnosis of lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia Helping to distinguish lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (low-grade B-cell lymphoma) from other subtypes Interpretation: Variant present or not detected; an interpretive report will be issued. Reference Values: Variant present or absent based on expected alteration polymerase chain reaction product size. Therefore, early diagnosis and initiation of appropriate immunosuppressant treatment is important to optimize the clinical outcome by preventing further attacks. Nerve conductions studies typically demonstrate a characteristic progressive sensory predominant mixed axonal and demyelinating neuropathy with reduced distal conduction velocities that are greater distally. Useful For: Evaluating peripheral neuropathy Evaluating paraproteinemic neuropathy Interpretation: A positive result is consistent with anti-myelin-associated glycoprotein neuropathy. In such settings, when used in conjunction with appropriate clinical and morphologic findings, flow cytometry immunophenotyping analysis can provide additional diagnostic information to help distinguish an underlying clonal hematopoietic neoplasm from a reactive or secondary response. In combination, the total number of abnormalities detected and the distinctiveness of the abnormalities themselves help determine the likelihood of specimen involvement by a clonal myeloid neoplasm. Due to this extramedullary presentation, the bone marrow may have a low number of myeloblasts due to a lack of bone marrow involvement. Generalized bone pain, anemia, limb numbness, or weakness, symptoms of hypercalcemia, and recurrent infections are all symptoms that may indicate multiple myeloma. As myeloma progresses, the malignant plasma cells interfere with normal blood product formation in the bone marrow resulting in anemia and leukopenia. This heterogeneity is manifested clinically by varying degrees of disease aggressiveness. Useful For: Aiding in the diagnosis of new cases of multiple myeloma or other plasma cell proliferative disorders using a fixed cell pellet derived from bone marrow Identifying prognostic markers based on the abnormalities found this test should not be used to track the progression of disease. Interpretation: A neoplastic clone is detected when the percent of cells with an abnormality exceeds the normal reference range for any given probe. The absence of an abnormal clone does not rule out the presence of neoplastic disorder. Antibodies to myeloperoxidase are most useful diagnostically to support myeloid lineage in acute leukemias. These antibodies also facilitate the detection of granulocyte precursors in myeloproliferative disorders and myelodysplastic syndromes. Useful For: A marker of myeloid lineage Interpretation: this test does not include pathologist interpretation, only technical performance of the stain. Lupus nephritis and Goodpasture syndrome, as well as Wegener granulomatosis may present with azotemia and progressive renal failure. It is not possible to distinguish among these diseases on the basis of clinical signs and symptoms; autoantibody testing may be helpful.