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Fundus examination demonstrated mild optic disc hyperemia bilaterally (figure e-1 on the Neurology Web site at In the early infection phase erectile dysfunction treatment new drugs buy himcolin with a visa, acute meningitis erectile dysfunction 30 order himcolin 30 gm with mastercard, meningovasculitis impotence of organic origin icd 9 buy cheap himcolin 30 gm line, and myelitis have been described. Cognitive impairment (general paralysis of the insane) and tabes dorsalis, characterized by sensory ataxia and lancinating pains, are seen in the late stages of the disease. This can pose a challenge in the immunocompromised patient since serology relies on the immune response to the infection. The recommended regimen is parenteral penicillin G administered as 3­ 4 million units every 4 hours (or continuous infusion of 18 ­24 million units per day), for 10 ­14 days. Williams serves on scientific advisory boards for Bausch Lomb, Novartis, Regeneron Pharmaceuticals, Inc. She reported 3 weeks of progressive clumsiness of the right limbs, weakness of the right leg, and an unsteady gait. She denied cognitive dysfunction, headache, bulbar or sensory symptoms, muscle stiffness/spasms, antecedent infection, fever, or other systemic complaints. Nine years earlier, the patient had experienced an episode of diplopia and unsteadiness which resolved spontaneously after 3 months. In the 1980s, a low vitamin B12 level (value unknown) was thought to have been an incidental finding; levels 500 ng/L have been maintained with a B12 supplement. General medical examination had normal results, including the absence of vitiligo. Funduscopic, pupillary, visual field, and monocular acuity examinations were unremarkable. Near card straightahead binocular acuity was 20/20, but only 20/50 in lateral downgaze due to oscillopsia. The eye movement abnormalities were saccadic pursuit, gazeevoked nystagmus, downbeating nystagmus maximal on lateral downgaze, and saccadic slowing but full range of the left adducting eye. There was 4 /5 right leg weakness (hip/ knee flexors, toe extensors), with hyperreflexia, downgoing plantar responses, and normal sensation. There was right-sided dysmetria, dysdiadochokinesia, loss of check, and exaggerated rebound. The patient could sit upright unsupported but required assistance to ambulate due to weakness and ataxia. Hashimoto thyroiditis was diagnosed several months after the second episode began. The hemiataxia and leg weakness may localize to the pontocerebellar and corticospinal tracts, respectively. While downbeat nystagmus, often seen in conjunction with saccadic pursuit and gaze-evoked nystagmus, traditionally localizes to the flocculus-paraflocculus, it is hypothesized to also occur with pontomedullary paramedian tract lesions. What is the differential diagnosis of a sporadic ataxia with or without brainstem features? The sporadic ataxias may also be split (imperfectly) into 2 groups according to their tendency to recur: 1) disorders that are either progressive or typically monophasic (but may recur) and 2) a smaller group that includes inherently recurrent conditions and recurrent stroke. Viral encephalitis can present as a unilateral brainstem syndrome, possibly recurrent, but typically with systemic symptoms. The recurrent ataxias include the episodic ataxias, relapsing multiple sclerosis, and strokes. Episodic ataxia 2 is characterized by episodes ranging from minutes to weeks but usually hours (vs seconds to minutes in type 1), with a typical age at onset from 5 to 15 years. Allelic to episodic ataxia 2, spinocerebellar ataxia 6 occasionally presents with episodic ataxia. Vitamins B12 (911 ng/L) and E, thyroid function tests, Lyme titer, and celiac and paraneoplastic panels (including amphiphysin) were normal or negative. Thyroperoxidase/ thyroglobulin, pancreatic islet cell, and gastric parietal cell/intrinsic factor antibody levels were also elevated. The clinical course was usually subacute but could be insidious, and only rarely relapsing. Acute/subacute onset or quick progression (although insidious onset/progression is not uncommon), late onset, relapses, prominent asymmetry/ unilaterality, and stiff-person phenomenon 2. While response to immunotherapy is often limited, significant improvement may occur. Less likely diagnostic possibilities include recurrent demyelination, stroke, Bickerstaff or viral brainstem encephalitis, or that the episodes were unrelated to each other. Silvers has received honoraria for educational activities from Teva Pharmaceutical Industries Ltd. Glutamic acid decarboxylase autoimmunity with brainstem, extrapyramidal, and spinal cord dysfunction. Antibodies against glutamic acid decarboxylase: prevalence in neurological diseases. Spectrum of neurological syndromes associated with glutamic acid decarboxylase antibodies: diagnostic clues for this association. Downbeating nystagmus and muscle spasms in a patient with glutamic-acid decarboxylase antibodies. Autoantibodies to glutamic acid decarboxylase in three patients with cerebellar ataxia, late-onset insulin-dependent diabetes mellitus, and polyendocrine autoimmunity. Cerebellar ataxia with anti-glutamic acid decarboxylase antibodies: study of 14 patients. Neurology 75 August 17, 2010 e33 181 Disorders presenting with headache, dizziness, or seizures Headache, dizziness, and seizure are 3 of the most common conditions for which neurologists are consulted. Headache and dizziness can be the presenting symptoms of both benign and potentially fatal conditions. Seizures may be due to idiopathic epilepsy syndromes or can be symptomatic of underlying neurologic or systemic pathology. Each symptom therefore requires a detailed history, neurologic examination, and evaluation to distinguish between the diverse potential etiologies of these common "chief complaints. The term dizziness can have diverse mean- primary headache syndrome or may be secondary to an underlying disease process. The differential diagnosis for secondary causes of headache is extensive and includes pathology of any cranial structure, as well as a variety of systemic diseases. Headaches that have any of the following "red flags" require thorough evaluation for underlying intracranial pathology: ings and may represent vertigo, light-headedness, unsteadiness, or even anxiety. The evaluation of a patient with dizziness therefore requires a search for potential systemic causes, and, if excluded, the primary task of the neurologist is to distinguish dizziness of peripheral etiology (due to pathology of the inner ear and/or vestibulocochlear nerve) from dizziness due to central pathology (due to pathology of the brainstem and/or cerebellum). This requires an intimate familiarity with subtle neuro-ophthalmologic and neuro-otologic examination maneuvers and the interpretation of their findings. New-onset seizures therefore require a thorough evaluation for an underlying trigger: intracranial pathology. The cases in this section depict the clinical approach to patients presenting with headaches, dizziness, or seizures. She was born to nonconsanguineous parents from Somalia at 415/7 weeks of gestation. The mother was group B streptococcus­positive and was appropriately treated with antibiotics during labor. Labor and vaginal delivery were uncomplicated (no history of prolonged rupture of membranes or birth trauma). The baby appeared to be well on the first day of life but began having seizures on the second day. On presentation to our facility, the patient exhibited rhythmic jerking movements of her extremities, consistent with myoclonic seizures. She also had multiple apneic episodes and was therefore intubated and mechanically ventilated. Neurologic examination revealed diffuse hypotonia with symmetrically hypoactive reflexes in all 4 extremities. Bedside funduscopic examination revealed normal Moro; suck and rooting reflexes were poor, but palmar grasp reflex was present bilaterally. There was no family history of neurologic or metabolic disorders (including seizures).

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Genes in conflict: the biology of selfish genetic elements / Austin Burt and Robert Trivers does erectile dysfunction cause infertility order 30gm himcolin otc. By logic erectile dysfunction brands proven 30 gm himcolin, situations of conflict erectile dysfunction caused by anabolic steroids cheap himcolin 30gm, in which natural selection acts in opposing directions simultaneously, are likely to generate strong coadaptations on each side. At the level of individuals interacting within (or between) species, this conflict is seen to generate a wide range of behaviors and structures. Conflict within an individual might be expected to generate similar kinds of genetic complexity. Empirically it has become apparent that some categories of selfish genetic elements. We taught a course together on the subject at the University of California at Santa Cruz in 1992 and we have taught the material since then at Rutgers University and Imperial College London. Throughout the book we have been guided by two principles: we have tried to include all types of selfish elements in all species (except bacteria and viruses) and we have tried to organize these examples logically by reference to the way in which natural selection acts on the selfish elements and, in turn, on the larger genomes they inhabit. We begin by expressing our gratitude to those who have provided the financial support that has permitted this work. For reading the entire book and giving us their comments, we are most grateful to Jim Bull, Juan Pedro Camacho, Alan Grafen, and David Haig. For reading individual chapters we are grateful to Camille Barr, John Brookfield, Brian Charlesworth, Kelly Dawe, Tom Eickbush, Steve Frank, Jody Hey, Hopi Hoekstra, John Jaenike, Neil Jones, Vasso Koufopanou, Benjamin Normark, Sally Otto, Brian Palestis, Carmen Sapienza, Jonathan Swire, William Zimmerman, and Elefterios Zouros. We are also grateful to David Haig for sharing over many years his extensive knowledge of the subject and to Vasso Koufopanou for advice and support throughout. We are most grateful to Elizabeth Collins for her copyediting, Margaret Nelson for drawing all of the figures, Darine Zaatari for help with the bibliography, and Anne McGuire and Jennifer Snodgrass for help with final editing and production. And we are especially grateful to our editor Michael Fisher for encouragement and unfailing support over many years. In animals, for example, some genes may want (or act as if they want) a male to produce lots of healthy sperm, but other genes in the same male want half the sperm to be defective. Some genes in a female want her to nourish all her embryos; others want her to abort half of them. Some genes in a fetus want it to grow quickly, others slowly, and yet others at an intermediate level. Some genes want it to become a male, others a female-and the reason they want it to be a female is so that a quarter of her fertilized eggs will be defective! Some genes want the plant to allow a particular pollen grain to fertilize an ovule, and others want to kill that pollen grain. Some genes want to protect chromosomes from damage, while others want to break them. Indeed, in the extreme case, some genes want to inactivate half of the genome, while the targeted half prefers to remain active. Or genes that want a female to judge her mate as more attractive than he really is versus genes that want the opposite effect. These conflicts arise because genes are able to spread in a population despite being harmful to the larger organism. Such genes give themselves a benefit but typically cause negative effects on other nonlinked genes in the same creature. They, in turn, select for nonlinked genes that suppress their activity, and thereby mitigate the harm. That is, the evolution of selfish genetic elements inevitably leads to withinindividual-or intragenomic-conflict. This occurs over evolutionary time, as genes at different locations within the genome are selected to have contradictory effects. It also occurs over developmental time as organisms experience these conflicting effects. In this sense, we speak of "genes in conflict," that is, genes within a single body that are in conflict over the appropriate development or action to be taken. Genetic Cooperation and Conflict We mammals have some 30,000 genes that together make an astonishingly complex creature. Most of these genes are beneficial to us, that is, they make a positive contribution to survival and reproduction most of the time. This is no accident: most genes have spread in populations precisely because they increase organismal fitness. All the complex adaptations that organisms show for survival and reproduction-that distinguish living from nonliving things, biology from chemistry and physics-exist because of natural selection; and for most genes, the selection of alternative alleles is based on how they contribute to organismal function. This cooperation arises in large part because most genes are transmitted from one generation to the next in a transparent, "fair" manner, with diploid individuals transmitting the 2 copies of each gene with equal likelihood, 50:50. Thus the gametes produced by an individual are usually a faithful reflection of the gametes from which it was derived, and the trans2 Selfish Genetic Elements mission of genes from one generation to the next does not in itself lead to a change in gene frequencies. It avoids the introduction of an arbitrary bias into the genome, one that would warp gene frequencies away from what is optimal for phenotypic function. Ignoring mutation and random effects, gene frequencies will change only if individuals of different genetic constitution reproduce to different degrees. Because different genes in the same organism are selected in the same direction (to increase survival and reproduction), they can evolve to cooperate in, say, the construction of an eye or leg. But some genes have discovered ways to spread and persist without contributing to organismal fitness. At times, this means encoding actions that are diametrically opposed to those of the majority of genes. As a consequence, most organisms are not completely harmonious wholes and the individual is, in fact, divisible. This book is about these alternative routes to genetic posterity, and the diverse array of adaptations and counteradaptations that such selection has produced. As we shall see, selfish genetic elements are a universal feature of life, with pervasive effects on the genetic system and the larger phenotype, diversified into many different forms, with adaptations and counteradaptations on both sides, a truly subterranean world of sociogenetic interactions, usually hidden completely from sight. While most genes in a sexual organism are transmitted to the Mendelian 50% of progeny, these selfish genetic elements manage to get into 60%, or 99%, or 50. Instead of being fair and transparent, the process of gene transmission in itself leads to an increase in gene frequencies. Genes inherited in a biased manner can spread in a population without doing anything good for the organism. This book documents all the many answers to this question that have been discovered to date, at least in eukaryotic organisms (that is, we exclude bacteria and viruses). Here we sketch in broad outline the 3 main strategies that have evolved over and over again. This strategy is the most blatantly selfish: getting ahead by disrupting the transmission of the alternative allele. For example, a gene in a diploid organism might sabotage the 50% of the gametes to which it is not transmitted, thereby increasing its own success. Or it might kill the 50% of offspring that do not carry a copy of it, thereby increasing survival of the 50% that do. This is the most diverse class of selfish genetic elements, and we shall see examples throughout the book (see Chaps. They have been found on autosomes, sex chromosomes, and in organelles; and the targets of their action can be genes, chromosomal regions, whole chromosomes, or entire haploid sets of chromosomes. Note that in all of these cases, a kind of "kinship" discrimination is being made, at least at a very local level. That is, the selfish element is able to direct behavior against others according to the chance it will be located in them. Other selfish genetic elements bias their transmission to the next generation by getting themselves replicated more often than other genes in the same organism. In the nucleus, complex mechanisms have evolved to ensure that most genes are replicated exactly once per cell cycle, but various classes of selfish element manage to circumvent these mechanisms. Such selection may account in part for the small size of organelle genomes (see Chap. Finally, genes can also increase in frequency within an organism by increasing the replication rate of the entire cell, if different cells are genetically distinct, thereby creating a selfish cell lineage (see Chap. The third strategy of drive is to move preferentially toward the germline, and away from somatic cells, when presented with the choice. For example, female meiosis in most species gives rise to 1 functional egg or ovule and 2 or 3 nonfunctional "polar bodies. Competition to be included in the egg and avoid the polar bodies has led to the evolution of "knobs" on the chromosomes of maize, which act as centromeres during meiosis and pull themselves along the spindle (see Chap.


  • Hemeralopia, congenital essential
  • Brachymesophalangy mesomelic short limbs osseous anomalies
  • Amelogenesis imperfecta hypomaturation type
  • Eem syndrome
  • Polydactyly
  • Fanconi pancytopenia
  • Leukomelanoderma mental retardation hypotrichosis
  • Ben Ari Shuper Mimouni syndrome

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In most transformation protocols optimal transformation is achieved at room temperature erectile dysfunction johns hopkins purchase 30 gm himcolin amex. An interesting aspect of Agrobacterium-mediated transformation of mammalian cells is that it occurred at 37°C after pre-growth of Agrobacterium at 28°C (Kunik et al erectile dysfunction wife generic himcolin 30 gm mastercard. The effect of pH during co-cultivation on the transformation frequency was tested in Agrobacterium-mediated transformation of C erectile dysfunction doctor sydney safe himcolin 30 gm. It was found that the optimal pH, leading to the highest transformation frequency, is between 5. The optimal pH also depends on the Agrobacterium strain used, as the pH requirements for optimal vir-gene induction are slightly different for the different Agrobacterium strains (Turk et al. For efficient Agrobacterium-mediated transformation cells are cocultivated on a solid support such as nitrocellulose filters, Hybond, filter paper, cellophane sheets, and polyvinylidene difluoride. Frequently used markers in both plants and non-plant systems are different antibiotic resistance genes from bacterial plasmids. Also an herbicide resistance gene (bar) has been used as a selection marker in fungi (Fang et al. It is important that these markers are controlled by a promoter active in the host organism. Uracil auxotrophy markers have also been used during Agrobacterium-mediated transformation of filamentous fungi (Gouka et al. However, their role in transformation of non-plant organisms is not well-established. On the other hand, Agrobacterium-Mediated Transformation of Non-Plant Organisms 659 Agrobacterium-mediated transformation of mammalian cells depends on the presence of the chvA and chvB genes (Kunik et al. Reversely, it was reported that inactivation of one of the chromosomal genes involved in the biosynthesis of cellulose fibrils increases the frequency of transformation of Aspergillus awamori (Michielse et al. Some information is also available on the role of the Vir proteins in the transformation of nonplant organisms. The virA, virB, virD and virG genes are essential not only for plant transformation, but also for transformation of non-plant organism such as the yeast S. Although inactivation of virE2 almost eliminates the ability of Agrobacterium to transform plants, transformation of S. Hooykaas approximately 50-100-fold more efficient than via non-homologous recombination in the yeast (Bundock et al. For the application of Agrobacterium-mediated transformation in functional genomics or biotechnology it is of great importance to improve the efficiency of integration via homologous recombination over non-homologous recombination. In yeast, and most likely also in mammals, this reaction relies on the Mre11 nuclease activity in a multiprotein complex consisting of Rad50, Mre11, and Xrs2 (Figure 18-2A). This has led to the proposal that this Rad52 binding channels to repair by homologous recombination instead of to non-homologous end joining. This process is influenced by other proteins such as replication protein A (Rpa), Rad52 and Rad54. Non-homologous end joining is initiated by a double strand break, followed by binding of the Ku70 and Ku80 proteins to the ends (Figure182b). With the help of other proteins the break is then sealed restoring the original sequence or with small deletions. Some plant homologs of the components of these complexes have already been found which show positive effects on or are differentially expressed during Agrobacterium-mediated transformation (Veena et al. It is expected that in the absence of the non-homologous end joining proteins Ku70 or Ku80 integration by homologous recombination will become relatively more frequent. This protein transfer is not restricted to plant cells, but it has been shown that the VirE2, VirE3 and VirF proteins can be transferred into cells from the yeast Saccharomyces cerevisiae as well (Schrammeijer et al. To study protein transfer from Agrobacterium to yeast, the Cre recombinase reporter assay for translocation has been used. Transfer of the Cre-Vir fusion proteins from Agrobacterium to yeast can be monitored by a selectable excision event resulting from site-specific recombination mediated by Cre on a lox-flanked transgene in yeast. This assay illustrates the potential of Agrobacterium to introduce genome modifying enzymes into eukaryotic cells. This property of Agrobacterium is promising for its application in protein therapy of both plant and non-plant cells. Especially for the transformation of various fungi, it has great advantages over other transformation methods. The efficiencies are much higher and the transformation protocols are relatively facile. Most likely, protein transfer occurs during the transformation of all host cells, irrespective of their origin. Because of this property, Agrobacterium has a great potential for use in protein therapies. As these key proteins are strongly conserved in other eukaryotes (from fungi to plants and animals) the knowledge obtained from yeast may be directly applicable in these other organisms to improve the frequency of targeted integration. Recently, it has been shown that expression in the plant Arabidopsis thaliana of the S. In the rhizosphere where vir inducers are readily available and numerous microorganisms are living in close proximity, it is very likely that Agrobacterium-mediated transformation of non-plant organisms is occurring. Future research has to show whether such horizontal gene transfer has contributed to evolution. Appl Environ Microbiol 66: 4510-4513 Cheney D, Metz B, Stiller J (2001) Agrobacterium-mediated genetic transformation in the macroscopic marine red alga Porphyra yezoensis. Mycol Res 105: 259-264 Dai Q, Sun Z, Schnabel G (2003) Development of spontaneous hygromycin B resistance in Monilinia fructicola and Its impact on growth rate, morphology, susceptibility to demethylation inhibitor fungicides, and sporulation. Annu Rev Genet 39: 431-451 Daniell H, Kumar S, Dufourmantel N (2005) Breakthrough in chloroplast genetic engineering of agronomically important crops. Trends Biotechnol 23: 238-245 De Block M, Schell J, Van Montagu M (1985) Chloroplast transformation by Agrobacterium tumefaciens. Nat Biotechnol 16: 839-842 Degefu Y, Hanif M (2003) Agrobacterium tumefaciens-mediated transformation of Helminthosporium turcicum, the maize leaf-blight fungus. J Microbiol Methods 58: 197-202 Dudasova Z, Dudas A, Chovanec M (2004) Non-homologous end-joining factors of Saccharomyces cerevisiae. Hooykaas Eckert M, Maguire K, Urban M, Foster S, Fitt B, Lucas J, Hammond-Kosack K (2005) Agrobacterium tumefaciens-mediated transformation of Leptosphaeria spp. Curr Genet 41: 183-188 Hoffman B, Breuil C (2004) Disruption of the subtilase gene, albin1, in Ophiostoma piliferum. Eukaryot Cell 3: 420-429 Ishida Y, Saito H, Ohta S, Hiei Y, Komari T, Kumashiro T (1996) High efficiency transformation of maize (Zea mays L. Plant Sci 166: 731-738 Kunik T, Tzfira T, Kapulnik Y, Gafni Y, Dingwall C, Citovsky V (2001) Genetic transformation of HeLa cells by Agrobacterium. Sheng Wu Gong Cheng Xue Bao 19: 419-423 Li M, Gong X, Zheng J, Jiang D, Fu Y, Hou M (2005) Transformation of Coniothyrium minitans, a parasite of Sclerotinia sclerotiorum, with Agrobacterium tumefaciens. Plant Physiol 138: 1318-1321 Malonek S, Meinhardt F (2001) Agrobacterium tumefaciens-mediated genetic transformation of the phytopathogenic ascomycete Calonectria morganii. Fungal Genet Biol 42: 904-913 Meyer V, Mueller D, Strowig T, Stahl U (2003) Comparison of different transformation methods for Aspergillus giganteus. Phytopathol 91: 173-180 Ninomiya Y, Suzuki K, Ishii C, Inoue H (2004) Highly efficient gene replacements in Neurospora strains deficient for nonhomologous end-joining. Microbiol Mol Biol Rev 66: 630670 Takahara H, Tsuji G, Kubo Y, Yamamoto M, Toyoda K, Inagaki Y, Ichinose Y, Shiraishi T (2004) Agrobacterium tumefaciens-mediated transformation as a tool for random mutagenesis of Colletotrichum trifolii. Curr Genet 45: 242-248 Tanguay P, Breuil C (2003) Transforming the sapstaining fungus Ophiostoma piceae with Agrobacterium tumefaciens. Hooykaas Tsuji G, Fujii S, Fujihara N, Hirose C, Tsuge S, Shiraishi T, Kubo Y (2003) Agrobacterium tumefaciens-mediated transformation for random insertional mutagenesis in Colletotrichum lagenarium. Nat Rev Mol Cell Biol 4: 435-445 White D, Chen W (2005) Genetic transformation of Ascochyta rabiei using Agrobacterium-mediated transformation. Gene Ther 10: 1791-1799 Zeilinger S (2004) Gene disruption in Trichoderma atroviride via Agrobacteriummediated transformation. From the early stages of genetic engineering legal frameworks were set up to ensure the safe development of this technology. These regulatory frameworks focus primarily on risks to human health and the environment, and the concepts of substantial equivalence and familiarity seem to be the two universally adopted principles on which risk assessments are based.

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One concern about plant-based pharmaceuticals is the potential for non-mammalian glycosylation patterns that might result in immune sensitization or loss of function (Bardor et al erectile dysfunction doctors in ct cheap himcolin 30gm without a prescription. However erectile dysfunction doctors in houston tx purchase genuine himcolin on-line, at least one plant-derived monoclonal antibody was found to impotence lexapro himcolin 30gm generic be functional despite differences in N-linked glycosylation (Ko et al. Other potential limitations of plant expression systems include low and/or variable yield (Chargelegue et al. Finally, contamination of food and feed crops with pharmaceutical crops, either in the field or postharvest, poses potentially serious health and public relations risks (Ma et al. By integrating an engineered marker gene, beta-glucuronidase, Barbara Hohn and coworkers have pioneered a strategy in which transgenic Arabidopsis has successfully been used to report enhanced rates of homologous recombination or point mutation due to heavy metal ions (Kovalchuk et al. Agrobacterium and Plant Biotechnology 97 Increasing levels of pollution resulting from global industrialization have focused attention on the possibility of phytoremediation: using plants to remove or inactivate pollutants from soil or surface waters. Theoretically, genetic manipulation of heavy metal accumulation in plants could be used to imbue a plant with any of these traits or to enhance an existing capability (Clemens et al. Introduction of bacterial genes has enabled the creation of transgenic Arabidopsis plants capable of converting the highly toxic contaminant methylmercury to the volatile and much less toxic elemental mercury (Bizily et al. Similar modifications have resulted in Arabidopsis and poplar able to process and sequester mercury ion (Rugh et al. To deplete arsenic contamination from groundwater, researchers have introduced bacterial genes that confer on Arabidopsis the ability to extract and accumulate in the leaf levels of arsenic that would normally poison the plant (Dhankher et al. Second generation phytoremediating plants will likely capitalize on the finding that overexpression of a yeast vacuolar transporter in Arabidopsis leads to enhanced accumulation, and hence tolerance, of heavy metals such as cadmium and lead (Song et al. The advent of basic plant molecular biology, made possible in large part by the availability of Agrobacterium-mediated techniques for introducing and knocking out plant genes, has dramatically augmented our understanding of how plant architecture and generation time are regulated, and these discoveries may enable further improvements in yield. Banta and Maywa Montenegro brassinosteroid levels resulted in a more erect leaf structure in rice, increasing yield under dense planting conditions (Sakamoto et al. Tissue-specific modulation of the growth hormone gibberellin catabolism in transgenic rice led to a semi-dwarf phenotype without a loss in grain productivity (Sakamoto et al. In other cases, yield may be enhanced by decreasing the time required for the plant to produce the edible portion. Dormancy in potatoes was controlled by expressing a bacterial gene that altered sprouting behavior (Farre et al. Other attempts to increase yield potential have centered on the photosynthetic process, and in particular the inefficiency of the carbon assimilation pathway in C3 plants, a group that includes many agronomically important crop plants. Using Agrobacterium-mediated transformation, Matsuoka and co-workers have expressed three key C4 enzymes in rice (a C3 plant) (Ku et al. However, grain production is tightly linked to nitrogen availability, and hence larger plants will not necessarily yield more grain unless soil nitrogen levels are sufficient (Sinclair et al. Among the approaches to mitigating these constraints are some that involve genetically modifying the crop plant. It is Agrobacterium and Plant Biotechnology 99 important to stress that there are also many highly successful nonbiotechnological practices that have been in use for centuries, including integrated pest and vector management, crop rotation, dissemination of pathogen-free plant material (Rudolph et al. Indeed, farming systems that combine careful land management with a diverse array of species and genetic backgrounds within a species can be highly productive even in the absence of modern varieties or biotechnology "improvements" (Brown, 1998). The lessons of such a holistic approach to agriculture are enjoying a resurgence of popularity among small and some medium-scale farmers in the industrialized world; for example, integrated production and organic farming guidelines are in practice on 85% of the farmland in Switzerland (Xie et al. Nonetheless, the predominant model of agriculture in much of the developed world is one of monocultures grown with high external inputs. At the other end of the spectrum, resource-poor farmers cultivating marginally arable land face myriad environmental constraints which, for a variety of reasons, have proven recalcitrant to the available integrated approaches. The following sections highlight some of the applications of Agrobacterium-mediated genetic modification of plants that may address these constraints and/or mitigate negative consequences of the conventional solutions. On the other hand, although biotechnology is anathema to most proponents of organic farming practices, it is likely that our ability to meet the growing challenge of adequate food production may benefit from open-mindedness and creative approaches that incorporate the genetic modifications described below into sustainable, ecosystem-centered cultivation systems. Engineering plants with enhanced capabilities to absorb micronutrients from the soil, by over-expressing nitrogen, potassium and phosphorus transporters and/or manipulating their regulation, could decrease the need for fertilizers (Hirsch and Sussman, 1999). For some nutrients, such as iron and phosphorus, the limiting factor is often solubility rather than abundance in the soil. Plants synthesize and secrete a variety of organic acids that can chelate insoluble compounds, allowing uptake of the complex (Guerinot, 2001). Banta and Maywa Montenegro maize and sorghum are particularly sensitive to low iron availability in alkaline soils, where iron is less soluble. Agrobacterium-mediated introduction of genes conferring enhanced biosynthesis of an iron chelator in rice resulted in improved growth and four-fold higher grain yields under conditions of low iron availability (Takahashi et al. The abundant metal aluminum normally exists as harmless oxides and aluminosilicates, but in acidic soils it is solubilized into the toxic Al3+, which inhibits root growth. Plants that tolerate otherwise toxic levels of Al3+, do so by secreting organic acids such as citrate or malate at the root apex that chelate the Al3+, in the soil and prevent uptake (Ma et al. Improved tolerance of zinc in transgenic plants has also been observed (van der Zaal et al. Metal contamination in the soil is but one of the abiotic stresses that constrain crop plant productivity. Growing global demand for food continues to force farmers onto marginally arable land where soil salinity, water deficits, and climatic challenges such as low or high temperatures limit cultivation (Bartels, 2001). Strategies to engineer enhanced tolerance to such adverse conditions fall into at least two categories: direct protection from the stressor(s), and enhanced resistance to the physiological damage caused by the stressor. In the latter category, a family of aldose-aldehyde reductases are activated in response to a wide variety of stresses (Bartels, 2001). Ectopic expression of an alfalfa aldose-aldehyde reductase gene via Agrobacterium-mediated transformation results in reduced damage upon oxidative stress, apparently by eliminating reactive aldehydes, and increased tolerance to salt, dehydration, or heavy metal stress (Oberschall et al. Osmolytes also confer stress tolerance by scavenging reactive oxygen species (Zhu, 2001). The non-reducing disaccharide trehalose stabilizes biological structures upon dessication in many bacteria, fungi, and invertebrates, but apparently does not accumulate naturally in plants (Penna, 2003). Transgenic tobacco and rice engineered to produce trehalose exhibit enhanced resistance to drought (Romero et al. Production of mannitol results in tobacco with enhanced tolerance to high salinity (Tarczynski et al. Other low-molecular-weight compatible solutes that accumulate in some plants to protect proteins from stress-induced damage include glycinebetaine, polyols and amino acids. Glycinebetaine accumulation confers on transgenic Arabidopsis an increased ability to withstand high temperatures during germination and seedling growth (Alia et al. In addition to small osmolytes, a number of proteins have also been shown to have stress protective activity, primarily in response to low temperature. A variety of plants produce antifreeze proteins, as do several fish and insects; these proteins function to inhibit growth of intercellular ice crystals (Griffith and Yaish, 2004). There have been a variety of attempts to introduce a gene encoding one of these anti-freeze proteins into tobacco, tomato, potato, and Arabidopsis, with the ultimate goal of lowering the freezing temperature, even by a few degrees, so that the plants could survive a light frost (Griffith and Yaish, 2004). At least one such experiment was successful; although the plant did not exhibit higher rates of survival upon freezing, the freezing temperature was indeed lowered (Huang et al. Another transgenic strategy to achieve freezing tolerance involves the introduction of bacterial ice nucleation genes, which permit slow dehydration that minimizes tissue damage (Baertlein et al. High soil salinity impedes plant growth, both by creating a water deficit in the soil and within the plant, as sodium ions impinge on many key biochemical processes. Strategies to increase salt tolerance involve limiting exposure of cytoplasmic enzymes to the salt and may include blocking Na+, influx, increasing Na+ efflux, and compartmentalizing Na+ (Zhu, 2001). Successful transgenic approaches are described in detail in Yamaguchi and Blumwald (2005). Many of these entail over-expressing the Arabidopsis vacuolar Na+/H+ antiporter, which enhances tolerance to soil salinity, with few or no detrimental effects on seed quality or plant growth, in canola (Zhang et al. Sequestration of cations in the Arabidopsis vacuole, resulting in enhanced salt and drought tolerance was also achieved by overexpressing the vacuolar H+ pyrophosphatase (Gaxiola et al. Increased expression of a plasma membrane Na+/H+ antiporter augmented salt tolerance by limiting Na+ accumulation (Shi et al. Finally, tolerance in rice, resulting from expression of a bacterial Na+/H+ antiporter, was accompanied by biosynthetic activation of the osmoregulatory molecule proline (Wu et al. Both freezing and high temperature cause damage to plant tissues and proteins, leading to diminished crop yield. A comprehensive listing of attempts to enhance plant thermo-tolerance through genetic modification can be found in Sung et al. One of the earliest reports of altered chilling sensitivity resulted from engineering the degree of fatty acid saturation in tobacco membranes (Murata et al. In concluding this section on engineering tolerance to environmental constraints, it is important to recognize that reductions in crop viability and yield are compounded by combinations of abiotic stresses.

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Pyridoxal phosphate Neurological indication Refractory epilepsy in infants (may be superior to erectile dysfunction operations cheap himcolin online visa pyridoxine) erectile dysfunction drugs trimix cheap generic himcolin canada. Pyridoxine (vitamin B6) Neurological indications Treatment of refractory epilepsy in infants (see b p erectile dysfunction under 25 order himcolin 30gm on-line. Preparation Tablets (10, 20, and 50 mg; can be halved, quartered, or crushed and dissolved in water), injection (50 mg/2 mL), liquid. Try not to make any other changes in anti-epileptics during this period to aid interpretation (see b p. The dose for optimal neurodevelopmental outcome may be greater than the dose that controls seizures. Dosing Starting doses and escalation regimen Movement disorder: over 12 yrs, 1 mg/24 h divided in 2 doses increasing at weekly intervals by 1 mg/24 h if required. Maintenance doses Movement disorder: over 12 yrs, up to 4 mg/24 h divided in 2 doses. Comments Use of antipsychotics to manage acutely disturbed behaviour should only be considered in extreme situations. Rufinamide Neurological indications Epilepsy, particularly Lennox-Gastaut syndrome. Dosing Child 4­18 years less than 30 kg: 100 mg bd increasing if required by 100 mg bd at 7­14-day intervals; max. Preparations 100, 200, and 400 mg tablets, which may be crushed and mixed with water. Important interactions and unwanted effects May raise phenytoin levels; metabolism inhibited by valproate. Comments A serious hypersensitivity syndrome has been reported in children after initiating therapy; consider withdrawal if rash or signs or symptoms of hypersensitivity syndrome develop. Stiripentol Neurological indications Anti-epileptic drug particularly for severe myoclonic epilepsy of infancy (Dravet Syndrome). Dosing Starting doses and escalation regimen Child 3­18 years: initially 10 mg/kg in 2­3 divided doses; titrate dose over minimum of 3 days to max. Comments Most commonly used in conjunction with valproate and/or clobazam in treatment of severe myoclonic epilepsy of infancy (see b p. Important interactions and unwanted effects Antimuscarinic effects; may cause agitation in low dose, hepatitis. Contraindications Vasospasm, previous cerebrovascular accident or transient ischaemic attack, peripheral vascular disease, hypertension. Important interactions and unwanted effects Taste disturbance, mild irritation or burning sensation in the nose or throat, heat, heaviness, pressure or tightness, flushing in any part of the body, dizziness, weakness, fatigue, drowsiness and transient increases in blood pressure. Other triptans are not direct equivalents: rizatriptan has a short half-life, and frovatriptan has a much longer half-life than sumatriptan. Important interactions and unwanted effects Interacts with metoclopramide: increased risk of dystonia. Dosing Starting doses and escalation regimen Over 12 yrs: 5 mg bd for 1 week po increased by 5­10 mg/24 h divided in 2 doses every 5­7 days. Maintenance doses 30­45 mg/24 h po divided in 2­3 doses as tolerated and according to response. Important interactions and unwanted effects Nausea, diarrhoea, sleepiness, tremor, rarely non-convulsive status epilepticus. Important interactions and unwanted effects Interacts with ciprofloxacin and phenytoin. Important interactions and unwanted effects Nausea, anorexia with weight loss, paraesthesiae. Dosing Starting doses and escalation regimen 3 months­18 yrs: 1­2 mg/24 h po in 1 or 2 divided doses incrementing by 1 mg/24 h every 3­7 days, divided in 3­4 doses according to response. Contraindications Intestinal obstruction, urinary retention, closed angle glaucoma, myasthenia gravis. Important interactions and unwanted effects Urinary retention, constipation, tachycardia, anhidrosis (and hyperpyrexia), dry mouth, blurred vision, confusion, agitation, hallucination. Gradual dose escalation can result in children tolerating comparatively high doses. Dosing Starting doses and escalation regimen Epilepsy and migraine: 10 mg/kg/24 h (>12 yrs 600 mg/24 h) divided in 2 doses increasing by 10 mg/kg/24 h (>12 yrs 200 mg/24 h) every 5­7 days. Maintenance doses 20­40 mg/kg/24 h divided in 2 doses, max 60 mg/24 h (adult 1­2 g/24 h, occasionally 2. Preparations Crushable tablet (100 mg) enteric-coated tablets (200 and 500 mg) controlled-release tablet (200, 300, and 500 mg), oral liquid (200 mg/5 mL), intravenous injection (100 mg/mL) modified-release granules (50, 100, 250, 500, and 750 mg, and 1 g). Impaired hepatic function leading rarely to fatal hepatic failure (some cases likely to be due to unidentified beta-oxidation or mitochondrial depletion (Alper) syndromes: avoid use if mitochondrial disease suspected). Teratogen causing distinct foetal valproate syndrome and/or neural tube defects, and possible adverse developmental outcomes in babies exposed in utero (see b p. Comments Routine monitoring of liver function in an asymptomatic child is not indicated. Carers should be taught to seek medical attention in case of unexplained nausea, vomiting, darkened urine or jaundice. Vigabatrin Neurological indications Treatment of infantile spasms particularly in tuberous sclerosis. Dosing Starting doses and escalation regimen Infantile spasms: 50 mg/kg/24 h increasing if required every 48 h to 100 mg/kg/24 h and then 150 mg/kg/24 h divided in 2 doses. Powder can be dispersed in 10 mL of water and the appropriate volume used to give small doses. Contraindications Pre-existing or potential for visual impairment (particularly visual field impairments). Contraindications Severe gastritis or ulcer, severe hypertension, bacterial endocarditis. Dosing Starting doses and escalation regimen 2 mg/kg/24 h increasing by 2 mg/kg/24 h divided in two doses every 1­2 weeks. Maintenance doses 8­18 mg/kg/24 h (very occasionally up to 30 mg/kg/24 h) divided in two doses. More rarely nephrolithiasis, (encourage reporting of back/abdominal pain or urinary symptoms), Stevens­Johnson syndrome, agranulocytosis, oligohydrosis and hyperthermia (beware in small children). Interactions of anti-epileptic drugs Enzyme-inducing drugs, such as carbamazepine, phenobarbital, phenytoin may lower plasma concentrations of clobazam, clonazepam, lamotrigine and active metabolites of oxcarbazepine, phenytoin, tiagabine, topiramate and valproate, and at times ethosuximide and zonisamide. It is intended as a resource to inform decision-making in communities, organizations, and states. This table summarizes the strategies in this technical package, as well as specific ways. The hope is that multiple sectors, including public health, health care, education, social services, and non-governmental organizations, will use this technical package to improve asthma control in all age groups. Commitment, collaboration, and leadership from numerous sectors can maximize the impact of this technical package. This technical package is ready for implementation now; it reflects the mature evidence base about how to control asthma effectively. Fast Facts about Asthma in the United States One in 13 people has asthma (more than 24 million Americans)1 In the United States, 50% of adults with asthma and 40% of children with asthma do not have control of their disease2 Each year, asthma accounts for approximately: ­ 439,000 hospitalizations1 ­ 1. It is a resource to inform decision-making in communities, organizations, and states, by identifying strategies with the greatest potential impact on controlling asthma. Similarly, commitment, collaboration, and leadership from numerous sectors, including public health, health care, education, social services, and non-governmental organizations, can maximize the impact of this technical package. Additional details on these strategies and approaches are available in the Appendix, along with summaries of supporting evidence and relevant sectors well positioned to lead implementation efforts. A technical package is a compilation of a core set of strategies to achieve and sustain substantial reductions in a specific risk factor or outcome. This technical package is a resource to guide and inform decision-making in communities, organizations, and states. Reducing impairment, which includes preventing asthma symptoms, reducing rescue medication use, maintaining lung function, and maintaining normal physical activity levels and attendance at work or school 2. If these study designs were unavailable because of feasibility or ethics, pre­post comparisons were acceptable. Regarding evidence from another country, consideration was given to the feasibility of implementation in a U.


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Cross References Hemiparesis; Paresis; Quadriparesis erectile dysfunction meds list purchase 30gm himcolin visa, Quadriplegia Setting Sun Sign the setting sun sign erectile dysfunction treatment lloyds pharmacy discount himcolin 30 gm without prescription, or sunset sign erectile dysfunction protocol jason order himcolin canada, consists of tonic downward deviation of the eyes with retraction of the upper eyelids exposing the sclera. Setting sun sign is a sign of dorsal midbrain compression in children with untreated hydrocephalus. Cross Reference Dementia Shin-Tapping A modification of the heel­knee­shin test or heel­shin test in which the heel is tapped repetitively on the shin before sliding it down to the foot, claimed to be a better test of motor coordination. Cross References Ataxia; Cerebellar syndromes; Heel­knee­shin test, Heel­shin test Sialorrhoea Sialorrhoea (drooling) is excessive salivation, possibly due to excess flow of saliva but more likely secondary to a reduced frequency of swallowing. Metallic poisonings (mercury, bismuth, lead) may also produce marked salivation (ptyalism). Recently, the use of intraparotid injections of botulinum toxin has been found useful. Botulinum toxin treatment of sialorrhoea: comparing different therapeutic preparations. Cross References Bulbar palsy; Parkinsonism Sighing Occasional deep involuntary sighs may occur in multiple system atrophy. Sighing is also a feature, along with yawning, of the early (diencephalic) stage of central herniation of the brainstem with an otherwise normal respiratory pattern. Recognition of single objects is preserved; this is likened to having a fragment or island of clear vision which may shift from region to region. Two types of simultanagnosia are described: Dorsal: An attentional limitation preventing more than one object being seen at a time; although superficially similar to apperceptive visual agnosia, with which it has sometimes been classified, patients with dorsal simultanagnosia can recognize objects quickly and accurately, but unattended objects are not seen. There may be inability to localize stimuli even when they are seen, manifest as visual disorientation. Ventral: A limitation in the number of objects which can be recognized in unit time, i. Ventral simultanagnosia is most evident during reading which is severely impaired and empirically this may be the same impairment as seen in pure alexia; otherwise deficits may not be evident, unlike dorsal simultanagnosia. This is thought to reflect damage to otolith-ocular pathways or vestibulo-ocular pathways. Skew deviation has been associated with posterior fossa lesions, from midbrain to medulla. Ipsiversive skew deviation (ipsilateral eye lowermost) has been associated with caudal pontomedullary lesions, whereas contraversive skew (contralateral eye lowermost) occurs with rostral pontomesencephalic lesions, indicating that skew type has localizing value. Skew deviation with ocular torsion: a vestibular brainstem sign of topographic diagnostic value. Dysarthria, facial paresis, hemiparesis with or without hemihypoaesthesia, and excessive laughing with or without crying were common accompanying features in one series. Smile­wink phenomenon: aggravated narrowing of palpebral fissure by smiling after lenticulocapsular stroke. Sensory nasal trigeminal afferents run to a putative sneeze centre, localized to the brainstem based on lesions causing loss of sneezing following lateral medullary syndrome and medullary neoplasm. Integration of inputs in this centre reaches a threshold at which point an expiratory phase occurs with exhalation, forced eye closure, and contraction of respiratory musculature. Cross Reference Lateral medullary syndrome Snoring Reduced muscle tone in the upper airway during sleep leads to increased resistance to the flow of air, and partial obstruction often results in loud snoring. Obstructive sleep apnoea­hypopnoea syndrome presenting in the neurology clinic: a prospective 5-year study. Cross Reference Hypersomnolence Snouting, Snout Reflex Sometimes used interchangeably with pout reflex, this term should probably be reserved for the puckering or pouting of the lips induced by constant pressure over the philtrum, rather than the phasic response to a tap over the muscle with finger or tendon hammer. Cross References Frontal release signs; Pout reflex; Primitive reflexes Somatoparaphrenia Ascription of hemiplegic limb(s) to another person. For example, flexor spasms in patients paraplegic due to upper motor neurone lesions are sudden contractions of the flexor musculature, particularly of the legs, either spontaneous or triggered by light touch. Spasm may also refer to a tetanic muscle contraction (tetany), as seen in hypocalcaemic states. Infantile seizures consisting of brief flexion of the trunk and limbs (emposthotonos, salaam or jack-knife seizures) may be known as spasms. This is usually a benign idiopathic condition, but the diagnosis should prompt consideration of an optic pathway tumour. Spasmus nutans-like nystagmus is often associated with underlying ocular, intracranial, or systemic abnormalities. The excessive resistance evident at the extremes of joint displacement may suddenly give way, a phenomenon known as clasp-knife (or, confusingly, clasp-knife rigidity). The amount and pattern of spasticity depends on the location of the lesion and tends to be greater with spinal cord than cortical lesions. Scales to quantitate spasticity are available (Ashworth, modified Ashworth, pendulum test of Wartenberg) but have shortcomings. Spasticity may also vary in distribution: for lesions above the spinal cord it typically affects the arm flexors and the leg extensors to a greater extent (hemiparetic posture). Slow, laboured speech, with slow voluntary tongue movements, may be referred to as spastic dysarthria, which may occur in the context of a pseudobulbar palsy. The pathogenesis of spasticity has traditionally been ascribed to damage to the corticospinal and/or corticobulbar pathways at any level from cerebral cortex to spinal cord. Treatment of severe spasticity, for example, in multiple sclerosis, often requires a multidisciplinary approach. Urinary infection, constipation, skin - 330 - Spinal Mass Reflex S ulceration, and pain can all exacerbate spasticity, as may inappropriate posture; appropriate management of these features may ameliorate spasticity. Drugs which may be useful include baclofen, dantrolene (a blocker of muscle excitation­ contraction coupling), and tizanidine (2 -adrenoreceptor agonist). Intrathecal baclofen given via a pump may also be of benefit in selected cases, and for focal spasticity injections of botulinum toxin may be appropriate. For painful immobile spastic legs with reflex spasms and double incontinence, irreversible nerve injury with intrathecal phenol or alcohol may be advocated to relieve symptoms. This, or a very similar, constellation of features has also been known as cortical dysarthria, aphemia, or phonetic disintegration. Speech apraxia has been associated with inferior frontal dominant (left) hemisphere damage in the region of the lower motor cortex or frontal operculum; it has been claimed that involvement of the anterior insula is specific for speech apraxia. The syndrome is thought to reflect disturbances of planning articulatory and phonatory functions, but is most often encountered as part of a non-fluent aphasia. If not deliberate, it presumably reflects a left hemisphere dysfunction in the appropriate sequencing of phonemes. A variant of this foraminal compression test involves rotation, side bend, and slight extension of the neck with the application of axial pressure to the head. Cross Reference Radiculopathy Square Wave Jerks Square wave jerks are small saccades which interrupt fixation, moving the eye away from the primary position and then returning. This instability of ocular fixation is a disorder of saccadic eye movements in which there is a saccadic interval (of about 200 ms; cf. Very obvious square wave jerks (amplitude > 7) are termed macrosquare wave jerks. Their name derives from the appearance they produce on electrooculographic recordings. Although square wave jerks may be normal in elderly individuals, they may be indicative of disease of the cerebellum or brainstem. In the strike phase, there is a characteristic slapping down of the foot, again a consequence of weak ankle dorsiflexion. Proprioceptive loss, as in dorsal column spinal disease, may also lead to a gait characterized by high lifting of the feet and also stomping (stamping with a heavily accented rhythm) or slapping of the foot onto the floor in the strike phase. This may lead to falls as a consequence of tripping over the foot, especially on up-hill gradients, and a characteristic pattern of wear on the point of the shoe. Whole areas of the body may be involved by stereotypies and hence this movement is more complex than a tic. Examples include patting, tapping, rubbing, clasping, - 333 - S Sternocleidomastoid Test wringing, digit sucking, body or head rocking or banging, grimacing, smelling, licking, spitting, and mouthing of objects. Stereotypies are common in patients with learning disability, autism, and schizophrenia.

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Saivin S erectile dysfunction cream 16 buy himcolin 30 gm with visa, Hulot T erectile dysfunction doctor new orleans order 30 gm himcolin visa, Chabac S icd 9 code of erectile dysfunction buy himcolin cheap online, Potgieter A, Durbin P, Houin G: Clinical pharmacokinetics of acamprosate. Linnoila M, Eckardt M, Durcan M, Lister R, Martin P: Interactions of serotonin with ethanol: clinical and animal studies. Alcohol Clin Exp Res 2000; 24:1041­1049 [B] Treatment of Patients With Substance Use Disorders 237 Copyright 2010, American Psychiatric Association. Edwards G, Brown D, Duckitt A, Oppenheimer E, Sheehan M, Taylor C: Outcome of alcoholism: the structure of patient attributions as to what causes change. J Stud Alcohol 1993; 54:652­666 [A­] Treatment of Patients With Substance Use Disorders 239 Copyright 2010, American Psychiatric Association. Robins E: the Final Months: A Study of the Lives of 134 Persons Who Committed Suicide. 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Degenhardt L, Hall W, Lynskey M: the relationship between cannabis use and other substance use in the general population. Copeland J, Swift W, Roffman R, Stephens R: A randomized controlled trial of brief cognitive-behavioral interventions for cannabis use disorder. Substance Abuse and Mental Health Services Administration: Results from the 2003 National Survey on Drug Use and Health: National Findings. Rockville, Md, Substance Abuse and Mental Health Services Administration, Office of Applied Studies, 2004. J Addict Dis 2002; 21:107­119 [D] Treatment of Patients With Substance Use Disorders 243 Copyright 2010, American Psychiatric Association. Srisurapanont M, Jarusuraisin N, Kittirattanapaiboon P: Treatment for amphetamine dependence and abuse. Yui K, Goto K, Ikemoto S: the role of noradrenergic and dopaminergic hyperactivity in the development of spontaneous recurrence of methamphetamine psychosis and susceptibility to episode recurrence. 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Once the T-complex enters the plant cell nucleus erectile dysfunction caverject injection order discount himcolin, it is further processed to erectile dysfunction help purchase himcolin 30gm amex integrate into the plant host genome by targeted proteolysis of Tcomplex-associated virulence proteins erectile dysfunction dr. hornsby buy himcolin 30gm overnight delivery, for instance (Tzfira et al. By performing extrachromosomal recombination assays, the T-strand has been shown to be imported into the plant cell nucleus as a single-stranded 444 Pieter Windels, Sylvie De Buck and Ann Depicker nucleoprotein complex (Tinland et al. In conclusion, and since experimental evidence has been provided for all the above hypotheses, these mechanisms might be not mutually exclusive. At the same time, these variables will affect tissue viability and regeneration potential, two parameters that are also very important in the recovery of transgenic plants. In most systems, selection for an efficient transformation system is essential, because the fraction of stably transformed cells is usually small (De Buck et al. Indeed, without selection for transformation competence, none of the 84 regenerants, obtained after Arabidopsis root transformation, were stably transformed. Nevertheless, the stable transformation frequency of co-cultivated tobacco (Nicotiana tabacum) protoplasts that regenerate into shoots was found to be relatively high: 18. The difference in competence for stable transformation can be based on several factors, such as the plant genotype, differentiation, physiology of the plant cells (De Buck et al. Because together with the nucleoprotein T-complex also other virulence proteins. As described above, it is wellestablished that the VirD2 protein could influence nuclear uptake (Ballas and Citovsky, 1997; Deng et al. Also, radiosensitive Arabidopsis ecotypes were shown to be recalcitrant for Agrobacterium infection (Nam et al. For instance, the rat5 mutants described above are recalcitrant to stable root transformation, but are easily transformed by flower vacuum infiltration (Mysore et al. Removal of the noncomplementary overhangs might result in the frequently observed truncated left and/or right borders. Polymerase slipping and extensive template switching explained more complex filler. An initial interaction between the right border and the plant target at a D-loop occurs after the left border end loops back to anneal to itself. This initial pairing is followed by removal of non-complementary overhangs by exo- or endonucleases. This second pathway was observed Agrobacterium Tumefaciens-Mediated Transformation 453 in approximately 50% of all analyzed junctions (Windels et al. Mutual microhomologies among the sequence motifs that constitute a filler segment were frequently observed. The maintenance of the information and structural organization of genomes is a prerequisite for the expression as well as for the transfer of this genetic information to next generations. As a consequence, prokaryotic as well as eukaryotic organisms dispose of mechanisms to deal with these kind of lesions. After transformation, circularized plasmids were recovered and the novel joints were sequenced. In Arabidopsis, deletions were on average larger than in tobacco and not associated with insertions (Kirik et al. However, a drawback was that many studies used Arabidopsis that is known for its very small genome with relatively few heterochromatic genome regions. Therefore, the observed preference for transcriptionally active gene-rich genomic loci could be explained as an artefact of the study design. However, it should be noted that most of the above described lines were obtained after selecting or screening for transgene expression, implying that transformants with integrations into genomic regions that suppress transcription of the selection marker will not be identified. The presence of VirD2 and VirE2 proteins has been implicated in conservation of the integrity of the T-strand. In the latter case, a number of variations have been observed: tandem repeat organization (upper), inverted repeat organization over the right border (middle), and inverted orientation over the left border (bottom). More complex integration patterns, consisting of a mixture of all the above have also been reported (not shown in figure). For each type of junction, a schematic overview is presented of the structure and the transformants in which they occur. Several possibilities have been put forward, but neither of them has been confirmed and contradictory results have been reported. The high frequency of inverted repeats has been attributed to the use of nopaline-type Vir functions (Jorgensen et al. The frequency of single-copy transformants is much higher after Arabidopsis root transformation than after floral dip transformation (De Buck et al. Transfer could erroneously start at the left border, proceeding through the binary vector, toward the right border, and finish only when the left border is encountered for the second time (Ramanathan and Veluthambi, 1995; van der Graaff et al. Alternatively, backbone Agrobacterium Tumefaciens-Mediated Transformation 467 transfer could be the result of initiation of transfer at the right border and readthrough over the left border (Kononov et al. The frequency of these backbone sequences might rise up to 75% or 80% from the transgenic population (Kononov et al. Recently, the presence of four copies of the left border repeat have been shown to positively prevent readthrough at the left border in rice transformants (Kuraya et al. Most of the target site deletions reported in the literature are smaller than 75 bp (Gheysen et al. Taken together, the process of illegitimate recombination usually introduces only small target site rearrangements. The most dramatic rearrangement was a 158-bp duplication of the plant target in combination with a 27-bp deletion. The final outcome of the chromosomal rearrangement would require two independent recombination events what is also improbable (Nacry et al. Essentially, two different models account for the paracentromeric inversion observed (Laufs et al. Mol Breed 6: 459-468 De Buck S, Jacobs A, Van Montagu M, Depicker A (1998) Agrobacterium tumefaciens transformation and cotransformation frequencies of Arabidopsis thaliana root explants and tobacco protoplasts. Plant Mol Biol 11: 365-377 Dillen W, De Clercq J, Kapila J, Zambre M, Van Montagu M, Angenon G (1997) the effect of temperature on Agrobacterium tumefaciens-mediated gene transfer to plants. Trends Plant Sci 8: 380-386 Fladung M (1999) Gene stability in transgenic aspen (Populus). Microbiol Mol Biol Rev 67: 16-37 Gheysen G, Angenon G, Van Montagu M (1998) Agrobacterium-mediated plant transformation: a scientifically intriguing story with significant applications. Bio/Technology 6: 185-189 Grunstein M (1997) Histone acetylation in chromatin structure and transcription. Plant Mol Biol 52: 761-773 Kirik A, Salomon S, Puchta H (2000) Species-specific double-strand break repair and genome evolution in plants. Plant Mol Biol 52: 247-258 Komari T, Ishida Y, Hiei Y (2004) Plant transformation technology: Agrobacterium-mediated transformation. Plant Cell 8: 873-886 Negruk V, Eisner G, Lemieux B (1996) Addition-deletion mutations in transgenic Arabidopsis thaliana generated by the seed co-cultivation method. In H Jones, ed, Plant Gene Transfer and Expression Protocols, Methods in Molecular Biology, Vol 49. Nature 388: 900-903 Tzfira T, Citovsky V (2002) Partners-in-infection: host proteins involved in the transformation of plant cells by Agrobacterium. Theor Appl Genet 105: 878-889 Valentine L (2003) Agrobacterium tumefaciens: the David and Goliath of modern genetics. J Mol Biol 86: 109-127 Zambre M, Terryn N, De Clercq J, De Buck S, Dillen W, Van Montagu M, Van Der Straeten D, Angenon G (2003) Light strongly promotes gene transfer from Agrobacterium tumefaciens to plant cells. Genetic transformation results from a complex interaction between Agrobacterium and host plant cells. However, we understand much less about the plant contribution to the transformation process. Plant species, and even varieties/ecotypes, differ markedly in their susceptibility to Agrobacterium. A genetic component underlies these differences, permitting scientists to identify specific host genes and proteins mediating transformation. In this chapter, I review what is known about the plant contribution to transformation, and the tools which scientists are using to reveal the mechanisms by which host genes and proteins function in various steps of the transformation process. However, our knowledge of the host contribution to the transformation process has lagged. It is clear that although Agrobacterium has an enormous host range encompassing species of numerous phylogenetic kingdoms, differences in susceptibility to Agrobacterium-mediated transformation abound among plant species.

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This chapter begins by describing the overarching principles for human genome editing adopted by the committee for this study homemade erectile dysfunction pump order himcolin pills in toronto, which are informed by those international instruments and national rules sublingual erectile dysfunction pills order himcolin master card, and which in turn inform the conclusions and recommendations presented in this report impotence causes and cures order himcolin 30gm free shipping. But while science is global, it proceeds within a variety of political systems and cultural norms. It is important to identify principles that can transcend these differences and divisions, while accommodating cultural diversity. Achieving consensus around overarching ethical principles to undergird specific recommendations for action can be difficult, whether because no one theory of ethics has been accepted by philosophers and theologians or because no one algorithm for deriving principles from those theories has been found. Utilitarians may agree on the need to evaluate overall beneficial consequences, but may disagree on whether to evaluate the consequences of a rule or of a specific act. It has also been dogged by long-standing debates about whether the best approach is high theory, from which all principles and specific actions flow, or anti-theory, in which deductive reasoning from specific cases leads to generalizable principles. And when bioethics is incorporated into public policy making, as opposed to individual clinical ethics analyses, it is necessary to incorporate a wider range of concerns about multicultural civil society, theories of democracy, and just distribution of burdens and benefits. It has helped shape influential statements and guidance documents across the globe. In its preamble, it states that "recognition of the inherent dignity and of the equal and inalienable rights of all members of the human family is the foundation of freedom, justice and peace in the world," and its very first provision reads, "All human beings are born free and equal in dignity and rights. Not every convention is legally binding in whole or part on every country, but even where not incorporated into domestic statutes or applied in domestic court cases, the principles underlying these conventions have become important elements of global norms and aspirations. Of particular relevance to genome13 14 Further discussion of these issues can be found in Arras (2016). Also of particular relevance is Guideline 7 on public engagement, needed not only to develop and legitimize good policy but also to help translate research into clinical benefit. These pillars of research ethics have been interpreted, expanded, deepened, and applied over the years and incorporated into the U. In practice, they have resulted in a focus on ensuring a reasonable balance between risk and hoped-for benefits, to the individual and to society, and on ensuring that both risks and benefits are equitably shared. These principles also have come to incorporate particular attention to the need for respect of individual autonomy, in the form of generally requiring informed and voluntary participation, and the need to provide special protection against coercion or abuse of those who are vulnerable because of incapacity or circumstances. Because both the science and the applications of genome editing will transcend national boundaries, the core principles for governance of human genome detailed below build on the foundations of these international and national norms. Some of these principles are generally relevant to biomedical research and care, while others are of particular importance in the context of an emerging technology, but all are foundational for the governance of human genome editing. In this context, the committee focused on principles that are aimed at protecting and promoting the health and well-being of individuals; approaching novel technologies with careful attention to constantly evolving information; respecting individual rights; guarding against unwanted societal effects; and equitably distributing information, burdens, and benefits. Differences in social and legal culture inevitably will lead to different domestic policies governing specific applications of genome editing. Thus, while the overarching principles presented here are aimed primarily at the U. Along with this promise comes the need for responsible and ethically appropriate approaches to research and clinical use. The following general principles are essential foundations for those approaches: 1. Promoting well-being: the principle of promoting well-being supports providing benefit and preventing harm to those affected, often referred to in the bioethics literature as the principles of beneficence and nonmaleficence. Responsibilities that flow from adherence to this principle include proceeding cautiously and incrementally, under appropriate supervision and in ways that allow for frequent reassessment in light of future advances and cultural opinions. Transnational cooperation: the principle of transnational cooperation supports a commitment to collaborative approaches to research and governance while respecting different cultural contexts. Responsibilities that flow from adherence to this principle include (1) respect for differing national policies, (2) coordination of regulatory standards and procedures whenever possible, and (3) transnational collaboration and data sharing among different scientific communities and responsible regulatory authorities. At the national level, regulation may be mandatory in all cases-for example, when the work is to be submitted to the U. Oversight also can proceed according to voluntary self-regulation pursuant to professional guidelines. In addition to national rules, individual states have at times issued rules on specific topics, such as embryo research, or attached restrictions to the use of state funds, such as for embryonic stem cell work. As a result, unlike some jurisdictions, such as the United Kingdom, in which work with embryos generally falls under a single statutory framework or regulatory body, the United States has individual rules related to stage of work and source of funding that overlap and interact in a manner that, in the end, provides fairly comprehensive coverage. Preclinical animal work is subject to regulation and oversight by institutional animal care and use committees pursuant to the Animal Welfare Act. Postmarket use may also encompass uses that go beyond the indications for which a therapy was approved. But outside of a study, "off label" use in clinical care is entirely legal, and has become a common practice among physicians with respect to drugs, and might be available for a gene transfer product using genome editing once it is approved. Physicians use their own expertise and sources of information, as well as the advice of professional societies. Table 2-1 provides a summary of the major steps in the anticipated regulatory pathway for the development of a new medical product created using genome editing. The individual steps and considerations listed in this table are discussed in greater detail in the remainder of the chapter. For most cells and tissues, an initial question is whether the donor can receive any kind of payment, in cash or kind. This has been a particularly sensitive issue with respect to gametes used in research, with debate being focused less on the ethics of research using gametes and more on the ethics of how they are obtained and whether it involves anything that resembles undue inducement. For embryos and fetal tissue, rules are influenced by broader legal regimes governing human reproduction and products of conception, to the extent such regimes exist in a given country. These rules can change, of course, when tissue is obtained from cadavers rather than live donors. This pattern of regulation is the same regardless of whether genome editing will be carried out on the tissues. This biosafety review is accomplished by assessing the appropriate physical and biological containment for the research and ensuring the researchers are adequately trained to conduct the work they are proposing safely. Human Tissue Use and Institutional Review Boards Laboratory-based research using human tissue may also trigger certain human subjects protections, even though all the work is done in vitro. The rules will change with respect to research using stored human specimens upon the effective date of the January 2017 revisions to the "Common Rule" that sets out the framework and requirements for human subjects research that is funded by most federal agencies and departments, or is otherwise subject to its jurisdiction. The common rule also applies to research conducted at institutions that have voluntarily extended its application to research funded by sources other than those listed above. Where the investigators do not have easy access to a key to break the code, the tissue will no longer have a personal identity that is "readily ascertainable. Furthermore, except when eligible for waiver, informed and voluntary consent is required from the research subject (in the present context, the person whose tissue is being used) or from a legally authorized representative. Additional Rules Governing Laboratory Research on Human Gametes and Embryos Basic science research on genome editing may entail experimentation on human gametes and embryos, with no intention of performing intrauterine transfer to establish a pregnancy in a woman (see Chapter 3). Indeed, such in vitro research on embryos has already proceeded in China (using nonviable embryos) and has been approved (with viable embryos) by the relevant regulatory bodies in Sweden and the United Kingdom. Work also is proceeding on understanding human germ cell development, research in which genome editing is one of many tools that can be used to explore the roles of specific genes (Irie et al. This laboratory research might take a number of forms, each raising slightly different ethical and legal issues. First, it might involve editing somatic tissue in such a way that gametes would or might also be affected. Second, it might involve editing an existing gamete or gamete progenitor, such as a spermatogonial stem cell, in vitro or in vivo. Its conclusions, reflect the view that embryos should be regarded as different from ordinary human tissue but nonetheless be used for some areas of research if in the service of important scientific knowledge that cannot be obtained with less controversial methods. Except in very limited circumstances, the panel called for use of only those embryos that, although originally created in the course of a reproductive effort, now would otherwise be discarded. Regulatory protections for human research subjects do not apply to the ex vivo embryo. The work can, however, be supported with funds from individual states and private sources, often with policies similar to those proposed by the 1994 embryo research panel. California, for example, has been funding embryo research and embryonic stem cell research for a decade using funds from a state bond issued during the years when federal funding was limited to a small number of older embryonic stem cell lines. The amendment has been attached to the annual appropriations bills for the Departments of Health and Human Services, Labor, and Education since 1996. A single-cell fertilized egg is treated as if it were an embryo for most relevant state and federal laws, and restrictions on the work or on the funding would apply. In addition, such a step would constitute making an embryo solely for research purposes (that is, without any intent to gestate the embryo and bring a fetus to term), and this has remained the most controversial form of embryo research in the United States.

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Banta and Maywa Montenegro for most important crop species are now available (Rommens impotence meme discount himcolin express, 2004) erectile dysfunction quiz buy himcolin online now. By selecting against backbone integration events as described above erectile dysfunction treatment protocol order himcolin amex, this approach can be used to generate completely marker-free transgenic plants at a frequency that is consistent with commercial scale production (Rommens et al. Expression levels of the transgene can be dramatically affected by the chromosomal context of the integration site, and insertional disruption of an active host gene can have unintended phenotypic consequences on the resulting plant (Kumar and Fladung, 2001). In at least one study, targeted insertions into the same site did result in reproducible transgene expression levels; however, in nearly half the insertion events, partial or complete silencing of the transgene was observed (Day et al. Such "position effects" are consistent with our growing appreciation for the striking variability and unpredictable nature of transgene expression levels, a ubiquitous phenomenon in almost all eukaryotes. In the face of repressive influences exerted on transgenes by neighboring genes or the surrounding chromosomal structure, the standard, albeit costly, approach has been to generate enough transgenic plants to find some with the desired level of expression (Hansen and Wright, 1999). In the context of plant transformation, transgene silencing also results from insertion of multiple copies or high-level expression from a constitutive promoter, and an introduced transgene can lead to silencing of a homologous host gene (Vaucheret et al. Multicopy transgenic loci, particularly those including binary vector sequences, appear prone to transcriptional silencing attributable to meiotically heritable epigenetic modifications, most often methylation and/or condensation of chromatin (Matzke and Matzke, 1998; Vaucheret et al. Strikingly, the enhancement is attributable to amplification of the gene copy number as well as increased transcription, and both changes are stably inherited (Borisjuk et al. Targeted transgene integration via site-specific recombination can be combined with a second recombination system that eliminates the selectable marker gene (Srivastava and Ow, 2004). A variety of approaches have been used to stabilize the insertion and prevent subsequent Cre-mediated excision (Gilbertson et al. Several refinements of this procedure enabled the first targeted disruption in a monocot, rice (Terada et al. Finally, it is plausible that recombination occurs more readily in the highly proliferative callus tissue typically used in rice transformation than in the plant tissues used in other transformations (Shimamoto, 2002). The gene targeted for disruption in this application was Waxy, which encodes granule-bound starch synthase. The success of this gene targeting process in rice paves the way for other gene knockouts in this important staple crop to study gene function or to alter nutritional or growth traits. Successful implementation of this approach was achieved using Agrobacterium-mediated transformation to introduce a synthetic zinc-finger nuclease that then created the double-stranded break (Lloyd et al. These zinc-finger nucleases consist of custom-made C2H2 zinc fingers, with each finger recognizing a specified three-nucleotide sequence, fused to a non-specific restriction enzyme. Using a heat-shock promoter to drive production of the zinc-finger nuclease in Arabidopsis, Lloyd et al. In a second approach to increasing the frequency of directed gene disruption or replacement, Shaked et al. On the bacterial side of the interaction, certain virulence loci including virC and virF are considered host range determinants (Yanofsky et al. Genes within the T-region can also affect the range of susceptible host species (Hoekema et al. The manipulation of host genes to improve transformation frequency is the subject of two recent reviews (Gelvin, 2003a; Gelvin, 2003b). Bacterial and plant contributions to host range are discussed in more detail in Chapters 1 and 13, respectively, in this volume. It is worth noting that there are almost certainly more factors yet to be identified that 88 Lois M. For example, maize root exudates contain a potent inhibitor of VirA/VirGmediated signal perception, leading to the possibility that bacterial mutants with enhanced resistance to this inhibition may prove useful in extending the transformation efficiency of maize (Zhang et al. One approach to circumvent host range limitations involves the use of Agrobacterium rhizogenes to generate composite plants, comprised of transgenic roots on wild-type shoots. This system provides a useful method to study transgene activity in the root in the context of a wild-type plant, and has been used successfully in species such as soybean, sweet potato and cassava, that are recalcitrant to A. Somewhat ironically, of all the advances in plant transformation described in this chapter, some of the most pronounced long-term impacts on plant biotechnology may result from an innovation that has the potential to obviate the requirement for Agrobacterium as a gene delivery vehicle. Motivated by the desire to "invent around" the myriad intellectual property constraints that limit use of Agrobacterium-mediated transformation by the public and the private sector, Broothaerts et al. Various tissues, and hence transformation mechanisms (floral dip for Arabidopsis, somatic tissue for tobacco and rice), were utilized in these experiments, and stable integration was confirmed by Southern blotting, sequence analysis of the insertion junctions, and Mendelian transmission of the transgene to progeny. This alternative technology may have Agrobacterium and Plant Biotechnology 89 profound implications for the plant biotechnology community for two reasons. First, this technology has been configured to be freely accessible and "open-source," with no commercial restrictions other than covenants for sharing improvements, relevant safety information, and regulatory data. Second, the exceptionally broad host range of the Rhizobium strain used, and the potential to extend the technology to additional bacteria species, make it likely that previously recalcitrant plant species may become transformable. As a plant pathogen, Agrobacterium elicits a variety of defense responses that can block any step of the transformation process, thereby limiting its host range. Protoplast transformation, although achievable through electroporation, microinjection, or polyethylene glycol fusion, proved to be inefficient because the regeneration of plants from protoplasts is time-consuming and non-trivial (Newell, 2000). This biolistic approach presents certain advantages over Agrobacterium-mediated gene delivery; many types of explants can be bombarded and yield fertile plants, and the gene to be delivered need not be cloned into a specialized transformation vector (Herrera-Estrella et al. These complex integration patterns can lead to genetic instability, due to homologous 90 Lois M. Banta and Maywa Montenegro recombination among the identical copies, and/or epigenetic silencing of the transgene (see section 2. The insertion events resulting from in planta VirD1/2-mediated processing and integration resemble those generated by traditional Agrobacteriummediated transformation (Hansen and Chilton, 1996). Heterologous genes can also be introduced into plants on viral vectors; because of the amplification associated with viral infection, transient expression of the transgenes can yield commercial-scale quantities of pharmaceutical proteins. Additional refinements of the viral vectors further enhanced the efficiency of the system, which was limited by the low infectivity of viral vectors carrying larger genes and apparently by nuclear processing of a viral transcript that normally never experiences the nuclear milieu (Marillonnet et al. By infiltrating whole mature plants with a suspension of agrobacteria carrying the encoded viral replicons, the bacteria take on the viral infection function, while the viral vector mediates cell-to-cell dissemination, amplification, and high-level expression of the transgene (Gleba et al. This "magnifection" process is rapid and scalable; the modular nature of the viral components facilitates adaptation to new transgenes, and the yield can reach 80% of total soluble protein (Marillonnet et al. Most applications to date have focused on field-grown plants, although recombinant proteins and metabolites can also be produced in plant cell cultures. In the future, functional genomics and combinatorial biochemistry are likely to increase dramatically the range of products that can be generated in genetically modified plant cell cultures (Oksman-Caldentey and Inze, 2004). Drawing on the natural ability of many bacterial species, including Ralstonia eutropha, to synthesize carbon storage products with plastic-like properties (Hanley et al. Further increases in yield, from 14% to as much as 40% of the plant dry weight, were achieved by using gas chromatography-mass spectrometry to screen large numbers of transgenic Arabidopsis plants for high levels of production; however, the high producing lines exhibited 92 Lois M. Banta and Maywa Montenegro stunted growth, loss of fertility, and significant alterations in the levels of various amino acids, organic acids, sugars, and sugar alcohols (Bohmert et al. Although these transgenic plants exhibit morphological alterations in chloroplast structure and in growth rate, additional engineering of the amino-acid biosynthesis pathways may permit economically viable levels of biodegradable plastic production (Conrad, 2005). If successful, the substitution of a renewable process (solar-driven carbon fixation) for conventional petrochemically derived plastic production technologies would have substantial positive environmental consequences, decreasing our reliance on finite petroleum resources, while reducing the accumulation of indestructible plastics (Poirier, 1999; Conrad, 2005). Terpenoids, also known as isoprenoids, are a family of more than 40,000 natural compounds, including both primary and secondary metabolites, that are critically important for plant growth and survival. Some of the primary metabolites produced by the terpenoid biosynthetic pathway include phytohormones, pigments involved in photosynthesis, and the ubiquinones required for respiration (Aharoni et al. Secondary metabolites, including monoterpenoids (C10), sesquiterpenoids (C15), diterpenoids (C20), and triterpenoids (C30), also provide physiological and ecological benefits to plants. Some function as antimicrobial agents, thus contributing to plant disease resistance, while other terpenoid compounds serve to repel pests, attract pollinators, or inhibit Agrobacterium and Plant Biotechnology 93 growth of neighboring competitor plant species. Additionally, many terpenoids have commercial value as medicinals, flavors, and fragrances. The terpenoid biosynthetic pathway and strategies for its manipulation have been reviewed recently (Mahmoud and Croteau, 2002; Aharoni et al. A comprehensive listing of transgenic plants with altered terpenoid biosynthetic properties is available elsewhere (Aharoni et al. Examples include expression of heterologous synthases in tomato, leading to enhanced aroma in ripening fruit (Lewinsohn et al. Other endogenous, plant-derived terpenoids with demonstrated pharmaceutical properties include the anti-malarial agent artemisinin, the diuretic glycyrrhizin, and the cancer drugs Taxol and perilla alcohol.


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