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Between April 2001 and March 2005, 1986 patients were diagnosed with primary stage 1 human African trypanoi i somiasis. Criteria for selection of the cohort retained for analysis is shown in Fig. 2. Of 692 patients discharged alive after treatment with pentamidine, 454 66% ; and 371 54% ; attended followi up at 6 months 2 months ; and one year 2 months ; , respectively. Overall, 652 patients 94% ; attended at least one. Although protoplasts from 'Just Right' turnip were stable and supported CaMV replication in various culture media, the most reproducible results were obtained when a modified Km8p medium Kao & Michayluk, 1975 ; was used in place of a simple salts mixture Takebe et al., 1968 ; . All the Brassiea protoplasts cultured in modified Km8p showed similar morphological changes with time which were independent of the PEG treatment used in virus inoculation. Upon isolation, leaf mesophyll protoplasts of turnip ranged in diameter from 10 Ixm to 30 ~tm. After culture in Km8p for 48 h these had swollen to approximately three times their initial size compare Fig. 1 c and 1f ; . Protoplasts developed many buds, some of which became as large as the original protoplast; protoplasts did not divide within the period of the experiments approx. 5 days ; . No correlation was observed between a particular protoplast type size, presence or absence of chloroplasts ; and the replication of CaMV; all protoplasts appeared to be able to accumulate CaMV. This also applied to subprotoplasts, present 72 h after inoculation, many of which contained either no large organelles, or only a nucleus. The possibility that anucleate subprotoplasts are able to synthesize mature CaMV particles is currently under study. Hybridization of C a protoplasts Using a quantitative spot hybridization assay for CaMV D N A was estimated that infected ceils in the inoculated and systemically infected leaves of 'Just Right' turnip contain an average of 2-6 x 103, and 104 to 105 CaMV genome equivalents infected cell respectively Maule et al., 1983 ; . Attempts to measure the amount of progeny CaMV in protoplasts infected in vitro were complicated by the retention of some of the inoculum by the protoplasts. However, the amount of virus inoculum, detected as CaMV DNA, in poorly or non-infected protoplasts decreased rapidly during protoplast incubation. Extrapolation of this decrease suggests that by 72 h postinoculation all the CaMV D N A detected by hybridization is progeny CaMV DNA. Based on this assumption each infected protoplast from 'Just Right' turnip contains an average of 104 to l0 s genome equivalents late in infection Fig. 2 ; , although there was variation between experiments, particularly with regard to the rate of viral D N A synthesis and the final level of CaMV D N A compare expt. 1 and expt. 2, Fig. 2 ; . The estimates of the yield of progeny viral D N A are less straightforward for the other species where the time course of D N synthesis appears to be different Fig. 2 ; , most noticeably with rape, where there is a significant increase in viral D N A post-inoculation. This was not accompanied by strong staining with fluorescent antibody, most virus accumulation occurring at 72 h post-inoculation. For chinese cabbage, cabbage and mustard, which all showed fewer infected protoplasts Table 1 ; , the average amount of progeny viral D N A was considerably lower than in turnip protoplasts. Hence effective comparisons of the kinetics of CaMV D N A synthesis between these species and either turnip or rape were not possible. DISCUSSION In many plant virus-host interactions the use of efficient protoplast systems has led to a better understanding of some aspects of virus replication and its implications for the host. Unfortunately in the study of CaMV, so far the contributions made using protoplasts have been few. This paper describes an alternative technique to that published Furusawa et al., 1980 ; , with which protoplasts from a range of Cruciferous species can be readily infected with CaMV following PEG-mediated uptake of virus particles. The routine infection of a high proportion of the protoplasts inoculated permits the measurement of viral nucleic acid produced following viral multiplication. The infection of S. alba and more particularly M. arvensis, in addition to several Brassica species, demonstrates the efficacy of the technique and its potential in extending the range of protoplast hosts to include those derived from susceptible and resistant plants. An estimate of the number of genome equivalents in each infected turnip protoplast, by spot hybridization, was similar to an estimate of the number of virus particles per protoplast made by electron microscopy Yamaoka et al., 1982b ; . These calculations indicate that leaf mesophyll protoplasts from mature turnip leaves are demonstrating a competence for supporting viral.

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Problem by learning how to count particles indirectly. They did this by measuring samples of the chemicals in particular ratios by their weights. To understand the means of doing this, we need to expand our concept of atomic weight to compounds in the form of the formula or molecular ; weight. a. Milligram Formula Milligram Molecular ; Weight. When atoms combine to form compounds, the atomic nuclei are not affected. There is no net loss of weight. Regardless of whether the particle formed is a molecule or an ion group, it will have a formula and a formula weight. The formula weight of a compound is the sum of the atomic weights of all the atoms that appear in its chemical formula. Consider, for example, carbon dioxide: Atoms: C + O CO2 molecule ; Atomic weights: 12 + 16 formula weight ; While we have arrived at a formula weight which is in terms of atomic mass units, it is much more useful to express it in terms of milligrams. This is known as the milligram formula weight. For the example above, CO2, the milligram formula weight is 44 mg. This is a quantity that we can measure and see, and thus can easily work with. It also represents a reacting unit of the compound. b. Molarity. A molar solution, or a one molar 1M ; solution, consists of onegram molecular weight GMW ; of solute dissolved in enough water to make 1 liter of finished solution. Molarity, then, is the number of GMWs dissolved in enough water to make a finished solution of 1000 ml. Molar solutions may have as a solute a solid, a liquid, or a gas. Later in this subcourse, we will use the concept of molarity to explain the measurement of acidity, called the pH. 1 ; Calculating the gram molecular weight. One-gram molecular weight of a substance is its molecular weight expressed in grams. Thus, a GMW of NaOH would be 40 grams, where the atomic weights are as follows: Na 23, O 16, and H 1. Thus, .5 GMW of NaOH would be 20 grams, and so forth. A mole is one-gram molecular weight of a substance. Thus, a mole of NaOH is 40 grams of NaOH; a halfmole .5 mole ; is 20 grams; two moles of NaOH are 80 grams, and so on. 2 ; Calculating the molarity of a solution. To find the molarity of a solution, we divide the number of gram molecular weights of solute by the number of liters of total solution. The formula may be written: Molarity no. of GMWs of solute no. of liters of solution and celestone.

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Mix. Sp. gr. 0.928, percentage of Ethyl Alcohol, by weight 45.5, by volume 53. The 1870 U. S. diluted Alcohol was made as follows and cerezyme. GUIDELINE FOR MANAGEMENT OF OTITIS MEDIA Statement: Physicians treating children should be aware of the increasing antibiotic resistance of the most common pathogens that cause acute otitis media AOM ; . PHP promotes prevention strategies, accurate diagnosis, and treatment of otitis media based upon the recommendations of the Centers for Disease Control and Prevention CDC ; and current literature. Definitions Acute otitis media AOM ; Bulging or opacification of the tympanic membrane, the mobility of which to pneumatic otoscopy was either absent or markedly decreased; accompanied by one or more signs of acute infection, such as fever, earache, irritability, ottorhea, anorexia, vomiting or diarrhea. Otitis media with effusion OME ; Fluid in the middle ear without signs or symptoms of an acute infection. Usually does not need antimicrobial treatment. Guidelines: 1. Prevention Strategies Complete immunizations including vaccine for Streptococcus pneumoniae, PnC7 PrevnarTM ; for all children 2 yrs. of age and high-risk children 5 yrs. of age. Influenza vaccine for all children ages 6 months - 2 years. Encourage families to eliminate the risk factors for AOM EBM: 1a, 3a, 5 ; A. Exposure to tobacco smoke B. Horizontal bottle feeding C. Use of pacifiers after 10 months of age D. Large day care settings if child has recurring AOM E. Eliminate, if possible, known allergens and treat allergies when appropriate 2. Medical Management A. B. C. Use pneumatic otoscopy to enhance the accuracy of the diagnosis of otitis media. EBM: 4, 5 ; Consider symptomatic treatment with analgesics and OTCs if first or sporadic episode of AOM in low-risk children 2 years old. EBM: 3a, 5 ; When the clinical decision is made to prescribe antimicrobials it is recommended that amoxicillin at at doses between 45-90 mg kg 24 bid be selected as the drug of choice. See table 1. If allergic to penicillin, then use 2nd generation cephalosporin or appropriate macrolide. EBM: 3a, 5 ; If within 72 hours, the child is not clinically improving, a reexamination is suggested. An alternative antibiotic is recommended. Consider the use of amoxicillin clavulanate Augmentin ; , or cefprozil Cefzil ; . See table and algorithm for details. EBM: 3a, 5 ; Follow up exam in 3 to weeks if otherwise improving. EBM: 3a, 5 ; If AOM recurs and more than 3 months has lapsed since the previous AOM, consider managing these sporadic episodes similar to a de novo infection. If the period is shorter between cases or if the child has been on antimicrobials for another reason, use a second line antimicrobial. See algorithm for details. EBM: 2a, 3a, 5.

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