Fijivirus and it is transmitted by the rice whitebacked planthopper (WBPH), Sogatella furcifera (Horváth). It was found that SRBSDV infected vectors might contribute negatively to the WBPH population, although the longer nymphal period might benefit viral acquisition, transmission and increase infection rate. The interaction between SRBSDV and its vector need to be further explored to gain better understanding of the dispersal of WBPH and the spread of virus disease, in particular the feeding and reproduction behavior of viruliferous WBPH.Southern rice black-streaked dwarf virus (SRBSDV) is a recently discovered member of the genus Newly hatched nymphs of WBPH were fed on healthy rice plant after feeding on SRBSDV-infected rice plants for 2 h, and newly emerged adults were numbered and tested. Feeding behaviors of WBPH adults were monitored electronically within a Faraday cage using a Giga-4 DC EPG amplifier. The newly emerged adults were paired, and the fecundity and egg hatchability were investigated. WBPH was molecularly identified for SRBSDV when they dead. According to the identification results, data on viruliferous and non-viruliferous WBPH were collected and analyzed.Feeding behavior of viruliferous WBPH was different from those of non-viruliferous WBPH. Frequency of phloem sap ingestion of viruliferous WBPH increased significantly, however the total feeding duration did not increase markedly. When both WBPH parents were infected with SRBSDV, their fecundity and hatchability of the eggs produced were significant lower than those of normal WBPH parents. However, if only one of the parents was viruliferous, fecundity and egg hatchability were only slightly affected.Viruliferous WBPH fed on the phloem more frequently than non-viruliferous WBPH and can thus contribute to virus transmission. When both vector parents are viruliferous fecundity and hatchability of the eggs were significantly reduced. However when only one of the parents WBPH was viruliferous, there were no significant effects. Southern rice black-streaked dwarf virus (SRBSDV) transmitted by the whitebacked planthopper (WBPH), Sogatella furcifera (Horváth), is a recently identified member of the genus Fijivirus in the family Reoviridae. The virus disease has caused disastrous damages in rice and maize production in China [in China ,2. It wain China ,4 and Jain China . In 2010in China ,4,6 whilin China . The losin China ,7.Since it was discovered, some studies had been conducted. Pu et al. (2012) showed that the frequency of viruliferous WBPH increased with the longer access time after 24 h virus feeding . VirulifThere are generally different effects of viruses on the biology of vector insect ,12. EcolIn this paper we report studies on the feeding behavior of viruliferous and non-viruliferous WBPH. In addition we also assessed the effects on the reproduction charactersistics of WBPH when both or one of the parents infected with SRBSDV. The results would help to further understand the relationship between SRBSDV and its vector to improve the management of WBPH and the SRBSDV disease.The EPG waveforms recorded from viruliferous and non-viruliferous WBPH were similar. There were seven distinctive waveforms, non-penetration (np), penetration initiation N1), salivation and stylet movement (N2), extracellular activity near the phloem region (N3), pathway phase (NC), intracellular activity in the phloem tissue (N4a), phloem sap ingestion (N4b), stylets in the phloem tissue (N4) (including N4-a and N4-b), and stylets in the xylem tissue (N5)  × ♂(−)), the fecundity of viruliferous pairs of male and females were significantly decreased by 76.25%. The hatchability of eggs from these pairs was also reduced by 30.72%. When only one of the pairs was viruliferous fecundity and egg hatchability were slightly affected and not significantly different from the control (Table Compared to control (♀Maize streak virus (MSV) were found in cells of phloem parenchyma, vascular bundle sheath and mesophyll tissue, and also in epidermal guard cells of plants infected with the maize strain of MSV. When the vector leafhopper Cicadulina mbila fed on MSV-infected plants, its feeding preferences was altered and the duration of feeding in mesophyll cell was prolonged. On healthy plants they primarily fed on the phloem. The feeding behavior changes benefited MSV by improving the acquisition and transmission efficiency of the vector [Circulifer tenellus (Baker), was also found to be related with the transmission of Beet severe curly top virus (BSCTV). Waveform D1, which was associated with phloem salivation, was the only waveform correlated with inoculation of BSCTV [Rice ragged stunt virus (RRSV)-carrying BPH preferred healthy plants to infected plants which might mean that plant viruses may alter host selection preference of vectors to enhance their spread and that of insects vectoring another virus to result in co-infection with more than one virus [Feeding behavior of the viruliferous vector may be altered by the plant virus. For instance the virus particles of e vector . Feedingof BSCTV . Our resne virus . HoweverOur earlier studies showed that the fitness of viruliferous WBPH was affected. The nymphal duration of the male was prolonged markedly, the weight and fecundity of the female decreased significantly and the average survival time was shortened at both test temeperatures of 26°C and 31°C . It was reported that the viruliferous WBPH laid significant less eggs than non-viruliferous hoppers. There were no significant differences in the hatchability of eggs laid by virulifierous and non viruliferous females . This stTomato yellow leaf curl virus (TYLCV) infested vectors Bemisia tabaci (Genadius) also have about for 17-23% shorter longevity and reduced fecundity of about 40-50% [Frankliniella occidentalis (Pergande), exposed to Tomato spotted wilt virus (TSWV) compared to the unexposed thrips reared on uninfected leaf discs, mean daily fecundity was reduced and the egg hatchability affected as well [Laodelphax striatellus , especially the male, infected with Rice striped virus (RSV) also had significant effects on the egg hatchability of the next generation [Nephotettix cincticeps (Uhler), were significantly increased when they were fed on Rice dwarf virus (RDV)-infected rice plants [Viriliferous vectors have reduced fecundities ,21. Tomat 40-50% . Similarneration . On the e plants .Tomato yellow leaf curl China virus (TYLCCNV) and its betasatellite can repress a-regulated defences of tobacco against invasive whiteflies Bemisia tabaci (Gennadius) and accelerate population increases of the insects [Furthermore, Co-infection of the begomovirus  insects . The triOur results showed viruliferous WBPH fed on phloem more frequently than non-viruliferous WBPH which might increase the probability of virus transmission. When only female or male WBPH was infected with SRBSDV, the fecundity and egg hatchability were lower than those of the non-infected pairs although not significant. If however both sexes were viruliferous, the fecundity and egg hatchability were adversely affected.indica hybrid rice in Wuyi county , Zhejiang province, China, where the SRBSDV disease frequently outbreaks. Germinated rice seeds were sown at the rate of 15 kg per hectare and transplanted at one seedling per hill spaced 20 by 20 cm apart. A total of 90 kg N, 90 kg P and 144 kg K per hectare was applied as base fertilizer before planting. Another 90 kg N was applied in two splits at tillering stage (60%) and reproductive stage (40%). Rice was naturally infected by immigrant viruliferous WBPH. Thirty days after sowing, rice plants with visible SRBSDV-infected symptoms were marked. The healthy and infected rice plants with typical disease symptoms were uprooted and transplanted individually in clay pots (Φ = 15 cm). After all arthropods were manually eliminated, potted plants were covered with a cage made of a transparent plastic cylinder with nylon mesh at the top for ventilation. All plants were molecularly identified following the method of Li et al. (2012) [Rice variety Y-liangyou1 was used in the experiment. It is a dominant . (2012) .A strain of WBPH were collected in experimental farm of China National Rice Research Institute in Hangzhou , China and maintained on healthy rice plants in laboratory. Fifty 5th instar nymphs of 2nd generation were collected and reared individually. After emergence, male and female WBPH were paired and transferred to a caged rice plant for oviposition. Adults were identified by RT-PCR to confirm whether they were viruliferous or not after they were died. All offspring of virus-free of original insects were maintained on healthy rice plants at 26 ± 1°C with photoperiods of 12 L: 12D. Newly hatched nymphs of next generation were used in this study.et al. (2011) [Following the methods described by He . (2011) , 45-day-et al. (2011) [9 Ω input resistance and an input bias current of less than 1 pA . Female adults, which emerged within 8 h were transferred into a glass tube  and provided water only (water-saturated cotton) for two hours before used. One end of a gold wire  was attached to the dorsal thorax of WBPH with water-soluble silver conductive glue. The other end of the wire was connected to the amplifier through the EPG probe. A copper wire  was inserted into the soil to serve as the plant electrode. The female attached to the gold wire was carefully placed onto the stem of healthy rice plant. The gain of the amplifier was set at 50 ×, and the plant voltage was adjusted to obtain an output voltage of between −5 and +5 V. EPG signals were analyzed with the PROBE 3.0 software . After EPG recording for 6 h, the females were identified for SRBSDV individually. According to the identification results, 22 and 18 replications were applied for viruliferous WBPH and non-viruliferous WBPH, respectively. All the EPG tests were conducted at 26 ± 1°C, 70 ± 5% RH and continuous light conditions.Following the methods described by He . (2011) , feedingEPG waveforms from WBPH recorded in the tests were characterized using the correlations found in Seo et al. (2009)  and He e(+) × non-viruliferous male ♂(−), non-viruliferous female ♀(−) × viruliferous male ♂(+), ♀(+) × ♂(+) and ♀(−) × ♂(−), respectively.Newly emerged adults were paired and transferred into a glass tube with two healthy rice tillers. They were allowed to mate and oviposit until death, and the fecundity was investigated as described below. Each tube was observed daily and newly hatched nymphs were counted and removed every day until no more nymphs hatched. Unhatched eggs were counted by dissecting under microscope. Death WBPH adults were collected and molecularly identified individually to confirm if they were infected with SRBSDV. According to the identification results, 15, 18, 10 and 30 replications were applied for the treatments viruliferous female ♀t-test. Single factor analysis of variance was conducted to analyze the fecundity and egg hatchability of WBPH . All statistical tests were performed using SPSS V18.0.The data on feeding behavior of viruliferous and non-viruliferous WBPH were analyzed using student’s The authors declare that they have no competing interests.HX and XH carried out the experiments and drafted the manuscript. XZ was involved in collecting WBPH and SRBSDV-infected rice plants. YY and JT helped with experimental procedures and manuscript preparation. ZL designed the study and critically revised the manuscript. All authors read and approved the final manuscript.


Viruses are known to be the most abundant organisms on earth, yet little is known about their collective origin and evolutionary history. With exceptionally high rates of genetic mutation and mosaicism, it is not currently possible to resolve deep evolutionary histories of the known major virus groups. Metagenomics offers a potential means of establishing a more comprehensive view of viral evolution as vast amounts of new sequence data becomes available for comparative analysis.Bioinformatic analysis of viral metagenomic sequences derived from a hot, acidic lake revealed a circular, putatively single-stranded DNA virus encoding a major capsid protein similar to those found only in single-stranded RNA viruses. The presence and circular configuration of the complete virus genome was confirmed by inverse PCR amplification from native DNA extracted from lake sediment. The virus genome appears to be the result of a RNA-DNA recombination event between two ostensibly unrelated virus groups. Environmental sequence databases were examined for homologous genes arranged in similar configurations and three similar putative virus genomes from marine environments were identified. This result indicates the existence of a widespread but previously undetected group of viruses.This unique viral genome carries implications for theories of virus emergence and evolution, as no mechanism for interviral RNA-DNA recombination has yet been identified, and only scant evidence exists that genetic exchange occurs between such distinct virus lineages.This article was reviewed by EK, MK (nominated by PF) and AM. For the full reviews, please go to the Reviewers' comments section. While viruses are known to be the most abundant organisms on earth, their collective evolutionary history, biodiversity and functional capacity is poorly understood -3. DespiThe known virosphere consists of three principal viral types; the RNA-only viruses, which do not require a DNA intermediate in the replication cycle, viruses with DNA-based genomes, and retroid viruses that require the reverse transcription of their RNA into DNA during the virus life-cycle . LateralWe report the discovery of a group of circovirus-like DNA genomes whose common ancestor appears to have incorporated a capsid protein (CP) gene known previously only in RNA viruses. The mechanism responsible for the integration of the RNA virus cistron into the DNA virus, and the point in evolutionary time at which it occurred, are unclear. Relatively low levels of sequence divergence between the homologous viral proteins indicate that the recombination event took place relatively recently. Moreover, it suggests that entirely new virus types may emerge via the lateral exchange of functional and structural modules from viruses of vastly different types, utilizing as yet unknown mechanisms.Archaea, Bacteria and several species of unicellular Eukarya sugg suggAuthnomenon  and trap. 5: The moniker “RNA-DNA hybrid virus” (RDHV) must go. This is a thoroughly misleading name. The authors show abundant evidence of a circovirus-like or nanovirus-like virus with single-strand DNA genome that, in the past, have acquired a capsid protein from an RNA virus. Nonetheless, it is a DNA virus now. This is not even the first example of that kind of mosaicism – BL1/BC1 proteins of bipartite geminiviruses are similar to the 30 K family of movement proteins of plant RNA viruses, but no one calls bipartite geminiviruses “DNA-RNA viruses” because of that. RNA genomes of closteroviruses encode homologs of cellular HSP70 proteins, but these viruses are not RNA-DNA viruses either. Descriptive name such as “Boiling Spring Lake Virus 1” or something of this kind should do just fine. Note that this objection to “RDHV” is not the nomenclature war, but rather aims at setting the molecular record straight.Author’s response: The moniker “RDHV” is mentioned in the text as provisional. We feel that a succinct descriptive name for this new virus genome-type is warranted, at least temporarily. Other conceivable names seem insufficient to describe a novel and probably wide-spread virus group and its ancestry, and would be significantly more confusing or excessively complicated (e.g. “a Boiling Springs Lake virus from the Sargasso Sea”). We completely agree that the genome discovered represents a DNA virus. Once we have identified the host and/or structure for the virus we will propose a taxonomically appropriate name through the ICTV (and let the nomenclature wars rage).p. 5 and later: I am sure that there is a straightforward sequence-similarity argument on the evolutionary relatedness of “RDHV” capsid protein and tombusviruses. I could obtain statistically significant similarity between the former and the latter by PSI-BLAST and HHPred approaches. I recommend that the authors do the same. Instead, we are reading “The predicted structure of the BSL RDHV capsid protein is congruent to the S-P domain double jelly-roll configuration found in the ssRNA Tomato Bushy Stunt (TBSV) and Melon Necrotic Spot (MNSV) tombusviruses ,13. AminAuthor’s response: This section has been extensively revised and Figures added .d Figures–6.p. 7: “The most parsimonious scenario” --- more parsimonious than which other scenarios?Author’s response: This section has also been revised. See reply to Krupovic in regard to the origin of linear and circular ssDNA viruses.pp. 7–8: Several mentions of satellite RNA viruses seem out of place – tombusviruses are not satellites and neither are fungal viruses discussed in the paper?Author’s response: These references have been clarified.pp. 8–9: (last paragraph of the paper) “Assuming that RNA viruses evolutionarily preceded all DNA virus groups,34, evidAuthor’s response: This final paragraph has been revised and clarified.


Drosophila, and may have important implications for tumorigenesis. While it is known that Dronc can induce the activity of Jun N-terminal kinase (JNK) for apoptosis-induced proliferation, the mechanistic details of this activation are largely unknown. It is also controversial if JNK activity occurs in dying or in surviving cells. Signaling molecules of the Wnt and BMP families have been implicated in apoptosis-induced proliferation, but it is unclear if they are the only ones. To address these questions, we have developed an efficient assay for screening and identification of genes that regulate or mediate apoptosis-induced proliferation. We have identified a subset of genes acting upstream of JNK activity including Rho1. We also demonstrate that JNK activation occurs both in apoptotic cells as well as in neighboring surviving cells. In a genetic screen, we identified signaling by the EGFR pathway as important for apoptosis-induced proliferation acting downstream of JNK signaling. These data underscore the importance of genetic screening and promise an improved understanding of the mechanisms of apoptosis-induced proliferation.Recent work in several model organisms has revealed that apoptotic cells are able to stimulate neighboring surviving cells to undergo additional proliferation, a phenomenon termed apoptosis-induced proliferation. This process depends critically on apoptotic caspases such as Dronc, the Caspase-9 ortholog in  spi, the EGF ligand in flies, as important for apoptosis-induced proliferation. We confirmed the involvement of EGF in a genuine apoptosis-induced regeneration system. These data underscore the importance of genetic screening and promise an improved understanding of the mechanisms of apoptosis-induced proliferation and regeneration.Work in recent years has revealed that apoptotic caspases not only induce apoptosis, but also have non-apoptotic functions. One of these functions is apoptosis-induced proliferation, a relatively recently discovered phenomenon by which apoptotic cells induce proliferation of surviving neighboring cells. This phenomenon may have important implications for stem cell activity, tissue regeneration and tumorigenesis. Here, we describe the development of a genetic model of apoptosis-induced proliferation and the use of this model for convenient and unbiased genetic screening to identify genes involved in the process. We tested mutants of our RNAi transgenic lines targeting the core components of the apoptotic pathway and of JNK signaling, a known mediator of apoptosis-induced proliferation. These assays demonstrate the feasibility of the system for systematic genetic screening and identified several new genes upstream of JNK that are involved in apoptosis-induced proliferation. Finally, we tested the model in a pilot screen for chromosome arm 2L and identified  Apoptosis is the major form of programmed cell death. It is used during development and under stress conditions to remove excess, unwanted or damaged cells. Deregulated apoptosis can give rise to malignancies including cancer and neurodegeneration Drosophila, only the initiator caspase Dronc and the two effector caspases DrICE and Dcp-1 have been implicated in apoptosis in imaginal discs Drosophila IAP1 (Diap1) binds to and inhibits Dronc, DrICE and Dcp-1 Caspases are very well conserved in the animal kingdom. Of the seven caspases in Drosophila imaginal discs can form a normal-sized and patterned organ even after more than 50% of their cells have been killed by X-ray treatment due to compensatory proliferation Drosophila, and later in hydra, Xenopus, planarians, newt and mice, has revealed that apoptotic caspases may be the driving force for compensatory proliferation in apoptotic tissue Developing organisms have the ability to compensate for massive apoptotic cell loss by inducing compensatory proliferation. For example, developing Drosophila. The first type of model takes advantage of the fact that another caspase inhibitor, the P35 protein from Baculovirus, specifically inhibits the effector caspases DrICE and Dcp-1, but not the initiator caspase Dronc p35-expressing cells triggers the apoptotic pathway down to Dronc, but cannot execute cell death because of inhibition of effector caspases by P35. These cells are referred to as ‘undead’ cells. Consequently, Dronc is functional in ‘undead’ tissues and can fulfill non-apoptotic roles including AiP which triggers overgrowth There are two commonly used experimental models that study AiP in larval imaginal discs, usually wing and eye discs in p53 and the stress-kinase JNK, encoded by basket (bsk) in Drosophilawingless (wg) and the TGFβ/BMP-family member decapentaplegic (dpp), both of which are potent mitogens p35-expressing conditions, JNK activity occurs only in ‘undead’ cells wg is not induced in all ‘undead’ cells and concern has been raised about the involvement of wg and dpp in ‘genuine’ AiP Much of what we know about AiP came from studies of ‘undead’ cells. In ‘undead’ cells, Dronc activates There are many other open questions in the field. For example, although it is well established that Dronc can stimulate JNK activity, the molecular mechanism of this interaction is not known. Furthermore, while JNK is best characterized for its ability to induce apoptosis Here, we present and test the feasibility of ‘undead’ and ‘genuine’ genetic models of AiP in eye imaginal discs. We identify additional components in the JNK pathway that mediate the activation of JNK by Dronc. We show that JNK activation occurs in dying cells as well as in neighboring surviving cells depending on the conditions used. We report the results of a pilot screen using the ‘undead’ AiP model that led to the identification of Spi/EGFR signaling as essential component for AiP. Finally, we demonstrate that Spi is at least partially required for regeneration in a ‘genuine’ AiP model of the eye disc.eyeless (ey) is a regulatory gene for eye development and is expressed during the growth phase of eye imaginal discs rd instar larvae, ey expression ceases in and posterior to the MF hid and p35 during the growth phase of the eye disc using ey-Gal4 (referred to as ey>hid-p35) may provide a convenient model to induce AiP. Correspondingly, in eye imaginal discs, the anterior portion of the eye disc is overgrown compared to controls forming an expanded head capsule due to increased cell proliferation dpp-lacZ expression puc-lacZ and TRE-dsRedey>hid-p35 model which therefore may represent a convenient AiP model for genetic screening.ey>hid-p35 model for genetic screening, we first examined if mutants and RNA interference (RNAi) of caspases and ark genetically modify the AiP phenotype. Heterozygosity of dronc and dronc RNAi strongly suppressed the AiP phenotype  and the Fos ortholog kayak (kay), also suppressed the ey>hid-p35 AiP phenotypes, although to a lesser extent  and MKK4  as AiP suppressor  antibody, a marker of Dronc activity Theoretically, it is possible that the suppression of AiP by these RNAi transgenes is an indirect result of suppression of apoptosis, as observed in the case of  or RNAi . To exclRho1 is the least well characterized regulatory component in the JNK pathway, we examined the effect of Rho1 knockdown on JNK activity in the AiP model. Loss of Rho1 suppresses puc-lacZ in ey>hid-p35 eye discs  AiP model, we repeated the experiments by Bergantinos et al. (2010) hid in a temporally and spatially controlled manner using ptc-Gal4 and tub-tsGal80 (tsptc>hid) by temperature shifts for various times. In control experiments, just expressing GFP in the ptc domain does not affect the puc-lacZ pattern  of hid expression followed by a 6 hours recovery period (ts6hR6h), an elevation of puc-lacZ activity was detected in dying cells and neighboring, surviving cells . This JNK activity was induced during the recovery period, because immediately after hid induction (ts6hR0h), no alteration of puc-lacZ expression was detected (hid expression was induced for a long period (16 hours) followed by 6 hours recovery (ts16hRh), puc-lacZ was strongly down-regulated in dying cells, likely as a result of apoptosis in these cells. Nevertheless, upregulation of puc-lacZ was detected in neighboring surviving cells  for modification of the AiP phenotype and identified four chromosomal segments as dominant AiP suppressors and seven deficiencies as dominant AiP enhancers ED1303 (spitz (spi) as a potential regulator of AiP  (Suppl. Drosophila orthologs of Ras (Dras) and MAPK (rolled (rl)) act as dominant suppressors of ey>hid-p35 ED1303  and led  compare . spi enc compare . We also) Suppl. . Downstr>hid-p35  suggesti>hid-p35  [26], [401068spi allele which is an enhancer trap insertion of lacZ into the spi gene (spi-lacZ) and can serve as a reporter for spi expression spi allele itself is a dominant suppressor of AiP: about 75% of the ey>hid-p35/01068spi flies show a weak AiP phenotype, while 25% are not suppressed and still show a moderately strong AiP phenotype (ey>hid-p35 eye discs (n = 28), we observed a strong induction of β-Gal labeling compared to control discs To further characterize the involvement of Spi/EGFR signaling for AiP, we took advantage of the henotype . Consistol discs . This pehenotype . The rempression . In addispi and bsk. Heterozygosity of 1spi dominantly suppresses the adult AiP phenotype of ey>hid-p35  tsGal80tsDE>hid. This model also induces GFP to label hid-expressing cells. Before and after the temperature shift, animals were incubated at 18°C  AiP model are also involved in ‘genuine’ (P35-independent) regeneration in the eye disc. To accomplish this we used a similar approach as previously described in wing discs  at 18°C  to inhib h later .GFP+ cells have died and have been replaced by new, GFP−, cells.In experimental discs immediately after the 30°C pulse (recovery 0 hours – R0 h), a strong apoptotic response is detectable ′ which cGFP+-labeling  tsDE>hid model. First, we examined if JNK activity is induced in the tsDE>hid model. Consistent with the ‘genuine’ AiP models in the wing TRE-dsRed as marker of JNK activity TRE-dsRed is confined to the GFP+ area, i.e. in the death domain  as a marker to reveal disc outline and thus assess the degree of regeneration. Control discs (tsDE>hid) at R72 h had completely regenerated  affects normal photoreceptor differentiation. However, eye discs treated in this way have a normal ELAV pattern  AiP and regeneration. Nevertheless, we developed the tsDE>hid model to verify genes identified in the ‘undead’ screen as being involved in ‘genuine’ AiP and regeneration.Apoptosis-induced proliferation (AiP) appears to be a mechanism by which developing organisms replace dying cells under stress conditions and initiate regenerative responses  known to activate JNK g clones . Neverthg clones  may supptsptc>hid), JNK activity is detectable in both dying and surviving cells. However, the surviving cells with increased JNK activity are also present in the ptc domain  AiP model  scored positive in the ey>hid-p35 model (data not shown). Therefore, at least in the eye disc, not every pathway involved in growth control is also involved in AiP. These observations stressed the necessity to perform unbiased genetic screens aimed at identifying the genes and mechanisms involved in AiP.We have tested signaling pathways known to be involved in growth control for modification of the ey>hid-p35 AiP model using deficiencies of chromosome arm 2L. We identified four deficiencies as suppressors and three as enhancers  may be the exclusive appearance of Cas3*-positive areas and areas with EGFR activity . Under normal conditions, the ablated tissue is replaced by new tissue due to compensatory proliferation. The authors then screened for mutants that fail to renew the lost tissue tssec5 screen tssec5–induced compensatory proliferation requires caspase activity in apoptotic cells Recently, a genetic screen has been reported aimed at identification and characterization of genes required for compensatory growth ey>hid-p35 model for genetic screening. We are confident that this model will close gaps in our understanding of AiP regulation under p35-expressing conditions and in p35-independent regeneration. Finally, it will have implications for the understanding of tumorigenesis by ‘undead’ as well as ‘genuine’ apoptotic tumor cells In summary, we have developed and tested the feasibility of the I29dronc; G8ark; 1spi; 01068spi; f2Egfr; ΔC40bras, 10arl, ey-Gal4; ptc-Gal4; DE-Gal4; tub-tsGal80; UAS-p35; UAS-hid; UAS-GFP; wg-lacZ; puc-lacZ; kek1-lacZ; spi-lacZ = 01068spi; TRE-dsRed; GMR-hid; GMR-Gal4 UAS-egr. UAS-based RNAi stocks of the following genes were obtained from various stock centers  and were tested for suppression of AiP: dronc, dcp-1, drICE; bsk, egr, wgn, PVR, dTraf1, dTraf2, Rho1, msn, slpr, Tak1, dMekk1, dAsk1, hep, dMkk4, Jra, kay, spi, Egfr. The exact genotype of ey>hid-p35 is UAS-hid; ey-Gal4 UAS-p35/CyO,tub-Gal80. Expression of tub-Gal80 in this stock suppresses the semi-lethality associated with ey-induced expression of hid and p35.The following mutants and transgenic stocks were used: rd instar larval stage. Genotype: hs-FLP/UAS-hid; UAS-p35/act>y+>Gal4 UAS-GFP; puc-lacZ/+.Larvae of the following genotype were heat shocked for 15min at 37°C, raised at room temperature for 48 h before they were analyzed at the late 3UAS-hid/+; ts/+ptc-Gal4 tub-Gal80; UAS-GFP/+  and ts>hidDE. Increase of PH3-positive cells in dorsal eye disc portions of ts>hidDE animals are consistently observed. PH3-positive cell numbers were counted in dorsal (GFP+) and ventral eye disc portions in selected discs. Size of the dorsal (GFP+) and ventral eye disc portions were measured through the “histogram” function in Adobe Photoshop CS. To compare the density of PH3+ cells in each disc portion, number of PH3+ cells were divided by size (in pixels) of the corresponding tissue which is used to calculate the number of cells in 100,000 pixels (density). Such normalized density of PH3+ cells in various portions of eye discs (mean ± SD) were used for the statistical chart. PH3+ cell numbers in 100,000 pixels is on average 88 in ts>hidDE dorsal eye discs compared to 62 in the control dorsal discs. Their statistical significance was evaluated through a two-tailed, unpaired Student's t-Tests (P<0.04). In contrast, the number of PH3+ cells are comparable in ventral disc portions of each genotype suggesting that increased proliferation mostly occurred in the dorsal part of the disc (at least at the time point of R24 h).Two rounds of experiments  were done for both rd instar larvae and stained using standard protocols. Antibodies to the following primary antigens were used: PH3 (Upstate), anti-cleaved Caspase-3 , β-GAL (Promega), ELAV and Wg (DHSB). Secondary antibodies were donkey Fab fragments from Jackson ImmunoResearch. Images were taken with either a Zeiss AxioImager or a confocal microscope.Imaginal discs were dissected from late 3Figure S1TRE-dsRed is induced in ‘undead’ ey>hid-p35 cells. Shown are (A) wild-type (wt), (B) ey>p35 and (C) ey>hid-p35 eye imaginal discs labeled for dsRed  and ELAV . Only ey>hid-p35 discs induce TRE-dsRed expression  and disrupt the ELAV pattern (C″).The JNK activity marker (TIF)Click here for additional data file.Figure S2RNAiUAS-dcp-1 and RNAiUAS-drICE stocks are functional. Combined expression of RNAiUAS-dcp-1 and RNAiUAS-drICE stocks suppresses both TUNEL-positive apoptosis  and eye-ablation of GMR-hid  suggesting that these stocks contain functional RNAi transgenes targeting dcp-1 and drICE.The (TIF)Click here for additional data file.Figure S3GMR-egr. (A) The unmodified GMR-Gal4 UAS-eiger (GMR>egr) eye ablation phenotype. (B–H) RNAi transgenes targeting the genes indicated above the panels suppress the eye ablation phenotype induced by GMR-Gal4 UAS-eiger (GMR>egr) suggesting that they are functional.Several UAS-RNAi transgenes of the JNK pathway suppress (TIF)Click here for additional data file.Figure S4ey>hid-p35 eye discs does not affect caspase activity. (A) A ey>hid-p35 disc labeled for Cas3* and ELAV. (B) dronc RNAi suppresses Cas3* and normalizes the ELAV pattern in ey>hid-p35 discs. (C–H) RNAi transgenes targeting the indicated JNK pathway components normalize the ELAV pattern, but fail to suppress Cas3* activity in ey>hid-p35 discs suggesting that they suppress AiP downstream of caspase activation.Inactivation of JNK pathway genes in (TIF)Click here for additional data file.Figure S5ey>hid-p35 discs are characterized by strong puc-lacZ  and wg  expression as well as disrupted photoreceptor pattern (ELAV).  RNAi targeting Rho1 suppresses puc-lacZ  and wg  expression as well as normalizes the ELAV pattern in ey>hid-p35 discs. Caspase activity is not affected suggesting that Rho1 acts downstream of Dronc and upstream of JNK.Rho1 acts upstream of JNK in the ‘undead’ AiP model.  (TIF)Click here for additional data file.Figure S6Egfr is required for AiP in a wing model. (A) A control wing disc expressing UAS-p35 under nubbin (nub)-Gal4 (nub>p35) control shows normal Wg expression (A′) and little to no Cas3* labeling (A″). (B) An experimental AiP disc expressing hid and p35 under nub control (nub>hid-p35) displays strong overgrowth with abnormal Wg pattern (B′) and strong Cas3* labeling (B″). Together with (D), these data suggests that nub>hid-p35 is a suitable ‘undead’ AiP model. (C) A nub-Gal4 UAS-p35 (nub>p35) control disc. puc-lacZ expression is detectable at low level. (D) Coexpression of hid and p35 induces strong JNK activity (puc-lacZ) in the enlarged nub domain. (E) RNAi targeting Egfr suppresses the overrepresentation of the nub domain, but leaves puc-lacZ intact. This result suggests that EGFR signaling is required for AiP in the wing disc and acts downstream of JNK.  Control disc expressing Egfr RNAi in the nub domain without hid, in the presence (F) or absence (G) of p35. The size of the nub domain is not significantly altered by Egfr RNAi compared to (C).(TIF)Click here for additional data file.Figure S7TRE-dsRed in tsDE>hid eye imaginal discs. (A–D) hid and GFP expression were temporally induced for 12 h by temperature shift to 30°C during early third instar larval stage as indicated in dsRed expression  was monitored at 0 h (A), 6 h (B), 12 h (C) and 24 h (D) recovery after the temperature shift. GFP  marks the DE domain. Blue is DAPI labeling to outline the discs. dsRed labeling is weakly detectable at R0 h, peaks at 6 h after recovery and fades off at R12 h. At R24 h, it is barely visible. (E) A tsDE>GFP control disc at 6 h recovery after the temperature shift, labeled for dsRed . JNK activity is not induced. GFP expression in (E″) is strong. Blue in (E) is DAPI labeling to outline the discs.Induction of the JNK activity marker (TIF)Click here for additional data file.Table S1ey>hid-p35 background.Chromosomal deficiencies tested in the AiP screen on 2L. Listed are the names of the deficiencies, the extent of the chromosomal deletions and the score in the AiP screen. Green marks suppressors and yellow marks enhancers. Deficiencies marked with * could not be scored, because they caused lethality in the (PDF)Click here for additional data file.


In May 2003, an outbreak of fatal hemorrhagic fever, caused by yellow fever virus, occurred in southern Sudan. Phylogenetic analysis showed that the virus belonged to the East African genotype, which supports the contention that yellow fever is endemic in East Africa with the potential to cause large outbreaks in humans. Flavivirus, family Flaviviridae) is a mosquitoborne virus endemic in tropical regions of Africa and South America, where it has been responsible for large epidemics. The virus is maintained in sylvatic transmission cycles involving nonhuman primates but uses humans as the sole vertebrate host in urban epidemics. In South America, the sylvatic vectors belong to the Haemogogus and Sabethes genera, and the urban vector is Aedes aegypti; in Africa, Aedes species mosquitoes serve as both sylvatic and urban vectors. The virus causes febrile disease with necrotic hepatitis in humans, and death rates can exceed 50%. Although an effective vaccine is available, the virus remains a major public health threat, particularly in Africa, where vaccination is limited by poverty, civil wars, and the inaccessibility of rural areas where outbreaks of the disease occur. The World Health Organization estimates 200,000 cases of the disease occur annually and 30,000 deaths, but because of underreporting, only a small percentage of these cases are recorded.Yellow fever virus , in Nairobi, where yellow fever virus was identified as the causative agent of the outbreak. Aliquots of specimens were submitted to the Special Pathogens Unit at the National Institute for Communicable Diseases in Johannesburg for confirmation of the diagnosis. A report dated June 19, 2003, from the Office of the United Nations Resident and Humanitarian Coordinator for the Sudan described a steady decline in the number of cases occurring during the 4 weeks before the report with a total of 162 suspected cases with 48 deaths in Torit County. However, the report also noted that the recording of suspected cases was inadequate, the figures quoted were an estimate, and final case numbers and deaths were uncertain. The details of the outbreak and laboratory investigations are reported in a separate publication.Virus isolates were obtained from three patients, two from Locomo village and one from Lofi village. The viral isolations were performed by injecting serum samples from all the patients into suckling mice. RNA was extracted from harvested mouse brain tissue by using Trizol reagent , according to the manufacturer's directions. A 670-bp region of the polyprotein gene, which included the 3´ end of the premembrane protein gene, the complete membrane protein gene, and the 5´ end of the envelope protein gene, was amplified by using reverse transcription–polymerase chain reaction (RT-PCR) . The seqAlignment of the nucleotide sequence data was performed by using DNASIS for Windows version 2.5 . The phylogenetic analysis was performed on a 572-bp region of the amplicons by using a weighted maximum parsimony method, transversion:transition weighting of 5:1, and Phylogenetic Analysis Using Parsimony (PAUP) software version 4.0b4a for Macintosh  shows th,Outbreaks of yellow fever virus have frequently been reported from areas within West Africa since the 18th century, with far fewer outbreaks being identified in East Africa. Serologic evidence for the presence of yellow fever virus in Sudan, Kenya, Uganda, and Tanzania was first reported in 1936 (


To determine which respiratory viruses circulate among confined dogs, we analyzed nasal and pharyngeal swab specimens from shelter dogs with acute respiratory disease. An unknown virus was isolated. Monoclonal antibody testing indicated that it was probably a pneumovirus. PCR and sequence analysis indicated that it was closely related to murine pneumovirus. Domestic dogs housed in close confinement, as in kennels or animal shelters, are often involved in outbreaks of acute respiratory disease . After 3 passages in culture, cells in some of the flasks showed subtle cytopathic changes. After continued passage, small foci of rounded cells developed, and rapid and progressive cell death throughout the flask ensued in a pattern uncharacteristic of the viruses commonly isolated from dogs. Testing these cultures with a panel of diagnostic reagents specific for common canine respiratory agents failed to identify a known virus. Immunofluorescence assays (IFAs) ultimately detected 13 positive cultures over a 4-month period when a monoclonal antibody (MAb) pool against human respiratory syncytial virus (RSV)  was used. We have commonly used this antibody preparation to detect bovine RSV. The staining pattern included filamentous membrane-bound and free-floating virions and cytoplasmic inclusions, typical of the pattern in RSV-infected cells.After obtaining IFA results, we attempted to amplify a fragment of the nucleocapsid gene (N) from the virus by using PCR primers designed on the basis of an alignment of human, bovine, and ovine RSV sequences. This attempt was unsuccessful (data not shown). Stocks of the individual MAbs in the anti-RSV pool and their specificities were obtained from the manufacturer and used for IFA . MAb 2G1Pneumovirinae (www.ncbi.nlm.nih.gov/blast/Blast.cgi) analysis showed the virus to be closely related to murine pneumovirus (MPV), traditionally known as pneumonia virus of mice. Two L-gene PCR products were found to be 96%–97% identical to MPV, and an N gene fragment was 96% identical , 55% (L fragment no. 1), and 72% (L fragment no. 2) when aligned with the same regions of human RSV, so finding cross-reactivity between human RSV MAbs and the newly isolated virus is not completely unexpected. Serologic cross-reactivity between RSV and MPV has been previously observed. Gimenez et al. (The isolation of a previously unknown virus from dogs does not imply disease causation. However, comparison with MPV leads to speculation that the virus isolated in this study may have pathogenic potential. MPV is commonly known to infect laboratory rodent colonies, and serologic evidence points to infection of several wild rodent species. However, little is known about MPV epidemiology, such as whether MPV has multiple natural hosts or whether closely related viruses are circulating in other species. Neutralizing antibodies to MPV in other mammalian species, including humans, were first reported in the 1940s (Questions remain as to whether this newly isolated virus commonly infects dogs and, if so, why it has not been previously isolated. Perhaps the strain that was circulating in these particular animal shelters is more easily isolated in culture. Or, because the initial cytopathic changes observed with these isolates were subtle, they could easily have been missed. Outbreaks of acute respiratory disease in dogs often involve multiple pathogens. As anticipated, other viruses, primarily canine influenza and parainfluenza viruses, were isolated during the study, often from the same animals that carried the pneumovirus. Work is ongoing to further determine pneumovirus prevalence among dogs and its involvement in acute respiratory disease of dogs.


Background. The most common chemotherapies in metastatic soft tissue sarcoma (mSTS) require intravenous (IV) administration. This often requires patients to make multiple outpatient visits per chemotherapy cycle, possibly impeding patients' daily activities and increasing caregiver burden and medical costs. This study investigated costs associated with IV cancer therapy administration in mSTS from the payer perspective of the health care system. Patients and Methods. From the Experian Healthcare database, 1,228 mSTS patients were selected. Data were analyzed on outpatient visits during 2005–2012 involving IV cancer therapy administration. Costs were estimated on a per patient per visit (PPPV) and per patient per month (PPPM) basis. Results. The mean (median) cost of IV therapy was $2,427  PPPV and $5,468  PPPM, of which approximately 60% was IV drug costs. IV administration costs averaged $399 PPPV and $900 PPPM, representing 16.5% of total visit costs. Anthracycline and alkylating-agents-based therapies had the highest PPPV and PPPM IV administration costs, respectively . Patients with managed care insurance had the highest IV administration costs . Conclusions. IV administration costs constitute a considerable proportion of the total costs of receiving an IV cancer therapy to treat mSTS. Soft tissue sarcomas (STS) are a rare, complex group of childhood and adult neoplasms with differentiation towards mesenchymal tissue, which may arise almost anywhere in the body . STS accThe most commonly used chemotherapies in metastatic soft tissue sarcoma (mSTS) are intravenously (IV) administered agents . AlthougThis study reports estimates of the total and component costs associated with IV cancer therapy administration in patients with mSTS from the payer perspective of the US health care system, based on an analysis of claims data from a large contract and claims management system.Data for this study were obtained from the Experian Healthcare (Experian) database, which maintains a contract and claims management system that supports 350 general/oncology clinics in the US. The database contains a complete history of diagnoses (ICD-9-CM codes), procedures, and drug therapies received by both publicly and privately insured patients within the clinics, as well as patient demographics  and insurance type . For every patient clinic visit, Experian records the service dates, total charged, total contracted payments, and total allowed, with individual services, procedures, and drugs broken out by line item . The Experian dataset used in this study covers the period from January 1, 2005, to April 30, 2012.This study employed a retrospective, longitudinal cohort design. The index date was defined as the date of the first IV cancer therapy infusion for mSTS treatment. Therapy windows were calculated as a patient-drug combination for the purposes of reporting study outcomes on a per month basis. The observation period for each patient-drug window begins at the index date and ends with either the last IV cancer therapy administration (if the patient only remains on one therapy) or the last visit of that IV cancer therapy before a patient switches to a different IV cancer therapy, plus a therapeutic effect. The therapeutic effect for patients that do not switch therapies is the average interval between administrations across the entire dataset for that particular IV cancer therapy. For patients who received a single IV therapy, the observation period length was calculated as the length of the therapy plus the average interval across the entire dataset for that particular IV cancer therapy. For patients that switch IV cancer therapies, the therapeutic effect is defined as the minimum of either the time between the last visit of that therapy and the start of the new therapy or average therapy interval across the entire dataset.a combination therapy of ifosfamide  and doxorubicin or liposomal doxorubicin  (both agents to be administered within 30 days), except if also diagnosed with osteosarcoma (ICD-9-CM: 170), nasopharyngeal carcinoma (ICD-9-CM: 147), lung cancer (ICD-9-CM:162.3-162.9), uterine cancer , breast cancer (ICD-9-CM:174), prostate cancer (ICD-9-CM: 185), renal cancer (ICD-9-CM: 189), or malignant neoplasm of lymphatic and hematopoietic tissue (ICD-9-CM: 200-208) at any time, orat least 1 medical claim for retroperitoneal or peritoneal cancer (ICD-9-CM: 158), except if also diagnosed with renal cell carcinoma , transitional cell carcinoma , mesothelioma (ICD-9-CM: 163), gastrointestinal stromal tumors , uterine cancer , ovarian cancer (ICD-9-CM: 183), or other and unspecified female genital-organ cancer (ICD-9-CM: 184) at any time.The selection of patients in the study sample is depicted in Treatment costs were estimated using the contracted allowed payment for a claim rather than the practice charges, based on adjudication of the claim by the patient's third-party insurance plan. A contracted payment is defined as the amount that the provider is eligible to receive from all parties, including primary and secondary payers and the patient, based on the contractual agreement with the payer. Because the contracted payment represents the actual payment to providers from payers, it depicts a more accurate and detailed view of the true economic burden to payers of IV cancer therapy administration in mSTS patients.Treatment costs included all costs incurred on eligible IV cancer therapy dates of service. Costs were categorized as (i) IV cancer therapy administration procedure costs, (ii) IV cancer therapy drug costs for mSTS, and (iii) costs associated with other visit-related drugs and services provided on the day of the IV cancer therapy visit. Claim codes used to identify IV infusion administration costs are consistent with those used in previous research –10.Treatment costs were calculated per patient per IV visit (PPPV) and per patient per month (PPPM). PPPM costs were calculated by first computing average per month treatment costs for each patient, defined as the sum of all costs on days with IV cancer therapy administrations during the observation period divided by the length of the observation period for each patient in person-months. Average PPPM treatment costs for the overall sample were then calculated as a weighted average of the average per month cost, using the length of the observation period as the weight. Reporting PPPM costs is an approach commonly used in nonexperimental study settings to account for different lengths of observation periods among study patients.Patient characteristics as of the index date were obtained from the Experian dataset. Age was standardized as patient age in 2009 and divided into five categories: less than 25, from 25 to 39, from 40 to 54, from 55 to 64, and greater than 64 years. Insurance type was defined at the time of the index mSTS diagnosis and was held constant during the study period. Patient characteristics were reported as frequencies and percentages for categorical variables and sample size, means, standard deviations, medians, and ranges for continuous variables.Medians, means, standard deviations, and quartile ranges of PPPV and PPPM treatment costs were reported for the overall mSTS patient sample. The total and component IV administration costs were further stratified by patient cohorts based on age, gender, region, insurance status, IV drugs, and number of administered drugs during the visit. Cost data was adjusted based on the medical consumer price index (CPI)  and repoA total of 1,228 patients with mSTS were identified. Baseline patient characteristics are presented in Per patient per IV visit costs (PPPV) in which an IV cancer therapy was administered, subdivided into IV drug costs, IV administration costs, and other visit-related service costs, are presented in Mean IV drug costs were $1,450 and represented 59.7% of the total cost of the visit. The remaining 23.8% of the visit costs ($578) were attributed to other visit-related services. Evaluation and management office visits, supplies and equipment, and other miscellaneous administration comprised a small part of these costs with a mean of $22 (3.7% of other visit-related services). The remaining 96.2% of other visit-related costs were other IV drugs and specially administered oral drugs. Drugs to treat the symptoms of the disease and side effects comprised the largest part of these other drugs: colony-stimulating factors mean cost of $160 per visit, antiemetic agents mean cost of $171 per visit, and antihypercalcemic agents mean cost of $30 per visit. The other large cost categories were off-label chemotherapy agents (mean cost of $50 PPPV) and monoclonal antibody agents (mean cost of $84 PPPV).IV visit costs on a PPPV basis, broken down by age, gender, region, type of insurance, IV drug used to treat mSTS, and number of IV drugs administered per visit, are presented in The largest differences in IV administration costs were found by insurance status. Managed care patients had the highest IV administration costs (mean $504 per visit) while Medicaid patients had the lowest (mean $92 per visit). The most expensive IV drugs in terms of IV administration costs were anthracycline-based therapies at $479 per visit and the cheapest were angiogenesis-based therapies at $301 per visit. Finally, costs of IV administration tended to increase with the number of IV drugs administered in a visit. Administering a single therapy cost an average of $304, increasing to $693 for three therapies and $936 for four therapies.Total mean PPPM costs, subdivided into IV drug costs, IV administration costs, and other visit-related service costs, are presented in IV visit costs on a PPPM basis, broken down by age, gender, region, type of insurance, IV drug used to treat mSTS, and number of IV drugs administered per visit, are presented in Large variations in PPPM IV administration and other visit-related service costs were observed by insurance status. The highest IV administration PPPM costs were found in managed care patients and indemnity patients at $1,121 and $1,052, respectively. These patients also experienced higher other visit-related service costs compared to other insurance types. As with the PPPV results, other visit-related service costs were highest among patients with workers' compensation, Tricare, or self-pay insurance. In contrast to the PPPV where anthracycline-based agents were reported to have higher IV administration costs, the alkylating-based agents had much higher PPPM IV administration costs compared to the other therapies. Finally, the step-like trend of increasing IV administration costs based on the number of administered IV drugs during the visit was also observed in the PPPM analysis.Tables In This retrospective analysis assessed costs associated with the administration of IV cancer therapies in mSTS patients from 2005 to 2012 using Experian data. IV administration costs accounted for about 16.5% of costs per IV visit among patients with mSTS. Other visit-related service costs accounted for about 23.8% of the cost PPPV. These results indicate that non-IV drug costs represent a considerable proportion of the total costs when receiving an IV cancer therapy to treat mSTS.Cost estimates stratified by patient characteristics indicate that some of the largest differences in IV administration costs across patients exist between patients with different types of insurance. Patients with managed care had the highest IV administration costs ($504 PPPV), while Medicaid patients had the lowest ($92 PPPV). IV administration costs also varied widely across regions, as well as by the type of IV cancer drug and number of IV cancer drugs administered.The share of total treatment costs related to IV administration reported in this study of mSTS patients is similar to estimates reported in both malignant and nonmalignant diseases where drugs are administered by IV infusion. A 2008 study of treatment costs in patients with metastatic breast cancer reported average total treatment costs of $2,477 PPPV, 10.2% of which were IV administration costs . In a seThis study did not account for indirect costs, such as those associated with patient and caregiver time  and lost productivity, which by their nature are more difficult to collect and suffer from greater variability. By only examining direct healthcare-system-related costs of IV cancer therapy administration in an mSTS sample, this study likely understates the true total cost of IV therapy to society.As with all retrospective studies using claims data, identification of mSTS patients relies on the accuracy of diagnosis coding. No clinical information was available to ascertain stage. Advanced STS was identified based on a claims algorithm that has not been validated. To address this concern, a sensitivity analysis was performed where costs were calculated using an alternative claims algorithm. Results of this sensitivity analysis were similar to those reported here. In addition, costs related to IV cancer therapy administration were determined only from clinic claims and were limited to services identified on the claim as an infusion administration cost. This method may underestimate true medical infusion costs since it does not capture costs related to late infusion reactions and complications that require medical care on days following the infusion. Clinic claims also exclude any additional costs of oral drugs or drugs administered in an inpatient setting.Other study limitations included limited information about patient enrollment and disease progression. Patients appear in the dataset through clinic visits, but the date of initial diagnosis of mSTS may have occurred earlier. In addition, it was not possible to determine why patients may have stopped visiting the clinic. It was also not possible to assess disease progression. To address these limitations, costs were estimated on the basis of actual clinic visits and reported both PPPV and PPPM.Using data from the Experian database, this study found that the mean cost of receiving IV cancer therapy in an mSTS patient population was $2,427 PPPV and $5,468 PPPM. IV administration costs accounted for 16.5% of these costs and other visit-related services accounted for 23.8% of these costs. Thus, nonstudy drug related costs are about 40% of the costs associated with administration of IV cancer therapy for mSTS patients. There was substantial variation in the cost of IV administration by insurance type, region, type of anticancer drug, and the number of anticancer drugs administered in a single visit.This study contributes to the understanding of IV therapy administration costs in mSTS patients. Further research is needed to investigate the association between IV administration costs and mSTS cancer stage and to estimate indirect costs of IV administration associated with mSTS patient and caregiver time.


Regional differences in the clinical care of Type 1 diabetes (T1D) in Brazil have been recently described. This study aimed to estimate the costs of T1D from the public health care system’s perspective across the regions of Brazil and to determine the components that influence these costs.This was a retrospective, cross-sectional and nationwide multicenter study conducted between December 2008 and December 2010 in 28 public clinics in 20 Brazilian cities. The study included 3,180 T1D subjects receiving healthcare from the National Brazilian Healthcare System (NBHCS) with a follow-up of at least one year. The direct medical costs were derived from the costs of medications, supplies, examinations, visits to the center, medical procedures and hospitalizations that occurred during the previous year. Clinical and demographic factors that determined the differences in the cost across four geographic regions  were investigated.per capita mean annual direct medical costs of T1D in US$ were 1,466.36, 1,252.83, 1,148.09 and 1,396.30 in southeast, south, north/northeast and mid-west regions, respectively. The costs of T1D in the southeast region were higher compared to south (p < 0.001) and north/northeast regions (p = < 0.001), but not to the mid-west (p = 0.146) region. The frequency of self-monitoring of blood glucose (SMBG) was different across the regions as well as the daily number of SMBG, use of insulin pumps or basal or prandial insulin analogs. Age, ethnicity, duration of diabetes, level of care, socioeconomic status and the prevalence of chronic diabetic complications differed among the regions. In a regression model the determinants of the costs were the presence of microvascular diabetes-related complications (p < 0.001), higher economic status (p < 0.001), and being from the southeast region (p < 0.001).The The present data reinforce the regional differences in the costs of T1D and in the socioeconomic profile and health care provided to the patients with T1D in specialized public centers in Brazil. Both factors influenced directly the costs of T1D and should be considered for discussing future health policies. Type 1 diabetes (T1D) is a chronic disease that carries a large risk of chronic disabling complications that have a negative impact in the costs of the disease and in the patient’s quality of life. Moreover, the incidence of T1D is increasing in many countries, including Brazil . DespiteBrazil is a large country with about 191 · 8 million inhabitants, according to the last population census conducted by the Brazilian Institute of Geography and Statistics (IBGE) . It is dThe direct costs of T1D has been recently estimated in Brazil . PreviouThe aim of our study was to estimate the costs of T1D from the public health care system’s perspective across the different regions of Brazil and to determine the regional differences of the components that influence these costs. We believe these data may provide important clues for reassessment of regional health policies in Brazil.This is a retrospective, cross-sectional and nationwide multicenter cost-of-illness study conducted between December 2008 and December 2010 in 28 centers located in 20 Brazilian cities in public clinics with secondary and tertiary care levels  , which eInformation on the diabetic treatment modalities, source of insulin pumps, medications and supplies for SMBG, frequency of SMBG, routine diabetes care and other data not available from medical records  were obtained through an interview with information provided directly by the patient or his caregiver. The following information was obtained from medical records assessed during the interview: the total number of HbA1c measurements performed in the prior year, fructosamine levels, fasting, and 2-h postprandial glycemia, total cholesterol, LDL-cholesterol, HDL-cholesterol, triglycerides, uric acid, plasma creatinine, plasma urea, sodium, potassium, liver enzymes , C-reactive protein (CRP), thyroid stimulating hormone (TSH), proteinuria, and albuminuria. The number of tests performed to screen for diabetes-related complications, such as electrocardiograms (ECGs), exercise stress tests, stress echocardiographs, calcium score tomographies, coronary artery angiographs, and fundoscopies, were analyzed. The frequencies of medical procedures, such as vitrectomy, laser therapy, hemodialysis, and hospitalizations due to diabetes decompensation or ketoacidosis were also evaluated.The direct medical costs were calculated using the costs of medications , SMBG supplies, blood analysis and other examinations, consultations with physicians, nurses and dietitians, medical procedures, and hospitalizations that occurred during the prior year. The drugs included in the analysis were those specific to treating T1D or its related comorbidities , and complications . The medication costs were obtained from the Brazilian Ministry of Health website  when theAll of the costs were converted into US dollars (at 1 US dollar = 1 · 9315 Reais) using the average of the 3 consecutive years (2008–2010).Statistical Package for the Social Sciences (SPSS) Version 17.0.The statistical analysis was performed using the Data are presented as mean [95% confidence interval] for continuous variables and as counts (relative frequencies) for discrete variables.Comparison of the direct medical costs across the four regions was performed using non- parametric test Mann–Whitney for every two regions. The statistically significant p-values were corrected using the Sidak procedure to control for type I errors (p < 0 · 01). Data on the north and northeast regions were evaluated together, so four geographic regions were compared . A regression analysis was performed to investigate the role of the geographic regions and other independent variables  in influencing the direct medical costs. In this regression analysis the geographic regions were introduced in the model as dummies variables.per capita mean [95% IC] direct medical costs of T1D were US$ 1,466 · 36 , US$ 1,252 · 83 , US$ 1,148 · 09 , and US$ 1,396 · 30  in southeast, south, north/northeast, and midwest regions, respectively. The costs of T1D in the southeast region were higher compared to the south (p < 0 · 001), and north/northeast regions (p < 0 · 001), but not to the mid-west region (p = 0 · 146). Midwest costs were statistically different from north/northeast (p < 0 · 001) but not from south regions (p = 0 · 292). South costs were higher than north/northeast (p < 0 · 001) .In the regression model using the direct medical costs as the dependent variable and geographic region, age, ethnicity, duration of diabetes, level of attention, socioeconomic status and the presence of chronic microvascular or macrovascular diabetes-related complications as the independent variables we observed that the independent determinants of the costs were the presence of microvascular diabetes-related complications , economic status , and being from the southeast region . This model explained only 5 · 4% of the variability of the costs  treatment . These rOther determinant of higher costs in T1D is the presence of chronic diabetes-related microvascular complications. In epidemiological studies, it is important to consider that the prevalence of chronic complications is proportional to the frequency of screening and the survival to acute complications of the disease. Patients from north/northeast presented lower prevalence of complications, which could reflect a survival bias. Intervention studies, such as the Diabetes Control and Complications Trial (DCCT), have shown that intensive treatment during the early stages of T1D reduces the risk of future microvascular complications . Informaper capita expenditure is higher in the southeast and lowest in the north/northeast regions .Executive steering committee: Marilia Brito Gomes (chair), Roberta Cobas, Sergio Atala Dib, Carlos Negrato.Principal investigators are indicated by an asterisk. Program coordinators are italic.Alessandra Matheus, Lucianne Tannus; Universidade Federal Rio de Janeiro: Melanie Rodacki*, Lenita Zadenverg; Hospital Geral de Bonsucesso: Neuza Braga Campos de Araújo*, Marilena de Menezes Cordeiro; Hospital Universitário Clementino Fraga Filho – IPPMG: Dr. Jorge Luiz Luescher*; Renata Szundy Berardo; Serviço de Diabetes da Disciplina de Endocrinologia e Metabologia do Hospital das Clínicas da Universidade de São Paulo: Marcia Nery*; Catarina Cani; Maria do Carmo Arruda Marques; Unidade de Endocrinologia Pediátrica da Santa Casa de Misericórdia de São Paulo: Luiz Eduardo Calliari*, Renata Maria de Noronha; Instituto da Criança do Hospital das Clínicas da Universidade de São Paulo: Thais Della Manna*, Roberta Salvodelli, Fernanda Garcia Penha; Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto – USP: Milton Cesar Foss*, Maria Cristina Foss-Freitas; Ambulatório da Faculdade Estadual de Medicina de São José do Rio Preto: Antonio Carlos Pires*, Fernando Cesar Robles; Associação de Diabéticos de Bauru: Carlos Antonio Negrato*, Maria de Fatima Guedes; Centro de Diabetes da Escola Paulista de Medicina: Sergio Atala Dib*, Patricia Dualib; Clínica de Endocrinologia da Santa Casa de Belo Horizonte Setor Diabetes Tipo 1: Saulo Cavalcanti da Silva*, Janice Sepulveda; Ambulatório Multiprofissional de Atendimento à Diabetes do Hospital de Clínicas da Universidade Estadual de Londrina: Henriqueta Guidio de Almeida*, Emerson Sampaio; Hospital de Clínicas da Universidade Federal do Paraná:Rosangela Roginski Rea*, Ana Cristina Ravazzani de Almeida Faria; Instituto da Criança com Diabete Rio Grande Sul: Balduino Tschiedel*, Suzana Lavigne, Gustavo Adolfo Cardozo; Hospital de Clínicas de Porto Alegre:Mirela Azevedo*, Luis Henrique Canani, Alessandra Teixeira Zucatti; Hospital Universitário de Santa Catarina: Marisa Helena Cesar Coral*, Daniela Aline Pereira; Instituto de Diabetes-Endocrinologia de Joinville: Luiz Antonio de Araujo*; Hospital Regional de Taguatinga, Brasília: Hermelinda Cordeiro Pedrosa*, Monica Tolentino; Flaviene Alves Prado; Hospital Geral de Goiânia: Dr Alberto Rassi: Nelson Rassi*, Leticia Bretones de Araujo; Centro de Diabetes e Endocrinologia do Estado da Bahia: Reine Marie Chaves Fonseca*; Alexis Dourado Guedes, Odelisa Silva de Mattos; Universidade Federal do Maranhão: Manuel Faria*, Rossana Azulay; Centro Integrado de Diabetes e Hipertensão do Ceará: Adriana Costa e Forti*, Maria Cristina Façanha; Universidade Federal do Ceará: Renan Montenegro Junior*, Ana Paula Montenegro; Universidade Federal de Sergipe: Naira Horta Melo*, Karla Freire Rezende; Hospital Universitário Alcides Carneiro: Alberto Ramos*; Hospital Universitário João de Barros Barreto, Pará: João Felício Soares*, Flavia Marques Santos; Hospital Universitário Getúlio Vargas, Hospital Adriano Jorge: Deborah Laredo Jezini*.Universidade Estado Rio de Janeiro: Roberta Cobas*,


To the Editor: Delhi, in the northern part of India, has had outbreaks of dengue caused by various dengue virus types in 1967, 1970, 1982, 1988, and 1996  of 42 samples processed for virus isolation . Of the All previous outbreaks in Delhi have occurred during the monsoon (rainy) season between August and November and subsided with the onset of winter. We recently reported the results of serologic testing during the 2003 outbreak, which also occurred from September to November, with a peak in mid-October 2003 (Dengue virus types 1, 2, and 3 have all been isolated during previous dengue outbreaks in Delhi, but a particular type has always predominated. During the 1996 outbreak of DHF/DSS, we had 26 isolates of dengue virus type 2, but only 1 isolate was identified as dengue type 1 (


RNAi experiments in insects are characterized by great variability in efficiency; for instance beetles and locusts are very amenable to dsRNA-mediated gene silencing, while other insect groups, most notably lepidopterans, are more refractory to RNAi. Several factors can be forwarded that could affect the efficiency of RNAi, such as the composition and function of the intracellular RNAi machinery, the mechanism of dsRNA uptake, the presence of dsRNA- and siRNA-degrading enzymes and non-specific activation of the innate immune response. In this essay, we investigate the evidence whether persistent infection with RNA viruses could be a major factor that affects the response to exogenous dsRNA in insects. The occurrence of RNA viruses in different insect groups will be discussed, as well as several mechanisms by which viruses could interfere with the process of RNAi. Finally, the impact of RNA virus infection on the design of dsRNA-based insect control strategies will be considered. This robustness actually resulted in the rise of the red flour beetle, Dicer-2 and Ago-2  and locusts (Orthoptera) injection of dsRNA can also trigger potent responses. In the cockroach, systemic RNAi has been used to unravel the ecdysone regulatory cascade that governs moulting and metamorphosis and the regulation of oogenesis , injection of dsRNA has also been applied successfully to achieve gene silencing. In adult mosquitoes, systemic RNAi has been used to knock-down and investigate the functional role of genes, for example in the immune response against parasites and in the olfactory response , injection of dsRNA has also resulted in gene silencing effects to analyze physiological and developmental processes such as sex determination, molting and the immune response . The analysis of the results deposited in the database led to the conclusion that RNAi is not an efficient process in Lepidoptera but failed to identify conditions that need to be met to achieve success in RNAi , a great variability in success rate exists in the literature with respect to systemic and environmental RNAi efficiency. In the silkmoth, several studies were reported in which successful gene silencing was achieved , which feed on plants through sap-sucking and for which no effective Bt toxins for control are known  in the tissues of the former. Unfortunately, very little is known with respect to differences in expression of the RNAi machinery among different insects. In Dicer-2 and Ago-2 in (adult) gonads could explain the lower RNAi efficiency in these tissues in Schistocerca gregaria -derived C6/36 has also been shown to have defective Dicer-2 expression which could explain the relatively robust growth of arthropod-borne viruses in this cell line  and silencing of sid-1-like genes in Tribolium did not interfere with systemic RNAi  revealed the involvement of scavenger receptors and the endocytosis pathway  series of mutants in C. elegans ; such exosomes can contain mRNAs and small RNAs and be transferred to recipient cells  and Hi5 (from Trichoplusia ni), it could not trigger RNAi, indicating a barrier in the presentation of dsRNA to the RNAi machinery and not in uptake per se  was demonstrated but not of siRNA  than in Manduca hemolymph (low systemic RNAi efficiency) , a non-specific DNA/RNA nuclease with dsRNase activity was purified from the midgut content that apparently is involved in the digestion of nucleic acids in the food  resulted in the induction of Hemolin, an immune response gene , which is characterized by a genome of linear dsRNA segments   can induce physiological effects in insect tissues. Injection of 10 μg of dsGFP in pupae of the Chinese oak silk moth Lutzomyia longipalpis  and that this induction requires Dicer-2 -ssRNA virus] . Reoviridae are characterized by a segmented genome of 10–12 linear dsRNA fragments , the accumulation of large amounts of viral RNAs and small RNAs that will overflow the RNAi machinery, the modulation of expression of host miRNAs and expression of viral miRNAs and the induction of a general antiviral state in the infected insects Figure .Drosophila C virus  and cricket paralysis virus   and the 1A proteins from Drosophila-derived S2 cells , O'nyong'nyong virus (vectored by Anopheles spp), Japanese encephalitis virus, West Nile virus (vectored by Culex spp), vesicular stomatitis virus (vectored by the sand fly Phlebotomus spp) and Bluetongue virus (vectored by biting midges in the genus Culicoides). More recently, however, a few publications appeared that, contrary to expectations discussed above, indicate that flaviviruses such as Dengue and West Nile virus can encode RNAi suppressors, such as the non-coding sfRNA and the non-structural protein NS4B . Another study, on the other hand, showed that Nora virus infection was not affected by Dicer-2, Ago-2 r2d2 and piwi mutations in flies  encodes an RNase III enzyme that is capable of degrading dsRNA and siRNA and is essential for infection  antiviral response pathway. The prevalence of the other antiviral pathway(s) may become very effective in controlling infection and subsequently render the RNAi pathway obsolete. In that case, maintenance of effective RNAi machinery against invading viruses could be selected against and mutations would start to accumulate in the RNAi machinery. Below follows a discussion of alternative mechanisms by which insects may control viral infections and that could lead to a selection against a functional RNAi machinery.Drosophila and mosquitoes  infection  mechanisms than RNAi, it is possible that the need for the maintenance of an active RNAi response against exogenous dsRNA disappears and that mutations accumulate in the RNAi machinery. For instance, frame-shift mutations in In another study of persistent infection of S2 cells by FHV, it was found that during establishment of viral persistence, RNA viruses were reverse-transcribed into cDNA by host reverse transcriptases    has been shown to produce several miRNAs (Singh et al., ie-1 and other important BmNPV genes and its decrease therefore results in the increase of virus accumulation (Singh et al., In case of persistent infection of baculovirus, it can therefore be expected that the miRNA profile in infected cells shows dramatic changes. Given the capacity of miRNAs to regulate many mRNA targets (Lim et al., To test whether persistent virus infection results in changes of RNAi efficiency, two complementary approaches can be used. In the first approach, persistent viral infections will need to be established in insects that have high RNAi efficiency and are virus-free. Examples of such insects are locusts, cockroaches and beetles, in which high RNAi efficiency was reported (Mané-Padrós et al., B. mori are persistently infected with the dsRNA virus BmCPV (Wu et al., Bombyx (Tamura et al., A second approach would consist of a population of persistently infected insects that becomes subsequently cleared from virus. For instance, it is known that many populations of D. virgifera, and the Colorado potato beetle, L. decemlineata, RNAi-based control methods offer a real alternative to other control methods (Gordon and Waterhouse, For coleopteran pests, such as the corn rootworm, C. elegans; Félix, As for every method for insect control, however, the rise of insecticide resistance is always a major issue. It has been argued that resistance against dsRNA or RNA hairpins might be difficult to occur because long dsRNAs can still function effectively, even when multiple mutations have accumulated in the target sequence (Swevers and Smagghe, The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The purpose of this research is to study any effects of managerial strategies on prevention of ketosis metabolic disorder in transition period in Shahroud commercial dairy farms. For this purpose, a questionnaire was prepared in order to obtain required information about the performance of these managerial strategies, performance costs, involvement situation with disorders relying upon clinical signs and treatment and health records, producing and economic situation, and fertility rate and its costs. The considered managerial guidelines include body condition score management or type evaluation in transition period, increase in dry matter intake close to parturition, using propylene glycol, using niacin, and high-quality feeding  in transition period. Finally and upon arrangement of data, it was possible to study any effects of mentioned managerial strategies on related variants through multiple linear regressions. Furthermore, in order to study any relation among variables, we considered Pearson correlation coefficients as well. Finally, it was revealed that any application of managerial strategies for prevention from Ketosis in transition period has a significant effect in betterment of managerial and economic parameters. The transition period, from 3 wk before to 3 wk after parturition, is critically important to health, production, and profitability of dairy cows. Most health disorders occur during this time. There are lots of serious changes for dairy cows in transition period from the end of pregnancy up to the beginning of lactation period. This period was really important for many years and for a lot of researchers due the effects on the health, production, and profitability by focusing on nutrition and management of dairy cows Drackley . Great nBoth clinical and subclinical forms of ketosis are accompanied with a reduction in production, reduction of milk protein, and a reduction in fertility level . There is an increase in suffering of ovary cysts and metritis due to ketosis Radostits et al., . The effRegarding the effect of niacin in prevention from ketosis, Fronk et al.  stated tRegarding Body Condition Score management as an effective strategy in ketosis prevention, Treacher et al.  concludeLinn and Otterby ; Mahana Also through a report published by Duffiled , it was Pickett et al.  confirmeOverton and Waldron ; Weiss aGhorbani and Asadi Alamuti  stated tRegarding the importance of feed quality, Mohebbi  stated tThe present research has been prepared within a time interval from January 2011 up to April 2011 for industrial herds of dairy cows at Shahroud city. There was a questionnaire for required data collection including different questions about 50 herds and analyzed accordingly. The mentioned questionnaire had some information about managerial strategies in first part for management cows in transition period to prevention from ketosis metabolic disorder including body condition score management (BCS), encouraging for consuming more dry matter (consuming dry matter management), using supplementary of propylene glycol, using niacin, and benefiting from high-quality feeding in transition period . Also the performance costs have been measured in the same art. Next part belongs to relevant information about ketosis incidence rate with relying upon clinical signs and therapeutic and health records and also treatment costs for each head. The next part is related to production and economic information including average production parameters per head and also gross income of milk sale per head in lactation cycle. The final part of questionnaire belongs to reproduction records, average fertility, and its costs. Upon data arrangement, we had an analysis through multiple linear regression (MLR) and Statistical Software (SAS) 9.1. Also, we could calculate correlation of effective managerial strategies and related variants through a correlation method as follows:This managerial strategy includes the body condition score control especially before parturition in a way that it is possible to make ready a cow with suitable BCS for parturition.P < 0.01), production average, gross income of milk sale, and average fertility (P < 0.05). But it has no more effects on fertility costs and artificial inbreeding from statistical view point (P > 0.05)  . The menP > 0.05). There was not any significant difference between treatments from treatment costs point of view (P > 0.05). Regarding the gross income, it is possible to say that there is not a significant difference among experimental treatments (P > 0.05). Also, there was not a significant difference between average fertility and artificial inbreeding (P > 0.01) (The obtained results out of average comparison show that there is not a significant relation from incidence rate between different groups ( > 0.01) .Following is relevant study of correlation coefficients between BCS management and other managerial and economic indices .r = −0.60 which was a positive effect of applied BCS in reducing of ketosis incidence rate. This means that any increase in application of BCS management may cause a reduction in ketosis incidence rate. We calculated any relation between BCS Management and ketosis incidence rate r = −0.54.Any relation between BCS and ketosis treatment costs. As it is expected any reduction in ketosis incidence rate in herds may cause required therapeutic costs. Therefore, with regard to the effect of BCS in reduction of ketosis rate, it is concluded that BCS management may cause a reduction in therapeutic costs. Correlation coefficient for such a relation is r = 0.31 which is a sign of positive effect of this managerial strategy on milk production average in herd.Any relation between BCS and average production. We calculated the correlation coefficient of BCS relation and average production equal to r = 0.32). It is possible to evaluate this relation as a direct relation of BCS and average production and also major role of average production in calculation of gross income in this research. As a result, it is expected to have betterment in gross income upon increasing the application of this managerial strategy.We evaluated and found out a positive relation between BCS management and gross income (r = 0.28), although this relation was now so much specific like other variants but it was considerable as well.Any relation between BCS and average fertility. In this part, we found out a positive Pearson correlation coefficient . But it has not a statistical significant effect on average fertility and fertility costs and artificial inbreeding (P > 0.05)  . These rP < 0.05). By comparing the averages, there is not a significant difference between the groups from therapeutic costs (P > 0.05). Although there are some differences for average production rates, gross income from milk sale, average fertility, and fertility costs, and artificial inbreeding, but it was not so much significant from statistical viewpoint (P > 0.05) (The results out of a comparison in averages in current research show that there is a significant difference between these two groups (executive group and a group with lack of attention) to Ketosis incidence rate from statistical point of view ( > 0.05) .The results out of Pearson correlation coefficients show the following relations among different variables .r = −0.67 was a sign of its effect in reducing of ketosis incidence rate.In order to study any relation between intake rate variable increase and ketosis incidence rate with calculated correlation coefficient, r = 0.43 was calculated as a sign of the importance of this parameter in milk production rate.Correlation coefficient for dry matter intake and average production, r = 0.42.Any relation between dry matter intake and gross income and considering the effect of this variable on production rate and direct relation of these two factors have been calculated by r = −0.15) as a sign of converse relation between these two factors with independent increase, dependent factor . Also r = 0.25 is the correlation coefficient for dry matter intake and average fertility.Any relation between fertility costs and current variable have been calculated by a negative digit (r = −0.55) is a sign of increasing the intake rate before parturition which may cause a reduction in relevant treatment costs of ketosis.Any relation between intake rate and treatment costs is a negative correlation. (P < 0.05)  . These rAlso after obtaining the mentioned results, Studer et al.  and FormP < 0.05). Regarding both variables of average production and gross income, there is a significant difference between both mentioned groups from the point of view of average properties (P < 0.05). Any comparison of the average of both groups did not prove any significant differences between fertility average and fertility costs (P > 0.05) (The results of comparing the average show that there is a significant difference between compared groups from the point of view of incidence rate and ketosis treatment costs ( > 0.05) .Also, we obtained the following results after calculating the correlation coefficients in this research .r = −0.71. This means that applying of propylene glycol may cause a considerable reduction in ketosis incidence rate.There was a considerable relation between using of propylene glycol and ketosis incidence rate in a way that correlation coefficient for these two variants was r = −0.73) in a way that cased a reduction in ketosis treatment costs.Regarding the positive effect of prevention, there was a considerable relation between application of propylene glycol and treatment costs . The results obtained by Bartlett et al. [P < 0.05). But there is not a significant difference between the mentioned groups from the viewpoints of production average, gross income, average fertility, and fertility costs (P > 0.05). Calculation of Pearson correlation coefficients (r = −0.74). Also there is a negative correlation and significant relation between niacin application and treatment costs (r = −0.79). Correlation coefficients between niacin application and average production, gross income of milk sale, fertility average, and fertility costs were, respectively, equal to 0.60, 0.60, 0.64, and −0.43.The findings out of variance analysis show that niacin application may have a positive effect on producing, economic, and managerial parameters such as average production, gross income, ketosis rate, treatment costs, and generation parameters (fertility average and its costs) , and fertility costs (P < 0.05). These findings are in compliance with the results of different researches made by Mohebbi [The obtained results out of regression and variance analysis revealed that there is a positive relation between performing this strategy and other related variants . This me Mohebbi  who stat Mohebbi  and Kead Mohebbi  who statP < 0.05). There was not a significant difference between both groups from statistical viewpoint and for treatment costs, average production, gross income, average fertility, and fertility costs (P > 0.05) (The obtained results out of comparing the averages show that there is a significant difference between occurrence rate parameter at probable level of 0.05 of both groups ( > 0.05) .r = −0.73). Furthermore, we calculated any relation between feed quality with treatment costs r = −0.62, production average r = 0.48, gross income r = 0.47, average fertility r = 0.38, and fertility costs r = −0.29.Also the correlation coefficient show that there is a considerable relation between quality of feeding and occurrence rate (P < 0.01). This means that any increase of costs resulted from applying of managerial strategies are effective in prevention from ketosis accompanied with betterment of functions of producing units and also upgrading of managerial and economic parameters. Generally, these strategies are reasonable from economic viewpoints.This variable includes a set of costs imposed on a cattleman for performing all above-mentioned factors . In this part, we will consider any relation as an independent variable with other related variants. The obtained results of regression performance and variance analysis  for thisP < 0.05). But from statistical viewpoint, there are not significant differences among mentioned groups as well (P > 0.05)  .We considered correlation coefficient for measuring any relation between performance costs of managerial strategies and other managerial and economic indices with the following results .r = −0.80. This means that any increase in mentioned costs in strategies performance will reduce the occurrence rate of ketosis in herds.Correlation coefficient for performance costs of strategies and ketosis rate was r = −0.82. As it was explained that parameter would reveal its effect on treatment costs by reducing the occurrence rate of ketosis.Correlation coefficient for performance costs of managerial strategies and ketosis treatment costs was r = 0.72 which is a sign of its positive effect on production.We calculated any relation between performance costs of strategies and production average as Correlation coefficient was calculated for gross income, fertility average, and inbreeding costs and fertility, respectively, as 0.71, 0.70, and −0.54.P < 0.01). It is in a way that any percentage increase of these strategies may cause betterment in managerial and economic conditions of training units.Calculation of this parameter is on percentage basis for total effective managerial strategies on ketosis prevention and as a sign of performance level of it for preventing this metabolic disorder in transition period. Regression results and analysis of variance  show thaP < 0.01), treatment costs (P < 0.05), average production, and gross income (P < 0.05). But they did not have any statistical difference for fertility average and fertility costs variants (P > 0.05).The results out of average comparisons among different groups effective on ketosis prevention show that considered groups had significant differences about the following variants like: occurrence rate (r = −0.98). Therefore, any increase in performing of managerial strategies has a considerable effect in prevention from ketosis as well. Also, these results confirm a positive relation of performance percentage of managerial strategies on treatment costs (r = −0.92). Correlation coefficient for above-mentioned variable for average production was r = 0.71, and for gross income was r = 0.71, for average fertility r = 0.62 and for fertility costs r = −0.44. As it is obvious in Tables r = −0.  80). Then, we have niacin application (r = −0.74), propylene glycol (r = −0.71), and increasing dry matter consumption (r = −0.67), respectively, with the highest rate of relation with this variable. Also regarding any relation with treatment costs, using niacin (r = −0.79) and propylene glycol (r = −0.73) have, respectively, highest rate of relation as well. Regarding the average production and gross income they have, respectively, highest rate of relation with propylene glycol (r = 0.67), niacin (r = 0.60), benefiting from highest-quality feed (r = 0.48), and propylene glycol supplement (r = 0.58).The results of correlation coefficients show that there is a close relationship between performance percentage of managerial strategies and ketosis occurrence rate ( r = −0.9. Correlar = −0.49) and niacin (r = −0.43) accordingly.Regarding the fertility costs and its relation with managerial strategies, there was exactly a contrast condition. It means that there is a more relation between application rate of propylene glycol (