Analysis of the A-U rich hairpin from the intergenic region of tospovirus S RNA as target and inducer of RNA silencing

Earlier work indicated that Tomato spotted wilt virus (TSWV) messenger transcripts, and not the (anti)genomic RNAs, are targeted by the RNA silencing machinery. Here, the predicted AU-rich hairpin (HP) structure encoded by the intergenic region (IGR) of the TSWV S RNA, and present at the 3' end of viral mRNAs, was analyzed as a target and inducer for RNA silencing. Virus-derived siRNAs (vsiRNAs) purified from virus infected plants were found to derive from all three genomic RNA segments but predominantly the ambisense M and S RNAs. Further profiling on the S RNA sequence revealed that vsiRNAs were found from almost the entire S RNA sequence, except the IGR from where hardly any vsiRNAs were found. Similar profiles were observed with the distantly related Tomato yellow ring tospovirus (TYRV). Dicer cleavage assays using Drosophila melanogaster (Dm) embryo extracts showed that synthetic transcripts of the IGR-HP region were recognized as substrate for Dicer. Transient agroinfiltration assays of a GFP-sensor construct containing the IGR-HP sequence at its 3' UTR (GFP-HP) did not show more rapid/strong silencing and profiling of the corresponding siRNAs, generated outside the context of a viral infection, still revealed relatively low levels of IGR-HP-derived siRNAs. These data support the idea that the IGR-HP is a weak inducer of RNA silencing and only plays a minor role in the amplification of a strong antiviral RNAi response

Molecular Diagnosis of Iris Yellow Spot Virus (IYSV) on Onion in Iran

Viral symptoms indicative of Iris yellow spot virus (IYSV) were observed on onion in several fields near Chenaran in Khorasan Razavi Province. Mechanical inoculation of herbaceous hosts with onion sap extracts from symptomatic plants showed similar symptoms to those described for IYSV. The mechanically transmitted virus reacted only with antisera specific to IYSV in DAS-ELISA but not with antisera specific to seven other tospoviruses. In RT-PCR, a DNA fragment approximately 822 bp in size was amplified from infected Nicotiana benthamiana by using primers specific to the nucleocapsid (N) gene of IYSV. After cloning and sequencing, the deduced N protein sequence of two isolates (GenBank accession no. HQ148173 and HQ148174) showed 98% amino acid identity with a Sri Lankan isolate, 96% with a Dutch isolate and 92% with a Brazilian isolate. To our knowledge, this is the first molecular characterization of IYSV in Ira

Strategic crossing of biomass and harvest index—source and sink—achieves genetic gains in wheat

To accelerate genetic gains in breeding, physiological trait (PT) characterization of candidate parents can help make more strategic crosses, increasing the probability of accumulating favorable alleles compared to crossing relatively uncharacterized lines. In this study, crosses were designed to complement “source” with “sink” traits, where at least one parent was selected for favorable expression of biomass and/or radiation use efficiency—source—and the other for sink-related traits like harvest-index, kernel weight and grains per spike. Female parents were selected from among genetic resources—including landraces and products of wide-crossing (i.e. synthetic wheat)—that had been evaluated in Mexico at high yield potential or under heat stress, while elite lines were used as males. Progeny of crosses were advanced to the F4 generation within Mexico, and F4-derived F5 and F6 generations were yield tested to populate four international nurseries, targeted to high yield environments (2nd and 3rd WYCYT) for yield potential, and heat stressed environments (2nd and 4th SATYN) for climate resilience, respectively. Each nursery was grown as multi-location yield trials. Genetic gains were achieved in both temperate and hot environments, with most new PT-derived lines expressing superior yield and biomass compared to local checks at almost all international sites. Furthermore, the tendency across all four nurseries indicated either the superiority of the best new PT lines compared with the CIMMYT elite checks, or the superiority of all new PT lines as a group compared with all checks, and in some cases, both. Results support—in a realistic breeding context—the hypothesis that yield and radiation use efficiency can be increased by improving source:sink balance, and validate the feasibility of incorporating exotic germplasm into mainstream breeding efforts to accelerate genetic gains for yield potential and climate resilience

FEMORAL MONONEUROPATHY: COMPLICATING A DIFFICULT PELVIC URETEROLITHOTOMY

Femoral mononeuropathy is a complication of gynecological and urological operations. Tlte objective of this citse report is to bring to the attention of fellow urologists the very existence of this complication and offer measures to prevent it. A 48 year old obese worman with a 2 year history of frequent left renal colics, anil a nonfunction left kidney on IVP underwent a retrograde ureterogram study anil a transvaginal ureterolithotomy, in an exaggerated lithotomy position. Postoperatively, a left leg paralysis required 3 weeks of care and encouragement to heal. To prevent this complication, avoid putting too long and too much pressure by retractors on the psoas muscles. During the abdomino-perineal procedures, frequent change of retractor site and monitoring of distal arterial pulses would he helpful. Also, do not use the exaggerated lithotomy position for an extended period of time

Does vesico-ureteral reflux in original kidney of transplant recipient increase the risk of infection or rejection?

A prospective study during 1990-1995 was conducted to assess the risk of rejection and infection in kidney-transplant recipients, who's original kidney had vesico-ureteral reflux (VER). Two groups of recipients with and without VER were followed for one month after transplant; 24 and 250 cases respectively. The calculated relative risks for the effect of VER showed a moderate increased risk for rejection in VER group; (RR=1.82, CI=1.12-2.95, P<0.05). No effect was found for VER being one-or two-sided. Hence, VER has a moderate effect on rejection, but no effect on infection

OUTCOME OF LIVE DONOR RENAL ALLOGRAFT TRANSPLANTATION FROM SINGLE VS MULTIPLE ARTERIES' GRAFTS

This study compare:.' [he results 0;,.1 outcome of live-donor transplantation between single-artery "',"' mull/pic-ana)' transplant kidneys. Cadaver kidneys with multiple vessels arc retrieved with a patch of the donor artery. 111is is not possible ill the !iI'C donation seuing. Therefore !i1'C donation of rcnal"nallografts with multiple arteries is lIot a straiglnjorward surgery. We studied 22 muttiplc-anery live donor renal allografts among 223 renal transplantations in a sequential. prospective mOllTlCr [or 3 ynJrs. One-year gra{! survival was l)(j.:V:(, ill single-anery group and 95.5":{, in tlns muliplc . arIer)' group. III the singleartery group the complications wae: dctavcd gm[l [unction ill 3.5'7;, rean astomosis o[ tlu: v-essels in 2,9':k, transient post-transplant dialysis in 1. 5 (X" graft nephrectomy ill 2,5';{, AT"' ill 1":'(" Urine leak in 2.5':{', renal anav stenosis in O.5S'(" and lvmpho cclc ill 1%. NOlie: o] thcsc occurred in the"nmultiptc-oncry group. This difference is statistically significant IX~ = 8.10). Cold ischemia time: l"'(lS significantly lunger in lilt' multiple . anery group (panastomosis was not siglliftcanl~"' dlffaelll among lht' 2,1,'Youps (I = 1.255). Ttu: totat tcngtli of tile operation IVas IOllga ill lhe mutsiptc-oncry group (p < O. 00(5). In conclusion it is appareIH snas t lu: intra-op crativc complications. posi-operati vc complications and one-year grafr survival are ccnnparabtc ill"nsingle - ane'Y' "'."'. mutsiptc - arrcry renal transplantation. tn other words, !i1'C - donor transptannuion with muliip!c . arIa)' reno! units is safe and has a good OI/lCO!1le

Significant pathological findings in routine urethrocystoscopy before open prostatectomy

Background: Over the years, Trans Urethral Resection of Prostate (TURP), as a treatment modality for obstructing benign prostatic hyperplasia (BPH), gained popularity throughout the world. It is considered the gold standard for the surgical management of BPH. However, there is not the same pattern in our country and open prostatectomy is more common. Specific evaluation should be done preoperatively. Cystoscopy is one of them that have special limited indications. The aim of this study was to define the rate of pathological findings in routine urethrocystoscopy before open prostatectomy.Methods: In a prospective study from 2003 to 2005 in Dr Shariati hospital 120 patients prepared for open prostatectomy underwent routine cystoscopy, without absolute indications for cystoscopy.Results: There was no special pathology observed in routine cystoscopy. The results of this study revealed that the mean of age in patients with meatal stricture was significantly lower than those without stricture (59.5±0.7 vs. 49.9±5.5 years, p=0.008). Also, the results of present study showed that in patients with meatal stricture serum creatinine level was significantly higher (2.2±1.34 vs. 1.2±0.42 mgr/dL, p=0.003). Furthermore, it was revealed that the mean of age in patients with bladder diverticulum was significantly higher compared with those without bladder diverticulum (76.3±3.1 vs. 68.4±4.9 years, p=0.001).Conclusion: Routine cystoscopy is not necessary before open prostatectom

First report of Moroccan pepper virus on Lisianthus in Iran and Worldwide

During the last decade, lisianthus (Eustoma grandiflorum) has been introduced in Iran in the horticultural cut-flower industry. This crop is currently produced in more than 800 small greenhouses on a surface of an estimated 0.8 km2 in the Pakdasht region (southeast of Teheran Province). Plants exhibiting virus-like symptoms were observed in several greenhouses in 2010. The infected plants produced yellow and necrotic spots on the leaves and became severely deformed because of a strong leaf curling and the production of shorter internodes. Flower breaking has not been observed in the blue flowering plants. Approximately 85% of the plants were apparently infected in the inspected greenhouses. Extracts of infected material inoculated onto some indicator plant species induced mosaic and leaf malformation on Nicotiana benthamiana, mottling on Capsicum annuum, necrotic lesions on Datura stramonium, chlorotic local spots on Vigna unguiculata, systemic necrotic spots on Emilia sonchifolia, chlorotic local spots on Cucumis sativus, and necrotic local lesions on Petunia hybrida. Back-inoculation of infected material on lisianthus seedlings resulted in several chlorotic spots on the inoculated leaves and a severe downward curling of the systemic infected leaves. No symptoms were observed after inoculation of Pisum sativum, Phaseolus vulgaris, Vicia faba, and Chrysanthemum spp. The virus could also be transferred from infected to healthy N. benthamiana plants by pricking leaves with a Pasteur pipette. Spherical tombusvirus-like particles of approximately 29 nm were found by transmission electron microscopy in leaf-dip and partially-purified preparations of infected N. benthamiana. Since Tomato bushy stunt virus (TBSV; genus Tombusvirus, family Tombusviridae) and Moroccan pepper virus (MPV) have been found in Iran, we studied by using ELISA whether our samples matched with TBSV. Since a negative response was obtained, two primers were designed on the basis of the available sequences of the coat protein in the GenBank (Accession No. EU27780) of an MPV isolate from soil in Fars Province, Iran. A reverse transcription (RT)-PCR of total RNA extract from infected lisianthus and N. benthamiana with the primers MPV-R (5'-TTACAACAATGTGGCATCATTG-3') and MPV-F (5'-ATGGCAATGGTAGTAAG AAAC-3') resulted in a DNA fragment of 1,176 bp. This fragment from N. benthamiana was cloned, sequenced (Accession No. HQ663881), and showed a 96% nucleotide and 99% amino acid identity with the coat protein of the soil isolate. MPV was originally found in pepper (1), tomato and pelargonium (4), pear tree (3), and surface water (2). To our knowledge, this is the first report of MPV on lisianthus in Iran and worldwide. This virus, which persists in soil, water, and plant debris, can be considered as a substantial threat for the lisianthus industry in Iran because farmers do not apply strict crop rotation or other sanitation measure

Molecular Identification of an Isolate of Peanut Mottle Virus (PeMoV) in Iran

Peanut plants showing mottling, yellow and necrotic spots on leaves were collected from peanut fields in Golestan province. Electron microscopic studies revealed the presence of flexuous filamentous particles ca. 700 nm in length, which was suggestive of a potyvirus infection. Healthy Nicotiana benthamiana plants mechanically inoculated with sap from infected peanut plants showed mottling, downward leaf curling, and wrinkling of the leaves. The virus was transmitted by Myzus persicae in a non-persistent manner to healthy N. benthamiana, on which symptoms were observed two weeks later. RT-PCR using an Oligo-dT and a NIb primer set resulted in a fragment of about 1093 bp, which comprised the complete coat protein (CP) gene and 3´-non-coding region. Analysis of its CP nucleotide and amino acid sequence revealed 98-99% similarity and 95-99% identity to those of Peanut mottle virus (PeMoV) isolated from other countries, respectively. The molecular data confirmed serological, vector transmission, and electron microscopic findings on the incidence of PeMoV in Iran. Additionally, sequence and phylogenetic analyses of the CP revealed clustering of Iranian PeMoV isolate with Asian/Australian isolates