Existence <Emphasis Type="Italic">in vivo</Emphasis> of the loop E motif in potato spindle tuber viroid RNA

Summary In vitro experiments have previously identified in potato spindle tuber viroid (PSTVd), the type member of the nuclear viroids, an element of local tertiary structure termed loop E. Here, by direct UV irradiation of PSTVd-infected tomato tissue and subsequent RNA analysis by denaturing polyacrylamide gel electrophoresis, northern blot hybridization and primer extension, we report that PSTVd (+) RNA also forms the loop E in vivo. These results provide strong support for the physiological relevance of this structural motif, which is involved in a wide range of functions including replication, host specificity and pathogenesis.     

One-step site-directed mutagenesis of viroid dimeric cDNA

The study of viroids (plant pathogens constituted by small non-coding RNA) has greatly benefited from the use of site-directed mutagenesis tools. However, compared to viral systems, this technique is complicated by the fact that, usually, infectious cDNAs carry two copies of the viroid genome. A simple method for a one-step site-directed mutagenesis of viroids is described and tested by estimating the rate of mutation incorporation of three random nucleotide substitutions in each Chrysanthemum stunt viroid (CSVd) and Chrysanthemum chlorotic mottle viroid (CChMVd). The protocol is essentially based on the original QuickChange Stratagene methodology; dimeric cDNA templates are amplified directly using Pfu DNA polymerase and self-complementary mutagenic primers. The reaction typically yields dimeric, but also monomeric clones which can be easily distinguished by electrophoretic analysis. The data show that approximately 50% of the dimeric clones carry the desired mutation in both viroid copies. Since the proposed protocol is simple technically and rapid compared to previous methods, it could be applied routinely for site-directed mutagenesis studies in viroids.     

Ecliptic method for the determination of backscatter into the beam monitor chambers in photon beams of medical accelerators

A new method to measure the effect of the backscatter into the beam monitor chambers in linear accelerators is introduced from first principles. The technique, applicable to high-energy photon beams, is similar to the well-known telescopic method although here the heavy blocks are replaced by a very small, centred block on the shadow tray, thus the name 'ecliptic method'. This effect, caused mainly by backscattering from the secondary collimators, is known to be an output factor constituent and must be accounted for when detailed calculations involving the machine's head are required. Since its magnitude is generally small, experimental errors might obscure the behaviour of the phenomenon. Consequently, the procedure introduced goes along with an uncertainty assessment. Our theory was confirmed via measurements in cobalt-60 beams, where the studied effect does not contribute to the output factor. Measurements were also performed on our Saturne 41 linear accelerator and the results were qualitatively similar to those described elsewhere. The collimation systems were studied separately by varying one jaw setting while keeping the other at its maximum value. In the light of these results, we deduced an algorithm that can correlate the former data with the effect of backscattering to the beam monitor chambers for any rectangular field within 0.5%, which is of the order of the experimental uncertainty (0.6%). As we show, the experimental procedure is safe, simple, not invasive for the linac and requires only basic dosimetry equipment.     

Endophthalmitis after intravitreal injections versus cataract surgery: a 15-year cohort

To compare the clinical features, visual outcomes and causative organisms between endophthalmitis secondary to cataract surgery or to intravitreal injections (IVI). Meir Medical Center, Kfar Saba, Israel. Retrospective, non-randomized comparative chart review. Medical records of patients with proven or suspected endophthalmitis admitted to the Ophthalmology Department at Meir Medical Center 2/2002–2/2017 were reviewed. Clinical characteristics including presenting and final visual acuity (VA) outcomes, causative organisms and time to admission were assessed. Among 84 patients in our study, 35 had preceding cataract surgery and 12 had preceding IVI. The post-cataract group showed a significant improvement in VA following treatment with a presenting and final VA (logMar ± SD) of (1.80 ± 0.54 and 1.39 ± 0.65, P < 0.01) as opposed to the post-IVI group (1.72 ± 0.26 and 1.81 ± 0.32, P = 0.692), while most patients in the cataract group exhibited some degree of VA recovery (70.96%). Patients undergoing cataract surgery were divided into two separate groups; those who underwent cataract surgery in a private center and those operated at a public center. Patients undergoing surgery at a private medical center showed improvement in VA outcomes following treatment (1.80 ± 0.57 and 1.13 ± 0.66, P < 0.01) as opposed to those operated on at our public medical center. Overall, patients with endophthalmitis following cataract surgery had better visual outcomes and were more likely to show a VA improvement following treatment when compared with endophthalmitis following IVI. Final VA outcomes of patients with endophthalmitis after cataract surgery performed in a private center were better than those operated on and treated in a public medical center.     

Carnosic acid ameliorates depressive‐like symptoms along with the modulation of FGF9 in the hippocampus of middle carotid artery occlusion‐induced Sprague Dawley rats

The increased survival rate of stroke patients has led to the higher incidences of post‐stroke depression. Carnosic acid has the ability to cross blood brain barrier with good neuro‐modulatory actions. Recently, inclined level of fibroblast growth factor 9 (FGF9) in the postmortem brain of the depressed patients was noted. Therefore, in the present study, the effect of carnosic acid on post‐stroke depression‐like behavior, and the expression of FGF9 were evaluated. After 3 weeks of middle carotid artery occlusion in Sprague Dawley rats, carnosic acid (20 and 40 mg/kg) was administered for 2 weeks. Sucrose preference test, forced swimming test, and open field test were performed and hippocampi were analyzed for FGF9 and FGFR‐3. In comparison to post‐stroke depressed rats, carnosic acid increased the sucrose preference, and reduced the immobility time of the rats by ~2×. The speed and distance‐covered were also increased. At 40 mg/kg, FGF9 was reduced by ~3× while FGFR‐3/Actin was increased by ~1.5×. Altogether results suggest anti‐depressant‐like activity of carnosic acid in post‐stroke depressed rats with decreased expression of hippocampal FGF9.     

Vertebral artery injuries in cervical spine surgery

Background contextVertebral artery injuries (VAIs) are rare but serious complications of cervical spine surgery, with the potential to cause catastrophic bleeding, permanent neurologic impairment, and even death. The present literature regarding incidence of this complication largely comprises a single surgeon or small multicenter case series.PurposeWe sought to gather a large sample of high-volume surgeons to adequately characterize the incidence and risk factors for VAI, management strategies used, and patient outcomes after VAI.Study designThe study was constructed as a cross-sectional study comprising all cervical spine patients operated on by the members of the international Cervical Spine Research Society (CSRS).Patient sampleAll patients who have undergone cervical spine surgery by a current member of CSRS as of the spring of 2012.Outcome measuresFor each surgeon surveyed, we collected self-reported measures to include the number of cervical cases performed in the surgeon's career, the number of VAIs encountered, the stage of the case during which the injury occurred, the management strategies used, and the overall patient outcome after injury.MethodsAn anonymous 10-question web-based survey was distributed to the members of the CSRS. Statistical analysis was performed using Student t tests for numerical outcomes and chi-squared analysis for categorical variables.ResultsOne hundred forty-one CSRS members (of 195 total, 72%) responded to the survey, accounting for a total of 163,324 cervical spine surgeries performed. The overall incidence of VAI was 0.07% (111/163,324). Posterior instrumentation of the upper cervical spine (32.4%), anterior corpectomy (23.4%), and posterior exposure of the cervical spine (11.7%) were the most common stages of the case to result in an injury to the vertebral artery. Discectomy (9%) and anterior exposure of the spine (7.2%) were also common time points for an arterial injury. One-fifth (22/111) of all VAI involved an anomalous course of the vertebral artery. The most common management of VAI was by direct tamponade. The outcomes of VAIs included no permanent sequelae in 90% of patients, permanent neurologic sequelae in 5.5%, and death in 4.5%. Surgeons at academic and private centers had nearly identical rates of VAIs. However, surgeons who had performed 300 or fewer cervical spine surgeries in their career had a VAI incidence of 0.33% compared with 0.06% in those with greater than 300 lifetime cases (p=.028).ConclusionsThe overall incidence of VAI during cervical spine surgery reported from this survey was 0.07%. Less experienced surgeons had a higher rate of VAI compared with their more experienced peers. The results of VAI are highly variable, resulting in no permanent harm most of the time; however, permanent neurologic injury or death occur in 10% of cases.     

Molecular and Parasitological Study of Cryptosporidium Isolates From Cattle in Ilam,West of Iran

Background

Cryptosporidiosis is one of the most important parasitic infections in human and animals. This study was designed for survey on the prevalence of Cryptosporidium infection in farms of Ilam, west of Iran, using parasitology method and genotyping by Nested PCR-RFLP.

Methods

Fecal samples of 217 cattle were collected fresh and directly from the rectum of cattle. All of the samples were examined by microscopic observation after staining with modified Ziehl-Neelsen (MZN). Genomic DNA extracted by using EURx DNA kit. A Nested PCR-RFLP protocol amplifying 825 bp fragment of 18s rRNA gene conducted to differentiate species and genotyping of the isolates using SspI and VspI as restriction enzymes.

Results

The prevalence of Cryptosporidium infection in cattle using both methods is 3.68%. Most of the positive cattle were calves under six months. Species diagnosis carried out by digesting the secondary PCR product with SspI that C. parvum generated 3 visible bands of 448, 247 and 106 bp and digested by VspI restriction enzyme generated 2 visible bands of 628 and 104bp. In this investigation all of the positive samples were Cryptosporidium parvum.

Conclusion

C. parvum (bovine genotype) detected in all positive cattle samples in Ilam, west of Iran. The results of the present study can help for public health care systems to prevention and management of cryptosporidiosis in cattle and the assessment of cattle cryptosporidiosis as a reservoir for the human infection.     

Outcomes of junctional ectopic tachycardia ablation in adult population—a multicenter experience

Purpose

Idiopathic junctional ectopic tachycardia (JET) is typically refractory to antiarrhythmic agents. Catheter ablation for JET is feasible but is associated with high risk of unintended atrioventricular (AV) block. There is limited data on the appropriate procedural technique and clinical outcomes with catheter ablation for idiopathic JET in adults.

Methods

This is a multicenter, retrospective study of all adult patients (age?≥?18 years) who underwent catheter ablation for idiopathic JET. Patient, procedural characteristics, and long-term outcomes were evaluated.

Results

Fifteen patients [radiofrequency ablation (RF)?=?14 and cryoablation?=?1) were treated with catheter ablation. The median age was 58 years with 67% males. All patients underwent mapping of the right atrium and the aortic cusps prior to energy delivery. The location of earliest activation in relation to the atrioventricular (AV) node was postero-superior in 73% (11/15), posterior in 13% (2/15), and superior in 13% (2/15) respectively. Acute success was 100%. Arrhythmia recurrence occurred in 53% (8/15) all of whom underwent a repeat ablation. High-grade AV block requiring permanent pacemaker occurred in 20% (3/15). At 12-month follow-up in the redo-ablation group, 37.5% (3/8) had recurrence of the arrhythmia two of which underwent a third ablation procedure.

Conclusion

Catheter ablation of idiopathic JET in adults is associated with a high rate of recurrence requiring multiple procedures and high risk of AV block requiring a permanent pacemaker. Mapping and ablation of the non-coronary cusp can be considered as the arrhythmia was controlled in 3 patients with no inadvertent AV block.