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Background: Pain is common in children with cerebral palsy. The purpose of this systematic review was to evaluate the evidence regarding assessments and interventions for chronic pain in children aged ≤2 years with or at high risk for cerebral palsy. Methods: A comprehensive literature search was performed. Included articles were screened using PRISMA guidelines and quality of evidence was reviewed using best-evidence tools by independent reviewers. Using social media channels, an online survey was conducted to elicit parent preferences. Results: Six articles met criteria. Parent perception was an assessment option. Three pharmacologic interventions (gabapentin, medical cannabis, botulinum toxin type A) and 1 nonpharmacologic intervention were identified. Parent survey report parent-comfort and other nonpharmacologic interventions ranked as most preferable. Conclusion: A conditional GRADE recommendation was in favor of parent report for pain assessment. Clinical trials are sorely needed because of the lack of evidence for safety and efficacy of pharmacologic interventions. [ABSTRACT FROM AUTHOR]

Aim: To evaluate the properties of the Infant Motor Activity Log (IMAL), a caregiver-report for frequency and quality of use of more affected upper extremity in infants with neurological and functional impairments.Method: This was a prospective cohort study of 66 children (34 females, 32 males) aged 6 to 24 months (mean age [SD] 13.7mo [5.3]) with neurological and functional impairments and a confirmed cerebral palsy diagnoses after 2 years, and 51 age-matched typically developing children. The IMAL was administered at baseline and 4 weeks later. Typically developing infants were tested with randomly assigned 'more affected' upper extremity. Psychometric properties were evaluated using Spearman's correlation coefficient, Cronbach's alpha, and Jonckheere-Terpstra tests.Results: In the children with impairments, the IMAL showed internal consistency (alpha≥0.88) for the How Well Scale (HWS) and How Often Scale (HOS). Test-retest reliability was 0.64 (HOS) and 0.70 (HWS), demonstrating stability over time. Correlation with Bayley Scales of Infant and Toddler Development, Third Edition more affected arm raw scores were 0.70 (HOS) and 0.72 (HWS) (p<0.001) demonstrating construct validity. Both scale scores decreased with increasing Gross Motor Function Classification System and Mini-Manual Ability Classification System (p<0.001) levels, supporting discriminative validity. Discrimination between typically developing infants and infants with impairments was high (HWS: area under the receiver operating characteristic curve [AUC] 0.96, 95% confidence interval [CI] 0.94-0.99 and HOS AUC=0.95, CI 0.92-0.99).Interpretation: The IMAL is a valid and reliable discriminative caregiver measure of upper limb performance and may complement measures of capacity in infants with neurological and functional impairments.What This Paper Adds: The Infant Motor Activity Log (IMAL) is a valid and reliable measure of caregiver perception of upper limb function. The IMAL fills a measurement gap for infant motor performance in children with impairments. The IMAL discriminates among motor function levels. [ABSTRACT FROM AUTHOR]

Upper extremity (UE) impairments in infants with cerebral palsy (CP) result from reduced quality of motor experiences and "noisy" sensory inputs. We hypothesized that a neuroscience-based multi-component intervention would improve somatosensory processing and motor measures of more-affected (UEs) in infants with CP and asymmetric UE neurologic impairments, while remaining safe for less-affected UEs. Our randomized controlled trial compared infants (6–24 months) with CP receiving intervention (N = 37) versus a waitlisted group (N = 36). Treatment effects tested a direct measurement of reach smoothness (3D-kinematics), a measure of unimanual fine motor function (Bayley unimanual fine motor raw scores), and EEG measures of cortical somatosensory processing. The four-week therapist-directed, parent-administered intervention included daily (1) bimanual play; (2) less-affected UE wearing soft-constraint (6 h/day, electronically-monitored); (3) reach training on more-affected UE; (4) graduated motor-sensory training; and (5) parent education. Waitlist infants received only bimanual play. Effectiveness and safety were tested; z-scores from 54 posttest-matched typically-developing infants provided benchmarks for treatment effects. Intervention and waitlist infants had no pretest differences. Median weekly constraint wear was 38 h; parent-treatment fidelity averaged > 92%. On the more affected side, the intervention significantly increased smoothness of reach (Cohen's d = − 0.90; p <.001) and unimanual fine motor skill (d = 0.35; p =.004). Using unadjusted p values, intervention improved somatosensory processing (d = 0.53; p =.04). All intervention effects referenced well to typically developing children. Safety of the intervention was demonstrated through positive- or non-effects on measurements involving the constrained, less-affected UE and gross motor function; unexpected treatment effects on reach smoothness occurred in less-affected UEs (d = − 0.85; p =.01). This large clinical trial demonstrated intervention effectiveness and safety for developing sensory and motor systems with improvements in reach smoothness, and developmental abilities. Clinical Trail Registration: ClinicalTrials.gov NCT02567630, registered October 5, 2015. [ABSTRACT FROM AUTHOR]

Over lOU years after its discovery, Tobacco mosaic virus (TMV) remains an economically important pathogen for producers of many vegetatively propagated crops including petunias (Petunia x hybrida). To directly address this concern, we have developed a robust system to determine efficacy of disinfectants for treating TM V-contaminated cutting tools using a combination of preliminary screens and replicated trials. Contrary to widely held beliefs, wild-type (wt) TMV and four additional tobamovirus species infected four petunia cultivars without producing obvious viral symptoms. In contrast, a petunia isolate of TMV with 99.0% (nucleotide) and 99.4% (amino acid) coat protein sequence identity to wt TMV produced symptoms on all but one tested cultivar. We also show that TMV transmission can occur up to the twentieth petunia plant cut following a single cutting event on a TM V-infected plant. Although many new products are now available, treatment of TMV-contaminated tools with a 20% (wt/vol) solution of nonfat dry milk (NFDM) plus 0.1% Tween 20 or a 1:10 dilution of household bleach (0.6% sodium hypochlorite), two "old standbys", completely eliminated TMV transmission to petunias. Treatment of contaminated tools with 1% (wt/vol) Virkon S or 20% NFDM also significantly reduced the incidence of infected petunias. Other treatments identified in the preliminary screens are candidates for the second phase of screening that simulates contamination during the process of taking cuttings. [ABSTRACT FROM AUTHOR]

Abstract: Deletion and alanine-substitution mutants of the Tomato spotted wilt virus NSm protein were generated to identify domains involved in tubule formation, movement and symptomatology using a heterologous Tobacco mosaic virus expression system. Two regions of NSm, G19-S159 and G209-V283, were required for both tubule formation in protoplasts and cell-to-cell movement in plants, indicating a correlation between these activities. Three amino acid groups, D154, EYKK205–208 and EEEEE284–288 were linked with long-distance movement in Nicotiana benthamiana. EEEEE284–288 was essential for NSm-mediated long-distance movement, whereas D154 was essential for tubule formation and cell-to-cell movement; indicating separate genetic controls for cell-to-cell and long-distance movement. The region I57-N100 was identified as the determinant of foliar necrosis in Nicotiana benthamiana, and mutagenesis of HH93–94 greatly reduced necrosis. These findings are likely applicable to other tospovirus species, especially those within the ‘New World’ group as NSm sequences are highly conserved. [Copyright &y& Elsevier]

Abstract: A Florida isolate of Tomato spotted wilt virus (TSWV) was able to complement cell-to-cell movement of a movement-defective Tobacco mosaic virus (TMV) vector expressing the jellyfish green fluorescent protein (GFP). To test for complementation of movement in the absence of other TSWV proteins, the open reading frame for the NSm protein was expressed from TMV constructs encoding only the TMV replicase proteins. NSm was expressed from either the coat protein or movement protein subgenomic promoter, creating virus hybrids that moved cell to cell in inoculated leaves of tobacco, providing the first functional demonstration that NSm is the TSWV movement protein. Furthermore, these CP-deficient hybrids moved into upper leaves of Nicotiana benthamiana, demonstrating that NSm can support long-distance movement of viral RNAs. Tubules, characteristic of the NSm protein, were also formed in tobacco protoplasts infected with the TMV–TSWV hybrids. The C-terminus of the NSm protein was shown to be required for movement. TMV–TSWV hybrids expressing NSm and GFP moved within inoculated leaves. Our combination of single-cell and intact plant experiments to examine multiple functions of a heterologous viral protein provides a generalized strategy with wider application to other viruses also lacking a reverse genetic system. [Copyright &y& Elsevier]

Summary Taxonomy: Tobacco mosaic virus (TMV) is the type species of the Tobamovirus genus and a member of the alphavirus-like supergroup. Historically, many tobamoviruses are incorrectly called strains of TMV, although they can differ considerably in sequence similarities and host range from each other and from TMV. Physical properties: TMV virions are 300 × 18 nm rods with a central hollow cavity (Fig. 1) and are composed of 95% capsid protein (CP), and 5% RNA. Each CP subunit interacts with 3-nts in a helical arrangement around the RNA. Virions are stable for decades; infectivity in sap survives heating to 90 °C. Hosts: The natural host range of TMV is limited; however, a broad range of weed and crop species, mostly Solanaceae that includes tobacco, pepper and tomato can be infected experimentally [Holmes, F.O. (1946) A comparison of the experimental host ranges of tobacco etch and tobacco mosaic viruses. Phytopathology, 36, 643–657]. TMV distribution is worldwide. No biological vectors are known. Useful website: http://www.ncbi.nlm.nih.gov/ICTVdb/ICTVdB/71010001.htm. [ABSTRACT FROM AUTHOR]

The physiological and molecular events of ethylene-induced abscission in mature fruit calyx, laminar and floral abscission zones of cv. Valencia orange were examined. Continuous exposure of fruit explants to 5 µl 1-1 ethylene for 2 to 40 h resulted in marked increases in endo-1,4-β-glucanase (cellulase) and polygalacturonase (PG) activities in calyx abscission zones. Two abscission-related cellulases and one PG were found. The major peak of cellulase activity corresponded to a pI of 8.0 and molecular weight of 51 kDa, whereas the minor cellulase peak had a pI of 5.5. The abscission polygalacturonase had a pI of 5.5. Calyx abscission zone RNA was amplified with degenerate primers based on sequence of the purified Valencia orange calyx abscission cellulase, and cloned. The two partial cellulase cDNA clones were 59% identical at the nucleotide level. Genomic Southern analysis suggested that Valencia orange contained two groups of cellulase genes. A full-length cDNA clone from each group was isolated from a cDNA library prepared from ethylene-induced calyx abscission zone mRNA. Both genes were expressed in ethylene-induced calyx, laminar and floral abscission zones, but were not expressed in non-induced abscission zones or mature leaves treated with or without ethylene, young bark or young fruit of Valencia. [ABSTRACT FROM AUTHOR]

The genetics and expression of pectinesterase (PE) genes were examined in Valencia orange. Degenerate primers based on partial amino acid sequence of a 36 kDa PE protein isolated from juice vesicles were used to amplify a 350 bp DNA fragment from cDNA prepared from juice vesicle total RNA. Two groups of 350 bp PE clones with 66% sequence identity were isolated. A clone from each group was used to screen a Valencia orange genomic DNA λ library. Two different lambda clones that contained complete PE coding sequence (CsPME1 and CsPME3) and a third lambda clone that contained partial PE sequence (CsPME2) were characterized. The CsPME1 gene contained two exons (1063 and 689 bp) interrupted by a 1452 bp intron, whereas the CsPME3 gene had two exons (844 and 686 bp) interrupted by a 771 bp intron. CsPME1 shared significant sequence similarity with the partial clone CsPME2, including the entire cloned region of the first exon, a large region in the 5′ portion of the intron and the 3′ portion of the second exon, but the 3′ portion of the intron and the 5′ portion of the second exon were dissimilar. Southern blots suggested that Valencia orange has two genes within each PE group. Full-length cDNA clones that shared 99% sequence identity with CsPME1 and CsPME3 were isolated. Both groups of PE genes were differentially expressed in tissues of Valencia orange, and in addition CsPME3 appeared to be ethylene-regulated. The deduced proteins of PE cDNA clones CsPME1 (63.5 kDa) and CsPME3 (56.3 kDa) were considerably larger than the PE protein we isolated from Valencia orange juice vesicles and also other mature plant PE proteins. The estimated size of group I (2.2 kb) and group II (2.0 kb) PE mRNAs also predicted a larger protein than was isolated from juice vesicles. Alignment of the mature tomato and mung bean PE proteins, the most N-terminal sequence we obtained from polypeptides derived from the 36 kDa PE isolated from juice vesicles and the deduced amino acid sequences of plant PE cDNA clones suggest that a post-translational cleavage event separates the variable N-terminus from the more conserved C-terminal domain of the mature PE protein. [ABSTRACT FROM AUTHOR]

Abstract: Tobamovirus replicase proteins, which function in replication and gene expression, are also implicated in viral cell-to-cell and long-distance movement. The role(s) of Tobacco mosaic virus (TMV) 126-/183-kDa replicase protein in the complex movement process are not understood due to lack of systems that can separate the multiple steps involved. We previously developed a bipartite TMV-defective RNA (dRNA) system to dissect the role of the N-terminal methyltransferase (MT) domain in accumulation and cell-to-cell movement of dRNAs [Knapp, E., Danyluk, G.M., Achor, D., Lewandowski, D.J., 2005. A bipartite Tobacco mosaic virus–defective RNA (dRNA) system to study the role of the N-terminal methyltransferase domain in cell-to-cell movement of dRNAs. Virology 341, 47–58]. In the current study we analyzed long-distance movement of dRNAs in the presence of helper virus in Nicotiana benthamiana. dRNAs expressing ∼50% of the MT domain (ΔHinc151) moved long-distances in more than half of the plants. dRNAs expressing ∼90% of the MT domain sequences (ΔCla151) predominantly failed to accumulate in upper leaves. The helper virus moved systemically when inoculated alone or with a dRNA. In inoculated leaves, more ΔHinc151-induced infection foci spread adjacent to class V veins compared to those of ΔCla151. Consequently, ΔHinc151 infected more class V veins than ΔCla151. ΔCla151 was only detected in bundle sheath cells, whereas ΔHinc151 could accumulate in bundle sheath and phloem parenchyma cells of class V veins. However, the latter accumulation pattern did not always result in systemic accumulation of ΔHinc151, suggesting that factors in addition to those affecting cell-to-cell movement played a role in long-distance movement. [Copyright &y& Elsevier]

Hosta virus X (HVX) is rapidly becoming a serious pathogen of commercially important hosta plants worldwide. We report here biological and molecular characterization of a U.S. isolate of HVX, HVX-37. HVX-37 infectivity was tested in 21 hosta cultivars over three growth seasons, and three types of responses were defined based upon the ability of the virus to cause local and/or systemic infections. Four cultivars resistant to systemic HVX infection were identified. The full-length sequence of the HVX-37 genome was determined, the first complete sequence of a U.S. HVX isolate. Comparison with the previously sequenced HVX-Korea (Kr) genome revealed a high level of sequence similarity, as well as some differences. Notably, a 105-nucleotide long, near-perfect direct repeat in the Kr isolate is absent in HVX-37. The accuracy of the HVX-37 genome sequence was confirmed by infectivity of in vitro transcripts synthesized from a full-length HVX-37 cDNA on Nicotiana benthamiana and hosta plants. [ABSTRACT FROM AUTHOR]

Foliar symptoms suggestive of virus infection were recently observed on the noxious weed tropical soda apple (Solanum viarum) in Florida. An agent was mechanically transmitted to Nicotiana benthamiana, and virions were isolated from systemically infected leaves. Rod-shaped particles ~300 nm in length were observed in the partially purified preparations by electron microscopy. The host range determined by mechanical inoculation with purified virions included all tested plants in the Solanaceae (16 species including the important vegetable crops, pepper and tomato) and Chenopodiaceae (2 species) but excluded all tested plants in the Amaranthaceae, Apocynaceae, Brassicaceae, Caryophyllaceae, Cucurbitaceae, Fabaceae, Lamiaceae, Malvaceae, and Tropaeolaceae, including several common virus indicator hosts. Comparisons of the coat and movement protein nucleotide and deduced amino acid sequences of this putative tobamovirus with recognized members of this genus, indicate that it is a novel tobamovirus that shares the highest level of sequence identity with Pepper mild mottle virus followed by other members of the Solanaceae-infecting subgroup of tobamoviruses. The virus, for which the name Tropical soda apple mosaic virus (TSAMV) is proposed, was found to be widespread in tropical soda apple in peninsular Florida during an initial survey. TSAMV contamination of seed from infected tropical soda apple plants was found, suggesting that seed transmission may be important for TSAMV dissemination and epidemiology. [ABSTRACT FROM AUTHOR]

Abstract: Plant viruses, in particular Tobacco mosaic virus (TMV), are model systems to study RNA and protein trafficking in plants. Although TMV cell-to-cell transport controlled by the 30-kDa movement protein (MP) has been intensively studied, it was only recently demonstrated that the 126/183-kDa replicase proteins are also involved in cell-to-cell movement. Elucidating the role(s) of 126/183-kDa proteins in movement is complicated because these proteins have multiple functions associated with replication and gene expression. To overcome these difficulties we developed a TMV helper virus-defective RNA (dRNA) system to study the role of replicase protein sequences in dRNA cell-to-cell movement. Artificially constructed dRNAs lacking sequences encoding the helicase and polymerase domains of the replicase proteins and portions of the MP were viable in protoplasts and plants in the presence of helper virus. Expression of at least ∼50% of the methyl transferase (MT) domain was required for efficient dRNA movement in Nicotiana benthamiana. dRNAs that encoded the N-terminal 64 replicase amino acids or lacked a translatable MT domain failed to move or moved poorly. TMV dRNAs expressing 258 amino acids of the replicase protein moved into all specialized non-vascular tissues, whereas dRNAs expressing replicase sequences beyond amino acid 258 were restricted to the epidermis and palisade mesophyll tissues. Furthermore, second-site mutations within the dRNA-encoded truncated replicase protein altered efficiency in dRNA cell-to-cell movement. [Copyright &y& Elsevier]

Tobamoviruses are among the best characterized and most studied plant viruses. Three subgroups of tobamoviruses correspond to viral genome sequence and host range to include those viruses infecting (i) solanaceous plants, (ii) brassicas, or (iii) cucurbits or legumes. We isolated a virus from Florida landscape plantings of the malvaceous plant hibiscus (Hibiscus rosasinensis) that appears to be a tobamovirus based upon its virion morphology, genome organization, and coat protein sequence. The experimental host range of this virus included five malvaceous species but excluded all tested brassica, cucurbit, and legume species and 12 of the 19 solanaceous species tested. The unique host range and comparison of coat protein gene and protein sequences with those of recognized tobamoviruses indicate that this is a novel tobamovirus. A limited survey revealed that this virus is widespread in hibiscus and related species in the Florida landscape. [ABSTRACT FROM AUTHOR]