Phototherapy and topical treatments for cutaneous graft vs. host disease in haematopoietic stem cell transplantation patients: a scoping review.

Skin is usually the first and most affected organ involved in graft-versus-host disease (GvHD), and treatment is still a clinical challenge. Although the need for skin-directed treatments such as physical treatments and topical medications are generally agreed on, what the gold standard treatment strategy should be remains open to debate. The aim of this scoping review was to synthesize the current knowledge on the topical and physical treatments of cutaneous GvHD in haematopoietic stem cell transplantation patients and to highlight the best evidence available so as to reduce the gap between 'what is known' and 'what is done' in the clinical practice. Twenty-eight studies were included in this qualitative synthesis. Photo-biomodulation with psoralen was not included in this review. Phototherapy (ultraviolet A or B or narrowband B) was the physical treatment most described in the literature in both acute GvHD and chronic GvHD. Topical calcineurin inhibitors such as tacrolimus ointment and pimecrolimus cream as well as corticosteroid creams such as clobetasol and triamcinolone are mainly used in case of chronic GvHD. In all of the studies included in the review, topical treatments were always associated with systemic therapy. None of the topical interventions identified in our review provided strong evidence supporting its use, and the topical approaches seemed to have an adjuvant role in the treatment of cutaneous GvHD.

Vomero-premaxillary joint: A marker of evolution of the species.

OBJECTIVE: According to evolutionary developmental (evo-devo) theory, the vomers are bones derived from the secondary palate. Growth of the palatine processes of the maxillae, including the precursors of vomer bones, results in midline fusion posteriorly to the primary palate, which forces the ascension of the vomer bones towards the primary nasal septum, formed by septal cartilage and the perpendicular plate of the ethmoid. According to this hypothesis, the anterior border of the vomer articulates with the posterior surface of the premaxilla in the incisive canal (IC). MATERIAL AND METHOD: The objective of this retrospective study was to measure the degree of impaction of the anteroinferior angle of the vomer in the IC on CT scans showing a non-deformed nasal septum. Thirty-two out of a series of 506 nasal sinus CT scans were used to obtain measurements on coronal sections of non-deformed septa through the IC. RESULTS: Thirty-one of the 32 vomers were impacted in the IC. In the case of a Y-shaped vomer (n=26), 43% of the length of the vomer was impacted in 41% of the length of the IC. In the case of I-shaped vomers (n=6), 34% of the length of the vomer was impacted in 41% of the length of the IC. The only vomer that did not impact into the IC was Y-shaped. CONCLUSION: Impaction of the vomer in the IC posteriorly to the premaxilla can be explained by the evo-devo concept of the formation of the nasal cavities. In contrast, the classical embryological description of the formation of the nasal septum cannot provide an explanation for impaction of the vomer.

Topological Crystalline Insulator in a New Bi Semiconducting Phase.

Topological crystalline insulators are a type of topological insulators whose topological surface states are protected by a crystal symmetry, thus the surface gap can be tuned by applying strain or an electric field. In this paper we predict by means of ab initio calculations a new phase of Bi which is a topological crystalline insulator characterized by a mirror Chern number nM = -2, but not a strong topological insulator. This system presents an exceptional property: at the (001) surface its Dirac cones are pinned at the surface high-symmetry points. As a consequence they are also protected by time-reversal symmetry and can survive against weak disorder even if in-plane mirror symmetry is broken at the surface. Taking advantage of this dual protection, we present a strategy to tune the band-gap based on a topological phase transition unique to this system. Since the spin-texture of these topological surface states reduces the back-scattering in carrier transport, this effective band-engineering is expected to be suitable for electronic and optoelectronic devices with reduced dissipation.

A naphthalene diimide dyad for fluorescence switch-on detection of G-quadruplexes.

A non-fluorescent naphthalene diimide (NDI) dimer, conjugating red and blue NDI dyes, becomes red/NIR emitting upon G-quadruplex binding. The fluorescence lifetime which is significantly different for the complexes, the G-quadruplex/dimer and the weakly emitting ds-DNA/dimer is the key feature for the development of new rationally engineered G-quadruplex sensors.

Comment on "Topological insulators in ternary compounds with a honeycomb lattice".

A Comment on the Letter by Zhang et al., Phys. Rev. Lett. 106, 156402 (2011).

Response to UV-C radiation in topo I-deficient carrot cells with low ascorbate levels.

In animal cells, recent studies have emphasized the role played by DNA topoisomerase I (topo I) both as a cofactor of DNA repair complexes and/or as a damage sensor. All these functions are still unexplored in plant cells, where information concerning the relationships between DNA damage, PCD induction, and topo I are also limited. The main goal of this study was to investigate the possible responses activated in topo I-depleted plant cells under oxidative stress conditions which induce DNA damage. The carrot (Daucus carota L.) AT1-beta/22 cell line analysed in this study (characterized by an antisense-mediated reduction of top1beta gene expression of approximately 46% in association with a low ascorbate content) was more sensitive to UV-C radiation than the control line, showing consistent cell death and high levels of 8-oxo-dG accumulation. The topo I-depleted cells were also highly susceptible to the cross-linking agent mitomycin C. The death response was associated with a lack of oxidative burst and there were no changes in ascorbate metabolism in response to UV-C treatment. Electron and fluorescence microscopy suggested the presence of three forms of cell death in the UV-C-treated AT1-beta/22 population: necrosis, apoptotic-like PCD, and autophagy. Taken together, the data reported here support a reduced DNA repair capability in carrot topo I-deficient cells while the putative relationship between topo I-depletion and ascorbate impairment is also discussed.

Phytoplasmas Associated with Grapevine Yellows Disease in Chile.

An extensive survey was performed from 2002 to 2006 to detect and identify phytoplasmas associated with Chilean grapevines. Nested polymerase chain reaction assays using phytoplasma universal primer pairs P1/P7 and R16F2n/R2 detected phytoplasmas in 34 out of the 94 samples tested (36%). Restriction fragment length polymorphism (RFLP) analyses, cloning, and sequencing allowed identification of phytoplasmas belonging to ribosomal subgroups 16SrI-B, 16SrI-C, 16SrVII-A, and 16SrXII-A. The 16SrVII-A phytoplasma represents a new finding in grapevine; moreover, variability of the RFLP profile was observed in some of the 16SrXII-A phytoplasmas, indicating possible new ribosomal subgroups. Mixed phytoplasma infections and infections of phytoplasmas together with one or more viruses also occurred.

Diagnostic markers for diseases: SELDI-TOF profiling of pig sera for PRRS.

Porcine reproductive and respiratory syndrome (PRRS) is a highly infectious viral disease causing severe losses to the pig industry. Most weaning piglets are likely to be exposed to the infection and show at least asymptomatic PRRS viremia strongly related to productive performance. The aims of this study were to set up experimental conditions for pig sera proteomic profiling and to identify biomarkers that differentiate weaning asymptomatic piglets positive to PRRS viremia from negative controls (PCR tested) with potential predictive value for the subsequent occurrence of clinical PRRS. Protein profiles were generated by SELDI-TOF MS using the Bio-Rad Chips WCX, IMAC30 and H50. The discovery phase revealed that a consistent number of highly significant protein peaks can be detected by the WCX and IMAC30 surfaces; however none of these peaks were statistically confirmed by the subsequent validation phase, highlighting that serum concentration of the contaminant and most abundant proteins is a crucial parameterfor SELDI-TOF MS studies. Current protocols are being furtheroptimized and adapted to pig sera to reduce the unfavourable effects of the most abundant proteins and to increase the number of potential detectable biomarkers. Furthermore, proteomic fingerprint profiling has been shown to be a promising diagnostic tool that, in the future, may be useful to provide also insights into the mechanisms of early viral infection in vivo.

Grapevine yellows in Northern Italy: molecular identification of Flavescence dorée phytoplasma strains and of Bois Noir phytoplasmas.

AIMS: Verify the presence and the molecular identity of phytoplasmas in Northern and Central Italy vineyards where yellows diseases are widespread. METHODS AND RESULTS: Phytoplasma presence and identity were determined by PCR/RFLP analyses on 16S ribosomal gene testing 1424 symptomatic samples. The 65% of samples resulted phytoplasma infected; in particular 256 samples were found positive to phytoplasmas belonging to group 16SrV (mainly Flavescence dorée associated), and the remaining 37% was infected by phytoplasmas belonging to ribosomal subgroup 16SrXII-A (Stolbur or Bois Noir associated). 16SrV ribosomal group representative strains were further typed for variability in SecY and rpS3 genes. The results showed the presence of phytoplasmas belonging to 16SrV-C, 16SrV-D and to a lesser extent, 16SrV-A subgroup. CONCLUSIONS: Possible relationships between genetic polymorphisms of phytoplasma strains belonging to subgroup 16SrV-C and their geographic distribution and/or epidemic situations were detected. SIGNIFICANCE AND IMPACT OF THE STUDY: Bois Noir and Flavescence dorée phytoplasmas are present in significant percentages in the areas under investigation. Molecular tools allowed to identify phytoplasma-infected plants and the genes employed as polymorphism markers resulted useful in distinguishing and monitoring the spreading of the diseases associated with diverse phytoplasmas belonging to 16SrV subgroup in vineyards.

Ribosomal protein gene-based phylogeny for finer differentiation and classification of phytoplasmas.

Extensive phylogenetic analyses were performed based on sequences of the 16S rRNA gene and two ribosomal protein (rp) genes, rplV (rpl22) and rpsC (rps3), from 46 phytoplasma strains representing 12 phytoplasma 16Sr groups, 16 other mollicutes and 28 Gram-positive walled bacteria. The phylogenetic tree inferred from rp genes had a similar overall topology to that inferred from the 16S rRNA gene. However, the rp gene-based tree gave a more defined phylogenetic interrelationship among mollicutes and Gram-positive walled bacteria. Both phylogenies indicated that mollicutes formed a monophyletic group. Phytoplasmas clustered with Acholeplasma species and formed one clade paraphyletic with a clade consisting of the remaining mollicutes. The closest relatives of mollicutes were low-G+C-content Gram-positive bacteria. Comparative phylogenetic analyses using the 16S rRNA gene and rp genes were performed to evaluate their efficacy in resolving distinct phytoplasma strains. A phylogenetic tree was constructed based on analysis of rp gene sequences from 87 phytoplasma strains belonging to 12 16Sr phytoplasma groups. The phylogenetic relationships among phytoplasmas were generally in agreement with those obtained on the basis of the 16S rRNA gene in the present and previous works. However, the rp gene-based phylogeny allowed for finer resolution of distinct lineages within the phytoplasma 16Sr groups. RFLP analysis of rp gene sequences permitted finer differentiation of phytoplasma strains in a given 16Sr group. In this study, we also designed several semi-universal and 16Sr group-specific rp gene-based primers that allow for the amplification of 11 16Sr group phytoplasmas.

First Report of Phytoplasmas in Grapevine in South Africa.

In April 2006, grapevine plants with typical symptoms of yellows (GY) were observed in some South African vineyards. The affected plants showed premature yellowing or reddening and downward rolling of leaves. In some cases, these symptoms were associated with extensive lack of cane lignification that was undistinguishable from yellows symptoms reported in grapevine in the major viticultural areas of the world. Nucleic acids were extracted separately from 0.1 g of fresh leaf midribs and cane phloem scrapes from three symptomatic and three asymptomatic grapevine plants, cv. Shiraz, and from three symptomatic plants, cv. Cabernet, collected from three different locations using Qiagen (Milan, Italy) DNAeasy Plant Mini Kit. A nested polymerase chain reaction (PCR) assay was employed for phytoplasma detection with 2.5 µl of the extracted DNA. Direct and nested PCR assays were performed with P1/P7 (2) and R16F2/R2 (1) universal primer pairs, respectively, obtaining the expected products only from phloem scrapes of the symptomatic plant samples cv. Shiraz. Restriction fragment length polymorphism (RFLP) analyses of R16F2/R2 amplicons with TruI and Tsp509I restriction enzymes, discriminating among phytoplasma ribosomal group and subgroups, showed profiles corresponding to those of "Candidatus Phytoplasma aurantifolia" (ribosomal subgroup 16SrII-B) in all three positive samples. A Stolbur phytoplasma profile (ribosomal subgroup 16SrXII-A) was also observed in one of those samples, indicating the presence of mixed phytoplasma infection (1). Sequencing of the obtained amplicons confirmed the RFLP phytoplasma identification; in particular 16SrXII-A could be the same phytoplasma associated with the 'Bois Noir' disease reported in grapevine; the 1601-bp sequence of 16SrII-B phytoplasma showed 98% similarity to U15442, i.e., to the phytoplasma associated with lime witches'-broom disease in Oman ("Ca. P. aurantifolia") confirming RFLP results. To our knowledge, this is the first report of phytoplasmas in grapevine in South Africa. References: (1) I.-M. Lee et al. Phytopathology 85:728, 1995. (2) B. Schneider et al. Pages 369-380 in: Molecular and Diagnostic Procedures in Mycoplasmology Vol. I. Academic Press Inc., 1995.

Identification of a 16SrII-E Phytoplasma in Calendula arvensis, Solanum nigrum, and Chenopodium spp.

Epidemiological surveys were performed in Northern Sardinia (Italy) in a 10-year-old vineyard affected by "Bois noir" disease. Samples collected between May and October 2003 from chlorotic and stunted weeds belonging to 14 different taxonomic groups were indexed molecularly for detection of phytoplasmas. Nested polymerase chain reaction (PCR) assays using primers specific for the phytoplasma 16SrDNA gene showed three of six Calendula arvensis, one of two Solanum nigrum, and one of seven Chenopodium spp. assayed positive. Restriction fragment length polymorphism analyses and sequencing of amplified 16SrDNA fragments identified a putative phytoplasma in the ribosomal subgroup 16SrII-E. Further characterization of the rps3 gene, coding a ribosomal protein, confirmed the identification. However, the weeds and leafhop-per species collected in the vineyard tested negative by PCR assays for the Stolbur phytoplasma, the causal agent of "Bois noir". This is the first report of a phytoplasma of the 16SrII-E subgroup infecting C. arvensis, S. nigrum, and Chenopodium spp.

Association of phytoplasmas and viruses with malformed clovers.

Plants of Trifolium spp. exhibiting two different kinds of symptoms--phyllody associated with yellowing/reddening, and dwarf growth habit without floral abnormalities--were observed in several areas of the Czechia. Nested polymerase chain reaction (PCR) with phytoplasma specific primers, and restriction fragment length polymorphism (RFLP) analyses of 16SrDNA revealed that phyllody of T. repens was associated with phytoplasmas belonging to the 16SrI-C subgroup. Similar symptoms in T. hybridum and T. pratense plants revealed the presence of phytoplasmas belonging to two subgroups: 16SrI-C and 16SrIII-B. Dwarf disease of cultivated T. pratense plants was associated with more than one agent: 11 of 20 plants examined by PCR/RFLP analysis revealed the presence of phytoplasmas belonging to four distinct subgroups: 16SrI-B, 16SrI-C, 16SrIII-B and 16SrX-A. Moreover, two kinds of bacilliform virions were observed in ultrathin sections of 15 T. pratense plants. Particles occurred mostly in the parenchymatous cells of vascular bundles and were located in the cytoplasm as aggregates within an extended network of membranous cisternae. Phytoplasmas and rhabdoviruses occurred singly, and both together or in co-presence with filamentous virus-like particles.

Variability and functional role of chromosomal sequences in 16SrI-B subgroup phytoplasmas including aster yellows and related strains.

AIMS: Partial genetic characterization of several chromosomal regions on 35 16SrI-B phytoplasma strains maintained in periwinkle and collected in different geographical areas from plants of diverse species. METHODS AND RESULTS: Genes coding for ribosomal protein rpL22, elongation factor EF-Tu and random cloned sequences amplified with primers AY19p/m, G35p/m and BB88F1/R1 after RFLP analyses showed a high degree of polymorphism among the strains studied. The ribosomal protein (rp) subgroups B and K, and an undescribed subgroup designated N, were identified. Amplicons obtained with primers AY19p/m and BB88F1/R1, revealed a high and a low degree of polymorphism, respectively. CONCLUSIONS: A probable spacer role could be attributed to the AY19p/m sequence and a possible coding function to the BB88F1/R1 sequence. No relationship was found among genetic polymorphisms, identified by statistical analyses, and epidemiological or biological parameters. SIGNIFICANCE AND IMPACT OF THE STUDY: The analyses of five different genomic sequences of the 35 strains belonging to subgroup 16SrI-B allowed a finer distinction among them, confirming that the polymorphism level of 16S rDNA is too low to be adopted as unique parameter for classification.

First Report of an Elm Yellows Subgroup 16SrV-C Phytoplasma Infecting Grapevine in Serbia.

During a 2002 survey in Serbia, samples of grapevine (Vitis vinifera) were collected from plants showing typical phytoplasma-like symptoms: leaf roll, leaf redness, vein chlorosis and necrosis, and absence of lignification. The material was collected from one viticultural region (Zupa Aleksandrovac), where the disease was recorded in 2000 and showed an increasing percentage of symptomatic plants every year. Total nucleic acid was extracted separately from leaf midveins and stem bark collected from 10 symptomatic and 2 asymptomatic plants. Phytoplasma infection was detected using polymerase chain reaction (PCR) assays with universal primer pair P1/P7 for the amplification of phytoplasma 16S rRNA gene, and primer pair FD9f2/FD9r followed by FD9f3/FD9r2 in nested PCR for specific amplification of the FD9 nonribosomal DNA fragment of the EY-group (1). Phytoplasmas were detected in 9 of 10 midvein extracts from symptomatic grapevines (three of cv. Plovdina, two of cv. Smederevka, and four of cv. Gamé). Also, 6 of 10 bark preparations representing stem collections from the same plants were positive (two samples of cv. Plovdina, both samples of cv. Smederevka, and two samples of cv. Gamé). Both collections of midveins and bark tissues from asymptomatic plants were negative. Fragments amplified with universal P1/P7 primers (16S-23S rDNA) were analyzed by restriction fragment length polymorphism with TruI and TaqI restriction enzymes. The phytoplasmas produced identical restriction profiles to those of 16SrV Elm Yellows group and 16SrV-C Flavescence doreé subgroup (2). To our knowledge, this is the first report of phytoplasma infecting grapevines in Serbia, and the first survey in progress to verify the presence of Scaphoideus titanus to determine if this grapevine yellows could be defined as Flavescence dorée. References: (1) E. Angelini et al. Vitis 40:79, 2001. (2) M. Martini et al. Mol. Cell. Probes 16:197, 2002.

Genetic variability among flavescence dorée phytoplasmas from different origins in Italy and France.

Flavescence dorée is a devastating disease of grapevine widespread in several countries in EU such as France, Italy and Spain. Genetic variability among 17 Italian and 3 French FD strains was investigated by RFLP analyses based on a fragment of the ribosomal protein operon and on the non-ribosomal DNA fragment FD9. RFLP analysis of the PCR amplified ribosomal protein fragment, coding for the 3' end of rpl22 and the entire rps3 genes, differentiated 4 rp-subgroups among the FD strains and 4 subgroups among the reference strains belonging to elm yellows group (16SrV). Sequencing and phylogenetic analysis of the same ribosomal protein DNA fragment validated the delineation of 4 distinct FD strain types derived by RFLP analyses. The results supported the differentiation based on analysis of the non-ribosomal DNA fragment FD9. The phylogenetic analysis further revealed relationships and a probable evolutionary trend among the FD strains and the other representatives of elm yellows group. All the FD strains together with the reference strains ALY, RuS and JWB formed a cluster very well distinct from the EY/ULW cluster. Moreover, ALY was shown to be more closely related to three FD strain types: the Lombardia/Piemonte, the French FD70, and the French FD88/Italian FD-D strain clusters.

Molecular identification of a new phytoplasma associated with alfalfa witches'-broom in oman.

ABSTRACT Alfalfa (Medicago sativa) plants showing witches'-broom symptoms typical of phytoplasmas were observed from Al-Batinah, Al-Sharqiya, Al-Bureimi, and interior regions of the Sultanate of Oman. Phytoplasmas were detected from all symptomatic samples by the specific amplification of their 16S-23S rRNA gene. Polymerase chain reaction (PCR), utilizing phytoplasma-specific universal primer pairs, consistently amplified a product of expected lengths when DNA extract from symptomatic samples was used as template. Asymptomatic plant samples and the negative control yielded no amplification. Restriction fragment length polymorphism profiles of PCR-amplified 16S-23S rDNA of alfalfa using the P1/P7 primer pair identified phytoplasmas belonging to peanut witches'-broom group (16SrII or faba bean phyllody). Restriction enzyme profiles showed that the phytoplasmas detected in all 300 samples belonged to the same ribosomal group. Extensive comparative analyses on P1/P7 amplimers of 20 phytoplasmas with Tru9I, Tsp509I, HpaII, TaqI, and RsaI clearly indicated that this phytoplasma is different from all the other phytoplasmas employed belonging to subgroup 16SrII, except tomato big bud phytoplasma from Australia, and could be therefore classified in subgroup 16SrII-D. The alfalfa witches'-broom (AlfWB) phytoplasma P1/P7 PCR product was sequenced directly after cloning and yielded a 1,690-bp product. The homology search showed 99% similarity (1,667 of 1,690 base identity) with papaya yellow crinkle (PapayaYC) phytoplasma from New Zealand. A phylogenetic tree based on 16S plus spacer regions sequences of 35 phytoplasmas, mainly from the Southern Hemisphere, showed that AlfWB is a new phytoplasma species, with closest relationships to PapayaYC phytoplasmas from New Zealand and Chinese pigeon pea witches'-broom phytoplasmas from Taiwan but distinguishable from them considering the different associated plant hosts and the extreme geographical isolation.

Identification of the B-cell tumor-specific molecular fingerprint using non-radiolabelled PCR consensus primers.

BACKGROUND: The complementarity determining region 3 (CDR3) of the immunoglobulin (Ig) heavy chain variable region (VH) is the most reliable molecular fingerprint for most if not all human B cells. The nucleotide sequence encoding for any B-cell tumor-specific VH CDR3 is currently identified by PCR sequencing based on procedures involving the usage of either radioactive materials, patient/family-specific primers, or bacterial cloning. PATIENTS AND METHODS: In six consecutive patients with follicular lymphoma we assessed the feasibility of a method that allows for identification of the tumor-specific VH CDR3 using consensus primers while avoiding both radioactive materials and bacterial cloning procedures. RESULTS: The tumor-specific VH CDR3 was successfully identified in all six patients in nearly half the time typically required by any other method currently utilized. The feasibility of the proposed method was not significantly affected either by the tumor-specific Ig isotype, or by the tumor infiltration in the original biopsy specimen. In the three patients for whom tumor specimen-derived hybridomas were available, the tumor-specific VH CDR3 was also found in at least 8 of 10 of them. CONCLUSIONS: The proposed method allows the ability to quickly identify the B-cell tumor-specific VH CDR3 using consensus primers while avoiding radioactive materials and bacterial cloning procedures.

Cranial nerve involvement in CMT disease type 1 due to early growth response 2 gene mutation.

Mutations in the gene coding for the Schwann cell transcription factor early growth response 2 (EGR2), which seems to regulate myelinogenesis and hindbrain development, have been observed in few cases of inherited neuropathy. The authors describe a unique combination of cranial nerve deficits in one member of a Charcot-Marie-Tooth 1 family carrying an EGR2 mutation (Arg381His). This finding further supports the role of EGR2 in cranial nerve development.