The Zaire ebolavirus Recombinant protein VP35 is an essential component of the viral RNA polymerase complex and also functions to antagonize the cellular type I interferon (IFN) response by blocking the activation of the transcription factor IRF-3. We previously mapped the inhibitory domain of IRF-3 within the C-terminus of VP35. In the present study, we show that mutations that disrupt the IRF-3 inhibitory function of VP35 do not disrupt viral transcription/replication, suggesting that the two functions of VP35 are separable.
Second, using reverse genetics, we successfully recovered recombinant Ebolaviruses that contained mutations within the IRF-3 inhibitory domain. Importantly, we showed that the recombinant viruses were attenuated for growth in cell culture and that they activated IRF-3 and IRF-3-inducible gene expression at higher levels than that of wild-type VP35-containing Ebola virus. In the context of Ebola virus pathogenesis, VP35 may function to limit the early production of IFN-β and other antiviral signals generated by cells at the primary site of infection, thereby slowing the host’s ability to stop the replication of the virus and induce adaptive immunity.
Purity: greater than 90% as determined by SDS-PAGE.
Target names: NP
Uniprot No.: P18272
Research area: others
Alternative Names: NP Nucleoprotein; PN Ebola; in P; nucleocapsid protein; Protein N
Note: We will preferably ship the format we have in stock, however, if you have any special requirements for the format, please remark your requirement when placing the order, we will prepare according to your demand.
Buffer
If the dosage form is liquid, the default storage buffer is Tris/PBS based buffer, 5%-50% glycerol.
Note: If you have any special requirements for glycerol content, please remark it when you order. If the administration form is a lyophilized powder, the buffer before lyophilization is Tris/PBS-based buffer, 6% trehalose, pH 8.0.
Reconstitution
We recommend that this vial be briefly centrifuged before opening to bring the contents to the bottom. Reconstitute protein in sterile deionized water at a concentration of 0.1-1.0 mg/mL. We recommend adding 5-50% glycerol (final concentration) and an aliquot for long-term storage at -20°C/-80°C. Our final default glycerol concentration is 50%. Customers could use it for reference.
Storage Conditions
Store at -20°C/-80°C upon receipt, need to be aliquoted for multiple uses. Avoid repeated cycles of freezing and thawing.
Shelf life
Shelf life is related to many factors, storage condition, buffer ingredients, storage temperature and the stability of the protein itself. Generally, the shelf life of the liquid form is 6 months at -20°C/-80°C. The shelf life of the lyophilized form is 12 months at -20°C/-80°C.
Delivery time: 3-7 business days
Notes: Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Cells and viruses.
Vero E6 and 293T cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) containing 100 U/ml penicillin, 100 U/ml streptomycins, and 10 mM HEPES buffer (complete) supplemented with fetal bovine serum (FBS) at 10% unless otherwise stated. Huh7 hepatocytes (obtained from Apath, LLC) were cultured in complete DMEM as described above and further supplemented with non-essential amino acids and 10% FBS.
The human monocyte cell line U937 (ATCC CRL-1593.2) was grown in a complete RPMI medium containing 10% FBS. Ebola virus infections were performed in complete DMEM containing 2% FBS. All work with infectious Ebola virus was done inside positive pressure suits in the biosafety level 4 laboratory at the Centers for Disease Control and Prevention. When necessary, samples were quenched for infectivity with 2 × 106 to 5 × 106 rads of cobalt-60 radiation from a Gammacell irradiator.
Western spots.
Cell lysates from the transfections were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using the DuPage bis-tris system (Invitrogen). Samples were loaded onto 10% bis-tris gels and electrophoresed for 50 min at 200 V. Proteins were transferred to nitrocellulose membranes using the X-Cell II transfer module (Invitrogen) and then probed with an anti-VP35 monoclonal antibody (1:2000). ) or an antiactin control (1:1,000) (Chemicon). Blots were developed using the Western Breeze Chemiluminescence Detection System (Invitrogen) and exposed to film. Protein expression levels were determined using the AlphaImager EC camera and densitometry software.
Most “neutrality tests” assess whether particular data sets depart from the predictions of a standard neutral model without recombination. For Drosophila, where nuclear polymorphism data routinely show evidence of genetic exchange, the assumption of no recombination is often unrealistic. Furthermore, although conservative, this assumption is made at the cost of a large power loss.
Perhaps, as a result, tests of the frequency spectrum based on zero recombination suggest a good fit of the Drosophila polymorphism data to the predictions of the standard neutral model. Here, we analyze the frequency spectrum of a large number of loci in Drosophila melanogaster Recombinant and D. simulans using two summary statistics. We use an estimate of the population recombination rate based on a laboratory estimate of the rate of crossing by physical length and an estimate of the effective population size of the species.
In contrast to previous studies, we found that about half of the loci deviate from the predictions of the standard neutral model. The extent of the output depends on the exact recombination rate, but the overall pattern that emerges is robust. Interestingly, these deviations from neutral expectations are not one-way. The large variation in results may be due to a complex demographic history and inconsistent sampling, or to the pervasive action of natural selection.
Statistical Summary
We use two summary statistics. The first was D, which compares two estimates of the neutral mutation parameter, 𝛉 (Tajima 1989a). D is negative when there is an excess of rare mutations, as would be expected with recent population growth (Tajima 1989b) or after selective screening (Braverman et al. 1995). It is positive when there are too many intermediate frequency variants, as in the presence of a balanced polymorphism or under certain models of population subdivision (Tajima 1989b). Our second summary statistic, Gη, is based on the differences between the observed and expected number of mutations in each frequency class (Fu 1996).
Coalescing simulations
The simulations assumed an equilibrium Wright-Fisher population, from which random samples were drawn. There was no selection, and each new mutation occurred at a previously unmutated site (Kimura 1969). In this implementation, we generate pedigrees and then place the observed number of mutations, S, in the tree (Hudson 1993). This approach departs from standard coalescent simulations that place mutations at a constant rate (of 𝛉/2) along each of the branches. We took this approach because we were able to observe the number of segregating sites while 𝛉 was unknown (cf. Hudson 1993).
Simulations have shown that the type I errors of various statistical tests under this ‘fixed S’ scheme is approximately equal to the nominal rejection probabilities (Kelly 1997; unpublished data). The parameters of the simulations were, for each locus, the sample size, the number of base pairs and the population recombination rate C = Camp (see below). Although our simulations are unorthodox, we refer to our set of assumptions as of the “standard neutral model” (SNM). To implement coalescence with recombination, we use a modification of a program kindly provided by R. Hudson (see Hudson 1983, 1990).
There is no gene conversion and the rate of crossing over is constant per base pair. Tracing the sample’s ancestral lineages back in time, there are two possible genealogical events. Lineages can merge in the usual way or they can split in two as a result of a crossover event. The result of a split event is that there are now two lineages, one to the left of the crossover point and one to the right. These two lineages then follow each other in time and may merge or split. At any given nucleotide site, there is a standard coalescing tree (no recombination). However, recombination causes different sites to have trees that differ from each other. These trees are not independent of each other, as they share parts of their genealogical histories.
Effect of population recombination rate
Without recombination, there is only one family tree at a locus or one extraction from the (highly stochastic) evolutionary process. With recombination, sites very close to each other are likely to have similar pedigrees, but sites farther apart will have quite different pedigrees. As the rate of recombination increases, so will the number of distinct trees in a location (Hudson 1983). Summaries of the frequency spectrum at neutral sites (such as D and Gη) reflect aspects of the underlying trees.
As the recombination rate increases, they reflect an average of more trees and thus will tend to take values closer to your expectations. In other words, its variance decreases as the recombination rate increases. Wall (1999) showed that a large number of statistical tests (based on both frequency spectrum and linkage disequilibrium summaries) have very little power to detect population structure if the simulations are run with C = 0 when the actual value of C is much higher. This is because the actual variances of the test statistics are smaller than the sample variances of the simulations without recombination.
However, it was unknown whether, in practice, many loci would show deviations from the null model for C > 0. These results suggest that in Drosophila, population recombination rates are high enough that alternative assumptions about C have a bearing. significant effect on one’s own conclusions. In particular, with our conservative estimate of the population recombination rate, Camp, we found that the standard neutral model was a poor predictor of the observed frequency spectra at many loci.
Recombinant populations were the basis of Mendel’s early genetic experiments and continue to be key to the study of genes, heredity, and genetic variation today. Genotyping several hundred thousand loci in a single assay by hybridizing genomic DNA to oligonucleotide arrays provides a powerful technique for improving precision linkage mapping. The genotypes of two Arabidopsis thaliana Recombinant accessions were compared using a 400,000-feature exon-specific oligonucleotide array.
Approximately 16,000 single feature polymorphisms (SFPs) were detected in approximately 8,000 of the approximately 26,000 genes represented on the array. Allelic variation at these loci was measured in a population of recombinant inbred lines, which defined the location of 815 recombination breakpoints. The genetic linkage map had a total length of 422.5 cm, with 676 informative SFP markers representing ~0.6 cM intervals. One hundred and fifteen single gene intervals were identified.
The recombination rate, SFP distribution, and segregation in this population are not uniform. Many genomic regions show a cluster of recombination events that include significant hot spots. The precise haplotype structure of the recombinant population was defined with unprecedented precision and resolution. The resulting linkage map allows further refinement of the hundreds of quantitative trait loci identified in this well-studied population. Highly variable recombination rates along each chromosome and extensive segregation distortion was observed in the population.
Purity: greater than 85% as determined by SDS-PAGE.
Destination Names: NLP7
Uniprot No.: Q84TH9
Research Area: Epigenetic and Nuclear Signaling
Alternative Names: NIN-like protein 7
Species: Arabidopsis thaliana (Mouse-ear cress)
Source: Baculovirus
Expression Region: 1-959aa
Mole Weight: 107.4 kDa
Protein Length: Total length
Tag Information: N-terminal 10xHis-tagged
Form: Liquid or Lyophilized Powder
Note: We will preferably ship the format we have in stock, however, if you have any special requirements for the format, please remark your requirement when placing the order, we will prepare according to your demand.
Buffer
If the dosage form is liquid, the default storage buffer is a Tris/PBS based buffer, 5%-50% glycerol. If the administration form is a lyophilized powder, the buffer prior to lyophilization is a Tris/PBS based buffer, 6% trehalose, pH 8.0.
Reconstitution
We recommend that this vial be briefly centrifuged before opening to bring the contents to the bottom. Reconstitute protein in sterile deionized water at a concentration of 0.1-1.0 mg/mL. We recommend adding 5-50% glycerol (final concentration) and an aliquot for long-term storage at -20°C/-80°C. Our final default glycerol concentration is 50%. Customers could use it for reference.
Storage Conditions
Store at -20°C/-80°C upon receipt, need to be aliquoted for multiple uses. Avoid repeated cycles of freezing and thawing.
Shelf life
Shelf life is related to many factors, storage condition, buffer ingredients, storage temperature and the stability of the protein itself. Generally, the shelf life of the liquid form is 6 months at -20°C/-80°C. The shelf life of the lyophilized form is 12 months at -20°C/-80°C.
Delivery time: 3-7 business days
Notes: Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Synopsis
One goal of many genetic studies is to discover the underlying genetic condition (the genotype) of a specific physical manifestation in an organism (the phenotype), such as diabetes in humans or leaf rust in cultivated wheat. One limitation to making such discoveries is the ability to resolve the genotype. Gene arrays carry representations of the genome, called features, in high density on a thumbnail-sized surface.
In this study, microarrays designed to measure gene expression were used to detect polymorphisms in the DNA sequence. DNA from two different Arabidopsis strains was hybridized to arrays representing almost the entire coding region of the genome. Differences in hybridization intensity indicated differences in DNA sequence. Sequence differences, termed single trait polymorphisms, were then analyzed in a population of 100 plants derived from inbreeding of the progeny of the two-parent strains.
The precise location of the genetic recombination breakpoints was defined for each line. As a result, Singer et al. were able to generate one of the first very high-resolution genotyping datasets in a multicellular organism that allowed the construction of a high-resolution genetic map of Arabidopsis. This map will greatly facilitate attempts to make definitive associations between genotypes and phenotypes.
Vegetal material.
Columbia/Landsberg RIL (Col/Ler) seeds (eight generations inbreeding) [13] and parental lines were kindly provided by the Arabidopsis Biological Resource Center (ABRC) at The Ohio State University, Columbus, Ohio, United States. The accessions used in this study correspond to the first set of 100 RIL (CS1899), the parental lines were Columbia (CS933; Col-4, named Col) and Landsberg erecta (CS20, named Ler). Eight plants for each RIL were grown in a pot under long-day conditions (16 h light, 8 h dark) and pooled for analysis.
DNA isolation, labelling and hybridization of microarrays.
Total genomic DNA was isolated from leaf tissue with the DNeasy Plant Mini Kit (Qiagen, Valencia, California, United States) according to the manufacturer’s instructions. Four and six biological replicates of Col (CS933) and Ler (CS20) were made, respectively. For RILs 1 (CS1900) to 57 (CS1957), a single replicate was available, except for lane 6 (CS1906) which was tripled.
RILs 58 (CS1958) to 100 (CS4686) were doubled, except that line 73 (CS1973) was tripled and lines 83 (CS1983), 84 (CS1984) and 89 (CS1989) were quadrupled. Probe intensity values for replicate RIL microarrays were averaged for genotyping. DNA was labelled by random priming with biotin14-dCTP (Bioprime DNA Labeling System, Invitrogen, Carlsbad, California, USA). Hybridization, washing, staining and scanning were carried out using the standard Affymetrix Eukaryotic protocol. GeneChip Suite was used for image acquisition.
Real-time reverse transcription adopted by polymerase chain response (RT-PCR) is essentially the most appropriate technique for the detection and quantification of mRNA. It affords excessive sensitivity, good reproducibility and a large quantification vary.
Today, relative expression is more and more used, the place the expression of a goal gene is standardised by a non-regulated reference gene. Several mathematical algorithms have been developed to compute an expression ratio, based mostly on real-time PCR effectivity and the crossing level deviation of an unknown pattern versus a management.
But all printed equations and accessible fashions for the calculation of relative expression ratio permit solely for the willpower of a single transcription distinction between one management and one pattern.
Therefore a brand new software tool was established, named REST (relative expression software tool), which compares two teams, with as much as 16 information factors in a pattern and 16 in a management group, for reference and as much as 4 goal genes.
The mathematical mannequin used is predicated on the PCR efficiencies and the imply crossing level deviation between the pattern and management group. Subsequently, the expression ratio results of the 4 investigated transcripts are examined for significance by a randomisation check. Herein, improvement and software of REST is defined and the usefulness of relative expression in real-time PCR utilizing REST is mentioned.
Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR.
New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae.
We have constructed and examined a dominant resistance module, for choice of S. cerevisiae transformants, which totally consists of heterologous DNA. This kanMX module accommodates the identified kanr open reading-frame of the E. coli transposon Tn903 fused to transcriptional and translational management sequences of the TEF gene of the filamentous fungus Ashbya gossypii. This hybrid module permits environment friendly choice of transformants resistant towards geneticin (G418).
We additionally constructed a lacZMT reporter module in which the open reading-frame of the E. coli lacZ gene (missing the primary 9 codons) is fused at its 3′ finish to the S. cerevisiae ADH1 terminator.
KanMX and the lacZMT module, or each modules collectively, had been cloned in the middle of a brand new a number of cloning sequence comprising 18 distinctive restriction websites flanked by Not I websites. Using the double module for constructions of in-frame substitutions of genes, just one transformation experiment is important to check the exercise of the promotor and to look for phenotypes because of inactivation of this gene.
To permit for repeated use of the G418 choice some kanMX modules are flanked by 470 bp direct repeats, selling in vivo excision with frequencies of 10(-3)-10(-4). The 1.Four kb kanMX module was additionally proven to be very helpful for PCR based mostly gene disruptions.
In an experiment in which a gene disruption was accomplished with DNA molecules carrying PCR-added terminal sequences of solely 35 bases homology to every goal website, all twelve examined geneticin-resistant colonies carried the appropriately built-in kanMX module.
We have developed a easy and extremely environment friendly technique to disrupt chromosomal genes in Escherichia coli in which PCR primers present the homology to the focused gene(s).
In this process, recombination requires the phage lambda Red recombinase, which is synthesized underneath the management of an inducible promoter on an simply curable, low copy quantity plasmid.
To display the utility of this method, we generated PCR merchandise by using primers with 36- to 50-nt extensions which can be homologous to areas adjoining to the gene to be inactivated and template plasmids carrying antibiotic resistance genes which can be flanked by FRT (FLP recognition goal) websites.
By using the respective PCR merchandise, we made 13 completely different disruptions of chromosomal genes. Mutants of the arcB, cyaA, lacZYA, ompR-envZ, phnR, pstB, pstCA, pstS, pstSCAB-phoU, recA, and torSTRCAD genes or operons have been remoted as antibiotic-resistant colonies after the introduction into micro organism carrying a Red expression plasmid of artificial (PCR-generated) DNA.
The resistance genes have been then eradicated by using a helper plasmid encoding the FLP recombinase which can also be simply curable. This process ought to be extensively helpful, particularly in genome evaluation of E. coli and different micro organism as a result of the process will be finished in wild-type cells.
One-step inactivation of chromosomal genes in Escherichia coli Okay-12 using PCR merchandise.
The MIQE tips: minimal info for publication of quantitative real-time PCR experiments.
BACKGROUNDCurrently, an absence of consensus exists on how greatest to carry out and interpret quantitative real-time PCR (qPCR) experiments. The drawback is exacerbated by an absence of enough experimental element in many publications, which impedes a reader’s means to judge critically the standard of the outcomes introduced or to repeat the experiments.
BACKGROUNDThe Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) tips goal the reliability of outcomes to assist make sure the integrity of the scientific literature, promote consistency between laboratories, and improve experimental transparency.
MIQE is a set of tips that describe the minimal info crucial for evaluating qPCR experiments. Included is a guidelines to accompany the preliminary submission of a manuscript to the writer.
By offering all related experimental situations and assay traits, reviewers can assess the validity of the protocols used. Full disclosure of all reagents, sequences, and evaluation strategies is critical to allow different investigators to breed outcomes.
MIQE particulars ought to be printed both in abbreviated type or as a web-based complement.
CONCLUSIONSFollowing these tips will encourage higher experimental observe, permitting extra dependable and unequivocal interpretation of qPCR outcomes.
Use of the real-time polymerase chain response (PCR) to amplify cDNA merchandise reverse transcribed from mRNA is on the best way to changing into a routine instrument in molecular biology to review low abundance gene expression.
Real-time PCR is simple to carry out, offers the mandatory accuracy and produces dependable in addition to fast quantification outcomes.
But correct quantification of nucleic acids requires a reproducible methodology and an ample mathematical model for information evaluation. This examine enters into the actual matters of the relative quantification in real-time RT-PCR of a goal gene transcript in comparability to a reference gene transcript.
Therefore, a new mathematical model is introduced. The relative expression ratio is calculated solely from the real-time PCR efficiencies and the crossing level deviation of an unknown pattern versus a management.
This model wants no calibration curve. Control ranges have been included in the model to standardise every response run with respect to RNA integrity, pattern loading and inter-PCR variations. High accuracy and reproducibility (<2.5% variation) have been reached in LightCycler PCR utilizing the established mathematical model.
A new mathematical model for relative quantification in real-time RT-PCR.
Accurate normalization of real-time quantitative RT-PCR information by geometric averaging of a number of inside management genes.
BACKGROUNDGene-expression evaluation is more and more necessary in organic analysis, with real-time reverse transcription PCR (RT-PCR) changing into the strategy of alternative for high-throughput and correct expression profiling of chosen genes.
Given the elevated sensitivity, reproducibility and enormous dynamic vary of this technique, the necessities for a correct inside management gene for normalization have develop into more and more stringent.
Although housekeeping gene expression has been reported to differ significantly, no systematic survey has correctly decided the errors associated to the frequent observe of utilizing just one management gene, nor introduced an ample approach of working round this downside.
RESULTSWe define a sturdy and revolutionary technique to establish essentially the most stably expressed management genes in a given set of tissues, and to find out the minimal variety of genes required to calculate a dependable normalization issue.
We have evaluated ten housekeeping genes from totally different abundance and purposeful lessons in varied human tissues, and demonstrated that the standard use of a single gene for normalization results in comparatively massive errors in a major proportion of samples examined.
The geometric imply of a number of rigorously chosen housekeeping genes was validated as an correct normalization issue by analyzing publicly out there microarray information.CONCLUSIONSThe normalization technique introduced here’s a prerequisite for correct RT-PCR expression profiling, which, amongst different issues, opens up the potential for finding out the organic relevance of small expression variations.
Il nostro corpo produce costantemente anticorpi specifici, destinati a combattere gli invasori patogeni, come virus o allergeni. Questo team dell’Università di Augusta (Georgia) ha scoperto il modo in cui la loro produzione è alimentata e mantenuta. Questi lavori presentati sulla rivista Nature Communications aprono la speranza di poter un giorno essere in grado di regolare questo percorso di immunità, per mantenerci in salute.
È un meccanismo protettivo essenziale che resta da capire meglio, specifica l’autore principale, lo scienziato Nagendra Singh, immunologo nel dipartimento di biochimica e biologia di Augusta: “Stiamo cercando di progettare piccole molecole in grado di bloccare o per attivare questo canale “.
Un percorso chiamato “ufmilazione”: in questo percorso, un polipeptide chiamato Ufm1 è noto per colpire altre proteine, connettersi ad esse e modificarne la funzione. Una di queste proteine è l’Ufbp1, una proteina essenziale per le cellule immunitarie chiamate cellule B naive per diventare plasmacellule che producono anticorpi e per intensificare la produzione di anticorpi protettivi. “Comprendere meglio come funziona questo meccanismo di protezione naturale contribuirà alla progettazione di vaccini migliori”, scrivono i ricercatori. In effetti, gli attuali vaccini aiutano le cellule B ad avviare la memoria di alcuni invasori patogeni, consentendo una risposta immunitaria più efficace e più rapida.
L’aumento selettivo del percorso di ufilazione potrebbe, ad esempio, portare a un attacco più mirato del virus dell’influenza; in caso di allergie, un intervento selettivo potrebbe, al contrario, bloccare la produzione di anticorpi contro gli allergeni; per trattare malattie autoimmuni, come il lupus e l’artrite, manipolando questo stesso percorso, sarebbe possibile ridurre drasticamente i livelli di anticorpi che il corpo produce contro se stesso. Gli scienziati stanno già considerando di testare un modello di topo lupus.
Quale processo? Lo studio mostra che Ufbp1 sopprime l’enzima PERK per aiutare le cellule B a differenziarsi. Le proteine devono essere opportunamente ripiegate affinché si verifichino tutte le funzioni cellulari o corporee e PERK fa parte della naturale “risposta proteica spiegata” del corpo. PERK promuove quindi la risoluzione di problemi relativi a proteine ripiegate in modo improprio che non funzionano correttamente e possono diventare tossiche per le cellule: quando le proteine prodotte si piegano in modo inappropriato, viene attivato PERK, che interrompe la produzione di nuove proteine e riduce il numero di proteine ripiegate male.
Ma a questo punto, gli scienziati dimostrano che Ufbp1 sopprime PERK per garantire una produzione sufficiente di plasmacellule. Pertanto, quando c’è un deficit di Ufbp1 nelle cellule B, scoprono che, anche se le cellule B sopravvivono, lo sviluppo delle plasmacellule è compromesso. All’interno delle plasmacellule, Ufbp1 è sovraregolato, in modo che il reticolo endoplasmatico, che è la base dell’impianto di produzione di una cellula, possa espandersi e la capacità di piegare le proteine possono anche aumentare. Al contrario, i ricercatori dimostrano che un deficit di Ufbp1 nelle plasmacellule ostacola l’espansione del reticolo endoplasmatico e la produzione di anticorpi.
In sintesi, il percorso di ufilazione è un percorso essenziale per la produzione di cellule che secernono molte proteine, come le plasmacellule. Gli anticorpi funzionano come missili a lungo raggio e le plasmacellule solitamente li estraggono dal midollo osseo. Anche le cellule B sono prodotte nel e dal midollo osseo, ma circolano anche alla ricerca di invasori. Quando ne individuano uno, devono andare nella milza o in un linfonodo vicino per diventare una cellula del plasma. Le plasmacellule ritornano quindi nel midollo osseo. La sopravvivenza e il mantenimento delle plasmacellule è un equilibrio continuo e delicato, che può diventare fatale se qualcosa va storto. Senza questo equilibrio, le plasmacellule possono sfuggire al controllo, diventare cancerose anziché protettive e causare mieloma multiplo. Tra i molti passi avanti per gli scienziati, uno sarà determinare se il targeting per Ufbp1 è la soluzione per progettare la prossima generazione di trattamenti per mieloma multiplo.
Alcune persone nascono con l’assenza di alcuni componenti chiave del percorso di ufilazione e se l’impatto di questa carenza rimane scarsamente compreso, sappiamo che può provocare encefalopatia o disturbi del sangue. Il cattivo ripiegamento delle proteine è anche un fattore noto in condizioni come il morbo di Parkinson e il morbo di Alzheimer.
Questo team del Karolinska Institutet (Svezia) ha appena scoperto che gli anticorpi di tipo IgG svolgono un ruolo inaspettato nell’aterosclerosi. Il loro studio sui topi e pubblicato sulla rivista Circulation mostra che questi anticorpi stabilizzano la placca che si accumula sulle pareti delle arterie, riducendo il rischio di rottura e formazione di coaguli di sangue. I ricercatori sperano che questi risultati possano portare a trattamenti migliori.
Perché l’aterosclerosi è la principale causa sottostante di infarti e ictus (ictus) e sarà la principale causa di morte in tutto il mondo a lungo termine. Circa un terzo dei pazienti non risponde alla terapia con statine. La malattia è caratterizzata da un restringimento delle pareti arteriose derivante dall’accumulo (ateroma) di lipidi e cellule, chiamato placca di ateroma. Quando la placca si rompe, possono formarsi coaguli di sangue e limitare il flusso di sangue agli organi vitali, come il cuore e il cervello. Per ridurre il numero di decessi per aterosclerosi, i ricercatori stanno lavorando per trovare modi per prevenire la formazione e la rottura della placca.
Le cellule B nel sistema immunitario producono anticorpi che aiutano a combattere le infezioni. Questi anticorpi possono anche aiutare a rimuovere i tessuti danneggiati, ad esempio sotto forma di placche aterosclerotiche. Gli scienziati sanno anche che il sistema immunitario svolge un ruolo nello sviluppo della placca, ma il processo rimane poco compreso. Qui, il team svedese ha quindi studiato lo sviluppo della placca di ateroma nei topi privi di anticorpi. I ricercatori hanno scoperto che la placca formata in un ambiente privo di anticorpi è eccezionalmente piccola, ha un aspetto diverso e contiene più lipidi e meno cellule muscolari. Ciò suggerisce, scrivono i ricercatori nel loro comunicato stampa, che la placca è instabile e più soggetta a rotture.
Anticorpi IgG stabilizzanti: gli anticorpi IgG, la classe più comune di anticorpi nel sangue, vengono quindi identificati per il loro ruolo nella stabilizzazione della placca. Altre analisi mostrano anche che le cellule muscolari lisce nell’aorta hanno bisogno di questi anticorpi per dividersi correttamente. E quando le cellule non riescono a dividersi correttamente, la placca sembra diventare più piccola e più instabile.
Questi anticorpi svolgono quindi un ruolo chiave nella formazione e stabilità della placca arteriosa. Dovremo ora identificare quale particolare tipo di anticorpo IgG riconosce i componenti della placca.
Una volta identificati, questi anticorpi specifici potrebbero costituire la base di un nuovo trattamento in grado di attenuare l’aterosclerosi, stabilizzando la placca dell’ateroma e, sperano gli autori, contribuire a ridurre il numero di decessi cardiovascolari.
Our team of researchers observes the beauty of nature and draws lessons from it, and with creativity and innovation transforms them into new products, natural and safe. In particular, for products for veterinary use, we have selected Pleurotus ostreatus, a mushroom with relevant medicinal properties, rich in nutrients and bioactive molecules, among the ingredients.
Mushrooms between history and legend Mushrooms became part of human life thousands of years ago. We find ancient evidence of their use: they have been consumed both for nutritional and medical properties, as well as in shamanic and religious rituals. Traditional Chinese medicine has been able to recognize its benefits for human health, giving way to what is now known as Mycotherapy. And it is from the twentieth century that interest has also grown in the West. The history of P. ostreatus is also ancient, with references that come to us from the Sung dynasty (420-479 BC), calling it “mushroom of celestial flowers”. The first modern cultivation dates back to Germany in 1917, which used it as a subsistence measure during the First World War (M. Hofrichter, 2010). Its medicinal properties have been studied worldwide, from Central Europe to Africa and South America.
Beneficial properties of the mushroom Pleurotus Pleurotus ostreatus is a saprophytic-parasitic mushroom, with the characteristic shape that led it to be commonly known as the “oyster” mushroom. It grows on old stumps, in temperate and tropical climates and is today the third most common edible mushroom in the world, after the champignon and Shiitake. In recent years, studies concerning its nutraceutical and medicinal properties have increased. It is a source of protein, essential amino acids, B vitamins and vitamin C, as well as minerals. It therefore falls into the category of so-called functional foods, that is, containing bioactive molecules capable of bringing benefits to human health and reducing risks of chronic diseases (Patel et al., 2012).
A mushroom with relevant medicinal properties The bioactive substances contained in the mycelium and fruiting bodies have immunostimulating, antineoplastic, antidiabetic, anti-atherosclerotic, anti-inflammatory, antibacterial and antioxidant properties (Golak-Siwulska et al., 2018). A noteworthy secondary metabolite is Lovastatin, a statin involved in the reduction of blood cholesterol levels, which gives Pleurotus a protective action against heart disease (Zainal-Abidin et al., 2017). It is also a source of β-glucans such as Pleurane, capable of stimulating the body’s innate and adaptive immune response. Preclinical studies have reported a positive action, such as that carried out by Nita and collaborators (2018) where the administration of the fraction obtained by ethanolic extraction of P. ostreatus resulted in an increase in the population of lymphocytes, in the antibody titer and in the levels of γ -globulin in the blood of mice.
La sindrome di Goldenhar è una condizione rara descritta inizialmente nei primi anni ’50. È caratterizzato da una combinazione di anomalie: cisti epibulbar cutanee, appendici auricolari e malformazione delle orecchie. Nel 1963, Gorlin suggerì il nome displasia oculo-aurico-vertebrale (OAV) per questa condizione e includeva anche anomalie vertebrali come segni della sindrome. L’eziologia di questa malattia rara non è completamente compresa, poiché si è dimostrata variabile geneticamente e di cause poco chiare. Questo lavoro riporta un caso della sindrome di Goldenhar in una donna di 11 anni, che presentava tutti i segni classici di questa rara condizione
Parole chiave: deformità, manifestazione, displasia oculo-aurico-vertebrale.
SOMMARIO
La sindrome di Goldenhar è una condizione rara che è stata descritta per la prima volta nel 1952 come una combinazione di anomalie che includevano tumori dermoidi epibulbar, appendici auricolari e malformazioni dell’orecchio. Nel 1963, Gorlin suggerì il termine displasia oculo-aurico-vertebrale (OAV) includendo anomalie vertebrali in questa entità clinica. La sua eziologia non è chiara, essendo geneticamente variabile e di causa molto eterogenea. Gli autori riportano un caso clinico di Sindrome di Goldenhar in una bambina di 11 anni, che ha caratteristiche classiche di questa sindrome come tumore epibulbare dermoide, appendici auricolari, ipoplasia mandibolare e labbro leporino.
INTRODUZIONE
La sindrome di Goldenhar è una rara condizione presumibilmente ereditata, che ha un’eziopatologia multifattoriale che include anche fattori nutrizionali e ambientali che possono provocare disturbi della blastogenesi (1). Esistono diversi termini usati per descrivere questa rara condizione nota come displasia oculo-aurico-vertebrale (OAV), tra cui la sindrome di Goldenhar e la microsomia emifacciale (2). Goldenhar descrisse per la prima volta questa condizione nel 1952 come una malattia che presenta una combinazione di diverse anomalie come tumori epibulbar cutanei, appendici peri-auricolari e malformazione delle orecchie. All’inizio degli anni ’90, questa condizione è stata meglio compresa ed è stato concordato che, oltre al quadro descritto da Goldenhar (1952) e Gorlin (1963), questa sindrome può anche presentare malattie cardiache e ipoplasia dello zigomatico, ossa mandibolari e mascellari (3). Alcuni autori hanno anche sottolineato l’ipoplasia dei muscoli facciali, le anomalie anatomiche e morfologiche della lingua, le anomalie vertebrali, le anomalie degli occhi (1), il labbro e il palatoschisi (3), i disturbi del sistema nervoso centrale e altre anomalie viscerali (4).
Non ci sono abbastanza informazioni per identificarne i fattori eziologici. Sono state identificate anomalie dei cromosomi (5). D’altra parte, un altro studio ha suggerito un disturbo delle cellule della cresta neurale come causa della malattia (6). Anche l’influenza di altri fattori, incluso l’ambiente, durante la gravidanza è stata incolpata. L’ingestione di alcuni farmaci come la cocaina, la talidomide, l’acido retinoico e il tamoxifene da parte della madre era anche correlata allo sviluppo della malattia (4). Il diabete materno è stato anche suggerito come fattore eziologico (7).
Vi è un consenso generale sul fatto che la diagnosi di questa malattia non deve basarsi solo sui risultati radiologici o di laboratorio. La diagnosi della sindrome di Goldenhar dovrebbe basarsi principalmente sull’aspetto clinico e associata a condizioni sistemiche e risultati radiologici (7). La maggior parte degli autori considera la presenza di anomalie dell’orecchio (microtia) e di appendici sull’orecchio necessarie per la diagnosi. Inoltre, si osservano asimmetria facciale o ipoplasia facciale e / o mandibolare, tumori epibulbar cutanei, alterazioni palpebrali, anomalie vertebrali, schisi facciali laterali e problemi renali (8,9). Inoltre, sia i test di laboratorio che quelli di immagine sono importanti per la diagnosi della malattia poiché le anomalie delle ossa scheletriche o facciali possono essere diagnosticate mediante diversi tipi di esami di immagine disponibili oggi. L’esame radiografico delle ossa zigomatiche mostra una carenza macroscopica e una simmetria dello sviluppo. C’è anche un possibilità di agenesia di queste ossa con mancanza di fusione dell’arco zigomatico e agenesi delle ossa palatine. La schisi palatale può essere osservata radiograficamente (10). L’esame oftalmologico e otorinolaringoiatrico sono importanti anche per la diagnosi finale.
CASO CLINICO
Una femmina di 11 anni bianca è stata esaminata presso la clinica orale e maxillofacciale della Facoltà di Odontoiatria dell’Università Federale di Bahia. L’esame clinico ha indicato la sindrome di Goldenhar. Il paziente presentava asimmetria facciale, ipoplasia della mandibola, tumore epibulbare dermoide sull’occhio sinistro ( Figura 1 ) e segni di nascita sul labbro superiore e sul palato ( Figura 2 ). Inoltre, è stato dimostrato che i polipi peri-auricolari sono stati rimossi chirurgicamente quando il paziente aveva otto mesi ( Figura 3 ). Nessun problema mentale è stato rilevato durante l’esame. Non ci sono stati segni di compromissione dell’udito o del labbro e palatoschisi. La madre ha riferito l’uso di un farmaco anti-convulsivo (Comital ®) a causa dell’epilessia, prima di conoscere la gravidanza (circa quattro settimane) quando il farmaco è stato cambiato in fenobarbitale (Gardenal ® ) un farmaco più adatto per l’uso durante la gravidanza. L’esame radiografico del cranio e della colonna vertebrale non ha mostrato anomalie. Tuttavia, l’esame ortopantomografico ha rilevato ipoplasia della mandibola sul lato sinistro ( Figura 4 ), assenza del processo coronoideo e ipoplasia del condilo mandibolare. Lo sviluppo dentale era normale.
DISCUSSIONE
Lo studio di questa condizione è ancora controverso a causa della sua complessità e ampi aspetti clinici. La microtia sembra rappresentare la sua manifestazione meno complessa, tuttavia, possono anche essere osservate diverse anomalie facciali e sistemiche (9).
Il paziente ha mostrato caratteristiche cliniche della sindrome AOV complessa e grave come precedentemente descritto (1,6), tra cui asimmetria facciale, ipoplasia della mandibola, tumore dermoide epibulbar sull’occhio sinistro, vestigia di labbro leporino e presenza di appendici periauricolari precedentemente rimosse . L’asimmetria facciale e l’ipoplasia della mandibola sono caratteristiche tipiche della sindrome OAV (11). D’altra parte, la presenza del tumore dermoide epibulbar è variabile (3). Sebbene il paziente abbia mostrato tracce di un labbro leporino, questa alterazione si osserva in circa il 5% dei casi (3). È importante osservare che quando sono presenti tumori dermoidi epibulbar c’è una tendenza allo sviluppo di appendici biliare peri-auricolari osservate in questo caso specifico.
Nonostante la frequenza riportata di alterazioni cardiovascolari che varia dal 5 al 58% (12), in questo paziente non sono state riscontrate alterazioni cardiovascolari. Non sono stati osservati disturbi dell’udito o malformazione del meato uditivo esterno (13) né disfunzione del nervo facciale, la cui prevalenza è elevata (14). Non sono stati diagnosticati problemi renali comunemente associati a malformazioni delle orecchie (15). Precedenti rapporti di 294 pazienti (6) hanno mostrato che queste anomalie sono rare e compaiono in meno del 10% dei pazienti.
Va sottolineato che non vi erano precedenti rapporti familiari di questa condizione e che la madre aveva usato un farmaco anticonvulsivo all’inizio della gravidanza. Questa mancanza di insorgenza familiare può suggerire che la sindrome di Goldenhar può essere un evento sporadico che si verifica all’inizio dell’embriogenesi. Alcuni agenti teratologici come vitamina A, primidone, talidomide (16) e cocaina (4) sono stati associati allo sviluppo di questa sindrome, nonché alla malnutrizione, al tabacco e agli erbicidi che sono in grado di produrre radicali liberi che possono rompere il DNA e conseguentemente danno luogo a malformazioni congenite (1,17).
Sebbene l’OAV presenti alcune somiglianze con la sindrome di Treacher-Collins, ora è considerata un’entità distinta a causa di alcune caratteristiche non riscontrabili in entrambe le malattie, inclusa la mancanza di prove di un’origine genetica della sindrome OAV (10). In questo caso, la sostituzione di Comital Ò con Gardenal Ò può spiegare lo sviluppo della sindrome senza la presenza di tutte le sue varianti.
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