Comparação entre dois protocolos de tratamento de ceratoconjuntivite seca experimentalmente induzida em coelhos / Comparison of two treatment protocols of experimentally induced keratoconjunctivitis sicca in rabbits

O objetivo deste estudo foi avaliar e comparar a eficácia de dois protocolos de tratamento de ceratoconjuntivite seca (CCS) experimentalmente induzida em coelhos: uma formulação oftálmica tópica composta por álcool polivinílico 1,4%, adicionado com acetilcisteína 10% e pilocarpina 1% (AAP), e outro protocolo com o uso do óleo de semente de linhaça (OL) tópico em forma de colírio, durante 12 semanas. Foram utilizados 15 coelhos machos, adultos, da raça Nova Zelândia, alocados aleatoriamente em três grupos: grupo C (controle), grupo AAP (formulação oftálmica) e grupo L (OL tópica). Os animais foram avaliados semanalmente pelo teste lacrimal de Schirmer, teste de fluoresceína e teste de Rosa Bengala; uma vez por mês, pelo exame de citologia esfoliativa ocular; ao final do experimento, pela análise histopatológica da córnea e conjuntiva. Os resultados demonstraram que houve um aumento maior na produção lacrimal quando utilizada a formulação oftálmica, e uma resolução mais rápida das úlceras de córnea, bem como diminuição no número de células desvitalizadas quando utilizado o óleo de semente de linhaça, além de aumento no número de células caliciformes em ambos os grupos de tratamento. A associação desses dois protocolos pode ser no futuro uma alternativa no tratamento da CCS.

The objective of this study was to evaluate and compare the effectiveness of two treatment protocol of experimentally induced keratoconjunctivitis sicca (KCS) in rabbits, a topical ophthalmic formulation composed by 1.4% povinilic alcohol added with 10% acetylcysteine and 1% pilocarpine (AAP) and another protocol with the topical use of the linseed seed oil (LO) in eye drop form f or 12 weeks. Fifteen male New Zealand white rabbits were aleatory allocated in 3 groups: Group C (Control), Group AAP (ophthalmic formulation) and Group L (LO topical). The animals were evaluated weekly using the Schirmer's tear test, fluorescein test and Rose Bengal test monthly for ocular cytology, and at the end of the experiment for histopathological analysis of cornea and conjunctive. The results demonstrated that there was a larger increase in the tear production when the ophthalmic formulation was us ed and a faster rapid resolution of corneal ulcers and decrease in the number of devitalized cells when linseed seed oil was used, besides an increase in the number of caliciform cells in both treatment groups. The association of those two protocols can be a future alternative in the treatment of KCS.

Toward a systems biology approach to investigate cellular networks in normal and malignant B cells.

In recent years, we experienced an increasing development of new technologies that aim to comprehensively dissect the molecular genetics of cellular phenotypes. Pioneering studies have been performed on leukemia and lymphoma and then extended to many other types of malignancies. Genome-wide technologies allow taking snapshots of defined cellular context from an unbiased angle highlighting a complexity that we still struggle to fully interpret. The increasing availability of technologies to detect genetic, transcriptional and post-transcriptional characteristics of cellular systems needs to be associated with the development of computational tools to fully investigate these data in an integrated way. The evolution of different genome-wide technologies as well as data mining and integration tools will be discussed following studies performed on normal and malignant human mature B cells.

Gene expression profiling suggests primary central nervous system lymphomas to be derived from a late germinal center B cell.

To characterize the molecular origin of primary lymphomas of the central nervous system (PCNSL), 21 PCNSLs of immunocompetent patients were investigated by microarray-based gene expression profiling. Comparison of the transcriptional profile of PCNSL with various normal and neoplastic B-cell subsets demonstrated PCNSL (i) to display gene expression patterns most closely related to late germinal center B cells, (ii) to display a gene expression profile similar to systemic diffuse large B-cell lymphomas (DLBCLs) and (iii) to be in part assigned to the activated B-cell-like (ABC) or the germinal center B-cell-like (GCB) subtype of DLBCL.

Prospective analysis of minimal bone marrow infiltration in pediatric Burkitt's lymphomas by long-distance polymerase chain reaction for t(8;14)(q24;q32).

The chromosomal translocation t(8;14)(q24;q32) represents a characteristic marker for Burkitt's lymphoma (BL). This translocation involves the MYC oncogene on chromosome 8 and the immunoglobulin heavy-chain (IgH) locus on chromosome 14. Since the translocation does not produce a fusion gene, we established a long-distance polymerase chain reaction (LD-PCR) assay that can detect the t(8;14) at the genomic level. The sensitivity of the LD-PCR was 10(-4). We used the LD-PCR assay to prospectively study 78 BL patients and found a specific PCR product in 52 of them. Among the 52 positive patients, we could test both the tumor and the bone marrow (BM) at diagnosis in 33 and determined the prevalence of minimal disseminated disease (MDD) at diagnosis. In 12/33 patients, BM was positive by LD-PCR and in 10 of them we conducted a study of minimal residual disease (MRD). Eight out of 10 children showed a clearance of MRD after one cycle of chemotherapy. The only two patients who did not achieve a negative MRD status died of disease progression. The comparative analysis of sensitivity of BM aspirate, BM biopsy and LD-PCR in t(8;14)-positive patients demonstrated a superiority of the molecular method in the assessment of MDD. The LD-PCR for t(8;14) is an important tool to study minimal BM infiltration at diagnosis and to determine its response kinetics in BL.

Detection and characterization of micronuclei in a murine liver epithelial cell line, by application of the in vitro cytokinesis block MN assay and PRINS.

The cytokinesis block micronucleus assay was applied to murine cell line C6, derived from fetal liver, after an optimal protocol had been designed. Micronucleus frequencies were assayed after exposure to three concentrations of colcemid or diepoxybutane. Two-colour primed in situ DNA synthesis (PRINS) was applied to simultaneously label telomeric and centromeric (minor satellite DNA) sequences. Both chemicals induced a highly significant increase in MN and the effect was dose dependent. Diepoxybutane did not appear to significantly increase the frequency of centromere-positive micronuclei. Colcemid, as expected, induced high frequencies of centromere-positive micronuclei at all concentrations tested; in addition a significant increase in centromere-negative micronuclei was observed at 10(-5) M. Many centromere-positive micronuclei carried three or four telomeres, thus indicating that a duplicated (non-disjoined) chromosome with two chromatids was contained in the micronucleus. This observation leads to the conclusion that micronuclei deriving from missegregation could be due to errors occurring before the onset of anaphase. The results obtained on C6 cells are in good agreement with those obtained on other cell systems, indicating that this cell line can be considered for in vitro aneuploidy evaluation.

Improved long-distance polymerase chain reaction for the detection of t(8;14)(q24;q32) in Burkitt's lymphomas.

The t(8;14)(q24;q32), involving MYC gene (8q24) and the immunoglobulin heavy chain (IgH) locus (14q32), represents about 75% of all translocations in Burkitt's lymphoma (BL). Due to the great variability of the breakpoint region, a standard polymerase chain reaction assay is not sufficient for the detection of this chromosomal translocation. The availability of new and more efficient DNA polymerases that allow the amplification of genomic fragments many kilobase-pairs long, makes it possible to identify the t(8;14) in BL cells by long-distance polymerase chain reaction (LD-PCR). We have established a simplified and efficient LD-PCR for the detection of t(8;14)(q24;q32) that relies on the use of one primer specific for MYC exon II combined, in different reactions, with four primers for the IgH locus: three for the constant regions Cmu, Cgamma, and Calpha, and one for the joining region (JH). We first studied seven BL cell lines and optimized LD-PCR reaction for analysis of tumor specimens. Five of seven cell lines were positive for the t(8;14), whereas two lines derived from endemic BL were negative, as expected. Of 15 biopsies obtained from pediatric BL and subsequently analyzed, 13 (87%) were positive for the translocation detected by LD-PCR and showed a product ranging in size from 2.0 to 9.5 kb. Cmu region was involved in 6 cases, Cgamma and Calpha in 2 cases each, and JH in 3 cases. Interestingly, 2 of the tumors positive for JH showed a second, larger PCR product with the Calpha- and Cgamma-specific primer, respectively. We established that our LD-PCR method could detect 10(-3) BL cells within a population of hematopoietic cells lacking the translocation. In conclusion, our LD-PCR method represents a fast, highly sensitive, and specific tool to study sporadic BL and to detect minimal disease and residual disease in patients affected by t(8;14)-positive lymphomas.

A Western apache writing system: the symbols of silas john.

At the outset of this article, it was observed that the adequacy of an etic typology of written symbols could be judged by its ability to describe all the emic distinctions in all the writing systems of the world. In conclusion, we should like to return to this point and briefly examine the extent to which currently available etic concepts can be used to describe the distinctions made by Western Apaches in relation to the writing system of Silas John. Every symbol in the Silas John script may be classified as a phonetic-semantic sign. Symbols of this type denote linguistic expressions that consist of one or more words and contrast as a class with phonetic-nonsemantic signs, which denote phonemes (or phoneme clusters), syllables (or syllable clusters), and various prosodic phenomena (2, pp. 2, 248). Phonetic semantic signs are commonly partitioned into two subclasses: alogographs (which denote single words) and phraseographs (which denote on or more words). Although every symbol in the Silas John script can be assigned to one or the other of these categories, such an exercise is without justification (21). We have no evidence to suggest that Western Apaches classify symbols according to the length or complexity of their linguistic referents, and therefore the imposition of distinctions based on these criteria would be inappropriate and misleading. A far more useful contrast, and one we have already employed, is presented in most etic typologies as an opposition between compound (composite) and noncompound (noncomposite) symbols. Used to break down the category of phonetic-semantic signs, these two concepts enable us to describe more or less exactly the distinction Apaches draw between "symbol elements put together" (ke?escin ledidilgoh) and "symbol elements standing alone" (ke?- escin doledidildaahi). The former may now be defined as consisting of compound phonetic-semantic signs, while the latter is composed of noncompound phonetic-semantic signs. Up to this point, etic concepts have served us well. However, a deficiency appears when we search for a terminology that allows us to describe the distinction between "symbols that tell what to say" and "symbols that tell what to do." As far as we have been able to determine, standard typologies make no provision for this kind of contrast, apparently because their creators have tacitly assumed that systems composed of phonetic-semantic signs serve exclusively to communicate linguistic information. Consequently, the possibility that these systems might also convey nonlinguistic information seems to have been ignored. This oversight may be a product of Western ethnocentrism; after all, it is. we who use alphabets who most frequently associate writing with language (22). On the other hand, it may simply stem from the fact that systems incorporating symbols with kinesic referents are exceedingly rare and have not yet been reported. In any case, it is important to recognize that the etic inventory is not complete. Retaining the term "phonetic sign" as a label for written symbols. that denote linguistic phenomena, we propose that the term "kinetic sign" be introduced to label symbols that denote sequences of nonverbal behavior. Symbols of the latter type that simultaneously denote some unit of language may be classified as "phonetic-kinetic" signs. With these concepts, the contrast between " symbols that tell what to say" and "symbols that tell what to do" can be rephrased as one that distinguishes phonetic signs (by definition nonkinetic) from phonetic-kinetic signs. Purely kinetic signs-symbols that refer solely to physical gestures-are absent from the Silas John script. The utility of the kinetic sign and the phonetic-kinetic sign as comparative concepts must ultimately be judged on the basis of their capacity to clarify and describe emic distinctions in other systems of writing. However, as we have previously pointed out, ethnographic studies of American Indian systems that address themselves to the identification of these distinctions-and thus provide the information necessary to evaluate the relevance and applicability of etic concepts-are in very short supply. As a result, meaningful comparisons cannot be made. At this point, we simply alack the data with which to determine whether the kinetic component so prominen in the Silas John script is unique or whether it had counterparts else-where in North America. The view is still prevalent among anthropologists and linguists that the great majority of American Indian writing systems conform to one or two global "primitive" types. Our study of the Silas John script casts doubt upon this position, for it demonstrates that fundamental emic distinctions remain to be discovered and that existing etic frameworks are less than adequatelyequipped to describe them. The implications of these findings are clear. On the one hand, we must acknowledge the possibility that several structurally distinct forms of writing were developed by North America's Indian cultures. Concomitantly, we must be prepared to aabandon traditional ideas of typological similarity and simplicity among thes systems in favor of those that take into account variation and complexity.