Validation of in silico prediction by in vitro immunoserological results of fine epitope mapping on citrate synthase specific autoantibodies.

In silico antibody-antigen binding predictions are generally employed in research to rationalize epitope development. These techniques are widely spread despite their technical limitations. To validate the results of these bioinformatic calculations evidence based comparative in vitro studies are necessary. We have used a well-conserved mitochondrial inner membrane antigen-citrate synthase to develop a model for comparative analysis of the predicted and the immunoserologically verified epitopes of circulating autoantibodies. Epitopes were predicted using accepted tools: the GCG Wisconsin package and TEPITOPE 2000. An overlapping multipin ELISA assay--covering 49% of the citrate synthase molecule--was developed to map autoantibody epitopes of individuals (healthy, systemic autoimmune, and heart transplanted) in different immunopathological conditions. From the 40 synthesized decapeptides 34 were predicted in silico and 27 were validated in vitro. Thirty-two percent of epitopes were recognized by majority of sera 47% by at least one sera. False positive predictions were 21%. There was major difference in the recognized epitope pattern under different immunopathological conditions. Our results suggest that special databases are needed for training and weighing prediction methods by clinically well-characterized samples, due to the differences in the immune response under different health status. The development of these special algorithms needs a new approach. A high number of samples under these special immunological conditions are to be mapped and then used for the "fine tuning" of different prediction algorithms.

Early effects of different cytostatic protocols for head and neck cancer on oncogene activation in animal experiments.

In vivo investigations on oncogenes and onco-suppressor genes may provide new findings on the potential carcinogenic effects of various cytostatic protocols inducing secondary tumours of the head and neck. Further surgeries are often necessary due to regional recurrence after the Cisplatin-supplemented BVM (Bleomycin, Vincristine, Methotrexate) protocol in the treatment of human head and neck tumours. Our earlier studies have illustrated the carcinogenic and mutagenic potential of Cisplatin. The effect of Cisplatin on the alteration of different onco- and suppressor genes has also been proven. Our present study aimed at investigating the early effects of the BVM and the CFu (Cisplatin, 5-Fluorouracil) protocols on early oncogene and tumour suppressor gene expressions in mice. Body weight equivalent amounts of cytostatics were administered intraperitoneally to 6- to 8-week-old, inbred, female CBA/Ca mice. Twenty-four, 48 and 72 hours after the treatment, RNA was isolated from the target organs and the quantitative expression of c-myc, Ha-ras and p53 genes were examined. The protocols caused detectable changes. A "short-term" in vivo test, the 24-hour examination of gene expression, is suitable for detecting early effects of carcinogen exposure. The alterations of gene expression, caused by the Cisplatin-containing protocol, draw attention to the probable role of Cisplatin in the development of regional recurrence and to the possibility of prevention.

Surgical closures of oropharyngocutaneous fistulas.

Of the 146 patients undergoing surgery for oropharyngeal cancer in our institution, 12 (8.2 percent) developed fistulas. As a first line of therapy, conservative measures were used, which consisted of debridement, Xeroform gauze packing, and nasogastric feeding. Seven fistulas closed after conservative treatment. Of the five patients who required surgery for fistula closure, three had large (more than 20 mm) and two had mid-size (5- to 20-mm) fistulas. In all cases, internal flaps were prepared from the healthy viable tissues surrounding the fistula, and sternocleidomastoid-trapezius-platysma myocutaneous flaps were used for external closure. None of the closures failed, and we obtained good functional and aesthetic results.

[Double flap reconstruction of extended facial defects]. / Kiterjedt facialis defektusok kettós lebenyes rekonstrukciója.

The authors report the reconstruction of large facial defects remaining after resection of two advanced facial cancer cases. In both cases double flap closures were carried out. In the first case the internal flap was created from the remaining part of the oral mucosa membrane, while in the second a forearm fascio-cutan free flap was used. For external closure in both cases platysma based transpositional flaps were used, prepared from the neck. On the basis of excellent aesthetic results and safe applicability the authors recommend the more frequent use of this method.

[Randomized study of cisplatin-based combination chemotherapy for the treatment of planocellular cancer of the head and neck region]. / Kombinált cisplatin-tartalmú kemoterápia randomizált vizsgálata a fej-nyak régióban lévó planocellularis rákok kezelésében.

Between January 1996 and November 1998 38 patients were treated with induction chemotherapy. Patients were distributed in two randomized groups, 19-19 patients in each, receiving (group N) either bleomycin, vincristine and methotrexate (BVM) or the previous medication plus cisplatin (BVCM) chemotherapy (group C). Side effects were low and reversible during the treatments. The clinical regression rate (RR) of the cases was 87% including complete regression 24%. There was no difference between the two groups. There was no difference in the pathological macroscopic regression, however group C was better in microscopic regression. During the 27.5 months of average follow-up time, group N had a significantly better result in the tumor-free survival, with a lower rate of metastatic recidives (N/C = 2/9). There was no significant difference in the overall survival rate, due to the radical neck dissections of recidive metastases. According to our experience we recommend the use of cisplatin in conjunction with neck radiotherapy.

Transgingival lag-screw osteosynthesis of alveolar process fracture.

Fracture of the alveolar process is a common injury. In some cases, traditional fixation may not be possible. The teeth needed for splinting or mandibulo-maxillary fixation may be missing. The fracture line and soft tissue injury may jeopardize the blood supply of the broken bone. In these extreme and rare situations, the best rehabilitation is needed to avoid the loss of hard and soft tissues, and a secondary reconstruction is required. Between January 2003 and December 2006, of 468 cranio-facial trauma patients studied, alveolar process fracture was reported in 28 (6%) cases. In six (1%) cases, the anatomy of the fracture lines, and the position and number of the remaining teeth made splinting and mandibulo-maxillary fixation impossible. Patients were treated with a transgingival lag-screw (TLS) osteosynthesis. All patients healed well with no complications. There was no bone or tooth loss in the surgical area, and broken fragments were not absorbed. The TLS technique is recommended for alveolar fractures when the blood supply is jeopardized and dental splinting or mandibulo-maxillary fixation is not possible. There is no need for flap reflection.

[Modified submental endotracheal intubation in concurrent orthognathic surgery]. / Modifizierte submentale endotracheale Intubation bei kieferorthopädischen Eingriffen.

BACKGROUND: In 1986 Altemir described a method of submental endotracheal intubation in order to avoid tracheostomy in maxillofacial trauma cases where short-term intermaxillary fixation was required. His method has become widely established for airway maintenance in midfacial fractures. CASE: We present a 21-year-old male patient with cleft lip and palate on the left side. The patient underwent Le Fort I maxillary osteotomy. Nasal intubation was impossible due to nasal malformation. In this case we used submental endotracheal intubation for airway maintenance. We introduced new methods: a sterile nylon guiding tube and our new "2-2-2 rule" incision to make the procedure easier. RESULTS: The occlusion could be checked easily. There were no complications during and after the operation. The submental wound healed nicely. DISCUSSION: The described case shows that the technique is easy to use compared to "alternative" intubation methods. Submental scarring is acceptable. We recommend the technique for orthognathic use.

Gene survey of the pathogenic protozoan Trypanosoma cruzi.

We have performed a survey of the active genes in the important human pathogen Trypanosoma cruzi by analyzing 5013 expressed sequence tags (ESTs) generated from a normalized epimastigote cDNA library. Clustering of all sequences resulted in 771 clusters, comprising 54% of the ESTs. In total, the ESTs corresponded to 3054 transcripts that might represent one-fourth of the total gene repertoire in T. cruzi. About 33% of the T. cruzi transcripts showed similarity to sequences in the public databases, and a large number of hitherto undiscovered genes predicted to be involved in transcription, cell cycle control, cell division, signal transduction, secretion, and metabolism were identified. More than 140 full-length gene sequences were derived from the ESTs. Comparisons with all open reading frames in yeast and in Caenorhabditis elegans showed that only 12% of the T. cruzi transcripts were shared among diverse eukaryotic organisms. Comparison with other kinetoplastid sequences identified 237 orthologous genes that are shared between these evolutionarily divergent organisms. The generated data are a useful resource for further studies of the biology of the parasite and for development of new means to combat Chagas' disease.