Othon Papadopoulos

Abstract: Adipose tissue is believed to constitute an ideal source of uncultured stromal stem cells. By optimizing the harvesting, storage, and transplantation of adipose tissue, long-lasting results can be obtained. Cell-assisted lipotransfer (CAL) is a novel approach to autologous fat transplantation in which adipose-derived stem cells are attached to the aspirated fat. The authors describe the cell processing methods, delivery systems, and clinical applications of CAL.

Abstract: Inoculation of embryonated chicken eggs is the standard method for the titration of infectious Bluetongue virus (BTV). Here, six RNA extraction methods coupled with optimised dsRNA denaturation and real-time RT-PCR were evaluated for the quantitation of BTV in blood samples from experimentally infected sheep and results were correlated to infectious virus titres. An exogenous dsRNA internal control (IC) from the closely related Epizootic hemorrhagic disease virus (EHDV) was used to assess the efficiency of BTV genome extraction, dsRNA denaturation, RT, and PCR amplification. Recovery rates of IC and BTV dsRNA copies from extracted blood samples were highly correlated. Adjustment of BTV concentrations according to the IC recovery reduced variation in sample analyses among the different extraction methods and improved the accuracy of BTV quantitation. The EID(50)/ml titre, determined in blood samples from sheep infected experimentally with BTV-1 or BTV-9, correlated highly with the assessed concentration of BTV dsRNA copies. However, this correlation was consistent only during the first 28 days post-infection. The optimised extraction methods and quantitative RT-PCR could be useful for experimental studies of BTV transmission, pathogenesis and vaccine efficacy, or adapted further for the detection and quantitation of EHDV, African horse sickness virus and other dsRNA viruses.

Abstract: The association between PRNP variation and scrapie incidence was investigated in a highly affected Greek goat herd. Four mutations were identified at codons 171Q/R, 211R/Q, 222Q/K and 240P/S. Lysine at codon 222 was found to be associated with the protection from natural scrapie (P=0.0111). Glutamine at codon 211 was observed in eight animals, all of them being scrapie-negative, indicating a possible protective role of this polymorphism although statistical analysis failed to support it (P=0.1074). A positive association (P=0.0457) between scrapie-affected goats and the wild-type Q(171)R(211)Q(222)S(240) allele is presented for the first time. In addition, a novel R(171)RQS allele, which is identical to the A(136)R(154)R(171) allele that has been associated with resistance to classical scrapie in sheep, was observed in low frequency. Resistant alleles that include K(222) and Q(211) are absent or rare in sheep and can provide the basis for the development of a feasible breeding programme for scrapie eradication in goats.

Abstract: One of the most difficult ethical dilemmas facing health care professionals working in oncology is whether, when, how and how much to tell terminal cancer patients about their diagnosis and prognosis. The aim of this article is to review the trends in this issue worldwide. While a majority of physicians in both developed and developing countries tell the truth more often today than in the past, the assumption that truth-telling is always beneficial to patients can be questioned. The issue of truth-telling is still approached differently in different countries and cultures and there is a need for an increased awareness of cultural differences to truth-telling among patients from ethnic minorities.

Abstract: Fibrokeratoma is a rare benign invasive tumour usually located on the digits. We report a 35-year-old patient with a giant acquired fibrokeratoma of the heel. Despite its large size and unusual location, the tumour was microscopically benign and was successfully excised.

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Abstract: Culicoides biting midges (Diptera: Ceratopogonidae) were trapped between 1999 and 2004 at 122 locations in mainland Greece and on most of the larger Aegean and Ionian islands, using OVI light traps, in order to determine the distribution and seasonal activity of bluetongue virus vectors and other Culicoides species. Thirty-nine Culicoides species were identified, six of which (C. furcillatus, C. impunctatus, C. paolae, C. pictipennis, C. riethi, and C. scoticus) were identified for the first time in Greece. Two of these (C. impunctatus and C. scoticus) may be of veterinary importance due to their role as vectors of bluetongue virus and related orbiviruses. In addition, C. imicola was detected for the first time in mainland Greece.

Abstract: INTRODUCTION: Nasal reconstruction after tumor extirpation is a necessity. The aim of the current study was to present the Greek experience in this field for a long period. MATERIALS AND METHODS: Charts of patients who underwent nasal reconstruction, from 1985 to 2006, were reviewed retrospectively. They were analyzed in relation to their age, sex, location of the defect, histologic diagnosis of the lesion, type of reconstruction, recurrence of the tumor, and final outcome. RESULTS: A total of 1585 patients underwent nasal reconstruction by the senior author (O.P.) during a period of 21 years in our department. A clear male preponderance was shown (845 or 53.3% vs 740 or 46.7%). Their age ranged from 13 to 97 years with a mean of 65.9 years. One thousand five hundred ninety-three different tumors had been resected during the studied period (some patients had >1 lesion). Basal cell carcinoma was the most common type, affecting 1399 patients (87.8%), followed by squamous cell carcinoma, which was identified in 109 patients (6.8%). Cutaneous melanoma was not a frequent diagnosis. Excision and primary closure represented the most frequent type of reconstruction, followed by flap reconstruction and any type of graft. Sidewalls were the usual location in the whole population. Recurrence rate was 3.4%. CONCLUSIONS: Nasal reconstruction remains a challenge for every plastic surgeon. Efficient diagnosis and appropriate reconstruction are prerequisites for the final desired outcome.

Abstract: PURPOSE: A high-dose interferon alfa (IFN-alpha) regimen as reported in E1684 was unique for the incorporation of an induction phase of maximally tolerated dosages of intravenous (IV) therapy for the initial 4 weeks. This is the only trial that has shown prolongation of overall survival and relapse-free survival (RFS) in comparison with observation. Analysis of the hazard curves for RFS and overall survival (OS) in E1684 revealed separation of the high-dose and observation arms, suggesting that the induction phase may represent a critical component of this regimen, although this has not been tested prospectively. PATIENTS AND METHODS: We conducted a prospective randomized study of IV induction therapy versus a full year of high-dose IFN, with primary end points of RFS and OS for patients with stage IIB, IIC, and III melanoma, within 56 days of curative surgery. Patients were randomly assigned to receive IFN-alpha-2b 15 x 10(6) U/m2 IV x 5/7 days weekly x 4 weeks (arm A) versus the same regimen followed by IFN-alpha-2b 10 x 10(6) U (flat dose) administered subcutaneously three times a week for 48 weeks (arm B). RESULTS: Between 1998 and 2004, 364 patients were enrolled (353 eligible: arm A, n = 177; arm B, n = 176). At a median follow-up of 63 months (95% CI, 58.1 to 67.7), the median RFS was 24.1 months versus 27.9 months (P = .9) and the median OS was 64.4 months versus 65.3 months (P = .49). Patients in arm B had more grade 1 to 2 hepatotoxicity, nausea/vomiting, alopecia, and neurologic toxicity. CONCLUSION: There were no significant differences in OS and RFS between the regimens of 1 month and 1 year of treatment.

Abstract: The SLC24A5 gene, the human orthologue of the zebrafish golden gene, has been shown to play a key role in human pigmentation. In this study, we investigate the prevalence of the variant allele rs1426654 in a selected sample of Greek subjects. Allele-specific polymerase chain reaction was performed in peripheral blood samples from 158 attendants of a dermatology outpatient service. The results were correlated with pigmentary traits and MC1R genotype. The vast majority of subjects (99%) were homozygous for the Thr(111) allele. Only two subjects from the control group (1.26%) were heterozygous for the alanine and threonine allele. Both of these Thr(111)/Ala(111) heterozygotes carried a single polymorphism of MC1R (one with the V92M variant and another with the V60L variant). Following reports of the rs1426654 polymorphism reaching fixation in the European population, our study of Greek subjects showed a prevalence of the Thr(111) allele, even among subjects with darker skin pigmentation or phototype.

Abstract: BACKGROUND: The role of combination chemotherapy regimens in the management of ovarian cancer patients with tumors previously exposed to platinum compounds and paclitaxel has not yet been defined. The present phase II study evaluated the activity and toxicity of a gemcitabine-ifosfamide-cisplatin combination in the aforementioned group of patients. Given the in vitro and in vivo synergism between the three agents, it was believed that using a three-drug combination would overcome tumor resistance to cisplatin. PATIENTS AND METHODS: Twenty-four patients were enrolled in the study. The median age was 56 years and the median performance status 1. Eight (34%) had potentially platinum-sensitive, 6 (24%) had primary platinum-resistant and 10 (42%) patients had secondary platinum-resistant tumors. Treatment consisted of gemcitabine 1 g/m(2) i.v. on days 1 and 8, cisplatin 75 mg/m(2) i.v. over 2 h fractionated over days 8 and 9, and ifosfamide 5 mg/m(2) i.v. over 1 h fractionated on days 8-9 with mesna uroprotection. Courses were administered every 3 weeks on an outpatient basis. Granulocyte colony-stimulating factor (G-CSF) was given at a dose of 5 microg/kg/day on days 10-14. A median of 4 cycles were administered with the delivered dose intensity at 85% of the planned dose for the three agents. RESULTS: Among 24 patients evaluable for response and toxicity, there were 8 partial responses with a response rate of 33% (95% confidence interval 16.4-55%). Stable disease was recorded in 6 (25.7) and progressive disease in 10 (42%) patients. Subgroup analysis revealed a response rate of 50% in potentially platinum-sensitive, 16.5% in primary platinum-resistant and 30% in secondary platinum-resistant tumors. The median response duration was 5 months (range 3-12 months), the median time to progression 6 months (range 3-16 months) and the median survival 12 months (range 3-24 months). Myelotoxicity was significant, with neutropenia grade 3 and 4 occurring in 35% and 20% of patients, respectively. Four episodes (3.5% of all cycles) of febrile neutropenia were documented and were well managed with oral antibiotics and G-CSF continuation until complete recovery. Grade 1, 2 and 3 peripheral neuropathy developed in 40%, 30%, and 10% of patients, respectively. CONCLUSION: The three-drug combination demonstrated a significant effectiveness in potentially platinum-sensitive tumors and a moderate efficacy in platinum-resistant tumors. The regimen, although myelotoxic, is tolerable with G-CSF support. Further investigation via comparative studies is required to define any superiority of the present regimen over doublets of the three agents in this group of patients.

Abstract: Implants are widely used and are not without complications. The most serious complication is infection with consequent exposure of the implant and is characterized by increased morbidity and mortality. Until recently, infected implants were removed and replaced with another one. These operations are accompanied by increased morbidity and might result in permanent disability. Coverage of the implant with a flap is an alternative solution. The objective of this article is to study the effectiveness of muscle or myocutaneous flaps in covering infected and exposed implants. A series of 17 patients with infected implants, which were treated with coverage of the implant with muscle or myocutaneous flap, is presented. Eight patients had an orthopedic implant, seven a vascular implant, and two had a pacemaker. The flaps ensured immediate coverage of the implant and the biologic ability of eliminating the infectious process with the increased concentration of antibiotics resulting from increased blood flow of the muscle. In all the patients, the implants were preserved and the infectious process was eliminated. The use of muscle and myocutaneous flaps in the treatment of infected implants is therefore effective and in some cases it is superior to other methods because of lower morbidity and cost.

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Abstract: There is no standard treatment for breast cancer patients whose tumors have been exposed both to anthracyclines and taxanes. Oxaliplatin shows synergism with 5-fluorouracil (5-FU) and capecitabine is an oral prodrug of 5-FU with known efficacy in pretreated patients. This phase II trial studied the efficacy and toxicity of the oxaliplatin-capecitabine combination as salvage treatment in breast cancer patients pretreated with anthracyclines and taxanes. Patients received oxaliplatin 80 mg/m(2) on day 1 followed by oral capecitabine 1800 mg/m(2) divided in two doses for 7 days every two weeks for a maximum of twelve courses or until disease progression. Twenty-eight patients were evaluable for efficacy and toxicity. Objective responses (all partial) were documented in 9 patients [32%; 95% confidence interval (CI): 13-51.2%]. Responses were documented at all metastatic sites. The median response duration was 5 months (range 3-9), median time to progression was 4.5 months (range 2-10) and median overall survival was 10 months (range 2-18). Myelotoxicity was minimal with grade 3 thrombocytopenia as the main toxicity. Hand-foot syndrome was well tolerated. The present regimen was well tolerated with a rather moderate effectiveness but very significant for this group of patients. Further studies where the combination could be compared with single agent capecitabine are warranted.

Abstract: Scalp reconstruction is a challenging area in plastic surgery. The susceptibility of potentially exposed brain tissue is almost always a concern along with the provision of adequate soft tissue coverage in cases of full-thickness defects. The applied reconstructive strategy also affects efficiency in malignant disease treatment providing both local control of the disease and vigorous monitoring for recurrence or metastasis. The general condition of the patients presenting with malignant lesions of the scalp is often impaired because of old age or concomitant disease. Therefore, demanding, long-lasting, or multistage procedures may often be undesirable. The double scalping flap comprises a 1-stage procedure, which can be used in most full-thickness defects of the vertex of the scalp. Some of the primary advantages of the double scalping flap procedure are its versatility and arc of rotation, its minimal donor site morbidity, being relatively simple, and being a short technique. Disadvantages that may be considered are the loss of hair-bearing skin at the occipital region and the poor color and texture match between scalp skin and the split-thickness skin graft that is used on the donor site. We present 2 patients with exemplary case of neglected malignancy of the scalp and their successful treatment using the double scalping flap. Comparison and contrast of alternative reconstructive procedures are also included to further investigate scalp reconstruction.

Abstract: Keloids are a frequent finding after physical trauma. Keloids of the pinna (helix and antihelix) as a complication of ear-piercing are the most difficult cases. Clinicians have tended to avoid the surgical approach, preferring local conservative treatment with corticosteroids or other treatments. We report use of intrakeloid resection and a form-pressure device to treat pinna keloids and avoid recurrence. The purpose is to maintain the form of the folds of the pinna. The recommendation for this therapy is to maintain the pressure at 24-30 mmHg, and the duration of the form-pressure therapy in our patients was about 25 weeks.

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Abstract: OBJECTIVES: Limited data exist about the risk factors of melanoma in the Greek population. We investigated the association of melanoma with phenotypic and solar indices in this darker skin population residing in an environment of high ambient ultraviolet radiation. METHODS: Our study included 200 sporadic melanoma cases and 200 age-, sex-matched control subjects. Information on history of sun exposure patterns and cutaneous reaction to sunlight was obtained and a clinical evaluation of pigmentary traits, pigmented lesions, and actinic keratoses was performed. RESULTS: In the multivariate analysis, fair skin (OR: 4.63, for fair skin versus light brown, 95% CI: 1.54-13.92), intermittent sun exposure during childhood (OR: 3.33, >2 weeks/year of sun exposure versus < or =2 weeks/year 95% CI: 1.37-8.09), and outdoor leisure activities (OR: 2.74, 95% CI: 1.28-5.89), but not skin phototype or sunburns, were positively related to the risk of melanoma. In addition to an elevated count of common melanocytic nevi (OR: 6.27, > or =10 nevi versus no nevi, 95% CI: 1.65-23.76) and the presence of clinically atypical nevi (OR: 2.84, 95% CI: 1.16-6.98), solar lentigenes were an independent risk factor of melanoma (OR: 4.33, 95% CI: 1.67-11.22). CONCLUSIONS: Intermittent sun exposure of moderate intensity during childhood/adolescence and outdoor leisural activities, in conjunction with a more resistant skin phenotype to acute sunburns and a strong association with nevi and solar lentigenes was a prominent determinant of melanoma risk in our population.

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Abstract: Chest wall reconstructions can be a complex and challenging procedure and may require a multidisciplinary approach. The most common indications for chest wall reconstruction are repair of defects due to tumor resection, infection, radiation necrosis, congenital deformities or trauma. The repair of complex chest wall defects presents a challenging problem for the reconstructive surgeon. Although the majority of such defects can be repaired with the use of local and regional musculocutaneous flaps, more complicated cases require increasingly sophisticated reconstructive techniques. As defects increase in size, microsurgical techniques are necessary to augment blood flow to pedicled flaps or to provide free flap coverage from distant sites. A better understanding of the respiratory mechanics and local anatomy is crucial in managing these complex defects.

Abstract: Tissue defects in the antihelix and the concha due to oncological resection and trauma can be successfully repaired with a subcutaneously based postauricular island flap. Alternative methods of regional reconstruction usually need two stages or may require grafts in some patients. We present the one-stage technique, as described by Masson, without grafts, to provide adequate reconstruction and aesthetic restoration of the area, illustrated by 62 patients. In all patients there has been a follow-up period of 12 months. This report provides evidence for the aesthetic superiority of this method. An excellent aesthetic outcome was achieved in 46 patients, an adequate outcome in 15 patients, and a poor result in only 1 patient. No flap necrosis was observed. The method has considerable advantages for the repair of anterior conchal and antihelical defects.

Abstract: The hemifrontal flap is an efficient way of reconstructing large unilateral defects that involve the hemiforehead. Six patients had their foreheads reconstructed after the excision of neglected tumours in the frontal region. The flap was designed to provide adequate repair of large defects including the periosteum. The width of the flap above the eyebrow is almost equal to the vertical height of the forehead in the midline. Efficient reconstruction of the primary defect including surface and depth was achieved in all patients. There were no complications with the viability of the flap. The hairline and the line of the eyebrows were preserved in their original sites, with no alteration in the aesthetic landmarks of the forehead in 3 cases, where there was no invasion of tumour.

Abstract: Divided naevus of the upper and lower lids is an infrequent congenital naevus of the human eyelids. It is also known as kissing naevus or split ocular naevus. The histological types of divided naevi may be melanocytic, cellular or compound. Until now about 30 such cases have been reported in the literature. In our clinic, in the last 20 years we have seen seven patients with divided naevus, four female and three male. Three patients, two children of 10 and 11 years and a 77-year-old nun, did not accept any treatment. The remaining four patients, three female and one male, accepted surgical treatment. None of them had received any treatment in the past. Three of these divided naevi were cellular and one was melanocytic. Treatment included a two stage reconstruction in three cases and a single stage procedure in one case. The lower eyelid was usually treated first, and upper eyelid full thickness grafts were mostly used. Lid shortening was common, particularly in the cellular naevi. The results were aesthetically and functionally good. No major complications were observed.

Abstract: BACKGROUND: Copper has been used in wound healing since ancient times, and copper insufficiency seems to impair wound healing. Much less is known about the local effects of copper application on wound healing. METHODS: The authors studied 30 male Sprague-Dawley rats weighing between 250 and 350 g for 5 days. Full-thickness random skin flaps, measuring 1.5 x 7.5 cm, were elevated on the epigastric region. The rats were randomized into two groups according to the local treatment that was used. In group 1 (20 animals), 1 ml of copper ointment was applied on the flap bed after flap suturing. In group 2 (10 animals), 1 ml of placebo ointment (petroleum jelly) was used. Intraperitoneal injection of 30 mg/kg body weight of Hypoxyprobe-1 was performed in each rat on the fourth postoperative day and all animals were killed 3 hours later. Histologic examination of the distal end of the flaps was used for detection of hypoxic tissue changes. Expression, density, and extent of vascular endothelial growth factor activity was detected with the aid of immunocytochemistry. RESULTS: All rats survived throughout the study period. This experimental study showed that local application of copper at the random flap bed may promote wound-related angiogenesis by inducing vascular endothelial growth factor expression. The authors demonstrated statistically significant flap survival after local copper application at the flap bed. CONCLUSION: The marked benefits of copper application in flap survival in this experimental model may guarantee its clinical application in flaps with questionable viability.

Abstract: We describe the application of local anesthesia with intravenous sedation for covering a large defect of the upper back with the use of the island vertical trapezius myocutaneous flap to an 85-year-old male. According to the anesthesiologist, the patient was graded as ASA IV. A 200-ml solution which consisted of 0.75 mg xylocaine, 2 mg adrenaline (1/2,000,000) and 10 mmol/l sodium bicarbonate in a physiologic saline solution was used. The total operating time was about 90 min. During the first postoperative 24 h, the flap pedicle was checked every hour by Doppler. The postoperative course of the patient was uneventful. We believe that in selected cases, when microsurgery is not indicated due to the general medical condition of the patient, the combination of the tumescent technique with sedation and the vertical trapezius myocutaneous flap can be considered a reliable and low-risk procedure.

Abstract: BACKGROUND: p53 has a common polymorphism at amino acid 72, encoding either arginine or proline. p53Arg and p53Pro exhibit differences in various biological activities, such as cell-cycle arrest and induction of apoptosis. Numerous epidemiological studies have examined the role of this polymorphism in several human malignancies, including cutaneous cancers, with contradictory results. OBJECTIVES: To investigate the germline frequency of p53 codon 72 polymorphism in malignant melanoma in a Mediterranean population, and to examine possible associations with various clinicopathological factors. METHODS: In this hospital-based case-control study we used allele-specific polymerase chain reaction for p53 codon 72 genotyping in blood specimens from 107 Greek patients with sporadic cutaneous melanoma and 145 healthy controls. RESULTS: After adjustment for age, sex and phototype the Pro/Pro genotype was associated with increased risk for cutaneous melanoma compared with the Arg/Arg genotype (adjusted odds ratio, OR 3.17, 95% confidence interval, CI 1.03-9.78). This correlation was more pronounced in subjects with phototypes III or IV (adjusted OR 9.56, 95% CI 1.56-58.46), dark skin (adjusted OR 10.96, 95% CI 1.64-73.28), dark eyes (adjusted OR 8.86, 95% CI 1.69-46.52) and dark hair (adjusted OR 3.17, 95% CI 1.01-9.95), and among noncarriers of melanocortin 1 receptor gene (MC1R) red hair polymorphisms (adjusted OR 2.99, 95% CI 1.02-8.78). CONCLUSIONS: p53 codon 72 Pro/Pro genotype could be a risk factor for the development of melanoma in the Greek population, especially in subgroups with darker skin pigmentation, as well as among noncarriers of the MC1R red hair polymorphic variants.

Abstract: Reconstructive surgery of the lips after resection of tumors requires a complete understanding of the anatomy of this region. Most lip cancers remain localized and grow slowly, with a propensity for superficial rather than vertical spread. From January 1983 to December of 2005, 899 patients underwent reconstructive surgery for skin tumors involving the lips. SCCs were the most frequent skin tumors on male patients whereas BCCs were most common on the female patients. The lower lip was the anatomic zone most frequently involved in our series. Preoperative evaluation of the patients was performed in all cases. In the last five-years, we have performed preoperative fine needle aspiration (FNA) biopsy of the tumor. The FNA sample was then examined by ThinPrep technique (Cytolyt; Cytyc, Co, Boxborough, MA, USA). In our series, from the 550 patients who had a five-year follow-up we observed 62 recurrences of the primary tumor. The five-year recurrence rate was 11.28%. The aim of this retrospective study is to report our experience in the treatment of lips tumors with selective combination of treatment modalities.

Abstract: BACKGROUND: In a prospective study, we evaluated the blood perfusion and the blood flow of anterolateral thigh (ALT) flap by both near-infrared spectroscopy and color Doppler ultrasonography. Moreover, we assessed the ability of the perforator flap to reconstruct infected wounds of the lower extremity in 11 patients. METHODS: Near-infrared spectroscopy showed excellent oxygen saturation, and Doppler ultrasonography documented excellent blood flow and decreased vascular resistance in the ALT flap postoperatively. RESULTS: All flaps were successful and all wounds healed uneventfully within 2 weeks without any signs of recurrences or persistent infection. All patients achieved acceptable gait function after rehabilitation. CONCLUSIONS: Apart from the mandatory role of thorough debridement, our results indicate that the ALT flap offers rich blood supply to the recipient area, thus contributing to the sterilization and healing of an infected wound within a short time period.

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Abstract: INTRODUCTION: The sentinel lymph node (SLN) biopsy in melanoma assesses reliably the status of the regional lymph node basins, provides valuable prognostic information, facilitates early therapeutic lymphadenectomy and identifies patients who are candidates for different adjuvant treatments. The current study was designed to evaluate the feasibility of cytological specimens being placed in PreservCyt as a practical collection methodology for performing evaluation of the SLN status in patients with melanomas. PATIENTS AND METHODS: From January 2004 to December 2006, 70 patients with histologically confirmed cutaneous melanoma underwent intraoperative FNA biopsy of the SLN. After identification of the SLN(s), FNA biopsy of the SLN was performed with a 0.6 mm (23 gauge) diameter needle. All the SLNs specimens were examined (using light microscopy 40 x and 200 x) by the same pathologist and cytopathologist, neither of had any knowledge of the medical history of the patient. The histological result of the excised SLN was considered as the final diagnosis. RESULTS: The unsatisfactory rate for TP cytology was 2.17%. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and overall accuracy (OA) for the TP technique were 92.31%, 100%, 100%, 97.06%, and 97.83%, respectively. Using TP cytology, there was greater intensity and distribution of the staining in comparison with immunohistochemistry. DISCUSSION: The accuracy of TP technique in the evaluation of the SLN status is comparable to those of the histological evaluation, and could be of paramount importance for the preoperative planning of treatment.

Abstract: The present study demonstrates a new experimental model to compare the efficacy of end-to-end and end-to-end neurorrhaphy in free muscle flap neurotization. Forty animals were used, divided into four equal groups named A, B, C and D. The peripheral stump of the thoracodorsal nerve was stitched end-to-end in groups A and C and end-to-side in groups B and D to the long thoracic nerve. Free functional muscle transfer was simulated by putting vascular clamps to the thoracodorsal artery (FD SS8R, F: 15 to 20 g) and vein (FD SS6R, F: 10 to 15 g) for 60 minutes and transecting and then restitching the origin and insertion of the latissimus dorsi muscle. Electromyographic and histological studies were performed 150 days following completion of the experiment. The results could indicate the possibility that end-to-side neurorrhaphy might be used in free functional muscle transfer as an alternative to end-to-end neurorrhaphy. We believe that the proposed experimental model is useful for the comparative study between end-to-end and end-to-side neurorrhaphy in free muscle flap neurotization, as these are pure motor nerves and innervate synergistic muscles, are in close approximation, and have similar diameters.

Abstract: BACKGROUND: Several investigators have described a seasonal variation in the diagnosis of cutaneous melanoma. Limited data exist on the seasonality of melanoma diagnosis in Southern European countries. PATIENTS AND METHODS: The seasonal pattern of diagnosis was analyzed in 404 Greek patients diagnosed with cutaneous melanoma (CM) between 1996 and 2004. A summer-to-winter ratio was determined overall and in relation to gender, age, anatomic site, histopathologic type, and tumor thickness. RESULTS: The summer-to-winter ratio was 1.53 for all patients (95% CI [confidence interval]: 1.15-2.02) with a ratio of 1.83 for women (95% CI: 1.20-2.78) and 1.28 for men (95% CI: 0.87-1.88). A seasonal pattern of melanoma diagnosis was observed for patients younger than 50 years of age (1.70, 95% CI: 1.05-2.74) and between 50 and 69 years (1.64, 95% CI: 1.05-2.56), for melanoma located on the upper or lower extremities (2.50, 95% CI: 1.12-5.56 and 2.23, 95% CI: 1.19-4.18, respectively), for superficial spreading and nodular melanomas (1.73, 95% CI: 1.12-2.69 and 1.52 95% CI: 0.96-2.41) and for melanomas with a tumor thickness of 1-2 mm (1.69, 95% CI: 0.91-3.12) and > 4 mm (2.13, 95% CI: 1.04-4.35). CONCLUSIONS: No major differences were seen in the seasonal distribution of CM diagnosis in a Mediterranean population compared to previously reported results. A better ascertainment of the skin during the summer and an increased awareness due to the melanoma screening campaigns are the more likely reasons for the seasonality of melanoma diagnosis in Greece.

Abstract: Advanced technologies and regulatory regimes have contributed to the availability of veterinary vaccines that have high quality and favourable safety profiles in terms of potential risks posed to the target animals, the persons who come into contact with the vaccine, the consumers of food derived from vaccinated animals and the environment. The authorisation process requires that a range of safety studies are provided to evaluate the products. The design and production of vaccines, and their safe use, are primarily assessed by using data gathered from extensive pre-marketing studies performed on target animals and specific quality tests. The current post-marketing safeguards include good manufacturing practices, batch safety testing, inspections and pharmacovigilance. In addition to hazard identification, a full benefit/risk evaluation needs to be undertaken. The outcome of that evaluation will determine options for risk management and affect regulatory decisions on the safety of the vaccine; options might, for example, include special warnings on package inserts and labels.

Abstract: Skin tumors comprise the largest group of malignancies of the head. Despite the accessibility of such lesions, the treatment of neglected, far advanced cancers, many of which have extended deeply into the facial bones and skull, is often required. The key to the cure of malignant tumors of the head is an accurate diagnosis and evaluation of the margins of an excised tumor. Reconstructive surgery of the head after resection of tumors requires a complete understanding of the anatomy of this region. From January 1986 to December of 2005, 31 patients underwent reconstructive surgery for nonmelanoma skin tumors involving the craniofacial region. Preoperative evaluation of the patients was performed in all cases. The results were estimated from the oncologic and functional point of view. The reconstruction, which was performed, included local, regional, and free flaps. In our series, the 5-year disease-free survival rate was 87%. The primary goal of surgical treatment of skin tumors with invasion of craniofacial bone structure is three-dimensional tumor resection with histologically clear margins. This goal has to be balanced, however, with an acceptable functional and aesthetic result. Resections are planned according to pathologic considerations rather than according to the anatomy involved.

Abstract: BACKGROUND: Cyclin D1 plays an important role in regulating the progression of cells through the G1-phase of the cell cycle. The aim of the study was to investigate the expression of cyclin D1 and Ki-67 in squamous cell carcinomas (SCC) and in some premalignant lesions of the penis and to correlate it with clinicopathological parameters and patient survival. MATERIALS AND METHODS: Formalin-fixed paraffin-embedded tissues from 21 SCC, 7 lichen sclerosus, 5 condyloma acuminatum and 2 erythoplasia of Queyrat were stained by immunohistochemistry for cyclin D1 and Ki-67. RESULTS: Cyclin D1-positive nuclear staining was overexpressed in 13/21 SCC (61.9%) and in one case of erythoplasia of Queyrat. Strong reactivity for Ki-67 was found in 16 (76.2%) SCC, in 3 condyloma acuminatum and in one case of erythoplasia of Queyrat. A tendency for an association between cyclin D1 expression and tumour differentiation (p = 0.07) but not the level of tumour invasion (p = 0.50) was found. The Ki-67 expression was notably increased with the advance of tumour grade, but the difference did not reach a statistically significant level (p = 0.46). A slight tendency towards a relationship between Ki-67 and cyclin D1 protein expression was observed (p = 0.32). Two patients relapsed and one died from the disease over a median follow-up period of 4.6 years (range 0.1-10.3 years). CONCLUSION: Ki-67 antibody and cyclin D1 overexpression seem to parallel each other, supporting the concept that cyclin D1 serves as a cell cycle activator. Cyclin D1 overexpression may be used as a prognostic factor of poor outcome in penile carcinoma.

Abstract: BACKGROUND: The role of combination chemotherapy regimens in the management of ovarian cancer patients with tumors previously exposed to platinum compounds and paclitaxel has not yet been defined. The present phase II study evaluated the activity and toxicity of a paclitaxel-ifosfamide-cisplatin combination in the aforementioned group of patients. Given the in vitro and in vivo synergism between these three agents, it was believed that using a three drug combination would overcome tumor resistance to cisplatin. PATIENTS AND METHODS: Thirty-five patients were enrolled in the study. The median age was 55 and the median performance status 1. Thirteen (37%) had potentially platinum sensitive, 12 (35%) had primary platinum-resistant and 10 (28%) patients had secondary platinum-resistant tumors. Treatment consisted of paclitaxel 175 mg/m2 as a 3 h i.v. infusion on day 1, cisplatin 75 mg/m2 i.v. over 2 h fractionated over days 1 and 2, and ifosfamide 5 mg/m2 i.v. over 1 h fractionated on days 1-2 with mesna uroprotection. Courses were administered every 3 weeks on an outpatient basis. Granulocyte-colony stimulation factor (G-CSF) was given at a dose of 5 microg/kg/day on days 4-10. A median of 4 cycles were administered with the delivered dose intensity at 85% of the planned dose for the three agents. RESULTS: Among 35 patients evaluable for response and toxicity, there were 10 partial responses with a response rate of 28.6% (95% confidence interval 12%-45%). Stable disease was recorded in 9 (25.7%) and progressive disease in 16 (45.7%) patients. Subgroup analysis revealed a response rate of 38.5% in potentially platinum-sensitive, 16.5% in primary platinum-resistant and 30% in secondary platinum-resistant tumors. The median response duration was 5 months (range 3-14 months), the median time to progression 6 months (range 3-18 months) and the median survival 12 months (range 3-44 months). Myelotoxicity was significant with neutropenia grade 3 and 4 occurring in 35% and 45% of patients, respectively. Eight episodes (5% of all cycles) of febrile neutropenia were documented and well managed with oral or i.v. antibiotics and G-CSF continuation until complete recovery. Grade 1, 2 and 3 peripheral neuropathy developed in 30%, 30% and 10% of patients, respectively. In conclusion, the three drug combination demonstrated a significant effectiveness in potentially platinum-sensitive tumors and a moderate efficacy in platinum-resistant tumors. The regimen, although myelotoxic, is tolerable with G-CSF support. Further investigation via comparative studies is required to define any superiority of the present regimen over doublets of the three agents in this group of patients.

Abstract: Soft tissue sarcomas (STS) are particularly rare malignancies that constitute less than 1% of all malignancies. In recent years, prognostic clinical factors have been defined that help to stratify patients regarding their risk for local and distant recurrence and death from disease. Tumor grade, size, depth, completeness of resection, and presentation status are among the independent prognostic factors. At present, the treatment of these tumors constitutes a wide or marginal excision, adequate primary reconstruction, and radiotherapy. Surgery has generally been recommended as the primary method of treatment for achieving local control. Modern reconstructive surgery, especially musculocutaneous, either pedicle or free flaps, has made more extensive resections possible, while providing acceptable cosmetic and functional results. This study deals with our experience in the treatment of resectable STS with selective combination of treatment modalities.

Abstract: Individuals with melanocortin 1 receptor (MC1R) gene variants have been shown to carry an increased risk for the development of melanoma. In this study, we investigated the relationship of MC1R gene variants and the risk of melanoma in 123 melanoma patients and 155 control subjects from Greece. The entire MC1R gene was sequenced for polymorphisms and the results were correlated with host factors and pigmentary characteristics. MC1R polymorphisms were present in 59.4% of melanoma patients compared to 37.5% of controls, yielding an odds ratio (OR) of 2.43 (95% confidence interval (CI) = 1.50-3.96, P < 0.001) for melanoma among MC1R carriers. The risk of melanoma was enhanced in individuals carrying multiple variant alleles (OR = 6.97; 95% CI = 1.86-26.12, P = 0.004). Only the Val60Leu, Arg142His, and Arg151Cys variants were significantly associated with melanoma risk. In stratified analysis, the risk of melanoma among MC1R carriers was not influenced by skin phototype, skin color, or hair color. No association was found between MC1R genotype and the age of onset of melanoma, the tumor location, or the tumor thickness. In conclusion, MC1R polymorphisms are a predisposing factor of melanoma in a southern European population with a relatively low incidence of the disease.

Abstract: BACKGROUND: Immunotherapy for advanced melanoma induces serologic and clinical manifestations of autoimmunity. We assessed the prognostic significance of autoimmunity in patients with stage IIB, IIC, or III melanoma who were treated with high-dose adjuvant interferon alfa-2b. METHODS: We enrolled 200 patients in a substudy of a larger, ongoing randomized trial. Blood was obtained before the initiation of intravenous interferon therapy, after 1 month of therapy, and at 3, 6, 9, and 12 months. Serum was tested for antithyroid, antinuclear, anti-DNA, and anticardiolipin autoantibodies, and patients were examined for vitiligo. RESULTS: The median duration of follow-up was 45.6 months. Relapse occurred in 115 patients, and 82 patients died. The median relapse-free survival was 28.0 months, and the median overall survival was 58.7 months. Autoantibodies and clinical manifestations of autoimmunity were detected in 52 patients (26 percent). The median relapse-free survival was 16.0 months among patients without autoimmunity (108 of 148 had a relapse) and was not reached among patients with autoimmunity (7 of 52 had a relapse). The median survival was 37.6 months among patients without autoimmunity (80 of 148 died) and was not reached among patients with autoimmunity (2 of 52 died). In univariate and multivariate regression analyses, autoimmunity was an independent prognostic marker for improved relapse-free survival and overall survival (P<0.001). CONCLUSIONS: The appearance of autoantibodies or clinical manifestations of autoimmunity during treatment with interferon alfa-2b is associated with statistically significant improvements in relapse-free survival and overall survival in patients with melanoma.

Abstract: INTRODUCTION: The objective of the current study was to analyze the potential value of immunocytochemical analysis on ThinPrep (TP)-processed smears, from fine needle aspiration (FNA) biopsies, of breast tumors for the determination of ER and PR content as compared with the immunohistochemical analysis performed on paraffin-embedded breast tumor specimens. PATIENTS AND METHODS: Percutaneous FNA biopsy of focal breast lesions in 119 female adult patients during a 31-month period was performed. Subsequently, these patients underwent surgical resection of the tumors. ER and PR status of the tumors was determined by immunocytochemical analysis on TP-processed smears and by immunohistochemical studies in paraffin-embedded sections. RESULTS: With the use of TP technique adequate material was observed in all cases. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and overall accuracy (OA) of the TP technique for the ER were 98.95%, 100%, 100%, 95.84% and 99.15% respectively. In addition, sensitivity, specificity, PPV, NPV and OA of the TP technique for the PR were 100%, 87.5%, 95.60%, 100% and 96.64%, respectively. CONCLUSIONS: ER and PR status can be evaluated in FNA material from breast carcinomas by using the TP technique. Sample collection and storage is simple and permits the assortment of the FNA sample for both morphologic diagnosis and ancillary studies. The accuracy of TP technique in the detection of ER and PR content is comparable to those of the histological evaluation, and could be of paramount importance for the preoperative planning of treatment.

Abstract: In order to evaluate the variability of encephalomyocarditis virus (EMCV), field isolates originating from different European regions and inducing different clinical pictures in pigs have been molecularly characterised. The regions targeted were the poly(C) tract, a part of the 5'-UTR (360 nucleotides), the Leader gene (201 nucleotides), the complete capsid coding region (2502 nucleotides), the 2A gene (403 nucleotides), the end of the 3D polymerase gene (305 nucleotides) and the 3'-UTR (123 nucleotides). Analyses have also been performed on a virulent field isolate, which had been subjected to serial passages in vivo and in vitro resulting, in the case of the in vitro passaged virus, in attenuation, as demonstrated by animal experiments. The present study shows that different clinical pictures, such as acute fatal myocarditis or reproductive failure, may not only be caused by EMCV isolates which are genetically diverse but also by the same isolate. Thus no correlation could be demonstrated between genotype and clinical disease. However, the European isolate which showed the highest genetic divergence also gave rise to a more complex clinical picture. Despite EMCV having been isolated from cases of acute fatal myocarditis in pigs in certain areas of the world for many years, clinical disease, including a variety of clinical pictures and pathogenicity, has only been recognised in Europe since 1986 and thus it can be considered an emerging disease in this region. These findings, associated with the reported phenotype changes of the virus under environmental changes (passages), along with its wide distribution among vertebrate species (including higher primates), shows the validity of considering EMCV as a potential pathogen for recipients in xenotransplantation.

Abstract: BACKGROUND: The extensor apparatus vascularization is less studied compared with flexor tendons. The aim of the present anatomical work was to study it in a more systematic way and find clinical applications. METHODS: The study was carried on 40 fingers from 10 fresh cadaver hands. Two catheters (16 gauge) were introduced into the radial and ulnar arteries on the volar side of the wrist. Acetone was used for arterial cleaning and color latex for the arterial system coloration. The authors proceeded with the dissections, always in the same manner: they first made the longitudinal dorsal incisions from the metacarpophalangeal to distal interphalangeal joints. Raising the skin, they identified the thin subcutaneous branches into the hypodermal plane; they followed their course until their origin, which allowed identification of the arterial branches of the extensor apparatus. For measurements, they used as a point of reference the emergence distance of those dorsal branches from the metacarpophalangeal and proximal and distal interphalangeal joints. RESULTS: The extensor apparatus vascularization was segmental, realized through the paratenon from the branches of a vascular cutano-tendino-osseous complex through a common trunk, issued from the propria digital artery. This complex nourished not only the extensor apparatus but also the dorsal digital skin and periosteum. Its location was consistent at the level of both proximal and middle phalanges. CONCLUSIONS: This consistency explains the anatomical basis for the realization of composite-free tissue transfer in cases of banked fingers. The osteotomies could be performed at well-defined distances from the joints.

Abstract: The blood flow and the blood perfusion of pedicled anterolateral thigh (ALT) flap was evaluated in 11 patients with skin and soft tissue defects by means of color Doppler ultrasonography and of near-infrared spectroscopy, respectively. Also, the authors assessed the ability of the pedicled perforator flap to cover the above defects. A proximally based flap was used to cover defects at the lower abdominal wall (two patients), the suprapubic area, the penis (two patients), the greater trochanter (two patients), the lateral gluteal area, and the ischial tuberosity. A distally based flap was used to cover the exposed knee joint (two patients). Near-infrared spectroscopy revealed excellent oxygen saturation and Doppler ultrasonography documented increased blood flow and decreased vascular resistance in the pedicled ALT flap, postoperatively. All flaps survived completely and the wounds healed uneventfully, resulting in excellent esthetic and functional results. The vascular anatomy of ALT flap facilitates the design of versatile pedicled flaps with tremendous vascularity, two pivot points and large arc of rotation, able to cover defects from the lower abdominal wall to the knee joint.

Abstract: Neglected skin carcinomas are not very often in our days. It is known that skin is the most common site of malignancy Squamous cell carcinoma (SCC) is the second most common malignancy of the skin. We report a combined surgical treatment in an unusual case of a squamous cell carcinoma (T4 n0 m0) localized on scalp, presented primarily as a frontal ulcerative bloody mass, with no palpable lymph nodes and a whole body negative computed tomography. Three months after the first surgical procedure, the tumour reappeared presenting left frontal lobe invasion. A combined surgical management (neurosurgery and plastic surgery) was realized, in order to remove the tumour and to cover the defect. Three months later the tumour recrudesced invading the left antrum, infiltration of remained parotid gland and an ipsilateral mandible osteolysis, besides a 2 cm size distal metastasis at the right upper lobe of the lung was revealed. This article, through case report and a brief review of the literature, offers criteria for the surgical treatment of SCC that arise on the scalp.

Abstract: BACKGROUND: There is now increasing evidence that a constitutive expression of cyclooxygenase (COX)-2 plays a role in the development and progression of malignant epithelial tumors. Expression of COX-2 is seen in 93% of melanomas, as determined by immunohistochemistry. Temozolomide (TMZ) has demonstrated activity against melanoma and has been investigated as single agent or in combination. We designed a phase II study to assess the efficacy and toxicity of the combination of TMZ and celecoxib (a COX-2 inhibitor) in patients with advanced melanoma. PATIENTS AND METHODS: From January 2003 to July 2004, 52 patients were enrolled in the study. Nineteen patients were M1a, six M1b and 27 M1c. Patients received TMZ 200 mg/m(2) per day p.o. for 5 consecutive days every 4 weeks and celecoxib 400 mg b.i.d. p.o. for a maximum of six cycles. Celecoxib was continued until progression. RESULTS: The median age was 63 years. There were 29 males and 23 females. Among 50 assessable patients, there were 11 (21.5%) objective responses including five complete responses and six partial responses. Twenty patients (38.5%) had stabilization of their disease, and 19 (36.5%) progressed. The median time to progression was 4.6 months and the median survival 9.5 months. Twenty-two patients (41.5%) completed all cycles of treatment. Median relative dose intensity of TMZ was 0.99 (range 0.6-1.2). Most commonly seen toxic effects included anemia (27.5%), neutropenia (17.5%), thrombocytopenia (33%), nausea/vomiting (75%), gastrointestinal (52%) and fatigue (46.5%). One patient discontinued due to severe toxicity. COX-2 was determined by immunohistochemistry and was expressed in all cases. CONCLUSION: The combination of TMZ and celecoxib is safe and potentially effective in the treatment of metastatic melanoma. Randomized studies are needed to explore the role of celecoxib in combination with chemotherapy or as maintenance treatment in these patients.

Abstract: Small ruminant lentivirus (SRLV) infections are widespread in Greece, but SRLVs have never been isolated and characterized. In this study, we present the sequence of a 574-nucleotide (191-amino acid) region of the gag gene of SRLV strains from four sheep and one goat from a single geographic area of Greece. All five sequences appeared to be closely related at both nucleotide (2.1-14.2% variation) and deduced amino acid (1.6-4.2% variation) level. Greek SRLV strains were closer to ovine prototypic strains (average divergence 16.8%) than to the caprine strain CAEV-Co (21% divergence). By amino acid composition, the Greek SRLVs were on the average more than twice as distant from CAEV-Co as from other ovine strains. Phylogenetic analysis suggested that Greek strains segregate into a unique group, separate from, but related to, other ovine prototype sequences.

Abstract: BACKGROUND: Sunlight precipitates a series of genetic events that lead to the development of skin cancers such as basal cell carcinoma (BCC) and squamous cell carcinoma (SCC). The p53 tumour suppressor gene, which plays a pivotal role in cell division and apoptosis, is frequently found mutated in sunlight-induced skin tumours. OBJECTIVE: To investigate the immunoreactivity of the p53 gene in non-melanoma skin cancers and to correlate its expression with apoptotic and cell proliferation markers. METHODS: We analysed 35 non-melanoma tumours including 19 BCCs and 16 SCCs from sun-exposed skin areas. p53 protein expression was studied immunohistochemically using the DO7 monoclonal antibody against wild-type and mutant p53 forms. The percentage of p53-immunopositive nuclei was measured by image analysis. Cell proliferation and apoptosis were also assessed by image analysis following Ki-67 immunostaining and application of the TUNEL method on paraffin sections, respectively. RESULTS: The percentage of p53-expressing cells varied from 3.5 to 90 in BCCs (median value 54.4%) and from 3.7 to 94 in SCCs (median value 40.3%). The mean value of Ki-67-positive cells was comparable in both groups of tumours with a mean value of 40.6% in BCCs and 34.6% in SCCs. Conversely, the TUNEL assay showed sporadic staining of apoptotic cells within the tumours with a mean value of 1.12% in BCCs and 1.8% in SCCs. p53 protein expression was correlated positively with cell proliferation (r = 0.75, P = 0.000001) and negatively with apoptosis (r = -0.23, P = 0.05). CONCLUSION: p53 immunoreactivity was high in the majority of the skin carcinomas examined and correlated positively with cell proliferation and negatively with apoptosis. The p53 protein overexpression appears to be related to an inactivated protein resulting from mutations of the p53 gene or other unclear molecular mechanisms.

Abstract: The sequence of the genome segment 10 (Seg-10) encoding NS3/NS3A was determined for 19 field isolates of Bluetongue virus (BTV) of serotypes BTV-1, BTV-4, BTV-9 and BTV-16, derived from epizootics in Greece in the years 1979 and 1998-2001. The aim of the study was to define the molecular epidemiology of the virus in this part of the Mediterranean basin. On the basis of the Seg-10 sequences, the isolates grouped into two distinct phylogenetic clusters. These were Greek group I of solely serotype BTV-4 viruses, and Greek group II of serotypes BTV-1, BTV-9 and BTV-16 viruses. The isolates in Greek group I clustered with the Corsican and Tunisian BTV-2 serotypes and US group II strains of BTV-10 and BTV-13 serotypes, while those in Greek group II with Chinese, Indian and Australian viruses of different serotypes suggesting that viruses derived from two distinct ecosystems have caused BT incursions in Greece over the last 25 years. The NS3/NS3A sequences of most of the BTV-4 isolates were identical, irrespective of the year of isolation, geographical location and host species or tissue origin. Maximum of 15-16% nucleic acid sequence variation, but only 4% deduced amino acid substitution, were observed between groups I and II. Furthermore, the clustering of the NS3/NS3A sequences was independent of the viral serotype, indicating the occurrence of genome segment reassortment during the course of evolution of the viruses.

Abstract: Repair of any defect in the eyelid depends on its size and position and the state of the surrounding tissues. Basal cell carcinoma (BCC) is the most common malignant tumour of the eyelids, and squamous cell carcinoma (SCC), mixed carcinomas or basosquamous cell carcinomas (BSC), and cutaneous melanomas (CM), also invade the eyelids and periocular zones. Reconstruction of the eyelids and associated orbital structures after resection requires a complete understanding of the anatomy. The adequacy of the reconstruction is judged by the quality of functional restoration and the aesthetic appearance. The purpose of this study was to document various, simple or complex reconstructive procedures that may be used after excision of malignant tumours of the eyelids and to assess the outcome of surgical treatment.

Abstract: From 1986 to 2001, 17 patients (aged 26-77 years) were treated using the vertical trapezius musculocutaneous flap. A two-stage procedure was used in 7 and a single-stage island flap in 10. The donor site was closed directly in all patients. Mean length of hospital stay was 16 days (range 12-25). There was no operative mortality. Complications were one partial flap necrosis and two seromas of the donor site, complicated by infection. With a minimum follow-up of more than two years, our study confirms the usefulness of the vertical trapezius musculocutaneous flap in head and neck reconstructive surgery. It is a reliable, thin flap of uniform thickness, which carries hairless skin. The length and thickness of its pedicle allows excellent mobility. The main disadvantage of the flap is the complete sacrifice of the muscle necessary for total mobilisation of the flap, and the intraoperative repositioning of the patient.

Abstract: Many decades have passed since the first kidney transplantation, which is now the most common organ transplant performed worldwide. Despite the impressive advances, some patients may develop posttransplant complications that require proper management and treatment. The plastic and reconstructive surgeon, among others, may be called on to help resolve a number of reconstructive problems present in the immunosuppressed kidney recipients. This study presents our experience with 41 kidney recipients who needed plastic surgical treatment. Patients were placed into one of three study groups according to the type of posttransplant surgical condition. Group 1 included 17 patients with posttraumatic wound healing problems; group 2, 17 patients with skin tumors; and group 3, 7 patients with other posttransplant surgical complications. Only two of these patients had early posttransplant wound dehiscence; the remaining patients suffered late complications. In conclusion, the kidney recipient can successfully undergo minor or major reconstructive procedures. The possibility of surgical problems arising during the early posttransplant period presents increased complication rates, possibly due to high immunosuppressive drug levels.

Abstract: Two experiments were performed to compare the severity of encephalomyocarditis virus (EMCV) infection in pigs. The pigs were challenged with the Greek myocardial strain, at different ages and with different doses. In the first experiment, nineteen susceptible pigs, 40 days old, were divided into three groups and were experimentally infected with 10(6) TCID(50), 10(4) TCID(50) or 10(2) TCID(50) of the Greek EMCV strain. In the second experiment, 10 susceptible pigs, of either 20 or 105 days, were divided into two groups according to age and were experimentally infected with 10(6) TCID(50) of the Greek EMCV strain. In addition, five piglets, each one the same age as its experimental group, were used as uninfected controls. No clinical signs were observed after infection, except a transient temperature rise in some pigs. Another important observation was the difference in mortality between groups. The survival rate of the 40-day-old pigs was inversely related to the viral dose. In these pigs, a positive association between the viral dose and the severity of macroscopical and histopathological lesions of the heart was also evident. Viral isolations from various organs of the challenged 40-day-old pigs increased with the increasing dose level. When challenged with 10(6) TCID(50) of EMCV, there was no difference in the fatality rate of the 20- and 40-day-old pigs, but none of the 105-day-old pigs died. The severity of the macroscopical and the histopathological heart lesions was inversely related to the age of the pigs. Furthermore, viral isolations from the various organs were higher in 20- and 40-day-old pigs than in the older ones. In 40-day-old pigs, neutralizing antibodies linearly increased as the dose increased. These antibodies were consistently lower in 20-day-old pigs. Viraemia, and nasal and faecal excretions were detected in all groups and lasted 1-3 days, except for the 105-day-old pigs whose symptoms lasted for an additional day.

Abstract: Due to the probable role played by rodents as a reservoir for the transmission of the EMC virus to pigs, the experiment reported here was performed in order to assess the transmission rate of EMCV within a rat population. Twenty-five eight-week-old Wistar rats housed in individual plastic cages were experimentally infected either with a Greek myocardial EMCV strain (5 rats with a 0.2 x 10(6) TCID50 dose per rat and 10 rats with a 0.5 x 10(4.5) TCID50 dose per rat, oronasally) or a Belgian myocardial EMCV strain (10 rats with a 0.5 x 10(4.5) TCID50 dose per rat, oronasally). Two to five days later, each inoculated rat was moved to a new clean cage and coupled with a contact rat to compare the pathogenicity of the two strains and to estimate the basic reproduction ratio R0, indicating the level of EMCV transmission. During the experiments, faecal virus excretion was measured as well as the serological response against EMCV. After euthanasia, virus isolation was attempted from different rat tissues. Neither strains produced mortality, nor clinical signs and only low titres of neutralising antibodies were found. All contact rats, however, were infected and the virus was isolated from their faeces and from various tissues. Both 10-pair experiments revealed a point estimate for the R0 of infinity (95%-CI for both the Greek and Belgian EMCV strains = 4.48 - infinity), as did the 5-pair experiment with a higher dose of the Greek strain (95%-CI = 1.83 - infinity). Combining the results from the two 10-pair experiments resulted in an estimate for R0 of infinity (95%-CI: 9.87 - infinity). These results indicate that the EMC virus can spread very easily within a rat population by horizontal rat-to-rat transmission (R0 >> 1).

Abstract: A total of 216 local crossbred sheep from 16 scrapie-affected Greek flocks and 210 purebred sheep of the milk breeds Chios and Karagouniko from healthy flocks were analysed for scrapie-linked polymorphisms in the prion protein (PrP) gene. Of the 216 sheep in this case-control study, 96 sheep were clinical cases, 25 subclinical cases (asymptomatic at the moment of euthanasia but positive by histopathology and/or ELISA detecting proteinase-resistant PrP) and 95 healthy controls (negative by all evaluations). Polymorphisms at codons 136, 154 and 171 were determined by denaturing gradient gel electrophoresis, followed by RFLP and sequencing. Scrapie, both clinical and subclinical, was associated with the genotypes ARQ/ARQ (88 of 110 sheep of that genotype), ARQ/TRQ (9 of 13), ARQ/AHQ (15 of 38) and VRQ/VRQ (9 of 17). Histopathological lesions were more severe in the clinical cases. Genotypes ARQ/ARR (26 sheep), ARQ/ARK (seven sheep), AHQ/ARR (one sheep), ARH/ARH (one sheep) and ARR/ARH (three sheep) were detected exclusively in healthy control sheep. In the purebred survey, four genotypes were present in the Chios sheep (ARQ/ARQ, ARQ/TRQ, ARQ/AHQ and ARQ/ARR) and four in the Karagouniko sheep (ARQ/ARQ, ARQ/AHQ, ARQ/ARR and ARQ/ARH).

Abstract: OBJECTIVE: To investigate the efficacy and toxicity of gemcitabine administration followed by the combination of fluorouracil (5-FU) modulated by folinic acid in patients with advanced, symptomatic pancreatic cancer. The main objective was to estimate tumour response and any improvement in patients' quality of life. PATIENTS: The study included 48 evaluable patients with metastatic disease with no prior chemotherapy. The study duration was 3 years. INTERVENTIONS: Gemcitabine 1000 mg/m(2) intravenously was given on days 1 and 8 followed by fluorouracil 350 mg/m(2) intravenously as a bolus biologically modulated by folinic acid 350 mg/m(2) intravenously given on days 1, 2, 8 and 9 in order to develop the conditions for any potential drug synergism. The regimen was administered every 3 weeks for 1 year or until disease progression. RESULTS: Objective partial responses were documented in ten (21%) patients (95% CI 10.5, 35). Twenty-two (46%) patients had stable disease while 16 (33%) patients had progressive disease. The median response duration was 8 months (range 4-20). The median time to progression was 6 months (range 2-24), while the median survival of the group was 7 months (range 3-36) and the probability of surviving beyond 12 months was 20%. Of the 44 patients with tumour-related symptoms who were considered evaluable for clinical-benefit response, 28 (70%) patients had pain improvement, 25 (52%) patients had improvement of their performance status, and nine (28%) patients experienced weight gain during treatment. Serum concentrations of cancer antigen (Ca-19-9) were decreased by more than 50% in 14 (37%) of the 38 assessable patients. Chemotherapy was well tolerated, with mild myelotoxicity. Gastrointestinal toxicity was moderate with mild mucositis. CONCLUSION: The regimen of gemcitabine and fluorouracil administered in this study was well tolerated and showed a moderate antitumour activity and a significant palliative effect on tumour-related symptoms. Because fluorouracil is a low toxicity combination agent for gemcitabine, other forms of the two-drug combination warrant further investigation.

Abstract: Sequence analyses of the non-structural protein gene NS3/NS3A of eight Greek bluetongue (BT) virus (BTV) field isolates from the 1979 and 1999-2001 epizootics provide preliminary molecular data on the epidemiology of BT in Greece. These isolates from infected sheep belonged to serotypes BTV-1, BTV-4, BTV-9 and BTV-16. Phylogenetic analysis of the NS3/NS3A gene segregated these Greek isolates of BTV into two monophyletic groups. The first group was formed by all isolates of BTV-4; all were identical in their sequences, regardless of the area and year of isolation in Greece, and clustered with strains from Tunisia and Corsica. The isolates of BTV-1, BTV-9 and BTV-16 segregated into a second monophyletic group and clustered with Asian strains, showing a high homology (97-99%). From an epidemiological point of view, these preliminary results infer that one group of isolates is Mediterranean, whilst the second appears to be of Asian origin.

Abstract: The problem of availability of immunological products in minor species and minor indications/uses is complex and should be well defined. Data for species are available, but not for minor uses. "Case-by-case" decisions should be avoided, as far as possible. The following categories can be recognized: (a) a vaccine has not been authorized anywhere; (b) a vaccine is authorized for major species/indications only; (c) an authorized vaccine exists in another Member State and (d) a vaccine has been withdrawn. Old, well established products, with limited data for an updated dossier could be the problem in categories "c" and "d". Proposed measures and incentives include: "cascade" (not recommended), human orphan-like drug policy, provisional authorization, acceptance of bibliographical data (if justified), procedural and financial assistance. Standards of quality, safety and efficacy cannot be minimized, but specific tests can be considered.

Abstract: The purpose of this study was the comparison of the diagnostic sensitivity between buffy coat (BC), peripheral blood (PB), lymph node (LN), bone marrow (BM) and short-term culture (P-D) cytology that has been based on the detection of Ehrlichia canis morulae, in the acute phase of canine monocytic ehrlichiosis (CME). Their cellular localization, total numbers and microscopic differentials were also investigated. The highest sensitivities were achieved after evaluating 1000 oil immersion fields (OIFs) in BC (66%) and an equal number in LN (60.9%) smears, separately or together (74%). The morulae were more often detected into lymphocytes than monocytes. The highest total number of morulae (n=143) were found in P-D smears. Finally, to avoid false positive diagnoses, platelets, lymphocytic azurophilic granules, lymphoglandular bodies and phagocytosed nuclear material should not be confused with the morulae.

Abstract: Six piglets aged 20 days were inoculated oranasally with 5 ml of a suspension (10(6) TCID(50)/ml) of a Greek myocardial strain of encephalomyocarditis virus (EMCV). The animals either died (n=2) or were killed for examination on days 1,2 or 3 post-inoculation (pi). EMCV was isolated from virtually all organs examined (heart, tonsils, palatine glands, pancreas, spleen, small intestine and mesenteric lymph nodes). Histopathologically, interstitial myocarditis, necrosis of cardiac muscle cells and Purkinje fibres, and necrotizing tonsillitis were detected in all inoculated piglets. Focal interstitial pancreatitis, necrosis of pancreatic acinar cells and Langerhans islet cells, and necrosis of germinal centre lymphocytes of the lymph nodes and Peyer's patches were detected in two piglets that died or were killed on day 3 pi. Immunohistochemically, viral antigen was detected in epithelial cells of all organs examined, including the tonsils, palatine glands, pancreatic interlobular ducts and small intestine. This suggests that EMCV is epitheliotropic, in addition to its known myocardial tropism. The frequent presence of intracytoplasmic EMCV in macrophages of the tonsils and spleen supports the hypothesis that macrophages play a role in viral replication and dissemination in the body.

Abstract: This phase II trial studied the efficacy and toxicity of full dose paclitaxel plus vinorelbine, as salvage chemotherapy in patients with metastatic breast cancer resistant to anthracyclines. Patients received vinorelbine (30 mg/m2) followed 1 hour later by full dose paclitaxel (175 mg/m2) every 3 weeks for a maximum of 8 cycles or until disease progression. Because of the heavy pretreatment of the patients, prophylactic granulocyte-colony stimulating factor (5 microg/kg) was administered daily for 5-10 days. To minimize potentially cumulative neurotoxicity due to both agents, amifostine was given prior to chemotherapy. Thirty-four patients: 8 with tumors primary resistant and 26 with tumors recurring within 3-6 months after anthracycline treatment, were evaluable for efficacy and toxicity. Objective responses occurred in 11 patients [32%; 95% confidence interval (CI): 16.3-47.7%), all partial responses. Responses were observed in lung and liver. The median response duration was 4 months (range 3-7), median time to progression was 5 months (range 3-9) and median overall survival was 8 months (range 4-24). Neutropenia was dose limiting (35% grade 3-4 toxicity). The left ventricular ejection fraction, measured and followed in 18 patients, fell less than 20% below baseline level in 9 patients (50%), but only one patient developed congestive cardiac failure. The paclitaxel-vinorelbine regimen was moderately tolerated and moderately effective in poor prognosis breast cancer patients with visceral metastases and tumors resistant to anthracyclines. The combination at these doses and schedules should be considered in the design of regimens for advanced breast cancer.

Abstract: The purpose of this study was to review our experience with the management of patients with complicated cardiothoracic problems by the use of pedicled myocutaneous or muscle flaps, and discuss the various methods of reconstruction. Over the last 11 years, we have treated 54 patients with complicated cardiothoracic problems by reconstruction with pedicled myocutaneous or muscle flaps. The underlying causes were chest wall tumours (n = 13), radionecrosis of the chest wall (n = 12), deep or chronic sternal infections (n = 25), and bronchopleural fistulas (n = 4). The most commonly used muscles for reconstruction were pectoralis major and the rectus abdominis. Our results compare favourably with those reported elsewhere. We conclude that although the use of pedicled myocutaneous or muscle flaps is not free of complications, it is an effective and reliable method for the management of patients with complicated cardiothoracic problems.

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Abstract: A total of 51 goats, including seven clinical cases, from the first herd in Greece reported to have scrapie was examined to discern an association between scrapie susceptibility and polymorphisms of the gene encoding the prion protein (PrP). Each animal was evaluated for clinical signs of the disease, histopathological lesions associated with scrapie, the presence of detectable protease-resistant PrP in the brain and PrP genotype. Eleven different PrP genotypes encoding at least five unique predicted mature PrP amino acid sequences were found. These genotypes included the amino acid polymorphisms at codons 143 (H-->R) and 240 (S-->P) and 'silent' nucleotide alterations at codons 42 (a-->g) and 138 (c-->t). Additionally, novel caprine amino acid polymorphisms were detected at codons 21 (V-->A), 23 (L-->P), 49 (G-->S), 154 (R-->H), 168 (P-->Q) and 220 (Q-->H) and new silent mutations were found at codons 107 (g-->a) and 207 (g-->a). The following variants were found in scrapie-affected goats: VV(21), LL(23), GG(49,) SS(49), HH(143), HR(143), RR(154), PP(168), PP(240), SP(240) and SS(240). All scrapie-affected animals carried the HH(143)RR(154) genotype, with the exception of two goats (HR(143)), both of which had detectable protease-resistant PrP but showed no clinical signs or histopathological lesions characteristic of scrapie.

Abstract: The Greek chlamydial strains FAS, FAG, VPG and LLG, isolated from aborted sheep or goat foetuses, had been previously characterized as divergent on the basis of mouse cross-protection experiments, with LLG and its homologous POS significantly different from the rest in inclusion morphology, polypeptide profiles and reactivity with monoclonal antibodies. To determine the genetic basis of their divergence the 16S-23S ribosomal intergenic spacer was analysed by RFLP analysis of PCR 16SF2/23R amplicons. Using the restriction enzymes BfaI, SfcI, HpaI, BclI, DdeI and AclI, the strains were classified as Chlamydophila abortus. However, digestion with RsaI made it possible to differentiate strains FAS, FAG and VPG from strains LLG and POS, generating DNA fragments of 530/55 and 585bp, respectively. By subsequent sequence analysis of the 23S domain I rRNA gene only strain FAS was identical to reference strain A22 of C. abortus. Strains FAG and VPG presented an identical nucleotide deviation at position 593 of signature sequences. Strains LLG and POS presented three identical nucleotide deviations at positions 156, 186 and 307. Variation within the domain I signature sequences for the examined abortion strains was < or =0.69%. In conclusion, substantial genetic and biological diversity among strains of C. abortus was demonstrated, suggesting that subspecies variation status for certain strains may be applicable. Our findings suggest that differentiation may be possible at a subspecies level by RFLP analysis.

Abstract: A total of 45 non-uremic dogs, with clinical signs indicating leishmaniosis, entered the study. Diagnosis was confirmed by indirect immunofluorescence assay (IFA) on serum and polymerase chain reaction (PCR) on bone marrow samples. The dogs were randomly allocated into Group A (n=37) that received allopurinol (10mg/kg B.W., per os, twice daily) for 4 consecutive months, and Group B (n=8) that were placebo-treated. Clinical signs were scored just before and at monthly intervals throughout the study period, in a blinded and independent fashion. Complete blood count, serum biochemistry profile, urinalysis, lymph node and bone marrow parasitology, IFA and enzyme-linked immunosorbent assay (ELISA) serology and bone marrow PCR were carried out at the beginning and at the end of the trial. A total of three Group A and one Group B dogs died of end stage kidney disease that developed during the trial. In Group A animals that endured the trial there was a significant improvement in the general body condition, conjunctivitis, peripheral lymphadenopathy, splenomegaly, masticatory muscle atrophy, ulcerative stomatitis, epistaxis, exfoliative dermatitis, cutaneous ulcerations, blepharitis and nasodigital hyperkeratosis. The same observation was made for anemia, lymphopenia, hyperproteinemia, hyperglobulinemia, hyperphosphatemia, increased alkaline phosphatase activity and the low albumin/globulin ratio. By contrast, no improvement of any kind was seen in Group B dogs. Lymph node and bone marrow parasite numbers were significantly decreased in Group A animals. In Group B, that occurred only in the lymph nodes. Apart from remission of clinical signs and restoration to normal of clinicopathological abnormalities, allopurinol did not eliminate Leishmania organisms, as the PCR result on bone marrow was still positive in all the dogs that finished the trial.

Abstract: Two groups of pregnant ewes were experimentally infected oronasally in midpregnancy. A faecal and an abortion-source isolate of Chlamydophila abortus were used. They were derived from a healthy ewe from a flock with no history of abortion, and from an aborted foetus in a farm with enzootic abortion. As assessed by modified Ziehl-Neelsen (MZN) staining, egg culture, antigen ELISA, the Clearview test and immunohistochemistry, inoculation resulted in placental and/or foetal infection in all ewes. Histopathology revealed placentitis in two and four ewes inoculated with the enteric or abortion-source isolate, respectively, in addition, these samples were immunohistochemically positive for chlamydial antigen. All six ewes infected with the enteric isolate and five of seven ewes infected with the abortion-source isolate showed evidence for a serological response by an indirect ELISA or CFT. Neither chlamydiae nor lesions were detected in the placentae and lambs of the uninfected control ewes, which remained seronegative. Our results suggest that enteric C. abortus can be associated with placental and foetal lesions in sheep.

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Abstract: A duplex reverse transcription polymerase chain reaction (RT-PCR) assay for the detection of bluetongue virus (BTV) in clinical samples was developed. This assay, which detects the highly conserved S10 region of BTV, was assessed for sensitivity and application as a rapid and dependable diagnostic tool by comparison with standard assays of virus detection, such as virus isolation in embryonated chicken eggs and cell culture. Simultaneous detection of BTV and host beta-actin RNAs minimizes the possibility of false negative results. The sensitivity of the assay was found to be equal to five cell culture infectious dose (CCID(50)) units and its specificity was confirmed as no RT-PCR product was detected with RNAs from two closely related orbiviruses, i.e. epizootic haemorrhagic disease virus (serotypes 1, 2 and 318) and African horse sickness virus, serotype 9, or RNAs from uninfected BHK-21 cells and blood samples from uninfected sheep or goats. In this study, 36 blood samples from naturally infected mixed flocks of sheep and goats were examined. Seventeen animals were identified as BTV-positive by RT-PCR, whereas only 13 were found positive by virus isolation in embryonated chicken eggs and nine by cell culture assays. These results indicate that the duplex RT-PCR could be a useful technique for monitoring BTV infection in the field.

Abstract: OBJECTIVE: To evaluate the ultrastructural changes and localization of encephalomyocarditis virus (EMCV) and viral pathogenesis in the myocardium of experimentally infected piglets. ANIMALS: Eight 20-day-old piglets. PROCEDURE: Six piglets were inoculated oronasally with 5 ml (10(6) median tissue culture infective dose/ml) of EMCV suspension, and 2 were used as uninfected controls. Piglets were euthanatized or died between postinoculation days 1 and 3. Samples of heart tissue from all piglets were evaluated histologically, by virus isolation, and by use of immunohistochemistry and electron microscopy. RESULTS: All infected piglets had gross or microscopic lesions of interstitial myocarditis. immunohistochemically, EMCV antigen was detected in the cytoplasm of cardiac muscle cells, Purkinje fibers, and endothelial cells and in the nucleus of cardiac muscle cells and Purkinje fibers. Ultrastructural lesions were characterized by degeneration and necrosis of cardiac muscle cells and Purkinje fibers. Virus was present intracytoplasmically in cardiac muscle cells, Purkinje fibers, and endothelial cells of capillaries and intranuclearly in cardiac muscle cells. The cell membranes of the Purkinje fibers and endothelial cells had distinct protrusions that contained virus particles. In control piglets, no lesions were found, and no EMCV antigen was detected. CONCLUSIONS: Localization of EMCV intracytoplasmically or intranuclearly in various myocardial cells may well reflect the sites of viral proliferation. The presence of virus particles in cell membrane protrusions and in vacuoles within the lumen of capillaries indicates that virus is released not only by disintegration of the host cell but also via exocytosis.

Abstract: The development of a multiplex polymerase chain reaction (PCR) method with amplification of capripoxvirus in a single-step procedure from skin biopsies using three primer pairs, two specific for capripoxvirus and one specific for alpha-tubulin is described. A sensitive multiplex PCR was achieved by optimization of parameters such as the primer concentrations, magnesium and dNTPs concentrations. False negative results that sometimes arise due to inhibitors of DNA amplification may be avoided by the inclusion in the assay of alpha-tubulin primers. The results reported on 42 skin biopsies from sheep suspected to have poxvirus infection, indicated that the assay could monitor simultaneously DNA extraction from skin biopsy samples and allow improved detection of capripoxvirus within 24 h of specimen receipt in the laboratory.

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Abstract: Seven 40-day-old piglets were inoculated with encephalomyocarditis virus (EMCV) strain 424/90, isolated from an outbreak of the myocardial form of the disease in Greece. Two non-infected animals were used as controls. Of the seven inoculated piglets, five died suddenly on day 1.5, 2 (two piglets), 2.5 or 4 post-inoculation (p.i. ). The remaining two and the control piglets were killed on day 8 p. i. EMCV antigen was detected immunohistochemically in endothelial cells of capillaries from 1.5 to 2.5 days p.i. only, but was found in cardiac muscle cells, Purkinje fibres and macrophages on all occasions up to day 8 p.i. In endothelial cells and macrophages, EMCV antigen was detected intracytoplasmically, but in cardiac muscle cells and Purkinje fibres it was observed intracytoplasmically or intranuclearly, or both. The frequent presence of EMCV antigen in Purkinje fibres suggests an explanation for the sudden death of the piglets.

Abstract: Bovine spongiform encephalopathy (BSE) is a prion-associated disease where the infectious agent is thought to be a host-encoded protein with a protease-resistant conformation (PrP(Sc)). Here, data are presented on the solubilization of purified murine BSE material, using guanidine-HCl as a denaturing agent. This treatment led to loss of infectivity, which was partially recovered on renaturation after dialysis to remove the chaotropic agent. The renatured product was then fractionated on an isopycnic sucrose-density gradient and the fractions were analysed for the presence of PrP(Sc), nucleic acids and infectivity. It was found that the major part of PrP(Sc) (>90%) and the endogenous nucleic acids did not contribute towards the formation of infectious particles on renaturation. Infectivity was distributed in the top three, low-density fractions. Among these, the presence of considerable infectivity in the fraction of lowest density, with barely detectable PrP(Sc), is of particular interest.

Abstract: Basal cell carcinoma of the skin is the most common neoplasia in humans. Previous studies have shown the existence of allelic imbalance (loss of heterozygosity and microsatellite instability) in BCC on several human chromosomes. Chromosome region 9p21-p22 harbors the CDKN2a/p16INK4a, p19ARF, and p15INK4b tumor suppressor genes. To determine the contribution of these genes to the development of basal cell carcinomas we looked for evidence of allelic imbalance in 67 sporadic basal cell carcinoma specimens from Greek patients and screened 28 of them presenting loss of heterozygosity at 9p21-p22 for germline mutations in p16INK4a and p19ARF genes. Chromosome regions 17q21 and 17p13 were also screened for allelic imbalance in all the 67 basal cell carcinoma specimens. Overall, 69% (46 of 67) of the specimens displayed loss of heterozygosity in at least one microsatellite marker, whereas only six of the 67 (9%) exhibited microsatellite instability. For the 9p21-p22 locus the overall frequency of loss of heterozygosity reached 55% (37 of 67) and is the highest reported. The overall frequency of loss of heterozygosity for the 17q21 locus is 34% (22 of 64) and for the 17p13 locus is 11% (seven of 65). Two of the 28 loss of heterozygosity positive cases were heterozygous for a previously described polymorphism, Ala148Thr, in exon 2 of the CDKN2a gene. This is the first demonstration of polymorphism in the CDKN2a gene in human basal cell carcinomas. No sequence variation in exon 1beta of the p19ARF gene was found. Our results provide evidence of a significantly high occurrence of loss of heterozygosity for the 9p21-p22 locus; however, lack of p16INK4a/p19ARF mutation suggests that these genes seem not to be implicated by mutational inactivation in the development of basal cell carcinoma. Other(s), yet unidentified, tumor suppressor gene(s) located in this locus may be related to this specific type of skin cancer.

Abstract: The duration of viraemia and the serological responses were studied in two breeds of sheep and two breeds of goats, experimentally infected with bluetongue (BT) virus serotype 4. Viraemia, detectable by cell culture and embryonated chicken egg inoculation, lasted from the third to sixth day until the 27th-54th day post infection (p.i.). Significant differences between sheep and goats were not recorded. Lesbos sheep and goats together appeared to have significantly longer viraemias (n = 9, mean 41.3 days) than east-Friesian sheep and Saanen goats (n = 10, mean 30.4 days, p = 0.0039). Serological response was studied by competitive ELISA (c-ELISA) and agar gel immunodiffusion (AGID) tests. The c-ELISA was more sensitive in detecting BT virus antibodies in all animals than the AGID tests. No significant differences were observed between sheep and goats or between breeds. The epidemiological significance of subclinical infection and the extended BT virus viraemias in Lesbos sheep and goats, in relation to the maintenance of the virus and to overwintering is discussed.

Abstract: A simple, rapid and specific diagnostic polymerase chain reaction (PCR) method was developed for sheep poxvirus identification. The primers used were from the sequenced genomes of the capripox viruses KS-1 and InS-1. Six different sheep pox isolates were tested against two orf (parapox) and three animal herpesviruses as controls. Material from uninfected cell cultures was also used as control. The sensitivity of the PCR was approximately equivalent with each of the two primers and for the six sheep pox isolates. All the negative control virus DNAs were negative and differed clearly from those of the sheep pox strains.

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Abstract: Thirteen susceptible piglets, aged 40 days, were divided into two groups and were experimentally infected either with a Greek (myocardial) or a Belgian (reproductive) encephalomyocarditis virus (EMCV) strain (total dose 5 x 10(6) TCID50, intramuscularly and intranasally). Six piglets were placed in the same rooms, 24 h later, as contact controls. The following criteria were studied: ante mortem: clinical signs, serum cardiac isoenzyme activities (CK-MB and LD-1), viraemia, nasal and faecal virus excretion and serological response. Post mortem (after death or euthanasia): gross lesions, virus isolation from tissues, RT-PCR, as well as histopathological and immunohistochemical findings. The Greek strain was more pathogenic, producing mortality, with high cardiac isoenzyme activities and pronounced macroscopic myocardium lesions. The Belgian strain was able to induce mild heart lesions, as detected only by cardiac isoenzyme activity and histopathologically. All contact pigs were infected, within the first 1-2 days of their introduction, that coincided with the period of viral excretion by the experimentally infected pigs (up to the 3rd day post infection). Disease was mild, with no mortality.

Abstract: In an effort to use antineoplastic drug combinations which are active in platinum resistant ovarian cancer or which can induce a second response after a platinum first-line treatment, we conducted a study on 30 ovarian cancer patients previously treated with carboplatin plus cyclophosphamide who were given ifosfamide 5 g/m2 i.v. divided over days 1 to 3 plus mesma combined with cisplatin 100 mg/m2 i.v. divided over days 1 to 3 every 4 weeks as second-line treatment. Eight patients had never entered remission with first-line chemotherapy while 22 patients had tumor recurrence within 6 to 18 months after the end of chemotherapy and their tumors were considered potentially platinum sensitive. Responding patients received 6 courses while palliative treatment for nonresponders was provided. Of the 22 patients with tumor recurrence, 8 patients responded with one partial response (PR) and 7 complete clinical responses (CCR). Two out of the 8 patients with platinum resistant disease demonstrated short lasting PR. Seven patients with CCR underwent second-look operation and in two a pathological CR was documented. Median time to progression was 6 mo (4-12). The median overall survival was 12 mo (4-20). Myelotoxicity despite G-CSF administration was significant with grade 4 leukopenia in 40% and grade 3 thrombocytopenia in 20% of patients. Central nervous system (CNS) toxicity was significant with 30% somnolence, 20% disorientation and an episode of grand-mal epilepsy ascribed to ifosfamide. With a 33% response rate the combination is as effective as new agents employed in relapsed ovarian cancer. Platinum-refractory disease may respond to a lesser degree. The most important determinant of response was the progression-free interval from first-line chemotherapy. Whether patients recurring after carboplatin plus cyclophosphamide have a greater chance to respond to cisplatin plus ifosfamide or vice-versa cannot be supported by the current data and therefore randomized studies should be performed to this end.

Abstract: Six piglets that had survived experimental infection with encephalomyocarditis virus (EMCV) were treated with dexamethasone for a period of 5 days. The virus had not been detected in excretions of putative carriers for a period of 13-20 days before the treatment. All piglets showed a rise in cardiac isoenzyme (CK-MB) activity, from the first day of treatment, suggesting myocardial damage. Antibody titres against EMCV remained stable or slightly decreased during treatment. EMCV was isolated from blood, nasal and faecal samples from all piglets on days 2 and 3 after initiation of treatment and from various tissues of three piglets. Four contact piglets, that were housed together with the dexamethasone-treated piglets, became infected, indicating that EMCV was shed by treated piglets. It is suggested that recovered pigs may play an important role in the dissemination of EMCV.

Abstract: Increasing evidence suggests that the pathological alterations observed in brains affected by neurodegenerative disorders such as Creutzfeldt-Jakob disease and Alzheimer's disease also involve changes in glycosaminoglycans (GAGs). In the present study, we have isolated, purified, and characterized total GAGs from brain stems of healthy cows or those infected with the bovine spongiform encephalopathy (BSE) agent and we report on the differences between the two groups. Purification of the GAGs was achieved by gel filtration after homogenization, delipidation, and sequential treatment with pronase, DNase, and alkali borohydride. Fractionation of the total GAGs by Superose 6 gel filtration and HPLC revealed four major fractions, with average molecular masses of 360, 180, 15, and 2.3 kDa, respectively, both in controls and infected tissues. Enzymatic characterization, using GAG-degrading enzymes, showed that in both infected and normal brain stems, the 360- and 180-kDa fractions correspond to hyaluronic acid, which was also the most abundant GAG, while the 15-kDa fractions correspond to chondroitin sulfates as well as heparan sulfate and dermatan sulfate, the latter being the least prominent GAG. Electrophoresis on cellulose acetate membranes revealed that the relative ratio of GAGs was not significantly modified in infected brain stems, compared to controls. However, total GAGs in infected brain stems was significantly decreased by approximately 40%, compared to controls, and this decrease applied equally to all of the above GAG fractions. The diminution observed in total GAGs in infected brain stems is in good agreement with the recently reported neuroprotective role of certain GAG molecules and offers an additional criterion for differential diagnosis of BSE-infected animals.

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Abstract: The pathogenicity of two porcine encephalomyocarditis virus (EMCV) isolates for sows in gestation and young piglets was studied. One virus originated from a case of reproductive failure in pigs in Belgium and the other from a case of acute myocarditis in pigs in Greece. Sows in the mid-gestation period and one- to two-month old piglets were inoculated with each isolate. The molecular relationship between both isolates was studied by determining the nucleotide sequence located across the junction of the 1C and 1D capsid-coding genes. Antigenic analysis was performed using a panel of 35 monoclonal antibodies raised against an Italian field isolate of EMCV. All three approaches revealed differences between both isolates and also confirmed that there was no link between the two outbreaks of disease.

Abstract: INTRODUCTION: The authors report three cases of "Hyalinosis cutis et mucosae"; the first and the second case concern two siblings children (brother and sister) while the third is reported on a 42-year-old woman. PRESENTATION OF CASES: The case history and the personal anamnesis, the morphology of the skin lesions, the symptoms indicating laryngeal mucous affection as well as the histopathological picture, are compatible with the "Urbach-Wiethe disease". Furthermore, in the third case, the electron microscopic findings confirm the diagnosis. DISCUSSION: The particularities of the present cases are discussed in connection with the corresponding bibliographical data.

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Abstract: Twenty eight C. psittaci abortion strains had been previously classified in to 4 immunologically distinct groups on the basis of cross-protection experiments in a mouse model. To identify the molecular basis of their immunological divergence 4 representative strains were investigated by cellular, molecular and immunological techniques. An identical pattern was obtained by Alul digestion of the amplified major outer membrane protein gene (MOMP) by the polymerase chain reaction (PCR) of the 4 strains. However, inclusion morphology and polypeptide profiles clearly distinguished one strain, named LLG, and its homologous strain POS from the other prototypes by the presence of a unique protein at 26.5 kDa and the absence of a polypeptide at 23 kDa. Six out of 10 monoclonal antibodies (mAbs) raised against abortion strains failed to react with inclusions of the 2 strains. All 6 mAbs reacted with the chlamydial outer membrane complex (COMC). Two of these mAbs, one against the MOMP and one against an antigen at 90 kDa, did not react with immunoblots of LLG and POS. The data provide direct demonstration of the existence of strain variation in the field and classify strains LLG and POS as a distinct C. psittaci serotype 1-subtype. The antigenic diversity among abortion strains should be taken into consideration when designing a subunit vaccine.

Abstract: Trauma is the main cause of lower limb soft tissue defects followed by tumor resection, vascular disease and osteomyelitis. 17 patients were operated on using microsurgical techniques during the last 10 years. 9 of the patients had traumatic defects, 5 had various malignant tumors, 2 patients had arterial insufficiency and finally 1 patient had chronic osteomyelitis. The latissimus dorsi either as muscle flap with a split thickness skin graft or as a musculocutaneous flap was used the most frequently in our series. Other flaps, including the radial forearm neurovascular flap, the rectus abdominis flap, the scapular flap, the groin and the dorsalis pedis flaps were used less often. Delayed primary cover was used, except in 1 case with an opened knee joint trauma, where emergency repair was performed. Complications included: 2 partial skin graft necroses which required revision and 2 flap necroses caused by venous thrombosis. With follow-up more than 4 years in all but 2 patients, functional results were found satisfactory. Cosmetic results, however, were less satisfactory.

Abstract: Partial or total lower lid reconstruction implies the repair of three layers: the skin, the tarsus, and the conjunctiva. The method of reconstruction must, therefore, aim at three objectives: ensuring the stability of the lid by repairing the tarsoconjunctival layer, achieving an aesthetic result with the repair of the skin, and taking into consideration the primary role of the upper lid in the ocular protective system. We recommend a musculocutaneous flap, taken from the upper lid, with a single lateral or medial pedicle combined with a chondromucosal alar graft. During the last 10 years, 41 patients were operated on with malignant skin tumors requiring marginal, partial, and total lower lid reconstruction. The results were quite satisfactory from a functional and aesthetic point of view. We noticed two cases of partial flap necrosis and two cases of ectropion needing revision. We believe that the procedure described can be a tool in the armamentarium of the plastic surgeon involved in the field of the orbitopalpebral area.

Abstract: Reconstructive surgery of the head and neck has particularly benefited from the rapid developments in microsurgery. Resection of tumours from the head and neck which are associated with serious physical and psychological problems, or which often require multiple reconstructive procedures, can now be performed as a single-stage procedure with good oncological and functional results. The present series reports on the treatment of 12 primary or recurrent malignant tumours of the head and neck using the latissimus dorsi musculocutaneous free flap. Complications consisted of two venous thromboses which required revision of the anastomoses. The oncological and functional results were good, while the aesthetic outcome was fair. We conclude that the latissimus dorsi muscle or musculocutaneous free flap is the method of choice to repair wide defects of the head and neck following the resection of malignant tumours.

Abstract: Different procedures have been described for facial and oral reconstruction after malignant tumor removal, such as skin grafts and local and distal staged flaps. Myocutaneous island or free flaps are the method of choice for these reconstructions. Our experience concerns the use of the platysma myocutaneous flap in 12 patients after intraoral and extraoral malignant tumor resection. Six patients had squamous cell carcinomas, i.e., 2 malignant melanomas of the skin and 4 squamous cell carcinomas of the oral mucosa. The complete tumor resection was controlled by frozen-section biopsy in all patients, but for 7 of them a functional or radical neck dissection was performed. We suggest including in the flap the external jugular vein. The results were evaluated from an oncological, functional, and aesthetic point of view. We noticed 2 recurrences in a minimum follow-up of 4 years. The functional and aesthetic results were quite satisfactory and no serious complications were noticed in our series. However, the primary disadvantage of this interesting flap is that the dominant vasculature derived from the submental branch of the facial artery is seldom visualized. In spite of this, it should be regarded as very useful for limited intraoral or surface coverage problems in an arc extending 10 cm from the level of the facial artery at the mandible. This arc includes the cheek, the lower lip, the chin, the floor of the mouth, and the mastoid area.

Abstract: The specialty which has benefited the most from the rapid development of reconstructive microsurgery over the last decade, has been plastic surgery. In particular, the authors refer to reconstructive microsurgery after malignant tumor resection. Resections which used to cause serious psychological problems or which required multiple reconstructive procedures, can now be performed as a single stage procedure with good oncological, functional and aesthetic results. This experience concerns the treatment of 14 primary or recurrent malignant tumors of the skin and soft tissues using microsurgical techniques. The free latissimus dorsi musculocutaneous flap was the flap most commonly used. The authors also used the groin flap, the scapular flap, the tensor fasciae latae flap, the dorsalis pedis flap and finally the jejunum combined with the second metatarsus to reconstruct the mandible and the floor of the mouth. Complications consisted of complete necrosis of a dorsalis pedis flap and two revisions were performed for venous thrombosis. With a minimum follow-up of 5 years, one patient has developed a local recurrence and another has developed regional lymph node metastases. The functional and aesthetic results were quite satisfactory. The authors believe that microsurgery is a very valuable technique to treat difficult reconstructive problems.

Abstract: The influence of dexamethasone and cyclophosphamide on the goat immune system was investigated. Seven goats, with a previous contact with caprine herpesvirus type 1 (CHV-1), were used. All had been vaccinated with live Mycobacterium paratuberculosis vaccine. Six goats were injected intravenously (i.v.) with dexamethasone daily for 5 days (2.5-4 mg/kg BW per day). Three also received 25 mg/kg BW of cyclophosphamide on day 0. The seventh goat was not treated. Dexamethasone alone caused depression, slight lymphopenia and fall in tuberculin reaction. Dexamethasone plus cyclophosphamide caused a severe clinical reaction, marked leukopenia (lymphopenia and polymorphopenia), fall in tuberculin reaction and significant increase in CHV-1 neutralizing antibody titres. M. paratuberculosis antibody reaction was variable and thus difficult to be assessed. CHV-1 was not isolated.

Abstract: The pathogenicity of caprine herpesvirus type-1 (CHV-1) in goat kids and lambs was studied. Two experiments were carried out. In the first, two Saanen goat kids and four lambs of a local breed were infected intravenously with the Swiss strain E/CH of the virus. Clinical reaction was severe in the kids, but it was very mild in the lambs. Virus was excreted from the kids in higher titres and longer than in lambs. Virus was also isolated from tissue specimens but only from a kid that died on post inoculation day 4 (PID 4). The gross- and histopathological lesions were more severe in kids. In the second experiment only lambs were used. They were dexamethasone treated and then virus inoculated. A very mild infection developed. The lambs did not shed the virus, neither the virus was isolated from their tissues collected at necropsy. Nevertheless histopathological lesions were seen. In both experiments the animals seroconverted on PID 10.

Abstract: Plastic surgery of the peri-orbital area after tumor resection is a special and interesting field of reconstructive surgery. Our experience concerns the surgical treatment of 299 malignant tumors of the skin of the peri-orbital area in 291 patients. Basal cell carcinomas were the most common but we also had 8 squamous cell and 4 mixed carcinomas and 1 malignant melanoma. Among these tumors, 277 were primary and 22 were recurrences: 5 after X-Ray therapy, 9 after surgery elsewhere, 4 after electrocautery and 4 after combined treatments. The lower eyelids and the inner canthus areas were the most frequently involved in our series with relative infrequency in the outer canthus and upper eyelids. We used direct closure and local flaps in the repair of most of our patients, but we also used some full thickness skin grafts by preference for the upper eyelid. Complications consisted of 8 cases of ectropion and 2 flap necroses for which a revision was performed in some cases. Concerning our carcinological results among the 123 patients with one year minimum follow up we noticed 4 recurrences which represents a percentage of 3.2%.

Abstract: The two-stage surgical treatment of an adult female patient who presented with a congenital divided nevus of the left upper and lower lid is described.

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Abstract: An experiment was designed for a preliminary investigation of the caprine herpesvirus BHV-6 latency. The experiment was carried out in two steps. In the first, the virus inoculated six kids and the control, showed a mild infection and excreted the virus for 4-12 days. The second part started about one month after the cease of all symptoms and the virus isolations of the first part. No immunosuppression and therefore no virus reactivation could be achieved after eight daily administered injections of dexamethasone (0.15-0.50 mg/kg BW), as it was judged by the hematological, virological, pathological and serological findings.

Abstract: Previous preliminary studies had shown that caprine herpesvirus (BHV-6) infections exist in many countries where goats play an economical role. The extensive serum survey made in Greece reveals that the virus must be widespread because more than 50% of the goats have antibodies. The bucks were found to have a higher evidence of infections than the female goats. The kids possessing maternal antibodies became seronegative at the age of 4 months and new antibodies appeared at the age of 7-8 months. Observations in 2 closed goat herds showed that the virus does not spread during the mating and lactating period. Neutralization titres increased or new infections in seronegative animals occurred after the summer when breeding took place. The virus is present in a latent state and recurrent infections are assumed to occur. In spite of that all our efforts to isolate the virus were unsuccessful, even after experimental immunosuppression.

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Abstract: The demand for the reversal of vasectomy in the treatment of sterility has been increasing over the past decade. Microsurgery is acknowledged to be the best non-traumatic procedure for this reconstruction. This article reviews 115 patients treated by microvasovasostomy over a period of six years. Evaluation of the results shows 76% of patients with a positive seminal analysis, and 52% informed the authors that their partners were pregnant. In addition to the technical aspects (i.e. the non-traumatic manipulation of tissues using microsurgical procedures) the titer of antisperm bodies in the patient's serum and seminal plasma seems to play some part in the recovery of fertility.

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Abstract: We herewith present a review of 138 basal-cell carcinomas, mostly on the face, that were treated by surgical excision and immediate repair. In a few cases, frozen sections were taken to ensure complete excision of the neoplasm. Local flaps were favored; distant flaps and full-thickness skin grafts were required in selected case. The recurrence rate with a minimum of 3 years of follow-up is 2.1%. Cosmetic results, evaluated by the patients themselves and by one of the authors (G.D.) who did not participate in the surgery, were judged "excellent," that is, without visible scars or deformities, in 61.4% of the cases, and "good" in 31.4%. The results are discussed.

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Abstract: The surgical treatment of basal cell epitheliomas of the nose is based on wide excision and histological control with preferably an extemporaneous examination. The decision to operate should be made by a group consisting of the radiotherapist, the surgeon, and the dermatologist. Repair is usually carried out immediately, once it is certain that ablation is complete. There are no true contra-indications due to the age of the patient and treatment is less costly than irradiation. Surgery is essential for reconstruction of the nasal pyramid destroyed partially or completely by the tumor and consists of a reconstructive rhinoplasty using a frontal flap. Lesions that are of moderate dimensions but are too large to be treated by electrocoagulation, especially if they are close to the nasal orifices, are a good indication for surgery. A graft, especially a total skin graft taken from behind the ear or from the subclavicular region, is usually used. However, local flaps using the excess skin at the root of the nose (glabellar flap) or from the cheek (nasogenial flap) give excellent results. The authors describe the results obtained in a series of 93 patients in whom relapses were observed in only 7.6%. The only 3 cases in which the recurrence was not able to be treated, all three patients eventually dying, were cases in which the tumors had been operated upon for recurrences after radiotherapy.

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