Margin rotation for tarsal buckling following ptosis surgery.

PURPOSE: To assess the clinical characteristics of tarsal buckling after ptosis correction and its management with margin rotation techniques. METHODS: Multicenter retrospective review of ten patients who developed upper eyelid entropion following ptosis correction. In all cases the tarsal deformity was corrected with margin rotational procedures with either a lid crease anterior approach or a traditional posterior approach. Data collection included patient demographics, type of ptosis surgery, and photographic documentation of the affected eyelids. RESULTS: Entropion occurred after a variety of different ptosis surgery techniques, including frontalis sling, levator advancement and supramaximal levator resection. A horizontal tarsal fold was detected in all eyelids, being in the upper third of the tarsus in 70% and in the central tarsus in 20% of the cases. Tarsal buckling was corrected in all cases with rotational surgery, with nine cases being operated through an anterior lid crease approach and 1, through the traditional posterior approach. The most reported complication was minimal residual ptosis. CONCLUSION: Tarsal buckling following ptosis surgery is associated with folds located in the upper part of the tarsus. Margin rotation techniques are effective in restoring the natural position of the eyelid margin in these cases.

[Neurological side effects of checkpoint inhibitors]. / Neurologische Nebenwirkungen von Checkpoint-Inhibitoren.

BACKGROUND: In recent years the treatment of many tumor entities has been revolutionized by the use of modern immunotherapies with checkpoint inhibitors; however, good response rates are contrasted by many immune-mediated side effects. Neurological immune-mediated side effects are rare but often severe complications of checkpoint inhibitor treatment. METHOD: A systematic search in the PubMed and Web of Sciences databases was carried out for case reports and studies on neurological side effects during checkpoint inhibitor treatment. RESULTS: A total of 42 articles on neurological side effects of checkpoint inhibitors with a total of 85 reported cases could be identified. The most frequently reported neurological side effects were myopathies, neuropathies, diseases of the neuromuscular endplates and encephalitides. Among those, encephalitides and myopathies with accompanying myocarditis were associated with the highest morbidity and mortality. CONCLUSION: Against the background of a rapidly increasing use of checkpoint inhibitors, this article provides an overview of currently available reports on the clinical courses of neurological side effects. Controlled studies on the treatment of neurological side effects are lacking. From case studies it can be assumed that early steroid treatment increases the probability of a complete remission of neurological symptoms. Typical symptom constellations must therefore be rapidly recognized and an immunosuppressive treatment must be initiated.

Mucoadhesive assessment of different antifungal nanoformulations.

Oral candidiasis is an important opportunistic fungal infection and polyenes and azoles are still the most used antifungal agents. However, the oral absorption resulting from most available treatments is generally poor and, consequently, a very high frequency of administrations of antifungal agents is strongly required. Therefore, the major challenge is to improve the retention of the antifungal agents in buccal mucosa, and the encapsulation into mucoadhesive systems may be considered as a possible strategy to achieve this objective. Three types of mucoadhesive polymeric nanoparticles (polylactic acid (PLA), polylactic-co-glycolic acid (PLGA) and alginate) were prepared using nystatin as model drug. The drug-loaded nanoparticles were then included in toothpaste, oral gel and oral films, respectively. The results demonstrated that the loaded nanoparticles were successfully produced, presenting a mean size between 300-900 nm and with a negative surface charge. Also, the determination of the encapsulation efficiency of all nanoparticles showed values above 70%. In terms of the in vitro mucoadhesion, the best formulation was the oral film loaded with the PLGA nanoparticles followed by the oral gel with PLA nanoparticles and thirdly the toothpaste with alginate nanoparticles. This was confirmed in an in vitro rinsing model with mucus producing HT29-MTX cells, where the percentage of nystatin retained to the cells after 40 min of simulated saliva flow was between 10-27% when formulations were used and only 4% for free nystatin. Further studies will include in vivo testing using animal models.

Evaluation of Lung Function in Liver Transplant Candidates.

INTRODUCTION: A wide variety of pulmonary conditions are found in cirrhotic patients and may compromise the pleura, diaphragm, parenchyma, and pulmonary vasculature, influencing the results of liver transplantation. OBJECTIVE: To evaluate the pulmonary function (lung capacities, volumes, and gasometric study) of patients with liver cirrhosis awaiting liver transplantation. PATIENTS AND METHODS: Cirrhotic patients, subdivided into 3 groups stratified by liver disease severity using the Child-Pugh-Turcotte score, were compared with a control group of healthy volunteers. In spirometry, the parameters evaluated were total lung capacity, forced volume in the first second, and the relationship between forced volume in the first minute and forced vital capacity. Blood gas analysis was performed. In the control group, arterial oxygenation was evaluated by peripheral oxygen saturation by pulse oximetry. RESULTS: Of the 55 patients (75% men, 51 ± 12.77 years), 11 were Child A (73% men, 52 ± 14.01 years), 23 were Child B (75% men, 51 ± 12.77 years), and 21 were Child C (95% men, 50 ± 12.09 years). The control group had 20 individuals (50% men, 47 ± 8.15 years). Pulmonary capacities and volumes by the parameters evaluated were within the normal range. Arterial blood gas analysis detected no hypoxemia, but a tendency to low partial gas pressure was noted. CONCLUSION: In this population of cirrhotic patients the parameters of spirometry were normal in relation to the lung capacities and volumes in the different groups. No hypoxemia was detected, but a tendency to hypocapnia in the blood gas was noted.

POSEIDON study: a pilot, safety and feasibility trial of high-dose omega3 fatty acids and high-dose cholecalciferol supplementation in type 1 diabetes.

The anti-inflammatory and immunomodulatory properties of high-dose omega-3 fatty acids and Vitamin D, and the initial encouraging results from case reports on the use of this supplementation in new-onset Type 1 Diabetes (T1D), support further testing of this combination strategy. This intervention appears to be well tolerated, affordable, and sufficiently safe to be further tested in randomized prospective trials to determine whether this combination therapy may be of assistance to halt progression of autoimmunity and/or preserve residual beta-cell function in subjects with new onset and established T1D of up to 10 years duration. In addition, the 1st PreDiRe T1D conference (Preventing Disease and its Recurrence in Type 1 Diabetes - see Editorial in this issue) was organized to discuss initial results and possible alternative/complementary strategies, for collaborative international expansion of these trials, to include strategies for disease prevention. Our POSEIDON clinical trial will test the use of high dose vitamin D3 and highly purified Omega-3 fatty acids in new onset and established T1D. The draft of the study protocol, in addition to the informed consent and assent, is now shared open access to facilitate its international implementation by interested physicians and centers that would like to further test this approach through clinical trials.

S100A7: from mechanism to cancer therapy.

Within the tumor, malignant and stromal cells support each other by secreting a wide variety of growth factors and cytokines, allowing tumor growth and disease progression. The identification and regulation of those key factors in this crosstalk has opened the opportunity to develop new therapeutic strategies that not only act on the tumor cells but also on the stroma. Among these factors, S100A7 protein has gained interest in the last years. With key roles in cell motility its expression correlates with increased tumor growth, angiogenesis and metastatic potential. This work aims to deepen in the role played by extracellular S100A7 in the tumor microenvironment, offering a new integrative insight of its mechanism of action on each cellular compartment (tumor, endothelial, immune and fibroblast). As a result, we demonstrate its implication in cell migration and invasion, and its important contribution to the formation of a proinflammatory and proangiogenic environment that favors tumor progression and metastasis. Furthermore, we define its possible role in the pre-metastatic niche formation. Considering the relevance of S100A7 in cancer progression, we have developed neutralizing monoclonal antibodies, reporting for the first time the proof of principle of this promising therapeutic strategy for cancer treatment.

Coal-Bed Methane Water: Effects on Soil Properties and Camelina Productivity.

Every year, the production of coal-bed natural gas in the Powder River Basin results in the discharge of large amounts of coal-bed methane water (CBMW) in Wyoming; however, no sustainable disposal methods for CBMW are currently available. A greenhouse study was conducted to evaluate the potential to use CBMW as a source of irrigation water for camelina [ (L.) Crantz]. We assessed the effects of three CBMW concentrations (0% [1:0], 50% [1:1], and 100% [0:1] tap water to CBMW) on selected soil properties, growth, seed oil, and fatty acid composition of three camelina cultivars: Blaine Creek, Ligena, and Pronghorn. The 100% CBMW reduced seed and estimated biofuel yields by 24 and 23%, respectively, but increased the oil content by 3%, relative to the control. Additionally, the 100% CBMW visibly affected soil through formation of surface crust due to elevated levels of sodium (653 mg Na kg). The 50% CBMW had no significant effects on the seed yield, estimated biofuel yield, and oil content, but the soil Na levels were still high (464 mg kg), which could pose a long-term impact on soil structure. The CBMW tended to reduce the total saturated fatty acid, but it had no significant effects on the total monounsaturated or polyunsaturated fatty acids of camelina seeds. Overall, CBMW diluted with an equal amount of good-quality water could be used to irrigate camelina in the short term. Afterward, only good-quality water would have to be used until the accumulated dissolved solids are flushed out.

S100P antibody-mediated therapy as a new promising strategy for the treatment of pancreatic cancer.

Despite progresses in diagnosis and treatment, pancreatic cancer continues to have the worst prognosis of all solid malignant tumors. Recent evidences suggest that the metastasis-promoting protein S100P stimulates pancreatic tumor proliferation, survival, invasion and metastasis progression through extracellular functions. Moreover, its expression is strongly correlated with poor prognosis in patients with several types of cancer although the entire molecular mechanism responsible for the diverse biological functions is not fully understood. We showed that extracellular S100P stimulates pancreatic carcinoma BxPC3 cell line by promoting cell proliferation. We also demonstrated that S100P induces, in this cell line, the phosphorylation of IκBα and the secretion of matrix metalloproteinase 9 (MMP-9). In addition, treatment with S100P protected cells from injuries induced by the cytotoxic agent Gemcitabine. On the basis of these results, we developed function-blocking anti-S100P monoclonal antibodies (mAbs) that abolished all of its in vitro activities. Furthermore, in vivo treatment with the candidate 2H8 antibody decreased tumor growth and liver metastasis formation in a subcutaneous and orthotopic BxPC3 tumor model. We conclude here that a therapeutic strategy blocking the extracellular activity of S100P by means of specific mAbs could be an attractive therapeutic approach as a single agent or in combination with target-directed or chemotherapeutic drugs to treat pancreatic cancer.

Sarcoma with true epithelial differentiation secondary to irradiated glioblastoma.

Glioblastoma multiforme rarely shows true, immunohistochemically confirmed, epithelial differentiation. Furthermore, radiotherapy may induce cerebral sarcomatous tumors, and postsurgery glioblastoma irradiation may give rise to secondary gliosarcomas. We report a case of a 48-year-old male operated on a primary glioblastoma, followed by radiotherapy. A local recurrence occurred 23 months later that was operated too, and a second diagnosis of a fibrosarcoma with true epithelial differentiation was made. Primary systemic neoplasms were largely excluded. The patient died shortly after, and postmortem showed another cerebral dural-attached mass corresponding to a sarcoma without epithelial differentiation, and leptomeningeal seeding composed of malignant epithelial elements only. Cytogenetics, however, disclosed the second tumor to be similar to the primary one.

Diagnóstico coproparasitológico de Fasciola hepática en ganado bovino en una empresa pecuaria cubana / Coproparasitological diagnosis of Fasciola Hepatica in cattle in a cuban livestock farm

Con el objetivo de comparar el método coproparasitológico de sedimentación de Benedek con el kit inmunoenzimático FasciDIG para el diagnóstico de Fasciola hepática en ganado bovino de una empresa pecuaria cubana, se realizó un estudio descriptivo transversal entre febrero y marzo de 2008. Se colectó muestras de heces a 254 bovinos Jersey, Holstein y mestizos. Se encontró 16 (6.3%) animales positivos mediante el FasciDIG y 10 (3.9%) por la técnica de sedimentación de Benedek. Se concluye que el método inmunoenzimático (FasciDIG) demostró mayor detectabilidad diagnóstica que el método de sedimentación.

The aim of the study was to compare the coproparasitological method of Benedek sedimentation against the immunoenzimeassay for the diagnosis of Fasciola hepaticain cattle from a Cuban livestock large farm. A descriptive cross-sectional study was conducted in February-March 2008 and faecal samples were collected to 254 Jersey, Holstein, and crossbreed animals. Sixteen positive cases (6.3%) were detected using FasciDIG whereas only 10 (3.9%) using the Benedek sedimentation method. It was concluded that the immunoassay method (FasciDIG) showed better diagnostic sensitivity than the Benedek sedimentation test.

Acceleration of the Increase of Endogenous Thyrotropin Hormone for Follow up Studies or Radioiodine Treatment in Patients with Differentiated Thyroid Carcinoma / Incremento rápido de tirotrofina endógena en pacientes con carcinoma diferenciado de tiroides para seguimiento o tratamiento con radioyodo

In follow up (F-U), ablation (A), or treatment (T) with radioiodine of patients with differentiated thyroid carcinoma (DTC), it is necessary to obtain elevated figures of serum TSH to assess hTg serum values or carry out 131I scanning. During the past few decades, the method employed was the withdrawal of hormonal treatment (WTH) for several weeks and its variants with the inconvenient symptoms of hypothyroidism, often restraining the use of this method. We aimed to obtain a rapid rice of serum TSH after a very short withdrawal of thyroid hormonal treatment (eight to nine days ) with the use of three or four intravenous application of TRH (200 mcg) during the first 6 days of withdrawal (TRH-St). One hundred determinations were carried out in 66 patients with DTC (ages19-80 y.o ), 20 males and 46 females. Sixty seven TRH-St were carried out for F-U, 20 for FU/T and 13 for A. In all cases the TSH values after the 3rd or 4th TRH application (samples 1 and 2) were over the value of 25 mIU/L and in the case of the second sample 99/100 determination were over the value of 30 mU/L. The values obtained were for the first sample 70.9 mIU/L ± 54.5 (range 25-310) and for the second sample 85.2 ± 61.3 (range 26-360), p<0.001. Patients considered that the symptoms and discomfort observed were mild when compared to those observed in patients submitted previously to the WTH method for 4/5 weeks. The results observed with TRH-St, allow us to consider the method as an alternative to the classic withdrawal method or the use of rhTSH with an adequate relation cost benefit.

Para efectuar ablación (A) , tratamiento con radioyodo (T) o seguimiento (S) en pacientes portadores de carcinoma diferenciado de tiroides (CDT) se hace necesario incrementar los valores de tirotrofina sérica (TSH) para elevar la sensibilidad del centellograma y la especificidad de la determinación de tiroglobulina sérica (hTg). Por años el método clásico fue la suspensión del tratamiento opoterápico (WTH) o sus variantes y ocasionalmente el uso de TSH de origen animal o , raramente, humana. Hace una década, la introducción de la TRH recombinante (rhTSH) significó evitar la desagradable sintomatología del hipotiroidismo que conllevaba el uso del método (WTH) y que en ocasiones impedía su utilización. Nuestro objetivo: el rápido ascenso de la TSH sérica después de muy breve WTH (ocho a nueve días) utilizando tres o cuatro aplicaciones intravenosas de la hormona liberadora de tirotrofina (TRH) durante los primeros seis días de WTH, método que denominamos TRH-St. Se efectuaron cien TRH-St en 66 pacientes: 20 masculinos, 46 femeninos, edades 19-80 años; 61 carcinomas papilares de diversas variantes anatomopatológicas, 4 foliculares y una variantes Hürthle. En todos los estudios después de la 3ra y cuarta aplicación de TRH (muestras 1 y 2 respectivamente) los valores de TSH fueron superiores a 25 mUI/L y con respecto a la cuarta TRH, 99/100 estudios ofrecieron valores de TSH superiores a 30 mUI/L. Los promedios obtenidos fueron: muestra 1 : 70.9 ± 54,5 mUI/L de TSH (rango 25-310); muestra 2: 85.2 ± 61.3 (rango 26-360): p < 0,001. Los pacientes consideraron que la sintomatología adversa del hipotiroidismo y el "disconformismo" fueron leves y sin comparación con los observados por aquellos pacientes sometidos anteriormente al método de supresión hormonal por 4/5 semanas.. Estas observaciones nos llevan a considerar que el método TRH-St , es una alternativa válida del método clásico de suspensión hormonal o del uso de rhTSH con una relación adecuada costo / beneficio.

Poorly differentiated and anaplastic thyroid carcinomas: chromosomal and oligo-array profile of five new cell lines.

Information on gene alterations associated to poorly differentiated (PDTC) and anaplastic thyroid carcinomas (ATC) is scarce. Using human cancer cell lines as a tool for gene discovery, we performed a cytogenetic and oligo-array analysis in five new cell lines derived from two PDTC and three ATC. In PDTC we evidenced, as important, the involvement of the MAPK/ERK kinase pathway, and downregulation of a group of suppressor genes that include E-cadherin. In ATC, downregulation of a specific group of oncosuppressor genes was also observed. Our ATC cell lines presented chromosomal markers of gene amplification, and we were able to identify for the first time the nature of the involved amplicon target genes. We found that the main molecular differences between the two cell line types were related to signal transduction pathways, cell adhesion and motility process. TaqMan experiments performed for five amplicon target genes and for two genes, which allowed a clear distinction between ATC and PDTC: CDH13 and PLAU corroborated array results, not only in the cell lines, but also in an additional set of primary 14 PDTC and three ATC. We suggest that our findings may represent new tools for the development of more effective therapies to the hitherto untreatable ATC.

Frequent 7q gains in flow cytometric multiploid/hypertetraploid breast carcinomas: a study of chromosome imbalances by comparative genomic hybridisation.

OBJECTIVE: To investigate underlying genetic events associated with complex DNA ploidy breast carcinomas. METHODS: Screening for chromosome imbalances was carried out using comparative genomic hybridisation (CGH) in 14 frozen samples of tumour from a series of 13 breast cancer patients with multiploid (n = 11) and hypertetraploid (n = 2) tumours. They had previously been analysed by DNA flow cytometry and also assessed immunohistochemically for p53 tissue expression. Ploidy status was determined on frozen samples using the Multicycle software program. RESULTS: The total number of copy gains (n = 242) was significantly greater than the number of copy losses (n = 51). The mean (SD) number of gains per sample was 17.3 (5.7), and of losses, 3.6 (4.2) (p = 0.0001). Gains of chromosomal regions at 1q (14/14; 100%), 7q (12/14; 85.7%), and 3q (11/14; 78.6%), as well as 1p, 2q, 5p, 8q, and 13q (10/14; 71.4%) were the most frequent aberrations in this series. Losses were most commonly found on 17p (5/14; 35.7%). Three patients dying of the disease had tumours with high level amplifications at 1q12-qter, 3q22-q25, and 8q22-q23 regions. Six cases had p53 overexpression, of whom four showed 12q gains and two showed 17p losses. CONCLUSIONS: There is a very high incidence of genetic aberrations, mainly related to chromosomal gains, in this subgroup of aneuploid breast cancer patients, associated with a poor clinical outcome. The 7q locus, not previously reported as showing frequent changes in breast cancer, was found to be a potential site for some candidate oncogenes.

PAX8-PPARgamma rearrangement is frequently detected in the follicular variant of papillary thyroid carcinoma.

CONTEXT: The clinicopathological characteristics and the molecular features of the follicular variant of papillary thyroid carcinoma (FVPTC) remain controversial. OBJECTIVE/DESIGN/PATIENTS: In an attempt to clarify such controversies and to find whether or not FVPTC cases share the molecular features of follicular tumors, we searched for the presence of PAX8-PPARgamma rearrangements, RAS mutations, and RAP-1, RAF-1, and BRAF mutations in a series of 40 FVPTCs as well as in 27 follicular thyroid carcinomas (FTCs) and 12 follicular thyroid adenomas (FTAs). Fluorescence in situ hybridization and RT-PCR were used to detect the PAX8-PPARgamma rearrangement and PCR, single strand confirmational polymorphism, and sequencing for searching the mutations. RESULTS: The frequency of PAX8-PPARgamma rearrangement was similar in FVPTCs (37.5%), FTCs (45.5%), and FTAs (33.3%). The same holds true regarding the frequency and type of RAS mutations: FVPTC, 25.0%; FTC, 22.2%; and FTA, 33.3%. BRAF mutations were only detected in FVPTC (10%); the BRAF mutations in these cases (K601E and G474R) are different from the typical BRAF(V600E) mutation of conventional PTCs. No mutations were detected in RAP-1 and RAF-1. In FVPTCs, the PAX8-PPARgamma rearrangement was significantly associated with multifocality and vascular invasion, whereas the RAS mutations were significantly associated with the large tumor size. There were three cases of FVPTC, three FTCs and one FTA, harboring both PAX8-PPARgamma rearrangement and RAS mutations; patients with such tumors were usually very young. CONCLUSIONS: We conclude that a subset of FVPTC shares some of the molecular features of follicular tumors. Further studies are necessary to clarify the putative clinical significance (e.g. association to blood-born metastases) of PAX8-PPARgamma rearrangement, RAS mutations, and BRAF(K601E) in FVPTCs.

Digestible energy requirements of Mexican donkeys fed oat straw and maize stover.

The limited availability of food, together with the constraints that traditional management systems impose on the natural foraging behaviour of donkeys, often results in severe under-nutrition. Few studies have been conducted into the digestibility of different forages and little information exists on nutritional requirements of donkeys. In order to measure digestible energy requirements of donkeys under tropical conditions, an experiment was carried out at the Centre for Research in Agricultural Science (CICA) and the Faculty of Veterinary Medicine of the Universidad Aut6noma del Estado de México located in the Toluca valley, Central Mexico. Thirty-two donkeys of a body condition typical for Central Mexico were divided into four groups of 8 animals each according to their sex and live weight: group 1 (G1) comprised male donkeys below the average body weight (102+/-5 kg); group 2 (G2) comprised male donkeys of average body weight (121.5+/-4 kg); group 3 (G3) comprised female donkeys below average weight (111.8 +/- 5 kg); and group 4 (G4) comprised female donkeys of average weight (127.6 +/- 5 kg). A diet of oat straw or maize stover and 15% alfalfa hay was offered to meet exact maintenance requirements. The donkeys were monitored for 13 months. The live weight of all animals was recorded daily in order to monitor whether maintenance requirements were being met. Mean daily digestible energy (DE) requirements were measured during the winter, spring, summer and autumn of 2003-2004. Digestible energy requirements of all four sex and liveweight groups were significantly (p > 0.05) higher during the spring than during the other seasons of the year (13.5, 18.0, 10.4 and 14.3 MJ DE per day during winter, spring, summer and autumn, respectively). Predicted DE requirements of donkeys with a live weight range betweenn 90 and 150 kg using the data from the present study were less than those predicted using scaled-down horse feeding standards.

Underexpression of peroxisome proliferator-activated receptor (PPAR)gamma in PAX8/PPARgamma-negative thyroid tumours.

The expression of peroxisome proliferator-activated receptor (PPAR)gamma in thyroid neoplasias and in normal thyroid (NT) tissues has not been fully investigated. The objectives of the present work were: to study and compare the relative expression of PPARgamma in normal, benign and malignant thyroid tissues and to correlate PPARgamma immunostaining with clinical/pathological features of patients with thyroid cancer. We analysed the expression of PPARgamma in several types of thyroid tissues by reverse transcription-polymerase chain reaction (RT-PCR), interphase fluorescent in situ hybridisation, real-time RT-PCR and immunohistochemistry. We have demonstrated that NT tissues express PPARgamma both at mRNA and at protein level. PAX8-PPARgamma fusion gene expression was found in 25% (six of 24) of follicular thyroid carcinomas (FTCs) and in 17% (six of 36) of follicular thyroid adenomas, but in none of the 10 normal tissues, 28 nodular hyperplasias, 38 papillary thyroid carcinomas (PTCs) and 11 poorly differentiated thyroid carcinomas (PDTCs). By real-time RT-PCR, we observed that tumours negative for the PAX8-PPARgamma rearrangement expressed lower levels of PPARgamma mRNA than the NT. Overexpression of PPARgamma transcripts was detected in 80% (four of five) of translocation-positive tumours. Diffuse nuclear staining was significantly (P<0.05) less prevalent in FTCs (53%; 18 of 34), PTCs (49%; 19 of 39) and PDTCs (0%; zero of 13) than in normal tissue (77%; 36 of 47). Peroxisome proliferator-activated receptorgamma-negative FTCs were more likely to be locally invasive, to persist after surgery, to metastasise and to have poorly differentiated areas. Papillary thyroid carcinomas with a predominantly follicular pattern were more often PPARgamma negative than classic PTCs (80% vs 28%; P=0.01). Our results demonstrated that PPARgamma is underexpressed in translocation-negative thyroid tumours of follicular origin and that a further reduction of PPARgamma expression is associated with dedifferentiation at later stages of tumour development and progression.

Construction of an immunotoxin with the pore forming protein StI and ior C5, a monoclonal antibody against a colon cancer cell line.

Sticholysin I (StI), a potent cytolysin isolated from the sea anemone Stichodactyla helianthus, was linked to the monoclonal antibody (mAb) ior C5. StI acts by forming hydrophilic pores in the membrane of the attacked cells leading to osmotic lysis. ior C5 is a murine IgG1, which recognizes the tumor associated antigen (TAA) ior C2. The cytolysin and the mAb were coupled by using the heterobifunctional cross-linking reagent sulfosuccinimidyl 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (SMCC). Two hybrid molecules composed by one ior C5 and one or two StI molecules were obtained (named conjugated I and II, respectively). The purified conjugates were evaluated by a binding affinity assay against an ior C2-positive colon cancer cell line (SW948). Both molecules were able to recognize the antigen (Ag) in the same way that unconjugated ior C5 does. The activity of both conjugates against human erythrocytes and SW948 cells was assessed. They lost most of their hemolytic activity but their residual activity was very similar. Nevertheless, when their cytotoxicity was studied on the SW948 cell line, only conjugate II killed efficiently the cells, indicating a specific mAb-Ag interaction. In this chimeric molecule the ratio between the cytotoxic and the hemolytic activity was larger than that of the free cytolysin. This fact indicates an increase of the specificity of the toxic effect toward the SW948 cell line and consequently an increase of the difference between its hemolytic and cytotoxic doses. The results herein support the feasibility of directing StI to the surface of cancer cells expressing ior C2 Ag via the mAb ior C5.

Prediction from ultrasonographic measurements of the expected delivery date in two species of bottlenosed dolphin (Tursiops truncatus and Tursiops aduncus).

Ultrasonographic measurements were made at least once a month during 14 gestations in seven Tursiops truncatus and 12 gestations in five Tursiops aduncus (bottlenosed dolphins). The 121 measurements of the fetal biparietal diameter and 139 measurements of the fetal thoracic diameter in T truncatus and the 97 measurements of the biparietal diameter and 97 measurements of the thoracic diameter in Taduncus were used to establish regression lines for the increases in the diameter of the head and thorax of the fetus with time. From these relationships an easy-to-use computer program was developed to predict the date of birth of the two species of bottlenosed dolphin, and its predictions were compared with the actual dates of birth of other calves of both species. The births occurred within the range of predicted dates, and even when only a few measurements were available, the program provided accurate predictions.

Chromosomal imbalances associated with anaplastic transformation of follicular thyroid carcinomas.

The genetic alterations that underlie the progression of follicular thyroid carcinoma towards anaplasia are still largely uncharacterised. We compared the Comparative Genomic Hybridization (CGH) profiles of 20 follicular (FTCs), 12 poorly differentiated (PDTCs) and seven anaplastic thyroid carcinomas (ATCs), in order to identify the chromosomal imbalances potentially associated with cancer progression. We found: (i) when considering that a 'direct' transformation of FTC towards anaplasia occurs, the defined significantly important alterations were the increase of gains at 3q (P<0.05) and 20q (P<0.01), and the increase of losses at 7q (P<0.05) and Xp (P<0.01); (ii) regarding poorly differentiated carcinomas as an intermediate independent entity in the anaplastic transformation of follicular cancers, evidenced as important alterations towards anaplasia, were the proportional decrease in copy sequences at 7p, 7q, 12q and 13q resulting from the significant decrease of DNA gains at 7p and 12q (P<0.05), and the significant increase of losses at 7q and 13q (P<0.05). These results unveil the chromosomal regions where genes of interest in thyroid anaplastic transformation are to be located, and demonstrate that different gene dosage copy sequence imbalances are associated to the 'direct' pathway of transformation of follicular into anaplastic cancers and to the progressive FTC --> PDTC --> ATC pathway.