The effect of varying concentrations of chicken plasma egg yolk and glycerol on the viability of canine sperm following short and long-term preservation.

Background: Plasma egg yolk (PEY), due to simple preparation and easier access, could be a suitable alternative to raw egg yolk for preserving canine semen. Aims: The present study investigated suitable concentrations of PEY and glycerol for preservation of canine semen. Methods: Semen was collected by digital manipulation (seven replicates from four dogs). Following initial raw semen evaluation, the semen was diluted in a tris-based extender supplemented with varying concentrations of chicken PEY (0, 20, and 40% v/v) and glycerol (3%; v/v). After cooling the specimen to 4°C within 1 h, the specimens were diluted with an equal volume of freezing extender consisting of similar concentrations of chicken PEY and 0 and 7% glycerol to reach the final glycerol concentration of 1.5 and 5% for short-term storage of canine semen. Samples with different concentrations of PEY and 5% glycerol were frozen. The sperm viability parameters including total motility, progressive forward motility, plasma membrane integrity, and live percentage of sperm were assessed following short and long-term storage. Results: Sperm viability parameters of semen extended in an extender supplemented with 20 or 40% chicken PEY with either 1.5 or 5% glycerol remained superior until 72 h after semen collection compared to the specimen that did not receive any PEY (P<0.05). Post-thaw sperm viability was also greater in samples extended in extender supplemented with either 20 or 40% PEY compared to 0% PEY. Conclusion: Tris-based extender supplemented with either 20% chicken PEY could be suitable for short and long-term preservation of canine semen.

Transfection of bovine spermatogonial stem cells in vitro.

Spermatogonial stem cells (SSCs) are the only stem cells in adults that can transfer genetic information to the future generations. Considering the fact that a single SSC gives rise to a vast number of spermatozoa, genetic manipulation of these cells is a potential novel technology with feasible application to various animal species. The aim of this study was to evaluate enhanced green fluorescent protein (EGFP) gene transfection into bovine SSCs via liposome carrier and assess the best incubation day in uptake exogenous gene by SSCs. Transfection efficiency of EGFP gene with lipofectamine 2000 was determined in days following each three day of transfection (day 4, 6 and 8 of the culture) by fluorescent microscope. Results showed that the transfected cells through lipofection increased significantly (P<0.05) in each three days of transfection in comparison with those of the control groups. The transfected SSCs were higher in comparison with those of the free exogenous gene carrier groups (P<0.05). In comparison with these three days, the rate of infected cells was higher when transfection proceeds at day four. It was concluded that lipofectamine can be used safely for direct loading exogenous DNA to SSCs particularly during the fourth day of culture.

Ultrasonography of the supramammary lymph nodes for diagnosis of bovine chronic subclinical mastitis.

Currently, somatic cell count (SCC) and bacterial culture are considered as the gold standard of detecting subclinical Mastitis. Mastitis leads to proliferation of lymphocytes in the supramammary lymph nodes and subsequent enlargement of ipsilateral lymph node. Ultrasonography can be used to survey these changes. A portable ultrasound machine with a 2-5 MHz convex transducer was used to identify the supramammary lymph node size in 35 cows in a herd with chronic Staphylococcus aureus mastitis. After pre-milking udder preparation, a California mastitis test (CMT) was performed and individual milk samples were taken from each quarter for bacterial culture and somatic cell count (SCC) in accordance with NMC recommendations. The mean length (range 5.77-12.90 cm) and width (range 2.07-7.41 cm) of the lymph node were 9.2 and 4.03 cm, respectively. There was a positive correlation between lymph node size (length and depth) and culture of milk samples on ipsilateral quarters. Also, there was a significant difference correlation between CMT or mean log SCC of each side and size of supramammary lymph node in the same side. This study showed significant changes in supramammary lymph node dimensions in mastitis cases, so ultrasonography of this lymph node is probably a useful method for mastitis detection, especially in situations that test on milk is impossible.

Effect of donor cell age on development of ovine nuclear transfer embryos in vitro.

The effects of the age of cell donor animal on in vitro development of ovine nuclear transfer (NT) embryos were investigated. Somatic donor cells were obtained from two different sources: (1) adult cells (adult fibroblast cells; AFC and adult cumulus cells; ACC); and (2) fetal fibroblasts (40-day-old; FFC-40 and 65-day-old; FFC-65). The fibroblast cell lines were used for NT procedures within 4-13 subpassages. While the cumulus cells were used as non-cultured (fresh) cells. The in vitro matured abattoir-derived oocytes were considered as recipients. No differences in the rates of fusion (75.7, 77.7, 76.3 and 86.7%) and cleavage (80.1, 84.3, 77.8 and 74%) were detected among couplets reconstructed with FFC-40, FFC-65, AFC and ACC, respectively. Blastocyst formation rate of those oocytes reconstructed with FFC-40 was higher (18%; p < 0.001) than those reconstructed with FFC-65 (13%) and AFC (10.9) and comparable with those reconstructed with ACC (17.5%). When the effect of passage number was analysed within groups (FFC-40, FFC-65 and AFC) there were no significant differences in fusion, cleavage and blastocyst rates between reconstructed oocytes. The present study demonstrates that the fetal and adult fibroblasts as well as fresh cumulus cells are comparable in their ability to attain cell fusion and embryonic cleavage. Moreover, the blastocyst formation rate is influenced by the age of the donor animal and the fresh cumulus cells have similar remodelling potential to that of fetal fibroblasts in term of blastocyst formation rate.

Effects of different concentrations of bovine follicular fluid and estrous cow serum on development of murine 2-cell embryos.

Murine 2-cells embryos were isolated from murine oviducts at laboratory and transferred into Ham's F-10 medium containing 0.1 mg mL(-1) streptomycin and 100 IU mL(-1) penicillin G and supplemented with 3 mg mL(-1) bovine serum albumin (BSA) or different concentrations of bovine follicular fluid (bFF) and estrous cow serum (ECS). Significantly higher (p<0.05) > or =4-cell embryos were developed when embryos were cultured 20% bFF (84.33%) comparing to 10 and 15% bFF (48.33 and 69.33%) as well as 3 mg mL(-1) BSA (65.66%). Morula rates were also lower in 10% bFF (22.33%) comparing to the other groups and were similar in 15 and 20% bFF (62.66 and 72.33% morula rates) as well as BSA containing media (55.33%). The highest (p<0.05) blastocyst rates were obtained in medium containing 20% bFF (64.33%) and the lowest belonged to 10% bFF (15%) comparing to 15% bFF (33.66%) or 3 mg mL(-1) BSA. When embryos were cultured in ECS, no significant different was observed in different culture media (76.66, 72.33, 82.5 and 65.66% > or =4-cell embryos in 10, 15 and 20% bFF and 3 mg mL(-1) BSA, respectively). Morula and blastocyst rates were also similar in all groups (32.33, 41.66 and 66.25 and 55.33% morula rates and 15.33, 27, 44.50 and 29.66% blastocyst rates for 10, 15 and 20% bFF and 3 mg mL(-1) BSA, respectively). The results of the present study demonstrated that 20% bFF could be substituted for BSA when in vitro culture of murine embryos is carried.

Changes in regional fetal cerebral blood flow perfusion in relation to hemodynamic deterioration in severely growth-restricted fetuses.

OBJECTIVES: To study regional cerebral blood perfusion with power Doppler ultrasound (PDU) imaging in appropriate-for-gestational age (AGA) fetuses and those with intrauterine growth restriction (IUGR) at different hemodynamic stages of fetal deterioration. METHODS: Brain blood perfusion was studied with PDU imaging, and the fractional moving blood volume (FMBV) was estimated in 56 growth-restricted and 56 AGA matched fetuses at 26-32 weeks of gestation. Fetuses with IUGR were classified according to progression of hemodynamic deterioration as follows: Group 1, abnormal umbilical artery (UA) pulsatility index (PI) (mean > 2 SD, n = 13); Group 2, abnormal UA-PI and middle cerebral artery (MCA) PI (mean < 2 SD, n = 15); Group 3, abnormal UA-PI, MCA-PI and ductus venosus (DV) PI (mean > 2 SD) but atrial (a-wave) flow present (n = 16); and Group 4, absent or reversed DV atrial flow (n = 12). FMBV was calculated in the complete mid-sagittal, frontal, basal ganglia and cerebellar regions. RESULTS: In all growth-restricted fetuses, FMBV was significantly increased in all regions. Fetuses in Group 1 showed considerable increments in FMBV values in the frontal, complete mid-sagittal and cerebellar regions, and a mild increase in the basal ganglia. From Groups 2 to 4, there was a steady reduction (compared with Group 1) in frontal FMBV values (F = 3.25, P = 0.027) together with a significant increment in the basal ganglia values (F = 11.61, P < 0.001). A trend for increasing FMBV values was also observed in the cerebellum, whereas a decreasing trend was noted in the complete mid-sagittal area. CONCLUSIONS: Brain perfusion in growth-restricted fetuses shows clear regional variations, which change with progression of hemodynamic deterioration. After an initial and early increase in the frontal area, progression of fetal deterioration was rapidly associated with a pronounced decrease in frontal perfusion, together with an increase towards the basal ganglia.

Evaluation of fetal regional cerebral blood perfusion using power Doppler ultrasound and the estimation of fractional moving blood volume.

OBJECTIVE: To standardize the evaluation of regional fetal brain blood perfusion, using power Doppler ultrasound (PDU) to estimate the fractional moving blood volume (FMBV) and to evaluate the reproducibility of this estimation. METHODS: Brain blood perfusion was evaluated in 35 normally grown fetuses at 28-30 weeks of gestation, using PDU. The following cerebral regions were included in the PDU color box: anterior sagittal, complete sagittal, basal ganglia, and cerebellar. Ten consecutive good-quality images of each anatomical plane were recorded and the delimitation of the region of interest (ROI) was performed off-line. FMBV was quantified in the ROI of all images and the mean considered as the final value. Differences between regions, variability, reproducibility and agreement between observers were assessed. RESULTS: Power Doppler images of the described anatomical planes were obtained in all cases, regardless of fetal position. The median time for the acquisition of the images was 7 (range 4-12) min. Mean (range) FMBV values were: anterior sagittal, 16.5 (10.7-22.8)%, inter-patient coefficient of variation (CV) 0.22; complete sagittal, 13.5 (8.8-16.1)%, CV 0.27; basal ganglia, 18.3 (10.7-27.6)%, CV 0.27; and cerebellar, 6.6 (3.0-11.0)%, CV 0.38. There were statistically significant differences in FMBV between cerebellar and complete sagittal ROIs with the frontal and basal ganglia regions. Reproducibility analyses showed an intraclass correlation coefficient of 0.91 (95% CI 0.67-0.97) and an interclass correlation coefficient of 0.87 (95% CI 0.70-0.94). Interobserver agreement showed a mean difference between observers of -0.2 (SD 2.7) with 95% limits of agreement -5.6 to 5.2. CONCLUSIONS: When the regions of interest are well defined, the FMBV estimate offers a method to quantify blood flow perfusion in different fetal cerebral areas. There appear to be regional differences in FMBV within the fetal brain.

Maintenance of pregnancy in repeat breeder dairy cows by CIDR administration after breeding.

The aim of this study was to assess the effect of two periods of P4 therapy by CIDR on the 1st service Conception Rate (CR) of repeat breeder dairy cows. The cows were selected on the following basis: absence of dystocia, retained placenta and endometritis after the last parturition. They were inseminated according to the AM/PM rule relative to estrus onset and randomly assigned into 3 groups: (A) CIDR on day 5 after insemination that was removed on day 9 of the cycle (n = 40); (B) CIDR on day 5 after insemination that was removed on day 19 of the cycle (n = 36) and (C) untreated controls (n = 40). Pregnancy diagnosis was conducted by rectal palpation 45-55 days after AI in cows not observed in estrus. The difference in CR among the groups was analyzed by chi square analysis. The CR in groups A, B and C was 55, 41.7 and 30%, respectively and was greater (p<0.05) in treatment (A) cows than in control cows. In conclusion, repeat breeder cows in groups A and B benefited from progesterone supplementation, but significant effects of treatment for improvement of conception rate was seen in short treatment period (4 days treatment).

Expression of genes related to activity of oxaliplatin and 5-fluorouracil in endoscopic biopsies of primary esophageal cancer in patients receiving oxaliplatin, 5-flourouracil and radiation: characterization and exploratory analysis with survival.

With a goal of identifying relations between gene expression and response (mucosal or pathological) or survival in esophageal cancer patients (stages II to IV) receiving oxaliplatin, 5-fluorouracil (5FU) and radiation, we measured in endoscopic primary tumor biopsies from 38 patients, the expression of seven genes (gammaGCS, gammaGT, MRP-2, ERCC-1, XPA, TS and DPD) prior to treatment, 1 week following oxaliplatin alone and at the end of the combined radio-chemotherapy cycle using real time QRT-PCR. A higher pretreatment level of XPA was related to shorter survival with a hazard ratio of 2.43 (90% confidence interval 1.09 to 5.43) using Cox regression modeling. However, multivariate analysis with a Cox model indicated low expression of XPA or TS and combined stages II and III had a higher probability of survival (for XPA: hazard ratio 3.0 and 90% C.I. of 1.3 to 6.9, with adjustment for stage included; for TS: hazard ratio is 1.98 with 90% C.I. of 0.94 to 4.20. The expression of TS, gammaGCS, ERCC-1 and MRP-2 declined from D 1 to the end of the cycle (p<0.05, sign test). A validation and further understanding of the findings need to be carried out in a larger study with a more homogeneous population of patients.

Management of locoregional stage esophageal cancer: a single center experience.

Therapeutic options for locoregional esophageal cancer (EC) include primary surgery, neoadjuvant or definitive chemoradiation and systemic chemotherapy. The role of surgery in these multimodal strategies has recently been debated and definitive chemoradiation is being offered as an alternative to surgery at many centers. We examined our results with multimodal therapy and surgery in this patient population. We conducted a retrospective analysis of 172 patients with locoregional (AJCC stages I-III) EC treated at RPCI between February 14, 1990 and September 20, 2002. Median age was 65 years (range, 36-95); there were 136 male patients. There were 100 regional (stages IIB-III), 69 local (stages I-IIA) and three in situ cases. Initial therapy was either combined modality (n = 122) or single modality (surgery) (n = 50). There was 0%, 30-day, postoperative mortality. Median survival for all patients was 25.3 months and was better for local stage with surgery alone (75 months) than with neoadjuvant (35.7 months) or definitive chemoradiation (19.1 months, P < 0.001). Survival for patients with regional disease treated with surgery alone, neoadjuvant or definitive chemoradiation was 21.5, 24.4 and 11.8 months, respectively (P = not significant). The associations of prognostic factors with overall survival were evaluated using Cox proportional hazards regression analysis and 2-sided Wald's chi-square test. On multivariate analysis, carefully selected patients treated with surgery alone had better outcomes compared with those treated with definitive chemoradiation (P < 0.001). Patients with locoregional esophageal cancer who are eligible for surgical resection either alone or as a part of multimodal therapy may have better outcomes than those treated with non-surgical approaches.

Cardiac function after chemoradiation for esophageal cancer: comparison of heart dose-volume histogram parameters to multiple gated acquisition scan changes.

In this paper we determine if preoperative chemoradiation for locally advanced esophageal cancer leads to changes in cardiac ejection fraction. This is a retrospective review of 20 patients treated at our institution for esophageal cancer between 2000 and 2002. Multiple gated acquisition cardiac scans were obtained before and after platinum-based chemoradiation (50.4 Gy). Dose-volume histograms for heart, left ventricle and left anterior descending artery were analyzed. Outcomes assessed included pre- and postchemoradiation ejection fraction ratio and percentage change in ejection fraction postchemoradiation. A statistically significant difference was found between median prechemoradiation ejection fraction (59%) and postchemoradiation ejection fraction (54%) (P = 0.01), but the magnitude of the difference was not clinically significant. Median percentage volume of heart receiving more than 20, 30 and 40 Gy were 61.5%, 58.5% and 53.5%, respectively. Our data showed a clinically insignificant decline in ejection fraction following chemoradiation for esophageal cancer. We did not observe statistically or clinically significant associations between radiation dose to heart, left ventricle or left anterior descending artery and postchemoradiation ejection fraction.

Asociación entre malformaciones congénitas del sistema nervioso central y epilepsia en pacientes pediátricos de México / A link between congenital malformations of the central nervous system and epilepsy in paediatric patients in mexico

Introducción. Al ser la epilepsia una de las causas más frecuentes de atención en Neurología pediátrica, es necesario considerar una de las causas asociadas a la misma, la presencia de malformaciones congénitas, como segunda causa generadora de epilepsia; por ello, sobre la base de su neurodesarrollo, se pueden identificar las diferentes variedades de defectos congénitos relacionados con las epilepsias en Pediatría. Objetivo. Conocer las diferentes malformaciones congénitas asociadas a la epilepsia en Pediatría. Pacientes y métodos. Se incluyeron 116 casos con diagnóstico de epilepsia asociado a malformaciones congénitas del sistema nervioso central, en los que se evaluaron los estudios de imagen, como la resonancia magnética y tomografía computarizada de cráneo, y se agruparon de acuerdo al desarrollo cronológico embrionario normal del ser humano. Resultados. Todos los casos se seleccionaron por edad, y el grupo predominante se detectó en los menores de un año y en el grupo de escolares, con los trastornos de migración, entre los que se incluye la lisencefalia, como principal malformación; el otro grupo fue el de los trastornos de proliferación; en cuanto a los tipos de epilepsia asociados, los dos grupos de síndromes epilépticos en la infancia más comunes fueron el síndrome de West y el de Lennox-Gastaut; tomando en consideración los tipos de crisis epilépticas encontradas, correspondió a las crisis parciales. Conclusiones. Lo anterior muestra como principal causa asociada a la epilepsia las malformaciones congénitas del sistema nervioso central, y el estudio de neuroimagen con mayor sensibilidad hoy día es la resonancia magnética, por lo que se sugiere la utilización de este procedimiento en los casos en los que no se encuentre una causa aparente, para afinar con mayor precisión esta entidad nosológica. A pesar de la causa multifactorial, las edades menores de 25 y mayores de 35 años en las madres embarazadas se consideran de mayor riesgo potencial, sin predominio de área geográfica (AU)

Introduction. Since epilepsy is one of the most frequent causes of visits in Paediatric Neurology, attention must be given to one of the causes linked to it, namely congenital malformation, which is the second most common cause of epilepsy. To this end, different forms of congenital defects related to epilepsy in Paediatric medicine can be identified according to their neurodevelopment. Aims. The purpose of this study was to determine the different congenital malformations associated to epilepsy in Paediatrics. Patients and methods. We took a sample consisting of 116 patients diagnosed as suffering from epilepsy associated with congenital malformations of the central nervous system, following an evaluation of imaging studies, magnetic resonance and computerised tomography brain scans; subjects were then grouped according to the normal embryonic chronological development of the human being. Results. From the total number of cases, a selection was made according to age, where the predominant group was found in those below one year of age and in the group of school-age children, and migration disorders, where the main malformation included was lissencephaly; the other group was made up of proliferation disorders. Similarly, the associated types of epilepsy were the most common childhood epileptic syndromes, West and Lennox-Gastaut syndrome. The types of epileptic seizures that were found were partial seizures. Conclusions. The study outlined above shows congenital malformations of the central nervous system to be the main cause associated to epilepsy and the most sensitive neuroimaging study currently available is magnetic resonance. For this reason we suggest the use of this procedure in cases in which no apparent cause can be found so that this nosological entity can be defined to a greater degree of precision. Despite its multifactorial causation, being below 25 years of age and above 35 at the time of pregnancy is considered to constitute a higher potential risk, while no geographic location was found to predominate (AU)

[A link between congenital malformations of the central nervous system and epilepsy in paediatric patients in Mexico]. / Asociación de malformaciones congénitas del sistema nervioso central y epilepsia en pacientes pediátricos de México.

INTRODUCTION: Since epilepsy is one of the most frequent causes of visits in Paediatric Neurology, attention must be given to one of the causes linked to it, namely congenital malformation, which is the second most common cause of epilepsy. To this end, different forms of congenital defects related to epilepsy in Paediatric medicine can be identified according to their neurodevelopment. AIMS: The purpose of this study was to determine the different congenital malformations associated to epilepsy in Paediatrics. PATIENTS AND METHODS: We took a sample consisting of 116 patients diagnosed as suffering from epilepsy associated with congenital malformations of the central nervous system, following an evaluation of imaging studies, magnetic resonance and computerised tomography brain scans; subjects were then grouped according to the normal embryonic chronological development of the human being. RESULTS: From the total number of cases, a selection was made according to age, where the predominant group was found in those below one year of age and in the group of school-age children, and migration disorders, where the main malformation included was lissencephaly; the other group was made up of proliferation disorders. Similarly, the associated types of epilepsy were the most common childhood epileptic syndromes, West and Lennox-Gastaut syndrome. The types of epileptic seizures that were found were partial seizures. CONCLUSIONS: The study outlined above shows congenital malformations of the central nervous system to be the main cause associated to epilepsy and the most sensitive neuroimaging study currently available is magnetic resonance. For this reason we suggest the use of this procedure in cases in which no apparent cause can be found so that this nosological entity can be defined to a greater degree of precision. Despite its multifactorial causation, being below 25 years of age and above 35 at the time of pregnancy is considered to constitute a higher potential risk, while no geographic location was found to predominate.

Should preoperative, post-chemoradiotherapy endoscopy be routine for esophageal cancer patients?

Chemoradiation therapy is used widely for locoregional esophageal cancer. Patients with persistent disease may benefit from surgery. Preoperative esophagoscopy can identify persistent tumor but its accuracy is uncertain. The primary objective of this study is to assess the extent of agreement between esophagoscopy and surgical pathology in patients treated with neoadjuvant chemoradiation. A retrospective chart review of patients who underwent chemoradiation, preoperative endoscopy and surgery from January 1996 to December 2002 was performed. Cohen's kappa statistic was used to measure the degree of agreement between findings at endoscopic biopsy and surgical pathology. Thirty cases were identified. All patients received chemoradiation followed by surgical resection. There was insufficient agreement between tumor size (kappa 0.25, standard error 0.17, P = 0.07) and appearance (kappa 0.19, standard error 0.18, P = 0.14). Preoperative endoscopy revealed atypia/inflammation in 15 cases and dysplasia in eight. Of these 23 cases, 11 were adenocarcinomas at surgery. Only nine patients had concurrence between surgical pathology and endoscopy. The positive and negative predictive values of esophagoscopy for identifying residual tumor were 100% and 11%, respectively. Our data suggests that after chemoradiation, esophagoscopy is unreliable for excluding residual disease. The roles of other modalities need to be explored.

Pregnancy rate following post-insemination intrauterine treatment of endometritis in dairy cattle.

Summary Of 1500 examined Holstein cows in milk, there were 315 cows with mucopurulent discharges at the time of insemination (21%). Twenty-four hours after insemination, the infected animals randomly received no treatment (group I, control, n = 93), intrauterine infusion of 1 g oxytetracycline (group II, n = 63) and intrauterine infusion of 5 million IU procain penicillin G sodium (group III, n = 159). First service conception rate following treatment was 48.3, 49.2 and 47.7% in groups I, II and III, respectively. In conclusion, treatment with either antibiotic had no advantage relative to the control on first service conception rate.

Ultrasonic characteristics of preovulatory follicle and ovulation in Caspian mares.

The Caspian breed of horses is believed to be the direct descendant of the earliest equine animals. Some special characteristics of Caspian horse differentiate this breed of horses from other breeds. In the current study the ultrasonically observed characteristics of a preovulatory dominant follicle and the lengths of estrus, diestrus as well as some related parameters were studied during 42 interovulatory intervals in 11 healthy Caspian mares. The preovulatory dominant follicle deviated from subordinate follicles and became the largest follicle in the ovaries at Day -8.7 +/- 0.53 (Day 0=ovulation). Every mare was a single ovulator with ovulations more frequent from the left ovary than from the right (65% versus 35%). Mean length of estrus, diestrus, and interovulatory interval were 8.3 +/- 0.86, 13.8 +/- 0.59, and 22.1 +/- 0.40 days, respectively. The time interval from ovulation until the time in which the mares were no longer in estrus was 1.9 +/- 0.42 days.

[Preservation of hepatocytes in UW and HTK solutions with added fluorocarbons]. / Preservación de hepatocitos en soluciones de UW y HTK con adición de fluorocarbonos.

BACKGROUND: One of the limitations for hepatocyte transplantation is the short survival of isolated hepatocytes. OBJECTIVE: We evaluated the utility of fluorocarbons (PFC) added to University of Wisconsin (UW) and histidine-tryptofane-ketoglutarate (HTK) solutions on hepatocyte preservation. METHOD: Rat hepatocytes were isolated by colagenase digestion technique, with a viability greater than 85% measured by tripan blue exclusion. We suspended 1.2 x 10(8) hepatocytes in 20 mL of: HTK solution, HTK with perfluor-n-octane (HTK + PFC), UW solution, and E. Williams medium as control group (CG). Five samples of each group were stored at 4 degrees C for 40 h. We measured Cell viability, large membrane bleb formation and extracellular LDH at 0, 12, 24 and 36 hours. RESULTS: Cell viability was lower in the UW at 12 h (p < 0.05) and 24 h (p < 0.05) compared to the rest of the study groups; at 36 h we found no differences in cell viability. There were fewer hepatocytes with large membrane blebs in UW + PFC; compared with the remainder of the solutions, but this difference was only reached statistical significance when compared to UW at 24 h (p < 0.05) and at 36 h (p < 0.05). When comparing groups with or without fluorocarbons LDH levels showed no difference at 0, 12, 24, and 36 h. CONCLUSIONS: The present study shows that the addition of fluorocarbons to UW solution diminishes large membrane formation (irreversible damage) and improves cell viability at 12 and 24 h. However, fluorocarbons added to both solutions failed to increase preservation time beyond 24 h.

Is glucagonoma of the pancreas a curable disease?

BACKGROUND: Glucagonomas are rare neuroendocrine tumors of the pancreas. Because of its rarity, its natural history is not well understood. AIM: We evaluated the natural history of glucagonomas treated at a tertiary care cancer center. METHODS: A retrospective analysis of 12 patients during 1970 to 2000 was performed. Six patients (50%) had a tumor located in the head of the pancreas. RESULTS: Abdominal pain (83%) and weight loss (75%) were the most common symptoms. Median tumor size was 6 cm (range 0.04-10). Seven patients (58%) had liver metastases. Five patients (42%) underwent curative resection. Overall median survival was 66 mo, and 5-yr overall survival was 66%. Five-yr overall survival was 83% for patients who had resection versus 50% for the non-resected patients (p = 0.04). Patients who were disease-free had a complete resection of the primary tumor and no liver involvement. CONCLUSIONS: Glucagonomas generally present with liver metastases at the time of diagnosis. Cure is only possible if the disease is localized and completely resected.

Iatrogenic esophagobronchial fistula arising in irradiated Barrett's esophagus.

A 47-yr-old male underwent a right upper lobectomy for stage IIB bronchoalveolar carcinoma followed by 4600 Gy of irradiation. One year later a fistula formed from an ulcerated region of Barrett's esophagus into the left main bronchus. Bronchotomy repair with onlay patch intercostal muscle flap and esophageal repair with serratus anterior muscle flap plus postoperative esophageal stent placement for stricture resulted in good functional results.