Ovarian cancer stem cells.

Because of its semi-solid character in dissemination and growth, advanced ovarian cancer with its hundreds of peritoneal tumor nodules and plaques appears to be an excellent in vivo model for studying the cancer stem cell hypothesis. The most important obstacle, however, is to adequately define and isolate these tumor-initiating cells endowed with the properties of anoikis-resistance and unlimited self-renewal. Until now, no universal single marker or marker constellation has been found to faithfully isolate (ovarian) cancer stem cells. As these multipotent cells are known to possess highly elaborated efflux systems for cytotoxic agents, these pump systems have been exploited to outline putative stem cells as a side-population (SP) via dye exclusion analysis. Furthermore, the cells in question have been isolated via flow cytometry on the basis of cell surface markers thought to be characteristic for stem cells.In the Vienna variant of the ovarian cancer cell line A2780 a proof-of-principle model with both a stable SP and a stable ALDH1A1+ cell population was established. Double staining clearly revealed that both cell fractions were not identical. Of note, A2780V cells were negative for expression of surface markers CD44 and CD117 (c-kit). When cultured on monolayers of healthy human mesothelial cells, green-fluorescence-protein (GFP)-transfected SP of A2780V exhibited spheroid-formation, whereas non-side-population (NSP) developed a spare monolayer growing over the healthy mesothelium. Furthermore, A2780V SP was found to be partially resistant to platinum. However, this resistance could not be explained by over-expression of the "excision repair cross-complementation group 1" (ERCC1) gene, which is essentially involved in the repair of platinated DNA damage. ERCC1 was, nonetheless, over-expressed in A2780V cells grown as spheres under stem cell-selective conditions as compared to adherent monolayers cultured under differentiating conditions. The same was true for the primary ovarian cancer cells B-57.In summary our investigations indicate that even in multi-passaged cancer cell lines hierarchic government of growth and differentiation is conserved and that the key cancer stem cell population may be composed of small overlapping cell fractions defined by various arbitrary markers.

Molecular cloning and functional characterization of endogenous recombinant common marmoset monkey (Callithrix jacchus) follicle-stimulating hormone.

BACKGROUND: Common marmoset monkeys (Callithrix jacchus) are readily used in biomedical research. However, superovulation for embryonic stem cell production and developmental research still remain difficult. Inexplicably, follicle-stimulating hormone (FSH) as key player in superovulation has to be administered in extremely high dosages in this non-human primate compared to human. METHODS: To evaluate whether marmoset FSH (cjFSH) is functionally more competent than its human homologue on the marmoset FSH receptor (cjFSHR), we established in vitro a homologous system characterizing homologous and recombinantly produced cjFSH. RESULTS: Upon stimulation of two cell lines stably expressing either the marmoset or the human FSH receptor (cj/hFSHR), respectively, the second messenger signaling of the activated receptor displayed no significant difference in ED(50) values. Thermostability of cjFSH was significantly prolonged by roughly 20% on both FSHRs. CONCLUSION: High FSH dosage in marmoset superovulation cannot be explained by enhanced biopotency of the natural animal's gonadotropin.

Genetic differences in HLA-DQA1* and DQB1* allelic distributions between celiac and control children in Santiago, Chile.

Celiac disease is a permanent gluten intolerance strongly associated with HLA class II antigens. The over presentation of particular HLA alleles and haplotypes has been described in several populations. Different lines of evidence obtained during the last years suggest that a particular HLA-DQ heterodimer, encoded by the DQA1*0501 and DQB1*0201 genes in cis or trans conformation, confers the primary disease susceptibility. We report the HLA class II allelic distribution and DQA1/ DQB1 genotypes in 62 Chilean celiac patients compared with 124 control subjects in Santiago, Chile. We found a pronounced increase of the "susceptible" alleles :DQA1*0501 (0.480 vs 0.169, Pc < 0.0005), DQB1*0302 (0.430 vs 0.242, Pc = 0.002) and DQB1*0201 (0.250 vs 0.125, Pc = 0.037) in celiac patients in comparison with control children. As for "protective" alleles, we detected a high frequency of DQA1*0101 (0.310 vs 0.160, Pc = 0.01), DQA1*0201 (0.105 vs 0.010, Pc < 0.0075) and DQB1*0301 (0.250 vs 0.100, Pc = 0.010) in controls. In relation to risk haplotypes, the main combination observed was the conformation DQ8 (DQB1*0302/DQA1*0301) over DQ2 (DQB1*0201/DQA1*0501). In conclusion, results show that celiac disease in Chilean patients is primarily associated with DQ8 conformation. This is concordant with the high frequency of DR4 alleles (in linkage disequilibrium with DQB1*0302) detected in Amerind groups in Chile, where DQB1*0302 is more frequent than DQB1*0201.

Changes in human infants' sensitivity to slow displacements over the first 6 months.

Sensitivity to slow movement improves substantially during the early postnatal months. Thresholds were measured for slow oscillatory displacements for 68 infants aged 6, 12, 18, and 24 weeks. Standing wave line stimuli were used; 6-week-olds were tested at 1.2 Hz, and infants in the three older age groups were tested at either 0.6 or 1.2 Hz. Six-week-olds as a group were very insensitive to these slow displacements. Sensitivity increased systematically across the three older ages. Thresholds were marginally lower at 1.2 Hz than at 0.6 Hz, and there was some indication that these thresholds may reflect a mixture of detection by position-sensitive and motion-sensitive mechanisms. Several factors are hypothesized to be responsible for this development: (1) improvements in spatial resolution; (2) improvements in temporal contrast sensitivity; (3) decreases in the size of second order motion integration mechanisms; and (4) increased neural connectivity in the motion pathways.

Spatial-frequency-tuned channels in early infancy: VEP evidence.

Spatial frequency (SF) adaptation of 3-, 6-, and 12-week-old infants produced changes in steady-state VEP amplitude to SF sweeps that closely resemble post-adaptation changes reported previously in adults. Following SF adaptation, VEP amplitude was attenuated to SFs near the adapting SF, and enhanced at SFs removed from the adapting SF in all age groups. This is interpreted as evidence for bandpass SF-tuned channels with coinhibitory interactions. The data presented here, in combination with VEP estimates of infant acuity, imply the existence of multiple bandpass SF-tuned channels with inhibitory relationships by 3 weeks of age. We hypothesize that neonatal spatial frequency filters are qualitatively adultlike.

[Anesthesia with oral ketamine]. / Anesthésie par la kétamine orale.

Oral ketamine (7.5 mg X kg-1) was used to induce general anaesthesia in fourty seven young women undergoing voluntary abortion. Additional anaesthesia was necessary eight times because pre operative consciousness remained normal, and nine times because of important intra operative reactions. A majority (78.8%) of patients appreciated this technique of anaesthesia. Absence of narcosis (17%) and intra operative analgesia (23%) and frequency of post operative nightmares may require some modifications of the method.

PIP: Ketamine was administered to 47 women aged 14-41 years who were about to undergo induced abortions. The patients received 2.5 mg of lorazepam 2 hours before the operation and 7.5 mg/kg of ketamine diluted in orange juice or water 45 minutes before. In all cases, sleep and transfer to the operating table were accomplished in calm and semidarkness. Intravenous tubes were inserted and .02 mg/kg of atropine sulfate was administered. Patients considered still conscious were given intravenous injections of ketamine 1 mg/kg. Perioperative evaluation of the quality of anesthesia was done using a 4-level scale based on reactions to stimuli. Ketamine .5 mg/kg was administered intravenously each time significant reactions were obtained. 45 minutes after oral ketamine administration, 8 patients were still conscious and received additional anesthesia. 9 of the 39 patients asleep at the preoperative evaluation required additional anesthesia during the procedure. The immediate postoperative period was calm in all cases, even though some patients later reported having had disagreeable hallucinations. 46.8% had vomiting. The frequency of vomiting declined from 54.4% to 28.8% when pure water was substituted for orange juice as the vector for the preoperative oral ketamine. Correct responses to simple questions were obtained an average of 12.4 minutes postoperatively, but all patients had periods of somnolence lasting 4.8 hours on average. 15 had partial recollections of the surgery. 78.8% of the patients stated that the anesthesia used was excellent or good. 8.5% felt it was average, and 10.6% felt it was poor. Very few publications mention oral use of ketamine. The failure rates of 17% during the preoperative evaluation and 23% during the operation were not negligible and were probably due to the very low bioavailability of ketamine administered orally and the variability of digestive absorption of ketamine from 1 subject to another. The method appears to be appropriate for use in induced abortions, but better management is required to reduce failure rates and control side effects of vomiting and disagreeable postoperative hallucinations.

False-positive radio-iodinated metaiodobenzylguanidine (123I-MIBG) accumulation in a mast cell-infiltrated infantile haemangioma.

Radio-iodinated metaiodobenzylguanidine ((123)I-MIBG) is used for the detection and staging of neuroblastoma, pheochromcytoma and other neuroendocrine tumours in diagnostic nuclear medicine. A specific uptake and storage mechanism provides the basis for imaging with (123)I-MIBG. Nevertheless, cases of false-positive (123)I-MIBG scintigraphy with accumulation in non-chromaffin tumours have been described. Here, we present a case of a false-positive (123)I-MIBG scan in a case of a mast-cell infiltrated infantile haemangioma and discuss the possible uptake mechanism.

Intracoronary photodynamic therapy reduces neointimal growth without suppressing re-endothelialisation in a porcine model.

OBJECTIVE: To examine the effects of intracoronary PhotoPoint photodynamic therapy (PDT) with a new photosensitiser, MV0611, in the overstretch balloon and stent porcine models of restenosis. METHODS: 28 pigs were injected with 3 mg/kg of MV0611 systemically 4 h before the procedure. Animals were divided into either the balloon overstretch injury (BI) group (n = 19) or the stented group (n = 9). After BI, a centred delivery catheter was positioned in the artery to cover the injured area, and light (532 nm, 125 J/cm(2)) was applied to activate the drug (n = 10). Control arteries (n = 9) were not activated by light. In the stented group, the drug was light activated before stent deployment. Serial sections of vessels were processed 14 days after treatment in the BI group and 30 days after treatment in the stented group for histomorphometric or immunohistochemical analysis. RESULTS: Intracoronary PDT significantly reduced intimal thickness in both BI and stented arteries (about 65%: 0.22 (SEM 0.05) mm v 0.62 (0.05) mm, p < 0.01; and about 26%: 0.40 (0.04) mm v 0.54 (0.04) mm, p < 0.01, respectively). PDT increased luminal area by

[Celiac disease in adolescents and young adults. A challenge for gastroenterologists of children and adults]. / Enfermedad celíaca en el adolescente y adulto joven. Un desafío para gastroenterólogos de niños y adultos.

BACKGROUND: Follow-up of celiac patients in Chile is often interrupted when adolescents are referred to adult gastroenterologists. AIM: To study the evolution of patients with celiac disease when they reach adolescence or young adulthood. PATIENTS AND METHODS: Current adherence to gluten-free diet and its relation to symptoms and circulating antiendomysial antibodies were evaluated in the 58 confirmed celiac patients older than 12 years of age controlled at 3 hospitals in Santiago. RESULTS: Mean age at the moment of this assessment was 17.8 +/- 5 years, 65.5% were women, 12.5% were at nutritional risk (-IDS) while 20% were overweight/obese. Although all patients declared themselves asymptomatic, a focused questionnaire revealed that 26% suffered some symptoms. Only 24.1% followed a strict gluten-free diet. Eight of 20 patients who ate gluten-containing diets had negative antiendomysial antibodies (EMA), three of whom turned positive within 6 to 9 months. In three of four (asymptomatic) cases that accepted a new jejunal biopsy, histology was abnormal. One patient who followed a strict diet had EMA (+) and normal histology. CONCLUSIONS: These results confirm that after childhood, symptoms abate significantly in celiac patients. The observed sensitivity and specificity of EMA makes necessary to maintain small intestinal biopsies as the gold standard for diagnosis and confirmation of the disease.

Celiac disease in a Chilean population carrying Amerindian traits.

BACKGROUND: Although clinical manifestations of celiac disease may change throughout life, clinical, histologic, immunologic, and genetic studies show that there are incomplete forms of this condition, making it difficult to define the disease at a given moment. Because there is no information published in the Latin American-Amerindian population, this study was conducted to assess relations between these parameters in Chileans with celiac disease and their first-degree relatives. METHODS: Sixty-two persons with confirmed celiac disease (mean age, 17.9 +/- 5.1 years; 78.3% females) and 126 relatives (mean age, 27.9 +/- 17.2 years; 65.1% females) were evaluated. Clinical manifestations, antiendomysial antibodies (EMAs), and human leukocyte antigen (HLA) haplotypes were studied in patients. Additionally, jejunal biopsy specimens were assessed (light microscopy) in EMA-positive (EMA+) relatives. RESULTS: Of the patients, 24.1% adhered to a strict gluten-free diet; 26% were oligosymptomatic, and none were malnourished; 45% were EMA+; 13.8% who ingested gluten were EMA-negative (EMA-); one patient consuming a strict gluten-free diet was EMA+. The DQA1*0501 allele was present in the highest frequency (48%, P < 0.0005), whereas combinations of DQ8 were predominant. Of the relatives, 4.8% were EMA+; they had a significantly higher frequency of diarrhea, weight loss, and anorexia (P < 0.03); and all had abnormal histology in biopsy specimens. CONCLUSIONS: After childhood, celiac disease is oligosymptomatic and is often unrecognized by patients. Disease in 13.8% of patients and in 4.8% relatives appeared as incomplete forms of celiac disease. Predominance of DQ8 HLA haplotypes reflects the genetic Spanish-Mapuche heritage of this population.

[Wilson's disease: a review apropos of a clinical experience in 16 patients]. / Enfermedad de Wilson: una revisión a propósito de experiencia clínica en 16 pacientes.

Wilson's Disease is an inherited disorder of copper metabolism. We report 16 patients (6 males) with the disease; 6 had hepatic involvement exclusively, 4 had neurological involvement, 3 had a neurological and hepatic involvement and 3 were asymptomatic. The age onset was 9 years for hepatic and 17 years for neurologic involvement. The mean delay in diagnosis was 14 months. Chronic hepatitis, cirrhosis and fulminant hepatic failure were the clinical forms of liver disease. Patients with neurologic disorders had behavioral disturbances and extrapyramidal manifestations such as dystonia and parkinsonism. Patients had a good response to penicillamine, except 3 that died of liver complications, in whom the treatment was delayed or discontinued. We conclude that this metabolic disease must be suspected in pubertal children and in adults of less than 30 years old with liver disease of unknown origin or behavioral alterations associated to an extrapyramidal syndrome.

Differentiation of VEP intermodulation and second harmonic components by dichoptic, monocular, and binocular stimulation.

Modulation by two temporal frequencies differentiates visual processing at the fundamentals (1Fs), second harmonics (2Fs), and second-order intermodulation components (IMCs), the latter created neurally as the sum or difference of the two modulation frequencies. Steady-state VEPs were recorded while stereo-normal adults viewed luminance or grating stimuli modulated by up to three temporal frequencies under dichoptic, monocular, or ordinary (binocular) viewing conditions arranged using liquid crystal light shutters. In Experiment 1, modulation of luminance by a single temporal frequency produced strong 1F and 2F VEP components, but modulation of gratings produced only 2Fs. Modulation by two temporal frequencies resulted in IMCs, often in the absence of evoked activity in the EEG at the 1Fs. IMCs were generally larger during pattern as compared to luminance modulation. Amplitudes of 1Fs and IMCs were smaller, but 2Fs were larger, during dichoptic as compared to ordinary viewing. Although the 2F to a single modulation presented to one eye was not reduced when a second frequency was added to the opposite eye, monocular IMCs were diminished when a frequency was added to the opposite eye. We conclude that 2Fs and IMCs are associated with different neural substrates. Results are consistent with a two pathway model with one pathway having a nonlinear filter prior to binocular combination, the other pathway having a nonlinearity following binocular linear summation. Implications of these data for binocular function are discussed.

Reparacao de ferimento cardiaco:ocorrencia de dois episodios no mesmo paciente. / Repair of cardiac injury: ocurrence of two episodes in the same patient

Os autores relatam o caso de um paciente que por duas vezes foi vitima de ferimentos cardiacos com perfuracao de ventriculo esquerdo, tendo em ambas as ocasioes sido submetido a cirurgia de urgencia em um hospital de interior, sem equipamentos especializados ou medicos treinados em cirurgia cardiaca. Houve pleno sucesso em ambas as cirurgias e o paciente manteve-se vivendo normalmente apesar das sequelas remanescentes que, mais tarde, foram reveladas pela cineangiocoronariografia (obstrucao da coronaria descendente anterior esquerdo a aneurisma de ventriculo esquerdo)