Evaluation of adult papillary thyroid carcinomas by comparative genomic hybridization and microsatellite instability analysis

To clarify the mechanism of tumorigenesis in papillary thyroid carcinoma (PTC) and ascertain whether genomic changes correlate with histologic features, we conducted a comprehensive molecular evaluation of PTC using comparative genomic hybridization (CGH) and microsatellite instability (MSI) analysis in a set of 17 histologically well-characterized PTC specimens. To our knowledge, this is the first study that evaluates chromosomal and nucleotide instability in the same PTC tumor specimens. Four of 15 samples (27%) had aberrations detected by CGH. All four had a partial or complete gain of chromosome 20, and 3 of 4 had a partial or complete loss of chromosome 13. No MSI was detected in any of the PTC samples (n=16), and all samples examined by immunohistochemistry (n=9) expressed the DNA repair enzymes hmlh1 and hmsh2. All PTC samples with abnormal CGH had vascular invasion or invasion of the thyroid capsule, and there was a significant correlation between the presence of chromosomal aberrations and capsular/vascular invasion (P=0.026). We conclude that although chromosomal and microsatellite instability are uncommon in PTC, tumors with chromosomal aberrations are more likely to be associated with invasion.     

Gastric outlet obstruction as a consequence of a duodenal web masquerading as gastrinoma in an adult

We present the case of a 24-year-old man with recurrent peptic ulcers and hypergastrinemia, in whom a multidisciplinary investigation for gastrinoma revealed a duodenal web. The affected duodenal segment was excised, and a gastroduodenostomy with highly selective vagotomy was performed. Postoperative serum gastrin levels returned to the normal range over the next 6 weeks. Congenital duodenal anomalies are unusual causes of gastric outlet obstruction in adults. Chronic gastric outlet obstruction secondary to an adult duodenal web can induce neurohumoral changes in gastric function, which enhance both acid output and gastrin secretion. This case reminds clinicians to consider congenital anomalies in adults presenting with recurrent peptic ulcers and hypergastrinemia.     

Localization and developmental regulation of 11beta-hydroxysteroid dehydrogenase-1 and -2 in the baboon syncytiotrophoblast

We previously showed that the messenger RNA and protein levels of the 11ss-hydroxysteroid dehydrogenase (11betaHSD) enzymes catalyzing glucocorticoid reduction (11betaHSD-1) and oxidation (11betaHSD-2) increased with advancing baboon gestation and concluded that the estrogen-regulated change in placental cortisol metabolism from reduction at midgestation to oxidation near term is not simply the result of a change in the relative concentrations of these two enzymes. Therefore, in the current study we determined whether 11betaHSD-1 and -2 are located in different regions of the baboon and human syncytiotrophoblast and whether there is a developmental change in their localization with advancing baboon gestation. Western blot analyses, immunofluorescence, and electron microscopic immunocytochemistry indicated that 11betaHSD-1 expression was abundant in microvillus membranes (MVM) juxta the maternal circulation, and their levels are significantly lower, but detectable, in more internal regions of the syncytiotrophoblast, including membranes contiguous with the basal membrane (BM(m)) facing the fetal vasculature in both the human and baboon. In contrast, in both species 11betaHSD-2 expression was limited in the MVM and extensive throughout the remainder of the syncytiotrophoblast, including the BM(m). In the baboon, the relative mean (+/-SE) concentrations (arbitrary densitometric units per microgram protein) of 11betaHSD-1 in the MVM were similar at mid (i.e. day 100; 38,859 +/- 3,484; n = 3) and late (i.e. day 180; 43,561 +/- 1,784; n = 3) gestation (term = day 184) and exceeded (P < 0.01) respective values for 11betaHSD-2 by approximately 16-fold. In contrast, levels of 11betaHSD-1 in the BM(m) declined (P < 0.05) by approximately 50% between mid (7,099 +/- 758) and late (4,013 +/- 738) gestation, whereas levels of 11betaHSD-2 in this fraction increased. Thus, the ratio of 11betaHSD-2 to 11betaHSD-1 in the BM(m) at midgestation (1.22 +/- 0.10) was increased (P < 0.05) 2-fold in late gestation (2.66 +/- 0.05). Collectively, these findings indicate that the 11betaHSD-1 and -2 enzymes are localized to different membrane fractions of the baboon and human placental syncytiotrophoblast. Moreover, we propose that the developmental increase in the ratio of 11betaHSD-2 to 11betaHSD-1 in membranes facing fetal blood near term is consistent with and perhaps the subcellular mechanism responsible for the previously demonstrated switch in transplacental glucocorticoid metabolism from reduction at midgestation to oxidation late in gestation and appears to be responsible for the activation/maturation of the fetal pituitary-adrenocortical axis.     

Estrogen regulates 11 beta-hydroxysteroid dehydrogenase-1 and -2 localization in placental syncytiotrophoblast in the second half of primate pregnancy

We recently demonstrated that the 11 beta-hydroxysteroid dehydrogenase enzymes catalyzing cortisol-cortisone reduction (11 beta-hydroxysteroid dehydrogenase-1) and oxidation (11 beta-hydroxysteroid dehydrogenase-2) are located in different regions of the baboon and human placental syncytiotrophoblast. Moreover, there was a 2-fold increase in the ratio of 11 beta-hydroxysteroid dehydrogenase-2 to 11 beta-hydroxysteroid dehydrogenase-1 in syncytiotrophoblast membranes contiguous with the basal membrane (BMm) between mid and late baboon gestation. Our laboratories have also shown that estrogen regulates syncytiotrophoblast functional differentiation. Therefore, the current study determined whether the change in the ratio of 11 beta-hydroxysteroid dehydrogenase-2 to 11 beta-hydroxysteroid dehydrogenase-1 in the BMm was regulated by estrogen. Placentas were obtained on d 165 of gestation (term = d 184) from baboons that were untreated or were treated daily beginning on d 100 with the aromatase inhibitor CGS 20267, which reduced uterine and maternal serum E2 by more than 95% or with CGS 20267 plus E2 benzoate. Western blot analyses and immunofluorescence confirmed that in untreated controls the expression of 11 beta-hydroxysteroid dehydrogenase-1 was abundant in the microvillus membranes and considerably less in the BMm. In contrast, expression of 11 beta-hydroxysteroid dehydrogenase-2 was abundant in more internal regions of the syncytiotrophoblast, including the BMm, but was not detected in the microvillus membranes. The 11 beta-hydroxysteroid dehydrogenase-2 protein level was significantly decreased in the BMm of placentas from estrogen-suppressed baboons, resulting in a 2-fold decrease in the ratio of these enzymes in membranes juxta the fetal blood, and these changes were partially restored by CGS 20267 and E2. In contrast, estrogen had no effect on the ratio of 11 beta-hydroxysteroid dehydrogenase-2 to 11 beta-hydroxysteroid dehydrogenase-1 in whole villous homogenate or the micro-villus membranes. Collectively, these results indicate that estrogen regulates the developmental increase in the ratio of 11 beta-hydroxysteroid dehydrogenase-2 to 11 beta-hydroxysteroid dehydrogenase-1 in syncytiotrophoblast membranes juxta fetal blood, providing the subcellular architectural mechanism responsible for the previously demonstrated estrogen-dependent switch in transplacental glucocorticoid metabolism that regulates maturation of the primate fetal pituitary-adrenocortical axis.     

Collaboration, facilities and communities in day care services for older people

Collaborative working in care for older people is often seen as a desirable goal. However, there can be problems with this approach. This paper reports on a single blind randomized controlled trial which was carried out to compare outcomes of rehabilitation in two settings: a day hospital and social services day centres augmented by visiting therapists. The subjects were 105 older patients. Principal outcome measures were the Barthel Index, Philadelphia Geriatric Centre Morale Scale and the Caregiver Strain Index. Two aspects of the trial are examined here. Firstly, we investigated whether trial patients were more disabled than regular day centre attendees. Levels of health and well being amongst trial patients were compared with those of a random sample of 20 regular attendees from both of the participating day centres and an additional voluntary sector day centre. Secondly, key staff from the different settings were interviewed to assess how well the day centre model had worked in practice. Trial patients were significantly more disabled than regular day centre attendees according to the Barthel Index (P < 0.001), but this difference was no longer significant after three months of treatment. The day centre model had several problems, principally discharge policy, acceptability, facilities and attitudes of staff and regular attendees. Positive aspects of the day centre model, as well as successful rehabilitation, included shared skills, knowledge and resources. This paper suggests that collaborative working in day centres requires multipurpose facilities. If health staff maintain a permanent presence, benefits can include improved joint working, easier access to health care and the use of rehabilitative therapy as a preventative strategy. Day care settings can be analyzed as representing different types of communities. Allowing older users a greater degree of choice in facilities may increase the acceptability of care.     

Phase I trial of gemcitabine and CPT-11 given weekly for four weeks every?six?weeks

Background:Our previous studies have shown that the in vitrocytotoxicity of gemcitabine and SN-38, the active metabolite of irinotecan (CPT-11), is synergistic in human tumor cell lines. Patients and methods:Twenty-four patients with solid tumors, refractory to standard chemotherapy or for whom no effective therapy existed (age range 31–74; 7 female, 17 male; ECOG PS 0 = 12, 1 = 11, 2 = 1), received gemcitabine and CPT-11 weekly for four weeks out of every six weeks. Fifty courses of treatment (median 2, range 1–8) were given through five dose levels of gemcitabine/CPT-11 (600/75, 800/75, 800/100, 1000/100, 1000/125 mg/m²). Results:Grade 3 and 4 neutropenia occurred in eight and two patients, respectively. Grade 3 and 4 thrombocytopenia occurred in one and three patients, respectively. Hematologic toxicity resulted in 2 missed doses of treatment in two out of six patients and was therefore dose limiting at gemcitabine 1000 mg/m² and CPT-11 125 mg/m². Grade 3 and 4 diarrhea occurred in two and one patients, respectively. Other moderate non-hematologic toxicities included alopecia, anorexia, fatigue, nausea, vomiting, and weight loss. Conclusions:The maximum tolerated dose for this study recommended for phase II testing is gemcitabine 1000 mg/m² and CPT-11 100 mg/m². A partial response was seen in transitional cell carcinoma.     

Immunotherapy of advanced malignancy by direct gene transfer of an interleukin-2 DNA/DMRIE/DOPE lipid complex: phase I/II experience.

PURPOSE: We have completed a phase I study, followed by three phase I/II studies, in patients with metastatic melanoma, renal cell carcinoma (RCC), and sarcoma in order to evaluate the safety, toxicity, and antitumor activity of Leuvectin (Vical Inc, San Diego, CA), a gene transfer product containing a plasmid encoding human interleukin (IL)-2 formulated with the cationic lipid 1, 2-dimyristyloxypropyl-3-dimethyl-hydroxyethyl ammonium bromide/dioleyl-phosphatidyl-ethanolamine (DMRIE/DOPE) and administered intratumorally. PATIENTS AND METHODS: Twenty-four patients were treated in the phase I study. Leuvectin doses were 10 microg, 30 microg, or 300 microg weekly for 6 weeks. In three subsequent phase I/II studies, a total of 52 patients (18 with melanoma, 17 with RCC, and 17 with sarcoma) were treated with further escalating doses of Leuvectin: 300 microg twice a week for 3 weeks, 750 microg weekly for 6 weeks, and 1,500 microg weekly for 6 weeks. RESULTS: There were no drug-related grade 4 toxicities and only one grade 3 toxicity, but the majority of patients experienced mild constitutional symptoms after treatment. In the phase I/II studies, 45 patients were assessable for response (14 with RCC, 16 with melanoma, and 15 with sarcoma). Two patients with RCC and one with melanoma have achieved partial responses lasting from 16 to 19 months and continuing. In addition, two RCC, three melanoma, and six sarcoma patients had stable disease lasting from 3 to 18 months and continuing. The plasmid was detected by polymerase chain reaction assay in the posttreatment samples of 29 of 46 evaluated patients. Immunohistochemistry studies on serial biopsy specimens showed increased IL-2 expression and CD8(+) infiltration after treatment in the tumor samples of several patients (12 and 16, respectively). CONCLUSION: Direct intratumoral injection of Leuvectin is a safe and possibly effective immunotherapeutic approach in the treatment of certain tumor types.     

Use of transdermal amitriptyline gel in a patient with chronic pain and depression

A man with severe inflammatory bowel disease suffered from chronic abdominal pain and depression. A transdermal amitriptyline gel preparation was compounded since he was unable to take drugs orally Serum concentrations of amitriptyline and its active metabolite nortriptyline were measured over 24 hours. Symptoms of depression were monitored before starting transdermal therapy and at the end of 6 weeks. Pain symptoms and amitriptyline adverse drug events were monitored daily Steady-state serum concentrations of drug and metabolite were within the therapeutic range over 24 hours. The patient reported that his mood was improved but his abdominal pain remained unchanged. Transdermal amitriptyline gel was well tolerated and is an alternative delivery system in patients unable to take drugs orally.     

Semihistone protein A24 replaces H2A as an integral component of the nucleosome histone core.

The semihistone protein, A24, was shown to be a stable minor component of purified salt-washed nucleosome core particles. A24 was also shown to become integrated into nucleohistone during reconstitution in a manner characteristic of the core histones. Purified A24 in solution was shown to exhibit the same specificity of interaction with histone H2B as is exhibited by histone H2A. We conclude that A24 in chromatin replaces H2A as a stable integral component of certain nucleosome histone cores.