Human small cell lung cancer NYH cells selected for resistance to the bisdioxopiperazine topoisomerase II catalytic inhibitor ICRF-187 demonstrate a functional R162Q mutation in the Walker A consensus ATP binding domain of the alpha isoform.

Bisdioxopiperazine drugs such as ICRF-187 are catalytic inhibitors of DNA topoisomerase II, with at least two effects on the enzyme: namely, locking it in a closed-clamp form and inhibiting its ATPase activity. This is in contrast to topoisomerase II poisons as etoposide and amsacrine (m-AMSA), which act by stabilizing enzyme-DNA-drug complexes at a stage in which the DNA gate strand is cleaved and the protein is covalently attached to DNA. Human small cell lung cancer NYH cells selected for resistance to ICRF-187 (NYH/187) showed a 25% increase in topoisomerase IIalpha level and no change in expression of the beta isoform. Sequencing of the entire topoisomerase IIalpha cDNA from NYH/187 cells demonstrated a homozygous G-->A point mutation at nucleotide 485, leading to a R162Q conversion in the Walker A consensus ATP binding site (residues 161-165 in the alpha isoform), this being the first drug-selected mutation described at this site. Western blotting after incubation with ICRF-187 showed no depletion of the alpha isoform in NYH/187 cells in contrast to wild-type (wt) cells, whereas equal depletion of the beta isoform was observed in the two sublines. Alkaline elution assay demonstrated a lack of inhibition of etoposide-induced DNA single-stranded breaks in NYH/187 cells, whereas this inhibition was readily apparent in NYH cells. Site-directed mutagenesis in human topoisomerase IIalpha introduced into a yeast Saccharomyces cerevisiae strain with a temperature-conditional yeast TOP2 mutant demonstrated that R162Q conferred resistance to the bisdioxopiperazines ICRF-187 and -193 but not to etoposide or m-AMSA. Both etoposide and m-AMSA induced more DNA cleavage with purified R162Q enzyme than with the wt. The R162Q enzyme has a 20-25% decreased catalytic capacity compared to the wt and was almost inactive at <0.25 mM ATP compared to the wt. Kinetoplast DNA decatenation by the R162Q enzyme at 1 mM ATP was not resistant to ICRF-187 compared to wt, whereas it was clearly less sensitive than wt to ICRF-187 at low ATP concentrations. This suggests that it is a shift in the equilibrium to an open-clamp state in the enzyme's catalytic cycle caused by a decreased ATP binding by the mutated enzyme that is responsible for bisdioxopiperazine resistance.     

The role of fine-needle aspiration cytology in the evaluation of metastatic clear cell tumors

BACKGROUND: Clear cell tumors (CCTs) occur as primary neoplasms in a number of anatomic sites. Due to their overlapping morphologic features, these tumors can be challenging for the cytologist, particularly when they present as metastatic lesions. METHODS: Forty-nine fine-needle aspirations (FNA) of metastatic CCTs from 46 patients (age range, 29-87 years; mean, 64 years) were reviewed retrospectively. In addition to the routine smears and cell block preparations, ancillary studies were performed in selected cases. Clinical and/or histologic follow-up was obtained for all patients. RESULTS: The sites of the 49 FNAs were the lung (12 cases), lymph nodes (9 cases), liver (7 cases), bone (7 cases), soft tissue (4 cases), pelvis (2 cases), adrenal gland (2 cases), pancreas (1 case), thyroid (2 cases), peritoneum (2 cases), and vagina (1 case). Twenty-seven patients had a previous history of a CCT and the FNA material in these cases was consistent with a metastasis. The primary anatomic sites in these cases were the kidney (20 cases), ovary (2 cases), salivary gland (1 case), and cervix (1 case). On light microscopy, these tumors had a similar appearance and often were indistinguishable. Nineteen patients did not have a prior history of malignancy; 12 of these patients had a concurrent renal mass and the diagnosis of metastatic renal cell carcinoma was made. The anatomic site of origin of seven of the ten remaining tumors (kidney [2 cases], lung [2 cases], ovary [1 case], germ cell [1 case], and endometrium [1 case]) was established through immunocytochemical studies of cytologic material and clinical follow-up. CONCLUSIONS: FNA plays an important role in the diagnosis of metastatic CCT. Cytologic examination, ancillary studies, and clinical information can establish the anatomic site of origin in the majority (95%) of cases, precluding the necessity of obtaining additional tissue. Cancer (Cancer Cytopathol) Copyright 1999 American Cancer Society.     

Residential traffic noise and mammographic breast density in the Diet,Cancer, and Health cohort

Objectives

Traffic is the most important source of community noise, and it has been proposed to be associated with a range of disease outcomes, including breast cancer. As mammographic breast density (MD) is one of the strongest risk factors for developing breast cancer, the present study investigated whether there is an association between residential exposure to traffic noise and MD in a Danish cohort.

Methods

We included women with reproductive and lifestyle information available from the Diet, Cancer, and Health cohort, who also participated in the Copenhagen Mammography Screening Programme (n?=?5,260). Present and historical addresses from 1987 to 2011 were found in national registries, and traffic noise was modeled 5 years before mammogram. Analyses between residential traffic noise and MD were performed using logistic regression.

Results

We found no association between residential road and railway noise exposure 5 years before mammogram, and having a mixed/dense versus a fatty mammogram, and no interaction with menopausal status, BMI, HRT use, and railway noise exposure, for analyses on road traffic noise.

Conclusion

The present study does not suggest an association between residential traffic noise exposure and subsequent MD in a cohort of middle-aged Danish women.

     

Lack of pancreatic body and tail in HNF1B mutation carriers

Aims Hepatocyte nuclear factor 1B (HNF1B) gene mutation carriers have a systemic disease characterized by congenital malformations in the urogenital tract, diabetes mellitus of maturity‐onset diabetes of the young type and dysfunction of the liver and exocrine pancreas. We aimed to investigate pancreatic structure and exocrine function in carriers of HNF1B mutations. Methods We studied five subjects from two families with the previously reported mutation R137_K161del and the novel mutation F148L in HNF1B. All patients underwent computed tomography (CT) and magnetic resonance cholangiopancreatography (MRCP). We measured faecal elastase and serum vitamins D and E. Results One of the mutation carriers reported abdominal symptoms. All five subjects had faecal elastase deficiency, three had vitamin D deficiency and two had vitamin E deficiency. Neither CT nor MRCP depicted tissue corresponding to the pancreatic body and tail in the five mutation carriers, indicating agenesis of the dorsal pancreas. The head of the pancreas was slightly atrophic but had normal X‐ray attenuation at CT in all patients. Conclusions Agenesis of the pancreatic body and tail and pancreatic exocrine dysfunction are parts of the phenotype in HNF1B mutation carriers. This strengthens the evidence for a critical role of HNF1B in development and differentiation of at least the dorsal pancreas.     

Cunninghamella bertholletiae infection in a bone marrow transplant patient: amphotericin lung penetration, MIC determinations, and review of the literature.

Infections caused by Cunninghamella bertholletiae, an opportunistic fungal organism, have an extremely high mortality rate. A fatal case of C. bertholletiae fungal pneumonia occurred in a man who had received an allogeneic bone marrow transplant. Aggressive debridement and high-dose liposomal amphotericin B failed to eradicate the infection. Right lung tissue samples obtained during lobectomy were assayed for amphotericin B concentrations by high-performance liquid chromatography, and minimum inhibitory concentration (MIC) determinations of amphotericin B against C. bertholletiae were determined by the macrobroth dilution method. The MIC for the isolate of C. bertholletiae was 4 microg/ml. Amphotericin B lung concentrations averaged 9.5 microg/ml (range 3.7-13.8 microg/ml), with a corresponding serum trough concentration of 0.9 microg/ml. To our knowledge, this is the first reported case of amphotericin B concentrations measured at the site of infection in a patient with a pulmonary Cunninghamella infection, together with a corresponding MIC of the organism. The patient's death, which occurred despite aggressive debridement and high amphotericin B lung concentrations, highlights the need for novel strategies to treat infections caused by invasive molds such as C. bertholletiae.     

Nephrotoxicity of cyclosporin A in patients with newly diagnosed type 1 diabetes mellitus

Renal function was studied in 18 patients with Type 1 diabetes mellitus. All were participating in the Canadian-European randomized placebo-controlled cyclosporin trial in newly diagnosed Type 1 diabetic patients, nine being randomized to placebo, and nine to cyclosporin A. During treatment for 12 to 18 months, cyclosporin A caused significant reductions in the glomerular filtration rate (before drug withdrawal, cyclosporin 97 +/- 18 vs placebo 125 +/- 16 ml min-1 1.73-m-2, p less than 0.05), renal plasma flow (454 +/- 83 vs 536 +/- 70 ml min-1 1.73-m-2, p less than 0.05), and lithium clearance (17 +/- 3 vs 28 +/- 5 ml min-1 1.73-m-2, p less than 0.05). The fractional proximal reabsorption was increased (0.82 +/- 0.03 vs 0.78 +/- 0.03, p less than 0.05), and the fractional distal sodium reabsorption reduced (0.88 +/- 0.03 vs 0.94 +/- 0.02, p less than 0.05). These results are in accordance with the hypothesis that the nephrotoxic effect of cyclosporin A results from a preferential constriction of afferent glomerular vessels. One year after withdrawal of the drug, all variables were similar in the two groups, except for blood glucose control which was worse in the cyclosporin A treated group. When corrected for differences in blood glucose control it appeared that in three out of nine patients glomerular filtration rate had not completely returned to the reference range of the placebo group. We conclude that the nephrotoxic side-effects of cyclosporin A treatment for 1 year are reversible. There are, however, signs of minor and perhaps chronic renal injury.     

Painful, nonhealing, tooth extraction socket

The oral signs and symptoms of acute leukemia include gingival hypertrophy, gingival hemorrhage, petechiae and ecchymoses, mucosal ulceration, paresthesia, tooth pain, and oral infections. This case was unusual in that the diagnosis of acute leukemia was made only after a biopsy specimen of tissue protruding from the socket of a recently extracted tooth disclosed malignant cells suggestive of leukemia or lymphoma. The diagnosis of epulis granulomatosa should be made only after clinical, radiographic, and microscopic studies have been performed to rule out a malignant process.     

Acquired diverticulosis of the small intestine: case reports and literature review

Eleven cases of small bowel diverticulosis are discussed. Four patients presented with perforation, five with other symptoms attributable to this condition and in two patients diverticulosis was considered an incidental finding. A review of the literature suggests that small bowel diverticulosis may be: present in up to 1.3% of the population; associated with symptoms in approximately 50% of patients, and associated with acute surgical complications in 10% of patients. This may be a disorder of intestinal motility associated with colonic diverticulosis and related to other disorders of smooth muscle and myenteric plexus. Small bowel diverticulosis should not be regarded as a rare, incidental and inconsequential finding.     

Interobserver variation in the radiographic diagnosis of duodenal ulcer disease

Interobserver variation was estimated in radiologists' detection of duodenal ulcer and deformity of the duodenal bulb. A consecutive series of 156 patients with upper abdominal pain had a double contrast barium examination. The films were first read by routine by a specialist in radiology. A second reading was performed by another specialist who did not know the result of the first reading. Overall agreement between the two readings in the detection of duodenal ulcer and deformity of the duodenal bulb was 0.91 and 0.94, respectively. The overall agreement was adjusted for the expected chance agreement and kappa values were calculated. Kappa was 0.63 for the detection of duodenal ulcer and 0.85 for the detection of deformity of the duodenal bulb.     

Clinically-oriented hints for the technic of integrated dosage modification in a partial irradiation field

The authors present a method allowing the application of doses differing from the reference dose in partial zones of an irradiation volume. The compensator-integrated modification is performed by a simultaneous dose harmonization in the reference plane. Three methods are described by which a topographic and anatomic assignment of field and partial field is possible on the basis of the irradiation features. The optimum application of each method in the individual irradiation schemes is discussed.