Carcinoid tumors of the lung: a diagnostic challenge in bronchial washings

Bronchial washings are used routinely in the diagnosis of lung tumors. However, unlike other tumors, the diagnosis of bronchial carcinoids on bronchial washings is difficult. We reviewed 17 cases of histologically proven bronchial carcinoids from the files of the cytology laboratory over a period of 15 yr (1986–2001). The bronchial washings and histology sections of all the cases were reviewed separately by two independent observers and the results tabulated. Two cases had inadequate bronchial washings for evaluation and were excluded from the study. A growth was identified on bronchoscopy in 13 of 15 cases. Initial cytologic diagnoses were ?adenocarcinoma/?carcinoid and suspicious of carcinoid in one case each. However, on review, tumor was identified in 10 of 13 cases initially considered to be negative. The possible reasons for a false‐negative report on initial cytology include the paucity of tumor cell fragments in the bronchial washings (5 of 12 cases showing only one to two tumor fragments) and their bland appearance, often being mistaken for benign columnar cells. This study highlights the potential pitfalls in the diagnosis of bronchial carcinoids on bronchial washings and underlines the importance of a diligent search in cases with high clinical suspicion and positive bronchoscopic findings. Diagn. Cytopathol. 2004;30:62–66. © 2004 Wiley‐Liss, Inc.     

Protective effect of adenosine against neuronal injury induced by middle cerebral artery occlusion in rats as evidenced by diffusion-weighted imaging

In the present study, adenosine, an inhibitory neuromodulator, was studied in male Wistar rats subjected to 2 h of transient middle cerebral artery (MCA) occlusion. Adenosine (500 mg/kg ip) was administered twice-once at the time of MCA occlusion and again at the time of reperfusion-and evaluated for its protective effect by using diffusion-weighted imaging (DWI) (30 min after reperfusion). After the DWI experiments, one group of animals was euthanized 2 h after reperfusion for the estimation of oxidative stress markers, while in another group, neurological deficit was assessed 24 h after MCA occlusion. In the adenosine-treated group, percent hemispheric lesion area (%HLA) in DWI was significantly attenuated (11.7+/-5.2) as compared to vehicle-treated group (21.4+/-4.7). The level of malondialdehyde (MDA) (301.8+/-22 nmol/g wet tissue) in the adenosine-treated group was significantly decreased as compared to that in the vehicle-treated MCA-occluded rats (420+/-20 nmol/g wet tissue). An insignificant change was observed in the levels of glutathione in both the vehicle-treated MCA-occluded and the adenosine-treated groups. The neurological deficit was significantly improved in the adenosine-treated group (1.8+/-0.06) as compared to the vehicle-treated (2.9+/-0.38) group. This is the first study to demonstrate the effectiveness of adenosine using DWI in the MCA-occluded rats.     

Inhibition of hepatic stellate cell collagen synthesis by N-(methylamino)isobutyric acid

The increased deposition of extracellular matrix by hepatic stellate cells following liver injury, in a process known as activation, is considered a key mechanism for increased collagen content of liver during the development of liver fibrosis. We report that N-(methylamino)isobutyric acid (MeAIB), a specific inhibitor of System A-mediated amino acid uptake, reduces the accumulation of collagen in CFSC-2G hepatic stellate cell cultures and in a rat model of liver injury and fibrosis. Rat CFSC-2G cells were cultured in 0-5mM MeAIB, and the accumulation and synthesis of collagen were measured by binding to Sirius red F3B and pulse-labeling with [3H]-proline, respectively. The effect of MeAIB on collagen accumulation in vivo was evaluated utilizing a rat model of hepatic fibrosis. MeAIB inhibited collagen accumulation in CFSC-2G cultures in a concentration-dependent manner with 5mM MeAIB reducing collagen 44.6+/-1.2% compared with the control. In CFSC-2G cultures, MeAIB selectively inhibited the incorporation of proline into cellular macromolecules by 43+/-4%, while the synthesis of proteins containing leucine was not affected. In vivo, oral administration of 160mg MeAIB/kg body weight per day to rats significantly reduced the hepatic collagen accumulation in response to 1 week of CCl(4)-induced liver injury. MeAIB reduces the accumulation of collagen in CFSC-2G hepatic stellate cell cultures and in a CCl(4)-induced rat model of liver injury and fibrosis.     

Clostridium difficile infections in HIV-positive patients

The prevalence of Clostridium difficile infections in HIV-positive patients with regard to the presence of its enterotoxin was investigated. Enzyme immunoassay (EIA, Meridian Diagnostic Inc) was used for the detection of C. difficile enterotoxin in stool specimens collected from 201 HIV-positive and 271 HIV-negative diarrheal patients. Culture was performed on cycloserine cefoxitin fructose agar. Chromosomal DNA types of C. difficile isolates were determined by pulsed-field gel electrophoresis (PFGE). In the HIV-positive group, C. difficile enterotoxin was found in 58.8% and 12.6% of diarrheal and non-diarrheal patients, repectively, whereas this toxin was found in 36.5% of HIV-negative-diarrheal patients. However, 13.6% of stool samples were negative by toxin assay, but were positive for C. difficile by culture and latex agglutination test. Among 11 isolates from both HIV-positive and HIV-negative patients, 6 patterns of PFGE type were observed: A, B, C, D, E and F.     

p53 mutations in bladder cancer: evidence for exogenous versus endogenous risk factors

Bladder cancer is associated with smoking, occupational exposures, and glutathione S-transferase (GST) M1 and N-acetyltransferase (NAT) 2 polymorphisms that may influence carcinogen metabolism, but somatic p53mutations are often CpG dinucleotide G:C-A:T transitions that can occur spontaneously. We conducted a case-control study to determine whether p53mutation characteristics might distinguish cases with environmental versus endogenous causes. p53exons 4-9 were amplified from 146 bladder tumors by PCR, screened by single-strand conformational polymorphism analysis, and sequenced. Thirty-one cases were p53-positive, and 112 were p53-negative (germ line or silent). G:C-A:T transitions were also subclassified as CpG or non-CpG. Cases and 215 clinic controls were interviewed. GSTM1, NAT1, and NAT2 polymorphisms were assayed from peripheral blood. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using logistic and polytomous regression. Case-control ORs for smoking, occupations, and NAT1*10genotype were similar for p53-positive and p53-negative cases. Associations with GSTM1-null and NAT2-slow genotypes were somewhat stronger for p53-positive [OR, 3.3; CI, 1.4-7.8 (GSTM1 null); OR, 1.8; CI, 0.8-4.0 (NAT2 slow)] than p53-negative cases [OR, 1.5; CI:0.9-2.3 (GSTM1 null); OR, 0.9; CI, 0.6-1.4 (NAT2 slow)]. Smoking was strongly associated with CpG G:C-A:T (OR, 15.3; CI:3.6-65) versus other G:C-A:T (OR, 1.8; CI, 0.3-9.8). NAT2 slow genotypes were also associated with CpG G:C-A:T (OR, 6.2; CI:0.7-52), whereas GSTM1 null was associated with non-CpG G:C-A:T (OR, 7.8; CI, 0.9-65). Associations were not substantially different for case subtypes defined by p53mutation status alone. Estimates for p53 subtypes were imprecise but support in vitro evidence that some CpG G:C-A:T transitions may be caused by smoking and other environmental mutagens.     

Inhibition of markers of hepatic stellate cell activation by the hormone relaxin

Hepatic fibrosis results from excess extracellular matrix produced primarily by hepatic stellate cells (HSC). In response to injury, HSC differentiate to a myofibroblastic phenotype expressing smooth muscle actin and fibrillar collagens. Relaxin is a polypeptide hormone shown to have antifibrotic effects in fibrosis models. In this study, activated HSC from rat liver were treated with relaxin to determine if relaxin can reverse markers of HSC activation. Relaxin treatment resulted in a decrease in the expression of smooth muscle actin, but had no effect on cell proliferation rate. The levels of total collagen and type I collagen were reduced, while the synthesis of new collagen was inhibited. Furthermore, relaxin caused an increase in the expression and secretion of rodent interstitial collagenase (MMP-13), but there was no effect on the gelatinases MMP-2 or MMP-9. Relaxin also increased secretion of TIMP-1 and TIMP-2. The effective concentration of relaxin to induce these effects was consistent with action through the relaxin receptor. In conclusion, relaxin reversed markers of the activated phenotype of HSC including the production of fibrillar collagen. At the same time, the activity of a fibrillar collagenase was increased. These data suggest that relaxin not only inhibits HSC properties that contribute to the progression of hepatic fibrosis, but also promotes the clearance of fibrillar collagen. Therefore, relaxin may be a useful approach in the treatment of hepatic fibrosis.