Alcohol metabolism in human cells causes DNA damage and activates the Fanconi anemia-breast cancer susceptibility (FA-BRCA) DNA damage response network

Background: We recently reported that exposure of human cells in vitro to acetaldehyde resulted in the activation of the Fanconi anemia–breast cancer susceptibility (FA‐BRCA) DNA damage response network. Methods: To determine whether intracellular generation of acetaldehyde from ethanol metabolism can cause DNA damage and activate the FA‐BRCA network, we engineered HeLa cells to metabolize alcohol by expression of human alcohol dehydrogenase (ADH) 1B. Results: Incubation of HeLa‐ADH1B cells with ethanol (20 mM) resulted in acetaldehyde accumulation in the media, which was prevented by co‐incubation with 4‐methyl pyrazole (4‐MP), a specific inhibitor of ADH. Ethanol treatment of HeLa‐ADH1B cells produced a 4‐fold increase in the acetaldehyde–DNA adduct and N²‐ethylidene‐dGuo and also resulted in the activation of the FA‐BRCA DNA damage response network, as indicated by a monoubiquitination of FANCD2 and phosphorylation of BRCA1. Ser 1524 was identified as 1 site of BRCA1 phosphorylation. The increased levels of DNA adducts, FANCD2 monoubiquitination, and BRCA1 phosphorylation were all blocked by 4‐MP, indicating that acetaldehyde, rather than ethanol itself, was responsible for all 3 responses. Importantly, the ethanol concentration we used is within the range that can be attained in the human body during social drinking. Conclusions: Our results indicate that intracellular metabolism of ethanol to acetaldehyde results in DNA damage, which activates the FA‐BRCA DNA damage response network.     

Increased MCP-1 and MIP-1beta in bronchoalveolar lavage fluid of chronic bronchitics.

CC-chemokines are chemotactic factors expressed in a wide range of cell types and tissues. The aim of this study was to evaluate the involvement of CC-chemokines in the airways inflammation of patients affected by chronic bronchitis. The study evaluated, with an immunoassay, the concentrations of monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-1alpha (MIP-1alpha) and macrophage inflammatory protein-1beta (MIP-1beta), in the bronchoalveolar lavage fluid (BALF) of 12 smokers affected by chronic bronchitis and 14 smoking, 15 nonsmoking and six exsmoking healthy subjects. MCP-1 was significantly increased in patients with chronic bronchitis ((mean+/-SD) 10.75+/-4.04 pg x mL(-1)) and in the smoker control group (12.39+/-5.87 pg x mL(-1)) compared with healthy exsmokers: (7.12+/-1.60 pg x mL(-1), p=0.035 and p=0.045, respectively) and nonsmokers (6.41+/-3.87 pg x mL(-1), p=0.003 and p=0.006, respectively). MIP-1alpha concentrations were undetectable. A significant difference was observed in MIP-1-beta levels in BALF of chronic bronchitics (8.11+/-5.97 pg x mL(-1)) compared to smoker (3.57+/-2.90 pg x mL(-1), p=0.018), exsmoker (3.43+/-0.68 pg x mL(-1), p=0.025) and nonsmoker (3.39+/-3.73 pg x mL(-1), p=0.008) control groups. A negative correlation was observed between MIP-1beta levels and forced expiratory volume in one second values (p=-0.64, p=0.035) in chronic bronchitics. An increase of monocyte chemotactic protein-1 is related to smoking habit and seems consistent with a lung inflammatory reaction. On the contrary, an increase in macrophage inflammatory protein-1beta levels is restricted to smokers developing chronic obstructive pulmonary disease. These data suggest a role of CC-chemokines in the pathogenesis of chronic bronchitis.     

Decreased T lymphocyte infiltration in bronchial biopsies of subjects with severe chronic obstructive pulmonary disease

BACKGROUND: Studies on the inflammatory process in the large airways of patients with mild/moderate COPD have shown a prevalent T lymphocyte and macrophage infiltration of the bronchial mucosa. However, bronchial inflammation in more severe disease has not been extensively studied. OBJECTIVE: The aim of the present study was to characterize the lymphocyte infiltration in the bronchial mucosa of subjects with severe, compared to mild, COPD, and to examine the relationship between airflow limitation and T lymphocyte numbers in the bronchial mucosa. METHODS: We examined bronchial biopsies obtained from nine smokers with severe airflow limitation, nine smokers with mild/moderate airflow limitation and 14 smokers with normal lung function. Immunohistochemical methods on cryostat sections were used to assess the number of CD3+, CD4+, CD8+ cells and the number of CD3+ cells coexpressing the chemokine receptor CCR5 (CCR5+CD3+) in the subepithelium. RESULTS: Subjects with severe COPD had lower numbers of CD3+, CD8+ and CCR5+CD3+ cells than mild/moderate COPD (P < 0.012, P < 0.02 and P < 0.02, respectively) and control smokers (P < 0.015, P < 0.005 and P < 0.015, respectively). In subjects with airflow limitation the number of CD3+ and CD8+ cells was inversely correlated with the degree of airway obstruction (r = 0.59, P < 0.015 and r = 0.52, P < 0.032, respectively). CONCLUSIONS: Bronchial inflammation in severe COPD is characterized by lower numbers of CD3+ and CD8+ cells and decreased numbers of CD3+ cells coexpressing the chemokine receptor CCR5. T lymphocyte infiltration is inversely correlated with the degree of airflow limitation.     

Analysis of a series of cases of liver metastasis of colorectal carcinoma

The Authors present a six years case-report concerning 135 patients operated on colo-rectal carcinoma (82 of them are still followed up). The incidence of hepatic metastases (synchronous and metachronous) has been 30.37% in all. The preoperative ultrasonography has shown an 11% about margin mistake to visualize secondary lesions of liver. They have acknowledged the intraoperative ultrasonography very useful to visualize occult metastases and to guide an exeresis operation. The resectability of hepatic metastases from colo-rectal cancer has been of 12.19%. Synchronous forms have been rarely operable (5.88%) because they are constituted by multiple and disseminated lesions. They have found an higher incidence of operable forms among the metachronous metastases (42.85%), whose average time of appearance from the primary operation has been about 13 months. The operative mortality and the complications following hepatectomy have been null. The average survival time from operation has been over 22 months. Therefore they recognized the importance of a careful follow-up to find precociously operable lesions.     

Increased circulating Interleukin-18 levels in centenarians with no signs of vascular disease: another paradox of longevity?

Interleukin (IL)-18 is highly expressed in macrophages from human atherosclerotic plaques, suggesting its involvement in ischemic syndromes. We evaluated IL-18 and IL-18 binding protein (BP) in healthy centenarians, as longevity is characterized by a reduced incidence of ischemic events. For comparison, patients with chronic ischemic syndromes (CIS) were evaluated. Serum IL-18 and IL-18BP levels were measured by non-cross-reacting ELISA in 16 healthy centenarians and in two age-control populations, each of 18 healthy individuals aged 55.9+/-1.43 and 74.3+/-1.35, respectively, as well as in 23 CIS patients, and another cohort of 23 healthy subjects that were age- and sex-matched with CIS patients. Centenarians displayed significantly higher total IL-18 serum levels compared to each control group. Elevated IL-18 levels were also present in CIS patients. However, centenarians had a significant higher level of IL-18BP compared to the cohort of 23 controls (P=0.0014), and compared to CIS patients (P=0.043); as a result centenarians exhibited a lower level of free IL-18 than CIS patients. The present results indicate that quenching of IL-18 by IL-18BP may explain the apparent paradox of elevated serum IL-18 with no vascular signs in centenarians.     

Lower platelet count in healthy centenarians correlates with dispersion of the QT interval

BACKGROUND AND AIMS: Centenarians display a lower incidence of vascular ischemic events. A high platelet count and increased QT dispersion (QTd) represents a risk factor for cardiovascular events. The aim of this study was to evaluate platelet count and QTd in healthy centenarians and to establish correlations between these two indices. METHODS: 16 healthy centenarians (4 males, 12 females, range 100-105 years) living in a municipality of Eastern Sicily, and 40 healthy control subjects, divided into two groups: group A (N=20), age range 45-65 years, 7 males, 13 females; and group B (N=20), range 65-85 years, 6 males, 14 females, were examined. Platelets were counted using a blood analyzer and QTd was measured in standard 12-lead electrocardiograms. Differences in platelet count were assessed by one-way analysis of variance (ANOVA) and the Bonferroni test. Correlation coefficients between platelet count and QTd were calculated with the Spearman rank test. RESULTS: Centenarians showed a lower platelet count compared with controls, which was significant with respect to older controls, group B (p<0.05). QTd values did not significantly differ between centenarians and controls. A significant correlation between QTd and platelet count was evident in centenarians but not in controls. This correlation became evident in control subjects with a platelet count < or = 240,000/mm3. CONCLUSIONS: We hypothesize that a reduced number of platelets and the maintenance of normal QTd may contribute to extreme longevity and protect centenarians from cardiovascular events.