DNA in rat hepatocyte nuclei: effects of treatment with low levels of carbon tetrachloride and (or) chlordane

1979 ◽  
Vol 57 (5) ◽  
pp. 1003-1009 ◽  
Author(s):  
D. C. Mahon ◽  
K. K. Nair ◽  
P. C. Oloffs
Keyword(s):  
Liver Cirrhosis ◽  
Carbon Tetrachloride ◽  
Dna Content ◽  
Hepatic Dysfunction ◽  
Chromatin Condensation ◽  
Nuclear Size ◽  
Rat Hepatocyte ◽  
Highly Sensitive ◽  
Dna Contents ◽  
Low Levels

The effects of low-level, chronic feeding of the insecticide chlordane on the cytochemistry of the hepatocyte nuclei of male nits, with or without liver cirrhosis, were determined. Chlordane was given orally at 0.1 mg kg−1 day−1. Cirrhosis was induced by exposure of the animals to carbon tetrachloride (CCl4) vapours twice weekly for 10 weeks. The animals were killed after 20 weeks and the Feulgen-stained hepatocyte nuclei examined microspectrophotometrically for nuclear size, DNA content, and the degree of chromatin condensation. The five experimental groups were: (A) control; (B) chlordane only for 20 weeks; (C) cirrhosis only (CCl4 weeks 0–10); (D) chlordane (weeks 0–20) plus cirrhosis; (E) cirrhosis (CCl4, weeks 0–10) followed by chlordane (weeks 10–20). All treatments significantly (p < 0.01) increased nuclear size. Although DNA contents were not affected by chlordane alone, in cirrhotic animals treatment with chlordane caused significantly higher DNA contents than that in cirrhotics not receiving chlordane. This suggests that hepatic dysfunction increases the liver's sensitivity to respond to chlordane. Chromatin condensation was decreased by chlordane in both healthy and cirrhotic animals; animals with cirrhosis, but not receiving chlordane, had more condensed chromatin than the controls.The results of this study show that computer analysis of images of Feulgen-stained nuclei is a highly sensitive technique to detect the effects of chemicals on the degree of chromatin condensation even at dosage levels below the no-effect levels previously reported.

scholarly journals Bioanalytical screening of low levels of dioxins and dioxin-like PCBs in pig meat (pork) for checking compliance with EU maximum and action levels using highly sensitive “third generation” recombinant H4L7.5c2 rat hepatoma cells

2021 ◽  
Vol 33 (1) ◽  
Author(s):  
Johannes Haedrich ◽  
Claudia Stumpf ◽  
Michael S. Denison
Keyword(s):  
European Union ◽  
Hepatoma Cell ◽  
Animal Origin ◽  
The European Union ◽  
Highly Sensitive ◽  
Commission Regulation ◽  
Pig Meat ◽  
Low Levels ◽  
Action Levels ◽  
Rat Hepatoma

Abstract Background Low maximum and action levels set by the European Union for polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (DL-PCBs) in pig meat (pork) have led to a demand for reliable and cost-effective bioanalytical screening methods implemented upstream of gas chromatography/high-resolution mass spectrometry confirmatory technology, that can detect low levels of contamination in EU-regulated foods with quick turn-around times. Results Based on the Chemically Activated LUciferase gene eXpression (CALUX) bioassay, extraction and clean-up steps were optimized for recovery and reproducibility within working ranges significantly lower than in current bioassays. A highly sensitive “3rd generation” recombinant rat hepatoma cell line (H4L7.5c2) containing 20 dioxin responsive elements was exposed to pork sample extracts, and their PCDD/Fs and DL-PCBs levels were evaluated by measuring luciferase activity. The method was validated according to the provisions of Commission Regulation (EU) 2017/644 of 5 April 2017 with spiking experiments performed selectively for PCDD/Fs and DL-PCBs and individual calibration for PCDD/Fs, DL-PCBs and the calculated sum of PCDD/Fs and DL-PCBs. The resulting performance parameters met all legal specifications as confirmed by re-calibration using authentic samples. Cut-off concentrations for assessing compliance with low maximum levels and action levels set for PCDD/Fs and DL-PCBs within a range of 0.50–1.25 pg WHO-TEQ/g fat were derived, ensuring low rates of false-compliant results (ß-error < 1%) and keeping the rate of false-noncompliant results well under control (α-error < 12%). Conclusions We present a fast and efficient bioanalytical routine method validated according to the European Union’s legal requirements on the basis of authentic samples, allowing the analyst to reliably identify pork samples and any other EU-regulated foods of animal origin suspected to be noncompliant with a high level of performance and turn-around times of 52 h. This was facilitated in particular by a quick and efficient extraction step followed by selective clean-up, use of a highly sensitive “3rd generation” H4L7.5c2 recombinant rat hepatoma cell CALUX bioassay, and optimized assay performance with improved calibrator precision and reduced lack-of-fit errors. New restrictions are proposed for the calibrator bias and the unspecific background contribution to reportable results. The procedure can utilize comparably small sample amounts and allows an annual throughput of 840–1000 samples per lab technician. The described bioanalytical method contributes to the European Commission's objective of generating accurate and reproducible analytical results according to Commission Regulation (EU) 2017/644 across the European Union.

Increased serum levels of quinolinic acid indicate enhanced severity of hepatic dysfunction in patients with liver cirrhosis

2013 ◽  
Vol 74 (1) ◽  
pp. 60-66 ◽  
Author(s):  
Imad Lahdou ◽  
Mahmoud Sadeghi ◽  
Hani Oweira ◽  
Gerhard Fusch ◽  
Volker Daniel ◽  
...  
Keyword(s):  
Liver Cirrhosis ◽  
Quinolinic Acid ◽  
Hepatic Dysfunction ◽  
Serum Levels

Serum Phospholipid Fatty Acid Composition in Cystic Fibrosis Patients with and without Liver Cirrhosis

2017 ◽  
Vol 71 (1-2) ◽  
pp. 91-98 ◽  
Author(s):  
Sławomira Drzymała-Czyż ◽  
Mariusz Szczepanik ◽  
Patrycja Krzyżanowska ◽  
Monika Duś-Żuchowska ◽  
Andrzej Pogorzelski ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Liver Cirrhosis ◽  
Fatty Acid ◽  
Phospholipid Fatty Acid ◽  
Body Height ◽  
Gas Chromatography Mass Spectrometry ◽  
Phospholipid Fatty Acid Composition ◽  
Total N ◽  
Z Scores ◽  
Low Levels

Background/Aims: Cystic fibrosis (CF) liver disease is the third most frequent cause of death in CF patients. Although it alters fatty acid (FA) metabolism, data concerning the profile of FA in CF patients with liver cirrhosis is lacking. This study aimed to assess the FA composition of serum phospholipids in CF patients with and without liver cirrhosis. Methods: The study comprised 25 CF patients with liver cirrhosis and 25 without it. We assessed Z-scores for body height and weight, lung function, exocrine pancreatic sufficiency and colonization with Pseudomonas aeruginosa. FAs' profile of serum glycerophospholipids was quantified by gas chromatography mass spectrometry. Results: In CF patients with liver cirrhosis, the levels of C16:0 were higher and the amounts of C20:2n-6, C20:3n-6, C20:4n-6, and all the n-3 polyunsaturated FAs (PUFAs) (C18:3n-3, C20:5n-3, C22:5n-3, C22:6n-3) were lower than those in CF subjects without liver cirrhosis. The n-6/n-3, C20:4n-6/C18:2n-6, total n-6/C18:2n-6, C20:5n-3/C18:3n-3 and total n-3/C18:3n-3 ratios did not differ between the 2 groups. Conclusions: Liver cirrhosis may associate with profound abnormalities in the composition of serum glycerophospholipids FAs in CF patients. None of the analyzed clinical factors could explain the greater prevalence of low levels of PUFAs in this CF subgroup.

EFFECTS OF NATURAL AND RECOMBINANT INTERFERON ON CARBON TETRACHLORIDE- INDUCED LIVER CIRRHOSIS

2001 ◽  
Vol 10 (2) ◽  
pp. 11-17
Author(s):  
Mohamed Hassan ◽  
Mona El-Azab ◽  
EI-Sayed El-Awady ◽  
Soad Abou-El-Ela
Keyword(s):  
Liver Cirrhosis ◽  
Carbon Tetrachloride ◽  
Recombinant Interferon

scholarly journals Genome Size Evaluations in Cockroaches: New Entries

2022 ◽  
Author(s):  
Manuela Monti ◽  
Carlo Alberto Redi ◽  
Ernesto Capanna
Keyword(s):  
Genome Size ◽  
Dna Content ◽  
Periplaneta Americana ◽  
Field Trapping ◽  
American Cockroach ◽  
Neural Cells ◽  
Blatta Orientalis ◽  
Leucophaea Maderae ◽  
Sperm Dna ◽  
Dna Contents

Abstract Background: Ten years ago the main Genome Size (GS) database contained records for 830 insects; although this number has now nearly doubled, 1645 (Gregory 2011 vs Gregory 2021 databases), the paucity of records highlights both the difficulty of animal field trapping and the time-consuming laboratory techniques to evaluate them. Thus, new entries are necessary to reach a satisfactory GS panorama for cockroaches. Results: We report GS values for nine cockroaches (order Blattodea, families Blattidae, Blaberidae and Ectobiidae, ex Blattelidae,), three of which are original additions to the ten already present in the GS database: the death’s head roach (Blaberus craniifer), the Surinam cockroach (Pycnoscelus surinamensis) and the Madeira cockroach (Leucophaea maderae). Three of our values confirm the existing data for the German (Blattella germanica), the oriental (Blatta orientalis) and the giant Mexican (Blabera fusca) cockroaches. Regarding the GS of the American cockroach (Periplaneta americana) the GS database contains two contrasting values (2.72 vs 3.41 pg). We suggest that the 2.72 pg value is likely to be the correct GS as it strikingly similar to our sperm DNA content evaluation (2.80 ± 0.11 pg). Finally, we suggest halving the published GS of the Argentine cockroach Blaptica dubia and the spotted cockroach (the gray cockroach) Nauphoeta cinerea as our estimates come from the evaluation of the sperm DNA content. The data already reported in the literature are based on DNA contents of neural cells (likely polyploid) obtained by grinding entire heads of animals.Conclusions: Although the paucity of the GS data does not allow firm considerations on the possible evolutionary role played by the GS in diversifying cockroach species, we offer two speculative hypotheses that need to be validated by increasing the available GS records: (i) the occurrence of a correlation between increasing 2N chromosome number and GS within the order Blattodea; and (ii) the possible occurrence of a polyploidization phenomenon doubling a basic GS of 0.58 pg of some termite families (superfamily Blattoidea, epifamily Termitoidae) up to the maximum GS value of 3.24 for the Blaberidae family within the order Blattodea (super-order Dictyoptera).

scholarly journals Splenomegaly and thrombocytopenia in patients with liver cirrhosis

2010 ◽  
Vol 67 (2) ◽  
pp. 166-169 ◽  
Author(s):  
Jelena Djordjevic ◽  
Petar Svorcan ◽  
Dusica Vrinic ◽  
Branka Dapcevic
Keyword(s):  
Risk Factors ◽  
Liver Cirrhosis ◽  
Platelet Count ◽  
Portal Hypertension ◽  
Hepatic Dysfunction ◽  
Spleen Size ◽  
Hepatitis Viruses ◽  
Decompensated Liver Cirrhosis ◽  
Splenic Sequestration ◽  
Frequent Finding

Backgroud/Aim. Splenomegaly is a frequent finding in patients with liver cirrhosis and portal hypertension and may cause hypersplenism. The occurrence of thrombocytopenia in those patients can be considered as an event with multiple etiologies. Two mechanisms may act alone or synergistically with splenic sequestration. One is central which involves either myelosuppression because of hepatitis viruses or the toxic effects of alcohol abuse on the bone marrow. The second one involves the presence of antibodies against platelets. It also depends upon the stage and etiology of liver disease. The aim of the study was to investigate a correlation between the platelet count and spleen size and the risk factors for thrombocytopenia in patients with liver cirrhosis. Methods. We studied 40 patients with decompensated liver cirrhosis who were hospitalized in the Department of Gastroenterohepatology. The liver function was graded according to Child Pugh score. Spleen size was defined ultrasonografically on the basis of craniocaudal length. Suspicion of portal hypertension was present when longitudinal spleen length was more than 11 cm. Thrombocytopenia was determined by platelet count under 150 000/mL. Results. We did not find any significant correlation between hepatic dysfunction and spleen size (p = 0.9), and between hepatic dysfunction and thrombocytopenia (p = 0.17). Our study did not find any significant correlation between spleen size and peripheral platelet count (p = 0.5), but we found a significant correlation between thrombocytopenia and etiology of cirrhosis - decreased platelet count was more common among patients with cirrhosis of alcoholic etiology than in other etiologies of cirrhosis (p = 0.001). Conclusion. According to our study, liver cirrhosis, portal hypertension and thrombocytopenia could be present even in the absence of enlarged spleen suggesting the involvement of other mechanisms of decreasing platelet account.

scholarly journals Mechanical phenotyping of acute promyelocytic leukemia reveals unique biomechanical responses in retinoic acid-resistant populations

2021 ◽  
Author(s):  
Brian Li ◽  
Annie Maslan ◽  
Aaron M Streets ◽  
Lydia L. Sohn
Keyword(s):  
Retinoic Acid ◽  
Acute Promyelocytic Leukemia ◽  
Dna Content ◽  
Trichostatin A ◽  
Chromatin Condensation ◽  
Promyelocytic Leukemia ◽  
Mechanical Responses ◽  
Nuclear Mechanics ◽  
All Trans Retinoic Acid ◽  
M Phase

While all-trans retinoic acid (ATRA) is an essential therapy in the treatment of acute promyelocytic leukemia (APL), an aggressive subtype of acute myeloid leukemia, nearly 20% of APL patients are resistant to ATRA. As no biomarkers for ATRA resistance yet exist, we investigated whether cell mechanics could be associated with this pathological phenotype. Using mechano-node-pore sensing, a single-cell mechanical phenotyping platform, and patient-derived APL cell lines, NB4 (ATRA-sensitive) and AP-1060 (ATRA-resistant), we discovered that ATRA-resistant APL cells are less mechanically pliable. By investigating how different subcellular components of APL cells contribute to whole-cell mechanical phenotype, we determined that nuclear mechanics strongly influence APL cell mechanical responses. By arresting APL cells in S-phase or M-phase in the cell cycle, we found cell pliability to be inversely related to DNA content. In addition to DNA content affecting cell pliability, we observed that chromatin condensation also affects nuclear mechanics: decondensing chromatin with trichostatin A is especially effective in softening ATRA-resistant APL cells. RNA-Seq allowed us to compare the transcriptomic differences between ATRA-resistant and ATRA-responsive APL cells and highlighted gene expression changes that could be associated with mechanical changes. Overall, we demonstrate the potential of physical biomarkers in identifying APL resistance.

scholarly journals Hepatic Encephalopathy: From the Pathogenesis to the New Treatments

2014 ◽  
Vol 2014 ◽  
pp. 1-16 ◽  
Author(s):  
Juan Cordoba
Keyword(s):  
Clinical Trials ◽  
Liver Cirrhosis ◽  
Hepatic Encephalopathy ◽  
Prospective Studies ◽  
Hepatic Dysfunction ◽  
Short Term ◽  
Multiple Factors ◽  
Short Term Mortality ◽  
Cirrhotic Patients ◽  
New Treatments

Hepatic encephalopathy is a frequent and serious complication of liver cirrhosis; the pathophysiology of this complication is not fully understood although great efforts have been made during the last years. There are few prospective studies on the epidemiology of this complication; however, it is known that it confers with high short-term mortality. Hepatic encephalopathy has been classified into different groups depending on the degree of hepatic dysfunction, the presence of portal-systemic shunts, and the number of episodes. Due to the large clinical spectra of overt EH and the complexity of cirrhotic patients, it is very difficult to perform quality clinical trials for assessing the efficacy of the treatments proposed. The physiopathology, clinical manifestation, and the treatment of HE is a challenge because of the multiple factors that converge and coexist in an episode of overt HE.

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