Aims: (i) To characterize serum levels of pro/anti-inflammatory cytokines in non-cirrhotics with ... more Aims: (i) To characterize serum levels of pro/anti-inflammatory cytokines in non-cirrhotics with hepatitis C; (ii) to correlate levels of these cytokines with degree of disease at baseline; and (iii) to characterize the immuno-modulatory effects of therapy with response. Methods: We studied 103 patients that were part of randomized, controlled, clinical trials. Serum cytokines were measured using enzyme-linked immunosorbent assay. Results: Using standard therapy in the presence and absence of ribavirin, the sustained responders had lower baseline tumor necrosis alpha (TNF-alpha) levels as compared to relapsed responders and non-responders. In patients receiving pegylated therapy, the degree of inflammation as determined by histology was paralleled by high TNF-alpha levels at baseline. In pegylated combination therapy with high dose ribavirin, lower levels of TNF-alpha, transforming growth factor beta (TGF-beta) and fibrosis scores were seen when comparing baseline with follow up. In sustained responders, regardless of therapy, the histological activity scores were lower at follow up as compared to baseline. Conclusions: Pegylated combination therapy reduces and sustains TNF-alpha levels and liver inflammation as shown by the histological activity index. In addition, it is able to reduce fibrosis as judged both by TGF-beta levels and fibrosis scores as compared to standard therapy.
In vitro, ethanol in combination with acetaminophen induces hepatocyte apoptosis resembling immun... more In vitro, ethanol in combination with acetaminophen induces hepatocyte apoptosis resembling immune-mediated fulminant hepatic failure in human beings. Intracellular pathways originating at the mitochondria are linked to apoptosis. I studied ethanol-induced apoptosis and hepatocytotoxicity after using an in vitro model of normal human primary hepatocytes that were exposed to 5 or 10 mM acetaminophen, 40 or 100 mM ethanol, 40 mM ethanol + 5 mM acetaminophen, or 40 mM ethanol + 10 mM acetaminophen, or nonexposed (control; plain medium). Transmission electron microscopy was performed at different time points after exposure to the various treatments. Apoptosis, as assessed by transmission electron microscopy, was increased in a time-dependent manner after exposure to ethanol + acetaminophen. In the ethanol + acetaminophen model, mitochondrial injury was associated with apoptosis of hepatocytes. Ultrastructural damage and induction of apoptosis were seen in response to N-acetyl-benzoquinone-imine plus ethanol, supporting the suggestion that the damage was due to the active metabolite of acetaminophen. The modulation of mitochondrial damage in vitro may have implications for the development of new therapeutic strategies to prevent apoptosis.
MG Neuman, GG Katz, IM Malkiewicz, P Mathurin, Hidekazu Tsukamoto, Masayuki Adachi, Hiromasa Ishi... more MG Neuman, GG Katz, IM Malkiewicz, P Mathurin, Hidekazu Tsukamoto, Masayuki Adachi, Hiromasa Ishii, A Colell, C Garcia-Ruiz, JC Fernandez-Checa, CA Casey Alcohol 28:22, 117-128, 10/2002. X 24180 Social poisons & drug abuse.
Ethanol is commonly used in cosmetic and pharmaceutical preparations. The present study aimed to ... more Ethanol is commonly used in cosmetic and pharmaceutical preparations. The present study aimed to assess ethanol-induced apoptosis and the possible repair by hyaluronic acid (HA) in vitro. In addition we aimed to determine the modulation of tumor necrosis factor (TNF-alpha) and interferon (IFN-alpha). We treated human A431 epidermoid skin cells and mouse fibroblasts with two concentrations of ethanol for 24h. HA obtained from umbilical cord excision was used at three concentration levels (2%, 4% and 8%) to determine its efficacy in the treatment. We measured cytotoxicity, TNF-alpha and IFN-alpha and visualized the cultures by electron microscopy. Treatment of cells with ethanol at 50mM and 100mM increased both the percentage of cells undergoing apoptosis, as well as the release of TNF-alpha into the culture medium. Ethanol may induce apoptosis in skin cells by enhancing the effects of TNF-alpha. HA in the 2% and 4% concentrations reduced TNF-alpha and morphological inflammation both in human A431 epidermoid skin cells and in mouse fibroblasts.
The necessity of maintaining a close balance between cell birth and cell death has caused multice... more The necessity of maintaining a close balance between cell birth and cell death has caused multicellular organisms to evolve into unforgiving societies where cells that are no longer needed are requested to commit suicide.
Ibuprofen is a widely used antipyretic and analgesic nonsteroidal antiinflammatory drug (NSAID). ... more Ibuprofen is a widely used antipyretic and analgesic nonsteroidal antiinflammatory drug (NSAID). With the aging of the population, there will be a significant increase in the prevalence of painful degenerative and inflammatory rheumatic conditions. This increase likely will lead to a parallel increase in the use of NSAIDs, including ibuprofen. The primary effect of the NSAIDs is to inhibit cyclooxygenase (prostaglandin synthase), thereby impairing the ultimate transformation of arachidonic acid to prostaglandins, prostacyclin, and thromboxanes. Although in the majority of cases it is safe, this NSAID, ibuprofen, can produce an unpredictable, idiosyncratic, type B reaction that may pose a major concern in clinical practice. Type B reactions are known to occur in susceptible individuals. The true hypersensitivity reaction (HSR) is a systemic disease defined by the triad of fever, rash, and internal organ involvement that starts 1 day to 12 weeks after the initiation of therapy. HSR has limited the therapeutic use of many drugs, including ibuprofen. Hypersensitivity syndrome associated with ibuprofen is a host-dependent drug reaction that is idiosyncratic in nature. This reaction likely is caused by a combination of metabolic and immunologic factors. Immune mediated components, such as T-cell and their products cytokines and chemokines, can exacerbate cellular responses and create complex pathways that lead to a variety of clinical manifestations. Our review presents an ibuprofen-induced clinical manifestation of hypersensitivity syndrome and the necessity of wisely monitoring the patients clinically and by laboratory investigations when prescribing this drug.
To evaluate whether caspases are involved in ethanol (EtOH)-induced apoptosis and if polyenylphos... more To evaluate whether caspases are involved in ethanol (EtOH)-induced apoptosis and if polyenylphosphatidylcholine (PPC) affects apoptosis, in vitro in Hep G2 cells. Cells were treated with 100 mmol/L EtOH for 24 h and with 2 doses of 100 mmol/L EtOH (1/24 h) in the presence of absence of 20 mmol/L of PPC or 50 micromol/L caspase 3 inhibitor (IDN). Cells were analyzed for apoptosis by transmission electron microscopy (TEM) 6000 cells/treatment, DNA fragmentation by ELISA and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (T dt-mediated d-UTP) nick-end-labeling, TUNEL. 100 mmol/L dose of EtOH resulted in 22 +/- 2.5% (p < 0.001) apoptosis (vs. control). Two consecutive doses of 100 mmol/L EtOH for 24 h each caused 36 +/- 3.0% (p < 0.001 vs. control and p < 0.05 vs. one dose). PPC significantly reduced apoptosis (vs. non exposed to PPC): 100 mmol/L -12 +/- 1.5% (p < 0.05) and 2 x 10(-)(0) mmol/L -20 +/- 2.0% (p < 0.001). Pretreatment with 50 micromol caspase inhibitor reduced EtOH-induced apoptosis in a similar proportion. PPC downregulates EtOH-apoptosis by a mechanism similar to caspase inhibition.
To evaluate cross reactivity between sulfonamide antimicrobials and celecoxib in patients with hi... more To evaluate cross reactivity between sulfonamide antimicrobials and celecoxib in patients with histories of allergies to sulfonamide antimicrobials. Immunocompetent patients with a history of sulfonamide antimicrobial allergy who were being considered for therapy with celecoxib were prospectively enrolled. Sulfamethoxazole and trimethoprim skin prick and intradermal testing and/or an in vitro lymphocyte toxicity assay were performed. If skin testing was negative, an oral challenge with sulfamethoxazole and trimethoprim was performed. Oral challenges with celecoxib were administered to all patients. Twenty-eight immunocompetent patients (26 female; mean age 60 years) were evaluated. History of sulfonamide antimicrobial allergy included urticaria (n = 7), cutaneous eruptions (n = 9), and other (n = 12). Four of the 28 patients who were skin prick tested were positive to sulfamethoxazole and two of the ten patients who underwent in vitro testing were positive to sulfamethoxazole. All 28 patients were administered celecoxib and tolerated the medication. Phone call follow up in 25 patients disclosed that 15 patients continued to take celecoxib, while five patients did not take celecoxib following the oral challenge, and five discontinued celecoxib due to adverse effects, lack of drug efficacy or physician preference. Confusion exists regarding the potential for cross reactivity between sulfonamide antimicrobials and other sulfonamide-containing compounds. The six sulfonamide-allergic patients tolerated celecoxib uneventfully. This pilot study supports the hypothesis that the potential for cross-reactivity between celecoxib and sulfonamide antimicrobials appears to be low. However, further investigations are required to confirm this.
To utilize cytokine levels to predict sustained response (SR) to alpha interferon (IFN alpha) the... more To utilize cytokine levels to predict sustained response (SR) to alpha interferon (IFN alpha) therapy in chronic hepatitis C patients, and to determine the relationship between serum tumor necrosis factor alpha (TNF alpha), interleukin (IL) IL 6, IL 8, IL 12, transforming growth factor beta (TGF beta 1) and the degree of liver damage as reflected by traditional markers. Serum cytokine levels were assessed using ELISA in 18 patients included in a controlled clinical trial of IFN alpha. Of the 18 patients, 27% were sustained responders (SR), 27% were response and relapse responders (RR), and 46% were non-responders (NR). Multivariate analysis showed that a low serum TNF alpha level and high serum IL 8 levels were independent factors associated with SR to IFN alpha therapy. Serum TNF alpha level highly correlated with viral load and genotype predictive values (p < 0.001). Therapy lowered the IL 6 and IL 12 profile. TGF beta 1 levels in serum are positively correlated with fibrinogenesis. IFN alpha therapy modulates immune response to hepatitis C virus, contributing to sustained response.
Aims: (i) To characterize serum levels of pro/anti-inflammatory cytokines in non-cirrhotics with ... more Aims: (i) To characterize serum levels of pro/anti-inflammatory cytokines in non-cirrhotics with hepatitis C; (ii) to correlate levels of these cytokines with degree of disease at baseline; and (iii) to characterize the immuno-modulatory effects of therapy with response. Methods: We studied 103 patients that were part of randomized, controlled, clinical trials. Serum cytokines were measured using enzyme-linked immunosorbent assay. Results: Using standard therapy in the presence and absence of ribavirin, the sustained responders had lower baseline tumor necrosis alpha (TNF-alpha) levels as compared to relapsed responders and non-responders. In patients receiving pegylated therapy, the degree of inflammation as determined by histology was paralleled by high TNF-alpha levels at baseline. In pegylated combination therapy with high dose ribavirin, lower levels of TNF-alpha, transforming growth factor beta (TGF-beta) and fibrosis scores were seen when comparing baseline with follow up. In sustained responders, regardless of therapy, the histological activity scores were lower at follow up as compared to baseline. Conclusions: Pegylated combination therapy reduces and sustains TNF-alpha levels and liver inflammation as shown by the histological activity index. In addition, it is able to reduce fibrosis as judged both by TGF-beta levels and fibrosis scores as compared to standard therapy.
In vitro, ethanol in combination with acetaminophen induces hepatocyte apoptosis resembling immun... more In vitro, ethanol in combination with acetaminophen induces hepatocyte apoptosis resembling immune-mediated fulminant hepatic failure in human beings. Intracellular pathways originating at the mitochondria are linked to apoptosis. I studied ethanol-induced apoptosis and hepatocytotoxicity after using an in vitro model of normal human primary hepatocytes that were exposed to 5 or 10 mM acetaminophen, 40 or 100 mM ethanol, 40 mM ethanol + 5 mM acetaminophen, or 40 mM ethanol + 10 mM acetaminophen, or nonexposed (control; plain medium). Transmission electron microscopy was performed at different time points after exposure to the various treatments. Apoptosis, as assessed by transmission electron microscopy, was increased in a time-dependent manner after exposure to ethanol + acetaminophen. In the ethanol + acetaminophen model, mitochondrial injury was associated with apoptosis of hepatocytes. Ultrastructural damage and induction of apoptosis were seen in response to N-acetyl-benzoquinone-imine plus ethanol, supporting the suggestion that the damage was due to the active metabolite of acetaminophen. The modulation of mitochondrial damage in vitro may have implications for the development of new therapeutic strategies to prevent apoptosis.
MG Neuman, GG Katz, IM Malkiewicz, P Mathurin, Hidekazu Tsukamoto, Masayuki Adachi, Hiromasa Ishi... more MG Neuman, GG Katz, IM Malkiewicz, P Mathurin, Hidekazu Tsukamoto, Masayuki Adachi, Hiromasa Ishii, A Colell, C Garcia-Ruiz, JC Fernandez-Checa, CA Casey Alcohol 28:22, 117-128, 10/2002. X 24180 Social poisons & drug abuse.
Ethanol is commonly used in cosmetic and pharmaceutical preparations. The present study aimed to ... more Ethanol is commonly used in cosmetic and pharmaceutical preparations. The present study aimed to assess ethanol-induced apoptosis and the possible repair by hyaluronic acid (HA) in vitro. In addition we aimed to determine the modulation of tumor necrosis factor (TNF-alpha) and interferon (IFN-alpha). We treated human A431 epidermoid skin cells and mouse fibroblasts with two concentrations of ethanol for 24h. HA obtained from umbilical cord excision was used at three concentration levels (2%, 4% and 8%) to determine its efficacy in the treatment. We measured cytotoxicity, TNF-alpha and IFN-alpha and visualized the cultures by electron microscopy. Treatment of cells with ethanol at 50mM and 100mM increased both the percentage of cells undergoing apoptosis, as well as the release of TNF-alpha into the culture medium. Ethanol may induce apoptosis in skin cells by enhancing the effects of TNF-alpha. HA in the 2% and 4% concentrations reduced TNF-alpha and morphological inflammation both in human A431 epidermoid skin cells and in mouse fibroblasts.
The necessity of maintaining a close balance between cell birth and cell death has caused multice... more The necessity of maintaining a close balance between cell birth and cell death has caused multicellular organisms to evolve into unforgiving societies where cells that are no longer needed are requested to commit suicide.
Ibuprofen is a widely used antipyretic and analgesic nonsteroidal antiinflammatory drug (NSAID). ... more Ibuprofen is a widely used antipyretic and analgesic nonsteroidal antiinflammatory drug (NSAID). With the aging of the population, there will be a significant increase in the prevalence of painful degenerative and inflammatory rheumatic conditions. This increase likely will lead to a parallel increase in the use of NSAIDs, including ibuprofen. The primary effect of the NSAIDs is to inhibit cyclooxygenase (prostaglandin synthase), thereby impairing the ultimate transformation of arachidonic acid to prostaglandins, prostacyclin, and thromboxanes. Although in the majority of cases it is safe, this NSAID, ibuprofen, can produce an unpredictable, idiosyncratic, type B reaction that may pose a major concern in clinical practice. Type B reactions are known to occur in susceptible individuals. The true hypersensitivity reaction (HSR) is a systemic disease defined by the triad of fever, rash, and internal organ involvement that starts 1 day to 12 weeks after the initiation of therapy. HSR has limited the therapeutic use of many drugs, including ibuprofen. Hypersensitivity syndrome associated with ibuprofen is a host-dependent drug reaction that is idiosyncratic in nature. This reaction likely is caused by a combination of metabolic and immunologic factors. Immune mediated components, such as T-cell and their products cytokines and chemokines, can exacerbate cellular responses and create complex pathways that lead to a variety of clinical manifestations. Our review presents an ibuprofen-induced clinical manifestation of hypersensitivity syndrome and the necessity of wisely monitoring the patients clinically and by laboratory investigations when prescribing this drug.
To evaluate whether caspases are involved in ethanol (EtOH)-induced apoptosis and if polyenylphos... more To evaluate whether caspases are involved in ethanol (EtOH)-induced apoptosis and if polyenylphosphatidylcholine (PPC) affects apoptosis, in vitro in Hep G2 cells. Cells were treated with 100 mmol/L EtOH for 24 h and with 2 doses of 100 mmol/L EtOH (1/24 h) in the presence of absence of 20 mmol/L of PPC or 50 micromol/L caspase 3 inhibitor (IDN). Cells were analyzed for apoptosis by transmission electron microscopy (TEM) 6000 cells/treatment, DNA fragmentation by ELISA and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (T dt-mediated d-UTP) nick-end-labeling, TUNEL. 100 mmol/L dose of EtOH resulted in 22 +/- 2.5% (p < 0.001) apoptosis (vs. control). Two consecutive doses of 100 mmol/L EtOH for 24 h each caused 36 +/- 3.0% (p < 0.001 vs. control and p < 0.05 vs. one dose). PPC significantly reduced apoptosis (vs. non exposed to PPC): 100 mmol/L -12 +/- 1.5% (p < 0.05) and 2 x 10(-)(0) mmol/L -20 +/- 2.0% (p < 0.001). Pretreatment with 50 micromol caspase inhibitor reduced EtOH-induced apoptosis in a similar proportion. PPC downregulates EtOH-apoptosis by a mechanism similar to caspase inhibition.
To evaluate cross reactivity between sulfonamide antimicrobials and celecoxib in patients with hi... more To evaluate cross reactivity between sulfonamide antimicrobials and celecoxib in patients with histories of allergies to sulfonamide antimicrobials. Immunocompetent patients with a history of sulfonamide antimicrobial allergy who were being considered for therapy with celecoxib were prospectively enrolled. Sulfamethoxazole and trimethoprim skin prick and intradermal testing and/or an in vitro lymphocyte toxicity assay were performed. If skin testing was negative, an oral challenge with sulfamethoxazole and trimethoprim was performed. Oral challenges with celecoxib were administered to all patients. Twenty-eight immunocompetent patients (26 female; mean age 60 years) were evaluated. History of sulfonamide antimicrobial allergy included urticaria (n = 7), cutaneous eruptions (n = 9), and other (n = 12). Four of the 28 patients who were skin prick tested were positive to sulfamethoxazole and two of the ten patients who underwent in vitro testing were positive to sulfamethoxazole. All 28 patients were administered celecoxib and tolerated the medication. Phone call follow up in 25 patients disclosed that 15 patients continued to take celecoxib, while five patients did not take celecoxib following the oral challenge, and five discontinued celecoxib due to adverse effects, lack of drug efficacy or physician preference. Confusion exists regarding the potential for cross reactivity between sulfonamide antimicrobials and other sulfonamide-containing compounds. The six sulfonamide-allergic patients tolerated celecoxib uneventfully. This pilot study supports the hypothesis that the potential for cross-reactivity between celecoxib and sulfonamide antimicrobials appears to be low. However, further investigations are required to confirm this.
To utilize cytokine levels to predict sustained response (SR) to alpha interferon (IFN alpha) the... more To utilize cytokine levels to predict sustained response (SR) to alpha interferon (IFN alpha) therapy in chronic hepatitis C patients, and to determine the relationship between serum tumor necrosis factor alpha (TNF alpha), interleukin (IL) IL 6, IL 8, IL 12, transforming growth factor beta (TGF beta 1) and the degree of liver damage as reflected by traditional markers. Serum cytokine levels were assessed using ELISA in 18 patients included in a controlled clinical trial of IFN alpha. Of the 18 patients, 27% were sustained responders (SR), 27% were response and relapse responders (RR), and 46% were non-responders (NR). Multivariate analysis showed that a low serum TNF alpha level and high serum IL 8 levels were independent factors associated with SR to IFN alpha therapy. Serum TNF alpha level highly correlated with viral load and genotype predictive values (p < 0.001). Therapy lowered the IL 6 and IL 12 profile. TGF beta 1 levels in serum are positively correlated with fibrinogenesis. IFN alpha therapy modulates immune response to hepatitis C virus, contributing to sustained response.
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