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Different modulation by antimalarial drugs of nitric oxide and TNF production by peritoneal mouse macrophages
DIFFERENT MODULATION BY ANTIMALARIAL DRUGS OF NITRIC OXIDE AND TNF P R O D U C T I O N
THE INFLUENCE OF PENICILLINS AND CEPHALOSPORINS ON
ELECTROPHORETICAL CHARACTERISTICS OF SERUM PROTEINS . . . . 1 , , I , v 2 MnTana Abramovtc, Jasmma Tornm, Zonca Mancev
BY PERITONEAL
Instttut of Medical Chemistry 1, Institute of Pharmacology and Toxicology, Faculty of Medicine 2, University of Ni~ Brade Taskovida 81, Nt~ Yugoslavia,
MOUSE
MACROPHAGES.
N. Basilico, E. Pagani, R. Panzeri, D. Taramelli and M. Ghione*. Institute of Medical Microbiology, University of Milan, Via Pascal 36 and *Sieroterapico, Soot. Coop.a R.L.Milan, Italy.
Knowing the transport role of serum proteins, and possibility of provisional incorporation of different substances in protein molecules (KraghHansen, U. 1981) it is possible to expect, during that period, the modification of their electrophoretical caracteristics (1). This especially refers to antibiotics from groups of peniclllins and cephalosporins, because of their specific polarity and high concentration in blood during the using time. Examinations were done in vitro with the serum of clinically healthy persons, aged I8-20 and nearly the same body mass. Different quantities of drugs, calculated on maximal daily quantity and twice and three times larger
Cure and prophylaxis of malaria infections are complicated by the continuos emergence of drug-resistant strains of P.falciparum. At present, the 8-aminoquinoline, primaquine (PQ), although quite toxic, is the only available drug against relapsing malaria and the 4-aminoquinoline, choloroquine (CQ), is still the drug of choice against non resistant strains. Artemisinin (ARM), a sesquiterpene endoperoxide and its derivatives represent an new class of potent antimalarial agents characterized by low systemic toxicity. It is known that CQ exerts an anti-inflammatory action by inhibiting lymphocytes activation and cytokine production. However, the interactions of PQ or ARM with host phagocytic ceils have not been fully characterized. In the present work, we analyzed the in-vitro effects of PQ, ARM and CQ on the production of inflammatory mediators by macrophages (MO). Peritoneal MO from CD1 mice were pretreated with different doses of drugs and then stimulated with 1 I~g/mlof LPS for 48 hr. Supernatants were assayed for the presence of TNF and Nitric Oxide (NO) using a bioassay and the Griess reagent, respectively. The results confirm that CQ (50-10 tLM)significantly reduced the production of NO and TNF by LPS-MO in a dose-dependent manner. In contrast, at the same doses, PQ increased both NO and TNF production, whereas ARM was ineffective. ARM impaired by 40% the production of NO only when it was used at 1001~M. The drugs were not toxic nor they induced by themselves the production of NO and TNF at any of the doses employed. It appears thus that CQ, PQ and ARM affect MO functions, each in a different way. Whereas ARM does not interfere with MO activation, PQ seems to augment the inflammatory response: this may be related to its relevant in vivo toxicity.
were incubated with 0.5 gl of human serum at room temperature, during the period of 20 min. The separation of serum proteins has been done electrophoretically on acetate-cellulose gel, in verunal buffer (pH - 9.2) at 200 V during 5 min. Tile identification of protein fractions, after dying elpherograms by Ponceau S (C.I.27195), was made on celomatic E,LSCI~IYr 3. Obtained results show that, in serums under the influence of penicillins
(Benzyl-penicRlin, Ampicillin, Amoxicillin, Oxacillin, etc), there are changes in mobility of proteins. BenzTl-penicillin causes special effects in relative quantities of protein fractions. The most intensive changes oecux in aIbumine field. With the larger quantities of antibiotics, values become statistical important by increasing the values of albumin fraction. ~V~SDfor albumin incubated during 20 rain with 1.6 M i.j. of
benzyl-penicillingives values 65.6:t6.8, and incubated with 4.8 M ij. gives 66.2_+6.9 (io<0.05). Some decrease in all globulin fractions has been observed, with noticeable reduction in y-globulin field. Similar changes in serumproteins behavior were found under the influence
of cephalnsporins (Cephalexin, Cephosporin, Cephotaxim, etc) especially cephotaxim - Tolycar. Based on the proteinogram we can conclude that Tolicar makes drug-protein complexes not only with albumin but also with proteins of c~2globulin fraction. In serum incubated with the highest concentrations of tolycar
(6.0 g) the biggest changes have been noticed on elpherogram. Prominenet changes in fraction of serum proteins incubated with some antibioticswarn us of the poss~ility of getting similarproteinograms in vivo oonditions.
Reference: Kragh-Hansen, U., Pharmacol.Rew.,1981, 33, 17-53.
URINARY AND BILIARY RECOVERY OF A NEW ANTIMYCOTIC COMPOUND, SPA-S-753, IN RATS. M.R. Galmozzi, G. Buffa, T. Bruzzese and A. Bonabello Pharmacology Research Laboratories, SPA-Societ& Prodotti Antibiotici S.p.A., Milan, Italy
C L I N I C A L S T U D Y OF DIFLUCAN* I N T H E O R A L CAVIT~ MUCOUS MEMBRANE PATHOLOGY, C O M P L I C A T E D BY C A N D I D I A S I S V.M. Bezrukov, E.I. Belyaeva, L.N. Guertchikov, N.A. N e c h a e n k o - Central Stomatological Institute, Moscow, Russia.
SPA-S-753 (N-dimethylaminoacetyl-partricin A 2dimethylaminoethylamide diaspartate) is a new water soluble derivative of partricin A. It has previously been shown that this compound has a strong fungicidal effect "in vitro" (1) as well as "in vivo" (2) and a quite long half-life in rats (3). The current study was undertaken to calculate the urinary and biliary excretion (expressed as a percentage of the dose) of SPA-S753 in rats after an intravenous (i.v.) administration. We used male S.D. tats weighing 200 to 225 g. The animals received a single {.v. bolus dose of SPA-S-753 1.25 mg/kg. Urine specimens were collected over a period of time of 72 h while bile samples were collected up to 10 h after i.v. treatment.The urine and bile concentrations of the test compound were determined with a new HPLC method. This procedure involves the addition of an internal standard to the s a m p l e , protein precipitation with methanol and direct injection of the supernatant into the chromatograph. During the post-treatment period of 72 h the complete urinary recovery of the Compound was around 1% of the administered dose, whereas the biliary recovery was about 10%, within the limited period of 10 h, so showing that bile is the preferred excretion route. From these results and from the previous kinetic data we can assume at any rate that the excretion of the test compound is very slow suggesting a possible concentrating mechanism in the peripheral tissues.
Diseases of the m o u t h cavity m u c o u s m e m b r a n e , accompanied by acute, subacute or chronic process, are often c o m p l i c a t e d b y candidiasis. T h e r e f o r e , the application of Diflucan in a complex therapy is entirely justified. Acute a n d recurrent a p h t h o u s stomatitis, lichen p l a n u s , glosalgia, a c c o m p a n i e d b y algesic syndrome, are the m o s t frequent forms of the m o u t h cavity m u c o u s m e m b r a n e infections. In the differential diagnosis candidiasis ranks high a m o n g the indicated pathology f o r m s . We have carried out a r a n d o m i z e d e x a m i n a t i o n a n d t r e a t m e n t of 30 patients w i t h t h e m o u t h cavity m u c o u s m e m b r a n e candidiasis as a complication of the m a i n diseases of the oral cavity. Diflucan was a d m i n i s t e r e d to 30 patients d u r i n g 14 d a y s b y a daily d o s a g e of 50 mg. T h e p o s i t i v e therapeutic effect ( no traces of candida in the culture f r o m m u c o u s m e m b r a n e of the m o u t h cavity) on the completion of treatment has been observed in 60% of cases. O n the grounds of the results obtained, Diflucan c a n b e r e c o m m e n d e d f o r t h e treatment of not only pure forms of candidiasis, but also those c o m b i n e d w i t h candidiasis of other infections of mucous m e m b r a n e of the oral cavity. * (PFIZERCO., BELGIUM)
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1. Strippoli V. et al., Eur Bull. Drug Res. 1, 113-8, 1992 2. Bonabello A. and Bruzzese T., XXVll Cong. Naz. SIF,1994 3. Galmozzi M.R. et al., XXVII Cong. Naz. SIF,1994
134--
THE INFLUENCE OF PENICILLINS AND CEPHALOSPORINS ON
ELECTROPHORETICAL CHARACTERISTICS OF SERUM PROTEINS . . . . 1 , , I , v 2 MnTana Abramovtc, Jasmma Tornm, Zonca Mancev
BY PERITONEAL
Instttut of Medical Chemistry 1, Institute of Pharmacology and Toxicology, Faculty of Medicine 2, University of Ni~ Brade Taskovida 81, Nt~ Yugoslavia,
MOUSE
MACROPHAGES.
N. Basilico, E. Pagani, R. Panzeri, D. Taramelli and M. Ghione*. Institute of Medical Microbiology, University of Milan, Via Pascal 36 and *Sieroterapico, Soot. Coop.a R.L.Milan, Italy.
Knowing the transport role of serum proteins, and possibility of provisional incorporation of different substances in protein molecules (KraghHansen, U. 1981) it is possible to expect, during that period, the modification of their electrophoretical caracteristics (1). This especially refers to antibiotics from groups of peniclllins and cephalosporins, because of their specific polarity and high concentration in blood during the using time. Examinations were done in vitro with the serum of clinically healthy persons, aged I8-20 and nearly the same body mass. Different quantities of drugs, calculated on maximal daily quantity and twice and three times larger
Cure and prophylaxis of malaria infections are complicated by the continuos emergence of drug-resistant strains of P.falciparum. At present, the 8-aminoquinoline, primaquine (PQ), although quite toxic, is the only available drug against relapsing malaria and the 4-aminoquinoline, choloroquine (CQ), is still the drug of choice against non resistant strains. Artemisinin (ARM), a sesquiterpene endoperoxide and its derivatives represent an new class of potent antimalarial agents characterized by low systemic toxicity. It is known that CQ exerts an anti-inflammatory action by inhibiting lymphocytes activation and cytokine production. However, the interactions of PQ or ARM with host phagocytic ceils have not been fully characterized. In the present work, we analyzed the in-vitro effects of PQ, ARM and CQ on the production of inflammatory mediators by macrophages (MO). Peritoneal MO from CD1 mice were pretreated with different doses of drugs and then stimulated with 1 I~g/mlof LPS for 48 hr. Supernatants were assayed for the presence of TNF and Nitric Oxide (NO) using a bioassay and the Griess reagent, respectively. The results confirm that CQ (50-10 tLM)significantly reduced the production of NO and TNF by LPS-MO in a dose-dependent manner. In contrast, at the same doses, PQ increased both NO and TNF production, whereas ARM was ineffective. ARM impaired by 40% the production of NO only when it was used at 1001~M. The drugs were not toxic nor they induced by themselves the production of NO and TNF at any of the doses employed. It appears thus that CQ, PQ and ARM affect MO functions, each in a different way. Whereas ARM does not interfere with MO activation, PQ seems to augment the inflammatory response: this may be related to its relevant in vivo toxicity.
were incubated with 0.5 gl of human serum at room temperature, during the period of 20 min. The separation of serum proteins has been done electrophoretically on acetate-cellulose gel, in verunal buffer (pH - 9.2) at 200 V during 5 min. Tile identification of protein fractions, after dying elpherograms by Ponceau S (C.I.27195), was made on celomatic E,LSCI~IYr 3. Obtained results show that, in serums under the influence of penicillins
(Benzyl-penicRlin, Ampicillin, Amoxicillin, Oxacillin, etc), there are changes in mobility of proteins. BenzTl-penicillin causes special effects in relative quantities of protein fractions. The most intensive changes oecux in aIbumine field. With the larger quantities of antibiotics, values become statistical important by increasing the values of albumin fraction. ~V~SDfor albumin incubated during 20 rain with 1.6 M i.j. of
benzyl-penicillingives values 65.6:t6.8, and incubated with 4.8 M ij. gives 66.2_+6.9 (io<0.05). Some decrease in all globulin fractions has been observed, with noticeable reduction in y-globulin field. Similar changes in serumproteins behavior were found under the influence
of cephalnsporins (Cephalexin, Cephosporin, Cephotaxim, etc) especially cephotaxim - Tolycar. Based on the proteinogram we can conclude that Tolicar makes drug-protein complexes not only with albumin but also with proteins of c~2globulin fraction. In serum incubated with the highest concentrations of tolycar
(6.0 g) the biggest changes have been noticed on elpherogram. Prominenet changes in fraction of serum proteins incubated with some antibioticswarn us of the poss~ility of getting similarproteinograms in vivo oonditions.
Reference: Kragh-Hansen, U., Pharmacol.Rew.,1981, 33, 17-53.
URINARY AND BILIARY RECOVERY OF A NEW ANTIMYCOTIC COMPOUND, SPA-S-753, IN RATS. M.R. Galmozzi, G. Buffa, T. Bruzzese and A. Bonabello Pharmacology Research Laboratories, SPA-Societ& Prodotti Antibiotici S.p.A., Milan, Italy
C L I N I C A L S T U D Y OF DIFLUCAN* I N T H E O R A L CAVIT~ MUCOUS MEMBRANE PATHOLOGY, C O M P L I C A T E D BY C A N D I D I A S I S V.M. Bezrukov, E.I. Belyaeva, L.N. Guertchikov, N.A. N e c h a e n k o - Central Stomatological Institute, Moscow, Russia.
SPA-S-753 (N-dimethylaminoacetyl-partricin A 2dimethylaminoethylamide diaspartate) is a new water soluble derivative of partricin A. It has previously been shown that this compound has a strong fungicidal effect "in vitro" (1) as well as "in vivo" (2) and a quite long half-life in rats (3). The current study was undertaken to calculate the urinary and biliary excretion (expressed as a percentage of the dose) of SPA-S753 in rats after an intravenous (i.v.) administration. We used male S.D. tats weighing 200 to 225 g. The animals received a single {.v. bolus dose of SPA-S-753 1.25 mg/kg. Urine specimens were collected over a period of time of 72 h while bile samples were collected up to 10 h after i.v. treatment.The urine and bile concentrations of the test compound were determined with a new HPLC method. This procedure involves the addition of an internal standard to the s a m p l e , protein precipitation with methanol and direct injection of the supernatant into the chromatograph. During the post-treatment period of 72 h the complete urinary recovery of the Compound was around 1% of the administered dose, whereas the biliary recovery was about 10%, within the limited period of 10 h, so showing that bile is the preferred excretion route. From these results and from the previous kinetic data we can assume at any rate that the excretion of the test compound is very slow suggesting a possible concentrating mechanism in the peripheral tissues.
Diseases of the m o u t h cavity m u c o u s m e m b r a n e , accompanied by acute, subacute or chronic process, are often c o m p l i c a t e d b y candidiasis. T h e r e f o r e , the application of Diflucan in a complex therapy is entirely justified. Acute a n d recurrent a p h t h o u s stomatitis, lichen p l a n u s , glosalgia, a c c o m p a n i e d b y algesic syndrome, are the m o s t frequent forms of the m o u t h cavity m u c o u s m e m b r a n e infections. In the differential diagnosis candidiasis ranks high a m o n g the indicated pathology f o r m s . We have carried out a r a n d o m i z e d e x a m i n a t i o n a n d t r e a t m e n t of 30 patients w i t h t h e m o u t h cavity m u c o u s m e m b r a n e candidiasis as a complication of the m a i n diseases of the oral cavity. Diflucan was a d m i n i s t e r e d to 30 patients d u r i n g 14 d a y s b y a daily d o s a g e of 50 mg. T h e p o s i t i v e therapeutic effect ( no traces of candida in the culture f r o m m u c o u s m e m b r a n e of the m o u t h cavity) on the completion of treatment has been observed in 60% of cases. O n the grounds of the results obtained, Diflucan c a n b e r e c o m m e n d e d f o r t h e treatment of not only pure forms of candidiasis, but also those c o m b i n e d w i t h candidiasis of other infections of mucous m e m b r a n e of the oral cavity. * (PFIZERCO., BELGIUM)
--
1. Strippoli V. et al., Eur Bull. Drug Res. 1, 113-8, 1992 2. Bonabello A. and Bruzzese T., XXVll Cong. Naz. SIF,1994 3. Galmozzi M.R. et al., XXVII Cong. Naz. SIF,1994
134--