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Inosamycin, a complex of new aminoglycoside antibiotics. I. Production, isolation and properties.

A strain of Streptomyces hygroscopicus No. J296-21 (ATCC 39150) was found to produce a complex of new antibiotics, called inosamycins, which consisted of components A, B, C, D and E. They are novel aminocyclitol antibiotics structurally related to neomycin, paromomycin and ribostamycin. The antibiotic complex and each component of inosamycin exhibit a broad antibacterial spectrum but they are inactive against most of the aminoglycoside-resistant organisms. The antibacterial activity of inosamycin A, the major component of the complex, is comparable to that of neomycin but its acute toxicity is significantly lower (ca. 1/3) than that of neomycin.

Aminoglycosides↗

Inosamycin, a complex of new aminoglycoside antibiotics. II. Structure determination.

Structures of the new aminocyclitol antibiotics, inosamycins A, B, C, D and E, have been determined by a combination of chemical degradation and spectroscopic studies. They are structurally related to neomycin, paromomycin and ribostamycin but differ in that all the inosamycin components contain 2-deoxy-scyllo-inosamine in place of 2-deoxystreptamine in the known aminoglycoside antibiotics.

Aminoglycosides↗

Mode of action of deoxypheganomycin D on Mycobacterium smegmatis ATCC 607.

Deoxypheganomycin D, a specific inhibitor of mycobacteria, inhibits the growth in vitro of Mycobacterium smegmatis ATCC 607 (M. 607) bacteriostatically at concentrations as high as 7 X 10(-5) M. It shows no cross-resistance to paromomycin, capreomycin, viomycin, streptothricin, kanamycin and streptomycin. Deoxypheganomycin D at 2.8 X 10(-7) M where the cell growth of M. 607 is only partially inhibited does not significantly inhibit DNA, RNA or protein synthesis but leads to marked decrease (13% of control) in [14C]glycerol-derived radioactivity in cell-walls. In the presence of 7 X 10(-6) M deoxypheganomycin D, the influx of leucine but not thymidine is affected while the reverse is true with efflux. The data suggest that the effect of deoxypheganomycin D on M. 607 may be related to both the cell membrane and specific mycobacterial lipid like components of the cell-wall.

Amino Acids↗

Double stage activity in aminoglycoside antibiotics.

Fourteen different aminoglycoside antibiotics (AGs) were challenged with aminoglycoside acetyltransferases (AACs) of actinomycete origin in order to examine their 'double stage activity' that is arbitrarily defined as antibiotic activity retainable after enzymatic modification. In kanamycin (KM)-group AGs tested [KM, dibekacin (DKB), amikacin and arbekacin (ABK)], ABK retained activity after acetylations by AAC(3), AAC(2') and AAC(6'). DKB also retained a weak activity after acetylation by AAC(2'). In gentamicin (GM)-group AGs tested [GM, micronomicin, sisomicin (SISO), netilmicin (NTL) and isepamicin], GM, SISO and NTL retained activites after acetylation by AAC(2'). In neomycin (NM)-group AGs tested [ribostamycin, NM, paromomycin], NM retained activity after acetylation by AAC(6') and AAC(2'). None of astromicin (ASTM)-group AGs tested (ASTM and istamycin B) retained activity after acetylation by AAC(2') and AAC(6'). The activities of acetylated ABK derivatives by AAC(3) and AAC(2') were distinctively high, compared to the others. Streptomyces lividans TK21 containing the cloned aac genes were markedly sensitive to AGs that retained activities after acetylation, indicating the substantial effect of 'double stage activity'.

Acetylation↗

Stimulatory and inhibitory properties of aminoglycoside antibiotics at N-methyl-D-aspartate receptors.

The effects of aminoglycoside antibiotics on N-methyl-D-aspartate (NMDA) receptors were studied using voltage-clamp recording of recombinant NMDA receptors expressed in Xenopus oocytes. A number of aminoglycosides were found to potentiate macroscopic currents at heteromeric NR1A/NR2B receptors, but not at NR1A/NR2A, NR1A/NR2C, NR1A/NR2D, or NR1B/NR2B receptors. The degree of potentiation had a rank order neomycin B > paromomycin > gentamicin C > geneticin > kanamycin A > streptomycin. Potentiation was not seen with kasugamycin and spectinomycin. The degree of stimulation paralleled the number of the amino groups in the aminoglycosides. The stimulatory effects of aminoglycosides were more pronounced at subsaturating concentrations of glycine and at acidic pH, similar to the stimulatory effects of spermine. We measured the effects of aminoglycosides at mutant NMDA receptors to determine which amino acid residues in NMDA receptor subunits are involved in stimulation. Mutations that reduced or abolished spermine stimulation also reduced stimulation by aminoglycosides. Several aminoglycosides produced a weak voltage-dependent block of NMDA receptors, but the degree of inhibition did not appear to correlate with the number of amino groups in the molecule. The results suggest that aminoglycosides having more than three amino groups have stimulatory effects that are mediated through the spermine-binding site on NMDA receptors.

Aminoglycosides↗

Case studies in international travelers.

Family physicians should be alert for unusual diseases in patients who are returning from foreign travel. Malaria is a potentially fatal disease that can be acquired by travelers to certain areas of the world, primarily developing nations. Transmitted through the bite of the Anopheles mosquito, malaria usually presents with fever and a vague systemic illness. The disease is diagnosed by demonstration of Plasmodium organisms on a specially prepared blood film. Travelers can also acquire amebic infections, which may cause dysentery or, in some instances, liver abscess. Amebiasis is diagnosed by finding Entamoeba histolytica cysts or trophozoites in the stool. Invasive amebic infections are generally treated with metronidazole followed by iodoquinol or paromomycin. Cutaneous larva migrans is acquired by skin contact with hookworm larvae in the soil. The infection is characterized by the development of itchy papules followed by serpiginous or linear streaks. Cutaneous larva migrans is treated with invermectin or albendazole. Case studies are presented.

Adult↗

[Drug sensitivity of Yersinia enterocolitica and Yersinia pseudotuberculosis].

Drug-sensitivity of 70 strains of Y. enterocolitica and 24 of Y. pseudotuberculosis including reference strains and isolates from men or animals were determined by the agar plate-dilution method. All the strains of Y. enterocolitica were relatively resistant to penicillins and cephalosporins, while all of Y. pseudotuberculosis were sensitive to both of the agents. Most of the strains of both species excluding 13 resistant ones were sensitive to streptomycin. The resistant strains, however, were sensitive to other aminoglycoside antibiotics, i.e. kanamycin, paromomycin, gentamicin, tobramycin, dibekacin and amikacin. All of the strains were sensitive to chloramphenicol. Most of the strains excluding 3 resistant ones were highly sensitive to tetracycline. The resistant strains were also resistant to doxycycline and methacycline but not to minocycline. All of them were resistant to erythromycin, to lincomycin and to novobiocin. Most of them excluding 7 resistant strains were sensitive to sulfisoxazole. All of them were sensitive to nalidixic acid, piromidic acid, and also to dihydroxymethylfuratrizine. A new synthetic agent, fosfomycin showed a relatively wide range of activity to the strains, but none of resistant strains were noticed. These resistant strains were all found in the isolates from men and animals but not in reference strains. Among these resistant strains, resistance patterns were as follows; TC, SM, SA in 3 of Y. enterocolitica, SM, SA in 3 of Y. enterocolitica and in 1 of Y. pseudotuberculosis, and SM in 3 of the both species, respectively. As for polypeptide antibiotics, polymyxin B and colistin, some strains of the both species showed an uncontinous susceptibility, i.e. inhibited growth at a certain lower concentration but growth at a higher concentration.

Amikacin↗

Autochthonous mucosal leishmaniasis in a hemodialyzed Italian patient.

A case of mucosal leishmaniasis in a 60-year old hemodialysis patient who had never lived outside Italy is described. The patient complained of fever, epistaxis and nasal obstruction. An anterior rhinoscopy disclosed a mass of two centimetres in diameter in the right nasal fossa. Histological examination revealed Leishmania amastigotes. Serology for Leishmania was positive with antibody titer of 1/320. A culture yielded a very slow growth of Leishmania infantum MON-24. In spite of a two-month treatment with oral itraconazole, the lesions progressively worsened. Treatment with topical paromomycin sulfate determined the complete resolution of the lesions within four months, with a residual perforation of the septum. This case demonstrates that localization of Leishmania spp must be considered in the differential diagnosis of mucosal lesions in hemodialyzed patients, even in countries not at risk for this parasite. Moreover, this case indicates the important role of the immune system in the evolution of the disease.

Animals↗

Topical treatment for cutaneous leishmaniasis.

Parenteral pentavalent antimonials remain the standard therapy for cutaneous leishmaniasis. More effective and patient-compliant topical treatments are an important alternative treatment for the localized self-limiting forms of this disease. Two paromomycin ointments are commercially available but their use is limited by either toxicity or lack of efficacy. Other topical formulations have been in clinical trials, but many results have been equivocal and no major breakthroughs have been achieved. The focus of this review is on recent developments in the field of topical treatment for cutaneous leishmaniasis and rational approaches to enhance topical drug absorption.

Adjuvants, Pharmaceutic↗

Topical treatment of persistent cutaneous leishmaniasis with ethanolic lipid amphotericin B.

A 1.5-year-old infant was referred because of spreading skin lesions diagnosed earlier as cutaneous leishmaniasis that did not respond to repeated courses with paromomycin-containing ointment. After consideration of the alternative therapeutic options, the infant was treated with a topical colloidal solution of amphotericin B for 3 weeks, This mode of therapy resulted in resolution of the skin lesions. No local or systemic side effects were observed. There were no signs of recurrence 3 months after cessation of the treatment.

Administration, Topical↗

[Management of American cutaneous leishmaniasis. Outcome apropos of 326 cases treated with high-dose pentamidine isethionate].

The purpose of this retrospective analysis was to evaluate the outcome of pentamidine isethionate treatment (4 mg/kg of Pentamidine by the intramuscular route on Days 1 and 3) of cutaneous leishmaniasis in 326 cases that occurred during an outbreak among French military personnel in French Guyana from 1998 to 1999. A great difference was found between the 205 patients treated in French Guyana (series G) and 32 patients treated at the Laveran Military Hospital in Marseille, France (series L). Failure rate, i.e. 25% in series L versus 5% in series G, was significantly correlated with the delay to treatment which was much longer in series L. Extensive rhabdomyolysis was observed in all cases tested: this side-effect has not been reported. Based on these findings and a review of the literature on pentamidine isethionate, the authors recommend prompt treatment using lower doses. Other treatment alternatives for American cutaneous leishmaniasis are also presented including two of the latest developments in the field, i.e., oral treatment using miltefosine and topical treatment using agents such as paromomycine.

Antiprotozoal Agents↗

[Recovery of facultatives and anaerobes from frozen specimens with a polymicrobial nature].

Microbiological examination of frozen specimens is sometimes carried out in clinical microbiology and the result is used as an aid of diagnosis and/or treatment of polymicrobial infections. The study was carried out to reevaluate the merit of freezing specimens in clinical microbiology. A total of 10 specimens with a polymicrobial nature were included in this study. Before and after freezing specimens, we isolated facultative and anaerobic bacteria using a set of primary isolation media, consisting of three aerobic agar plates (MacConkey agar, blood agar and chocolate agar) and four pre-reduced anaerobic agar plates (HK Blood agar, HK blood agar with paromomycin (PM) and vancomycin (VM), phenyl ethyl-alcohol (PEA) agar and Bacteroides bile esculin (BBE) agar). All the procedures were done in a properly controlled anaerobic chamber. The number of isolates before and after freezing was 79 and 70, respectively. Among the strains isolated before freezing, 33 strains were recovered on the same kin of media artery freezing, without a remarkable decrease in the quantity. But 26 strains were not recovered and 2 strains were recovered with a remarkable decrease. Among 26 strains, 15 strains could be successfully backed up on the different kind of media. In conclusion, an anaerobic technique with an anaerobic chamber and a set of isolatin plates including blood agar, chocolate agar, HK blood agar, PEA blood agar, HK blood agar with PM and VM enable us to estimate the bacteriology before freezing from frozen specimens.

Bacteria, Anaerobic↗

Cutaneous leishmaniasis: recognition and treatment.

Cutaneous leishmaniasis is a parasitic disease occurring throughout the Americas from Texas to Argentina, and in the Old World, particularly the Middle East and North Africa. It is spread by the female sandfly. The condition is diagnosed every year in travelers, immigrants, and military personnel. Physicians in the United States must be alert to the diagnosis of leishmaniasis in travelers returning from endemic areas. Physicians working for short periods in endemic areas often must make the diagnosis and should be aware of local disease patterns. When faced with a possible leishmanial skin lesion, a skin scraping with microscopic analysis is the best test. Punch biopsies with tissue-impression smears also can be diagnostic. Needle aspiration of tissue fluid from the margin of a lesion can yield fluid for culture to isolate the organism and identify the species. Immunologic tests are being developed, including a highly sensitive polymerase chain reaction test. The treatment mainstay is pentavalent antimony (e.g., sodium stibogluconate). Not all patients require treatment; many lesions heal spontaneously. Antimonials have a high incidence of reversible adverse effects. Other medications used for treatment include amphotericin B, pentamidine isethionate, paromomycin, and antifungals. This disease must be considered in at-risk patients, and family physicians should know the basics of diagnosis and where to go for more help.

Animals↗

Leishmaniasis in patients with chronic renal failure: a diagnostic and therapeutic challenge for the clinician.

BACKGROUND: The role of leishmaniasis in dialyzed or transplanted patients for chronic renal failure is generally neglected. In this study, the authors present a series of three cases of leishmaniasis (one visceral, one mucous and one muco-visceral) in patients with end-stage renal failure characterized by an atypical presentation and/or resistance to therapy. CASE DESCRIPTION: Two patients had an atypical infection: the first patient demonstrated a mucosal form, while the second had visceral and mucosal involvement. These two presentations are very rare and, to the best of our knowledge, other autoctonous disease cases have never been described in Italy. In the first patient, a cycle of oral itraconazole was scarcely effective and poorly tolerated, while treatment with 15% topical paromomycin sulfate was successful. Patients two and three failed to respond to meglumine antimonate and amphotericin B lipid complex. A second cycle with liposomal amphotericin B was effective in both cases. In addition, a superior safety profile for liposomal amphotericin B in comparison with the lipid complex amphotericin B was observed. CONCLUSIONS: These three cases highlight the problem of leishmaniasis in both renal transplanted and dialyzed patients and suggest that this infection could be far from infrequent in addition to being resistant to therapies. Leishmaniasis should be considered in the differential diagnosis of fevers of unknown origin and mucosal lesions in these patients, even in countries not at risk for mucosal leishmaniasis.

Amphotericin B↗

[Canine leishmaniasis: evolution of the chemotherapeutic protocols].

Dogs are the domestic reservoir for Leishmania infantum (syn.: L. chagasi), the parasite causing zoonotic visceral leishmaniasis (ZVL) in both the Old and New Worlds. In foci of canine leishmaniasis (CanL), symptomatic disease occurs in less than 50% of infected dogs, and is characterized by chronic evolution of viscero-cutaneous signs. Among strategies recommended to control ZVL, detection and drug treatment of infected dogs are usually employed in the endemic countries of southern Europe. However, the conventional antileishmanial drugs successfully used in human therapy, such as pentavalent antimonials, amphotericin B, pentamidine or miltefosine, have low efficacy in the treatment of CanL. In dogs, these drugs induce only temporary remission of clinical signs, do not prevent occurrence of relapses, and often cause severe side effects. Leishmaniotic dogs may be classified into 4 groups: 1) Asymptomatic resistant dogs ("contacted dogs"), 2) Asymptomatic dogs (preclinical), 3) Dogs with minimal signs of leishmaniasis (oligosymptomatic dogs? Chronic form of leishmaniasis?), 4) Dogs suffering from different forms of clinical leishmaniasis (symptomatic dogs). The dog's immunological status and the associated clinical signs may influence the efficacy of antileishmanial drugs. Subjects belonging to groups 2, 3 and 4 should be always treated, in order to reduce their parasite load. Parameters that must be considered before starting the antileishmanial treatment are hemogram, renal and hepatic functions, electrophoretic protein pattern, antileishmania antibody titres, and bone marrow and lymph node parasite load. The most common antileishmanial drugs currently used in Italy to treat CanL are pentavalent antimonials (meglumine antimoniate) and allopurinol, alone or in combination. Other used drugs are aminosidine (syn.: paromomycin), pentamidine, metronidazole and spyramicin. Each drug regimen has different duration, from a few weeks (aminosidine), to a few months (meglumine antimoniate) or several months (allopurinol). One of the most recent drug used in human VL is liposomal amphotericin B (AmBisome--L-AMB), a powerful antileishmanial drug in both experimental murine models and in VL patients. In Italy, L-AMB is now considered the drug of choice for the treatment of human cases. However, in HIV co-infected patients high doses of L-AMB are ineffective in obtaining a radical cure. In dogs, L-AMB treatment rapidly leads to clinical recovery but is uneffective to eliminate the parasites. Drugs containing amphotericin B should not be used in veterinary practice in order to avoid selection of parasites resistant to the drug, as it already occurred for the pentavalent antimonials. Currently, there is not a standard protocol for CanL treatment in Italy, as there is an extreme variability of proposed dosages. Clinical studies on immunotherapeutics and new antileishmanial drugs, such as miltefosine and its derivates, are in progress.

Animals↗

Hydrophilic interaction chromatography combined with tandem-mass spectrometry to determine six aminoglycosides in serum.

A specific and automated method was developed to quantitate the aminoglycosides amikacin, gentamicin, kanamycin, neomycin, paromomycin, and tobramycin simultaneously in human serum. Samples were prepared with an automated solid phase extraction (SPE). The hydrophilic interaction chromatography (HILIC) was used for separation of analytes from endogenous compounds and baseline separation. The aminoglycosides were detected with electrospray ionisation tandem mass spectrometry (ESI-MS-MS). Using a volume of 500 microl biological sample the lower limits of quantification were 100 ng/ml or better. The described HILIC-MS-MS method is suitable for therapeutic drug monitoring and for clinical and pharmcokinetical investigations of the aminoglycosides.

Aminoglycosides↗

Comparison of Cryptosporidium parvum development in various cell lines for screening in vitro drug testing.

This study describes the development of Cryptosporidium parvum in MDCK, MA-104, Hep-2 and Vero cell lines. Differences in susceptibility, infectivity, and the methodology of excystation were determined. Various solutions were considered to determine the factors which enhanced the excystation (eg with and without sodium hypochlorite, trypsin or sodium taurocholate). It was shown that the sporozoites could be excysted in media either with or without trypsin and sodium taurocholate, but the number of sporozoites in the latter solution was less than the former one. Only oocysts digested by sodium hypochlorite and trypsin can enter the culture cells. Numerous meronts and oocysts were demonstrated and persisted for 9 days. Asexual stages were not observed in MA-104. Only few oocysts could be detected 1-3 days post-inoculation. There was a significant difference between the number of oocysts, which invaded MDCK, MA-104, and Hep-2 cells. MDCK gave the highest susceptibility to oocyst invasion among the three cell lines and asexual stages were also found. Among the 25 isolates, which had been cultivated, 23 isolates could infect MDCK and Hep-2. Only 2 isolates could not infect the MDCK cell. These 2 isolates could infect the Vero cell and yielded high numbers of trophozoites. Praziquantel (PZQ), doxycycline, and paromomycin (PRM) were tested on the infecting parasites. The drugs were added either with the inoculum or 24 hours after inoculation. None of them was effective, including PRM, which had been previously reported as effective.

Acquired Immunodeficiency Syndrome↗

Rifaximin, a nonabsorbed oral antibiotic, in the treatment of hepatic encephalopathy: antimicrobial activity, efficacy, and safety.

The nonabsorbed (< 0.4%) oral antibiotic rifaximin, which has been available for enteric bacterial conditions for more than a decade in several countries outside the United States, was recently introduced in the United States for the treatment of travelers' diarrhea, and is being evaluated in clinical trials for possible introduction for hepatic encephalopathy and other conditions involving enteric bacteria. This article discusses the antimicrobial activity, efficacy, and safety of rifaximin in hepatic encephalopathy. Rifaximin is a nonsystemic antibiotic with antibacterial activity against enteric pathogens for gastrointestinal infections. In 15 studies, several of which were adequately powered to assess efficacy, rifaximin was at least as effective as lactulose/lactitol (the current mainstay of pharmacologic treatment for hepatic encephalopathy) and neomycin and paromomycin (the antibiotics most commonly prescribed for hepatic encephalopathy) in improving neurologic signs and symptoms and reducing blood ammonia levels. The results of studies employing small samples are similar to those of the larger studies. Finally, rifaximin has a good tolerability profile in patients with hepatic encephalopathy, and thus appears to constitute a promising new option for this disorder. The current database on rifaximin would be strengthened by results of placebo-controlled studies to quantify more precisely the benefits of therapy.

Anti-Infective Agents↗