Streptomycin Solution Research Articles

Metabolism of Isolated Leaves: I. Changes in the Protein, Soluble Nitrogenous Compounds, Sugars, and Organic Acids in Tobacco Leaves in Light and Dark

Degradation of protein level in detached leaves has long been known and the accumulation of free amino acids and amide at the expense of protein breakdown has been evident from the time of Schulze (1898) followed by Chibnall (6), Mothes (cf. 6), Yemm. (42), Vickery et al (33), Mc Kee (18), Wood et al (41), and Viets et al (38). However the changes in individual amino acids remained uninvestigated because of the lack of proper methods. During recent years paper chromatography has been widely applied as a tool for biochemical investigations and recently an attempt was made to study the amino acid metabolism of leaves after removal from the plant under conditions of light and dark (23, 24). difficulty in detecting some of the overlapping amino acids by the horizontal migration method necessitated examination employing two-dimensional chromatography of amino acids. Since protein metabolism is closely related to other metabolic activities (6, 12) and hence is difficult to interpret without knowledge of the other processes, it was of interest to follow the changes in the other two important fractions, the sugars and the acids, simultaneously occurring in detached leaves. In this way it was hoped to obtain a clear picture of metabolic events operative during the culture of detached leaves. Nicotiana tabacumi L. was selected as the working material, for its metabolic behavior is known through the work of Vickery and his colleagues (33), and it has been found to exhibit characteristic metabolism not only in relation to protein, amino acids, and amides but also with respect to acids. Although the loss o.f malic acid and gain of citric acid has repeatedly been confirmed for Connecticut tobacco leaves cultured in the dark (33, 34, 35, 36), it becomes, however, difficult to explain these results if the normal citric acid cycle (16) is considered operative in these leaves. At the most it could explained by assuming that in detached leaves cultured in the dark the rate of synthesis of citric acid from malic acid exceeded that of the decomposition of citric acid. How darkening of detached leaves would foster these changes is a matter of speculation. Organic acids of the tricarboxylic acid cycle are not normally consumed for meeting the energy requirements of the tissue, they being intermediate oxidation products. Chibnall (6), while calling it an organic acid phase in the metabolism of starving tobacco leaves, strongly felt that such a behavior would most unlikely if the normal Krebs cycle is considered operative. In the latter case the levels of the respective acids should have remained unchanged. Regarding the possibility of an acid cycle participating in Nicotiana leaves to meet its energy requirements, Chibnall (6) himself wrote: The interpretation that I have given to these five sets of starvation experiments is admittedly speculative in that it is based on the hypothesis of an acid cycle in respiration for which there is no ad hoc proof at present and perhaps there never can be (p. 241). This conclusion was, of course, due to lack of the knowledge of pathways of malic-citric conversion involving enzyme systems other than those in the Krebs cycle at that time. Experiments have been conductecl to elaborate the pathway of malic-citric conversion in Nicotiana leaves. success has been mainly due to the finding that the leaves cultured in streptomycin and penicillin solution in dark failed to show the conversion, which ANicotiana tabacumn var. Cheroot leaves carried out when cultured in water in the dark. This observation made it necessary to examine whether the acid metabolism of variety S-20 leaves which (lid not show this conversion, could also altered by culturing on antibiotic solution, and whether the enzymes for malic-citric conversion of the regular citric acid cycle were operative in these leaves. latter has been demonstrated by the simple method of feeding the leaves with acid salts (succinate and citrate).

Influence of streptomycin solutions on the interaction between the agglutinating sera and the corresponding red blood cell receptors.

WORK on the influence of different chemical compounds on the reaction between blood group receptors of red cells and the corresponding antibodies has been published; but the action of antibiotics on this reaction, however, has not yet been fully examined except by Neter et al. 1, who described the effect of antibiotics on enterobacterial lipopolysaccharides utilizing haemagglutination and haemolysis reactions. Our chance discovery of the inhibiting effect of a streptomycin solution on the reaction between anti-D antibodies and D-positive erythrocytes led us to study the effect of different streptomycin concentrations on the antigen-antibody interaction in blood group systems. For our experiments we used streptomycin of Czechoslovak origin (‘Streptomycinum sulphuricum’, Penicillin Works, Prague). The different streptomycin concentrations were prepared by diluting 1 gm. of streptomycin in 2, 5, 10, 20, 50, 80 and 100 ml. of saline. Agglutinating sera of the systems A 1 A 2 BO, MN and Rh/Hr were chosen for the reaction; the red blood cells of the corresponding blood groups were washed three times in saline before use.

STUDIES ON THE UPTAKE AND TRANSLOCATION OF STREPTOMYCIN BY PEACH SEEDLINGS

Measurements of the antibiotic activity in the foliage of peach seedlings, kept with their roots in mineral nutrient solutions containing streptomycin, showed that streptomycin passed into the foliage and sometimes reached concentrations above those of the solutions surrounding the roots. The amount of the antibiotic in leaves varied with the time that roots were kept in the solutions and with the streptomycin concentrations of the solutions. Lower leaves contained much more than upper leaves.Adding macerated leaf tissue to streptomycin solutions decreased the amount of streptomycin detectable in the supernatant liquid more than could be accounted for by dilution or alteration of pH.Streptomycin first affected plants by decreasing apical growth, but at higher concentrations it caused chlorotic and necrotic areas on leaves and stems.

Absorption of antibiotics by plant cells. II. Streptomycin

When cells of the alga Nitella clavata are placed in a streptomycin solution, the antibiotic enters rapidly and accumulates within the cells. The effects of variations in external concentration, pH, temperature, competitive ions, respiratory inhibitors, and glucose on the rate of streptomycin uptake suggest that the process is an active transport involving an expenditure of energy by the cell.

Studies on the Streptomycin for Plants

It was found that a 0.3mcg/ml water solution of streptomycin at first inhibits the growth and later causes the death of Ps. solanacearum, while a concentration of 1mcg/ml is required to inhibit the growth of Ps. tabaci which can be sometimes resumed later. Both bacteria were killed at once by 5mcg/ml solution.And the writers have studied surface absorption of streptomycin in tobacco plants from various points of view, namely absorption in various plant organs, absorption amount and concentration, absorption and surface active agents, absorption and light, absorption rate and streptomycin injury to the plant.It was found that amount of streptomycin absorbed by the lower surface of a leaf is twice as large as that absorbed by the upper surface. The higher the concentration the larger amount of streptomycin is absorbed but from a certain limit upwards the injury inflicted upon the plant increases. By adding as a surface active agent 0.1% Tween 20 the absorption was improved and the difference between the lower and the upper leaf surface became smaller. The maximum of absorption was reached 8 hours after spraying. There was no correlation between light and absorption. The plants could recover when the injury was not severe.

Studies on the Streptomycin for Plants

Streptomycin would be diluted in the course of growth of the developing leaf but it remained over 5 weeks in the fully developed leaf. When applied to the stem, it moved to the phloem and pith, reaching to the next leaf above the place of absorption, and later proceeded gradually to the upper leaves. It could not be traced in the xylem. It could not traverse the midrib and move from one longitudinal half to the other, neither did it move from the basal to the apical part of the leaf or vice versa.When inoculation with Pseudomonas tabaci was carried out one day after spraying the leaves with 220, 110 and 73mcg/ml water solutions of streptomycin, the disease was found to be under complete control even 10 days after inoculation. A concentration of about 110mcg/ml can be recommended. The yellow halo around the spots of infection will turn green when sprayed with such a solution. In the case of Ps. solanacearum, 5 days after inoculation the pathogenic organism was present due to the dilution of streptomycin in the tissue, which should be controlled by making the repeated spraying.

Contamination of streptomycin solutions with resistant bacteria.

In the early 1990s a multicentre survey on asthma was performed on the young adult population (European Community Respiratory Health Survey - ECRHS). This study is to be repeated in order to estimate changes in the prevalence of asthma-like symptoms during the last decade and to assess the social and economic costs of the disease and their variations among countries. The self-administered questionnaire devised for this purpose is a two-page questionnaire. The first page contains the same items as those used in the first survey with four additional questions related to: 1) the frequency and severity of asthma attacks; 2) the presence of chronic bronchitis; 3) smoking habits; and 4) a visual analogue scale assessing perception of outdoor pollution. The second page aims to collect information regarding the direct and indirect costs of asthma. The influence of the length of the questionnaire on the response rate was assessed in a pilot study in Italy. Two random samples of 150 subjects received either the one-page questionnaire (first page) or the two-page questionnaire. The response rate was compared with that obtained from the first postal wave in the 1991-1992 survey. Although the response rate was unchanged when using the one-page questionnaire (45% versus 45%), it decreased by 7% when the two-page questionnaire was used (38% versus 45%). On the basis of these results, no problem should arise if four more questions are added to the one-page questionnaire. The slight reduction in the response rate of the two-page questionnaire is worrying but could be corrected by the use of telephone interviews.

STREPTOMYCIN, CAUSE OF DERMATITIS VENENATA IN A NURSE

Streptomycin may cause toxic erythema, 1 flushing, 2 urticaria, 2 erythema nodosum. 2c pruritus, 3 conjunctivitis, 4 exfoliative dermatitis, 5 maculopapular 6 and hemorrhagic 7 cutaneous eruptions. At the site of the injection swelling, erythema, heat, pain 3 and burning 2a as well as regional adenitis 8 and thrombosis 2b have been reported. Dermatitis venenata 3 has been noted in workers manufacturing this antibiotic. Since Paine and his group 7 found in a review of 291 papers on streptomycin that aforementioned reactions did not follow its ingestion, inhalation or local application, it is important to present this case of dermatitis venenata in a nurse who made and administered solutions of streptomycin. REPORT OF A CASE R. E. B., a 34 year old graduate nurse, was admitted to the department' of dermatology and syphilology on May 5, 1947, complaining of a dermatitis of the fingers of three weeks' duration as well as

Isolation of a Streptomycin-Resistant Organism Capable of Utilizing Streptomycin for Growth

ConclusionsThe isolation and cultural characteristics of a streptomycin-resistant organism capable of multiplying in aqueous solutions of streptomycin has been described.

Contact Dermatitis from Topical Tyrothricin and Associated with Polyvalent Hypersensitivity to Various Antibiotics : Report of a Case

In contrast to penicillin and other antibiotics, tyrothricin has to our knowl” edge not previously been reported as a sensitizing agent. We therefore wish to report the detailed study of a case of contact dermatitis following the topical application of a solution of tyrothricin. A 43 year old white woman developed an ulcer on the medial aspect of the left ankle tw° years previously. Various types of treatment for a period of 6 months were of no avai until application of tyrothricin solution compresses healed the lesion. The ulcer recurred two and one half months prior to admission to the hospital. Tyrothricin solution was again used and the ulcer began to heal. After about 6 weeks of this treatment an acute eczematoid dermatitis of the lower leg developed. Penicillin ointment was then used without relief and with perhaps further aggravation of the dermatitis. The past history was essentially negative except for a bilateral thrombophlebitis of the legs which followed a pregnancy 20 years before. There were no previous episodes of eczematoid dermatitis, no known fungous disease of the feet and, prior to the application of tyrothricin, no use of any antibiotic. The only physical findings were those relating to the acute dermatitis involving the lower one third of the left leg and the middle portion of the plantar surface and entire dorsum of the left foot. The skin of these areas was edematous, erythematous and eczematoid in character. There was also a moderately severe erythematous papulo-vesicular eruption of the upper extremities. There was no clinical evidence of fungous disease of the feet. Re-examination after 6 weeks likewise revealed no evidence of this type of infection. At the present time, the patient remains free from her dermatitis. The following patch tests were done: Material Concentration Result Tyrothricin solution 25 mg./cc. -1-1-I-{- Tyrothricin ointment 0.5 mg./gm. Penicillin ointment 1000 units/gm. Penicillin G solution 500 Units +-f Streptomycin ointment 5000 units/gm. - Streptomycin solution 25 mg. - Gliotoxin cream 1:1000 ++ + Gliotoxin ointment 1:1000 + + + Trichophytin solution 1:30 - Coccidioidin 1:1000 Histoplasmin 1:10 - Oidiomycin Undiluted - 243 244 THE JOURNAL OF INVESTIGATIVE DERMATOLOGY Material Concntraiion Remit Bacitracin 1200 units/cc. -\-}--f Ammoniated mercury ointment 3% . - There was no reaction to repeated applications of adhesive. The following intradermal tests were done: Material Concentration Results Penicillin G. solution 500 units Histoplasmin 1:100 Trichophytin 1:30 + + + H- Oidiomycin 1:100 + + + + Streptomycin solution 25 mg. + Bacitracin 120 units +h t Coccidioidin 1:100 - Later eczematoid reaction also. f Two spontaneous flare-up reactions (++) two weeks, and three weeks afterwards. DISCUSSION Because of the enthusiasm with which penicillin was accepted and used, tyrothricin, until recent periods, has not been used as extensively as a topical agent. Its hemolytic properties and low water solubility have further limited the use of tyrothricin to topical medication. Some workers feel that this remedy possesses some advantages and has not received the full consideration it deserves. It is our opinion that topical tyrothricin is less effective than topical penicillin or even topical streptomycin or topical bacitracin. In the case presented, we believe that we are dealing with a polyvalent hyper-sensitivity associated with therapy with (and initiated by) topical tyrothricin. There is no reason to suspect that this patient, prior to the demonstration of her sesitivities, was sensitive to the agents to which she later reacted. A prior sensitization to a common fungus or yeast-like antigen could not explain all the sensitivities, for hypersensitivity to tyrothricin and bacitracin was demonstrated. There is no ready explanation, also, for the flare-up reactions to bacitracin. In the past history, it will be recalled that the patient gave a history of thrombophlebitis of both legs 20 years previously. It is interesting to note the relative ease with which the skin of the lower extremity with underlying circulatory disturbances becomes sensitized in contrast to the normal skin (1). The reaction of sensitization is to be differentiated from the “grease reaction7' on the lower extremities. This problem is under studjr at the present time. CONCLUSIONS Contact dermatitis following the use of tyrothricin solution in a case of stasis syndrome is described. There was a wide range of eczematoid and tuberculin type sensitivities to various antibiotics including bacitracin demonstrated by skin test.

Topical use of streptomycin in wounds

1. 1. For topical application to wounds, a solution of streptomycin of the proper concentration should be used, not the powder. Freshly wounded tissues are not damaged further and wounds heal without interference when the concentration of streptomycin is at 200 units or micrograms per cc. Granulations are not damaged by 1,000 units per cc. At these concentrations, streptomycin is the best non-toxic antibiotic that has been found to date for gram-negative bacilli. 2. 2. Penicillin, up to 1,000 units, or sulfamylon 5 per cent should always be combined with streptomycin when it is used topically because streptomycin does not have a complete bacterial spectrum at its proper concentration and some susceptible bacteria rapidly acquire resistance to streptomycin alone. 3. 3. Conversely, streptomycin should always be used topically with penicillin because streptomycin kills gram-negative bacilli that penicillin is unable to destroy and these elaborate penicillinase that in turn destroys penicillin. 4. 4. Gram-negative bacilli are almost always present at some time in the evolution of the bacterial flora of wounds even though only a single strain of bacteria may be isolated at one time. Gram-negative bacilli usually dominate the flora of chronically infected wounds, accounting in part for the failure of topical application of penicillin alone in this type of wound. 5. 5. Gram-negative bacilli definitely interfere with the healing of wounds despite arguments that have been advanced that they are mere contaminants of pus. 6. 6. Because of its stability, sulfamylon 5 per cent makes a better combination with streptomycin than with penicillin. Sulfamylon has a wider bacterial spectrum than penicillin though it is not always as powerful and it acts rapidly in the presence of pus. It can be used with any form of parenteral therapy. This mixture encounters few resistant strains of bacteria. 7. 7. Topically, the solution of streptomycin and penicillin or sulfamylon will prevent infection in wounds better than it will hasten the resolution of established infection, except in certain instances in which no slough is present. 8. 8. In the presence of crushed tissue, prevention of infection can be obtained experimentally until three hours after wounding by washing the wound and injecting about 20 cc. of the solution in the surrounding tissues. With débridement of crushed tissue, prevention of infection has been effected as late as forty-eight hours after wounding when the solution is employed in the same manner. 9. 9. All devitalized tissue cannot be débrided from traumatic wounds nor can the process always be done in time; therefore, topical application of the solution of streptomycin and penicillin or sulfamylon is recommended in the treatment of traumatic wounds as a safety factor to prevent infection. 10. 10. The solution of streptomycin and penicillin or sulfamylon will decontaminate the clean-contaminated operative wound. The solution is applied topically before the wound is closed as one would an antiseptic to the skin. 11. 11. To hasten resolution of the established localized infection in the wound, adjunct chemotherapy in addition to antibacterial chemotherapy is required. This adjunct chemotherapy must liquefy slough and cause the antibacterial substances to penetrate and kill bacteria in slough and leukocytes. 12. 12. A mixture of acid, glycerine and thymol has been used for this adjunct chemotherapy. It is successful when sloughing fascia is present but it is not successful in the presence of osteomyelitis. 13. 13. Streptomycin alone can rid wounds of persistent gram-negative bacilli infection when no slough is present in the wound. For example, granulations that will not take skin grafts because they harbor a flora of the gram-negative bacilli can be freed of these bacteria by the local application of streptomycin and then skin grafts will take. 14. 14. No otic complications have been observed with the low concentration of streptomycin used topically. However, topical therapy has not been used as frequently nor over such long periods of time as parenteral therapy.

Streptomycin in Experimental Ocular Infections.

Summary1. The penetrability of streptomycin through the cornea may be increased by abrasion, inflammation, ion-transfer and wetting agents. 2. No local toxic effects were noted when saline solutions of streptomycin containing up to 10,000 μg per ml were used. With concentrations of 50,000 μg or as a dry powder, delayed healing occurred. 3. Intraocular injection, in amounts up to 1,000 μg of streptomycin in 0.1 ml saline were well tolerated. Smaller amounts (25 to 300 μg) were therapeutically effective up to 6 to 8 hours against a virulent strain of Str. pyogenes, though transient or negligible vitreous opacities occurred with these concentrations. 4. Experimental corneal ulcers produced by injection of B. pyocyaneus were prevented when treatment was started within 6 hours after inoculation by 3 applications at 2-hour intervals of a saline solution of streptomycin containing 10,000 μg per ml.

Further Studies on the Testing of Sterility of Concentrated Streptomycin Solutions

1. Further studies with semi-carbazide and hydroxylamine confirm earlier findings that for most of the bacterial species tested the former substance mol for mol is less bacteriostatic than the latter. 2. The use of either carbonyl reagent in sterility tests of concentrated streptomycin solutions, however, may lead to false negative findings. 3. Three strains of microörganisms have been isolated from streptomycin solutions which failed to grow out from similar material inactivated with either carbonyl reagent. 4. These organisms isolated from the streptomycin solutions grow poorly in thioglycolate broth, and one species was found to die out in this broth on incubation prolonged beyond 6 days. 5. The demonstration of psychrophilic organisms in streptomycin solutions suggests the need for incubating such sterility tests at more than one temperature.

Effect of Rubber Tubing Upon the Stability of Penicillin and Streptomycin Solutions

Effect of Rubber Tubing Upon the Stability of Penicillin and Streptomycin Solutions

Effect of Rubber Tubing Upon the Stability of Penicillin and Streptomycin Solutions

Effect of Rubber Tubing Upon the Stability of Penicillin and Streptomycin Solutions

EFFECT OF ZEPHIRAN CHLORIDE, TYROTHRICIN, PENICILLIN AND STREPTOMYCIN ON CILIARY ACTION

THIS study was made to determine what effect, if any, solutions of zephiran chloride, tyrothricin, penicillin and streptomycin have on the ciliary activity of the mucous membranes of the upper respiratory tract. MATERIALS AND METHODS The Animal Used .—Twenty normal healthy rabbits were used for the entire study. Five were utilized for each of the four drugs in question. Preparation of the Mucous Membranes .—The ciliated mucous membranes were studied by two methods. In one they were extirpated and placed in the solutions in question. In the other they were left intact and bathed with the solutions that were being tested. For removal of the ciliated mucosa the animal was given a lethal dose of either pentobarbital sodium or ethyl carbamate U.S.P. (urethane) intravenously. A midline incision of the skin of the nose was made, following the bridge throughout its entire length. The periosteum was elevated and the bones covering the nasal

A Procedure for Testing Sterility of Concentrated Streptomycin Solutions

Summary1. Details are given for a method of inactivating streptomycin with semicarbazide-hydrochloride in order to test the sterility of concentrated streptomycin solutions. It has been shown that although several carbonyl reagents inhibit bacterial growth, it requires from 533 to >11,000 times more semicarbazide-hydrochloride (one of the least toxic of the group) to cause this inhibition than is required of streptomycin.2. Spores of Bacillus sp. No. 290, when added to a streptomycin solution containing 28,700 units per ml, were able to grow out when diluted in thioglycolate broth. When a similar spore suspension-streptomycin mixture was treated with semicarbazide-hydrochloride (3, 2 or 1 y carbonyl reagent per unit (or y) of streptomycin) and then diluted in thioglycolate broth, growth occurred at a dilution.