scholarly journals STUDIES ON THE MECHANISM OF THE FORMATION OF THE PENICILLIN ANTIGEN

1961 ◽  
Vol 114 (6) ◽  
pp. 875-940 ◽  
Author(s):  
Bernard B. Levine ◽  
Zoltan Ovary
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Penicillin G ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Human Beings ◽  
Mixed Disulfide ◽  
Diastereomeric Mixture ◽  
Potassium Penicillin

An excess of D-benzylpenicillenic acid (BPE) was reacted with human γ-globulin, human serum albumin, gelatin, and poly-L-lysine in aqueous solution buffered at pH 7.5–8.0. Under these conditions, BPE reacted predominantly with lysine ϵ-amino groups of the proteins to form the mixture of diastereomers of ϵ-N-(D-α-benzylpenicilloyl)-lysine groups (Di-BPO-Lys). BPE reacted also, but to a considerably smaller extent, with cystine disulfide linkages of human γ-globulin and human serum albumin to form D-benzylpenicillenic acid-cysteine mixed disulfide groups (BPE-SS-Cys). Conjugates containing large numbers of BPE or D-penicillamine mixed disulfide groups were prepared by reaction of BPE or D-penicillamine with thiolated human γ-globulin under mild oxidizing conditions. Anti-penicillin antibodies were produced in rabbits by immunization with either potassium penicillin G (PG) or a preincubated mixture of PG with normal rabbit serum (PG-NRS) in complete Freund's adjuvant. Specific precipitation analyses in aqueous and gel media (Ouchterlony), PCA analyses, and specific inhibition of these reactions with haptens were carried out on the rabbit anti-PG and anti-(PG-NRS) sera, using the above conjugates as antigens. The anti-penicillin antibodies were found to be directed against the diastereomeric mixture of N-(D-α-benzylpenicilloyl) groups, predominantly the Di-BPO-Lys groups. By these techniques, no antibodies directed against the BPE-mixed disulfide or the D-penicillamine mixed disulfide groups were detected. Three out of six patients with histories of allergic reactions to PG responded with wheal-and-erythema reactions to the N-(D-α-benzylpenicilloyl) (BPO) groups contained in BPE-human gamma globulin conjugate. Another such patient exhibited serum antibodies specific for the BPO group. One patient being treated with 25 gm per day of PG showed the presence of non-dialyzable antigenic BPO-conjugates in his serum. These results demonstrate that the diastereomeric BPO groups (predominantly Di-BPO-Lys groups) are major antigenic determinant groups responsible for PG hypersensitivity in rabbits and human beings. The possible clinical usefulness of multivalent Di-BPO conjugates and univalent Di-BPO haptens is discussed.

98 SUCCESSFUL PREGNANCIES FOLLOWING TRANSFER OF VITRIFIED PORCINE EMBRYOS DERIVED FROM IN VITRO-MATURED OOCYTES

2006 ◽  
Vol 18 (2) ◽  
pp. 157 ◽  
Author(s):  
K. Hiruma ◽  
H. Ueda ◽  
H. Saito ◽  
C. Tanaka ◽  
N. Maeda ◽  
...  
Keyword(s):  
Calf Serum ◽  
Developmental Competence ◽  
Penicillin G ◽  
Cell Stage ◽  
Epidermal Growth ◽  
Potassium Penicillin ◽  
The One ◽  
Parthenogenetic Embryos

To date only in vivo-produced embryos have successfully produced live piglets after cryopreservation. In this study, we aimed to produce piglets from vitrified embryos derived from in vitro matured (IVM) oocytes. Cumulus-oocyte complexes collected from ovaries obtained at a local slaughterhouse were matured for 44 to 45 h in NCSU23 MEDIUM supplemented with 0.6 mM cysteine, 10 ng/mL epidermal growth factor, 10% (v/v) porcine follicular fluid, 75 �g/mL potassium penicillin G, 50 �g/mL streptomycin sulfate, and 10 IU/mL eCG/ hCG. These IVM oocytes were either activated for parthenogenesis or in vitro-fertilized (IVF). For IVF, oocytes were incubated with 5 � 106/mL of cryopreserved epididymal sperm in PGM-tac medium (Yoshioka et al. 2003 Biol. Reprod. 69, 2092-2099) for 20 h. Embryos were treated for removal of cytoplasmic lipid droplets (delipation; Nagashima et al. 1995 Nature 374, 416) at the 4- to 8-cell stages, around 50 to 54 h after activation or insemination. After culture in NCSU23 for 15 h, they were vitrified by the minimum volume cooling (MVC) method. Embryos were equilibrated with equilibration solution containing 7.5% (v/v) ethylene glycol (EG), 7.5% (v/v) dimethylsulfoxide (DMSO), and 20% (v/v) calf serum for 4 min, followed by exposure to vitrification solution containing 15% EG, 15% DMSO, 0.5 M sucrose, and 20% calf serum. Embryos were then loaded onto a Cryotop (Kitazato Supply Co., Tokyo, Japan) and immediately plunged into liquid nitrogen. Vitrified embryos were examined for viability in vitro and in vivo after warming. Their in vitro developmental competence was compared to that of corresponding control (nonvitrified) embryos. Vitrified 4- to 8-cell stage embryos, both parthenogenetic and IVF, showed developmental competence into blastocysts comparable to that of control embryos (parthenogenetic: 46.8%, 36/77 vs. 51.7%, 31/60; IVF: 40.0%, 30/75 vs. 44.3%, 35/79). Of four surrogate gilts that received a total of 251 vitrified parthenogenetic embryos, three became pregnant and had 20 fetuses (8.0%, 22 to 23 days old). Three surrogates gilts that received 267 vitrified IVF embryos all became pregnant. Of those, the one that received 47 embryos was confirmed to have eight fetuses (17.0%, 22 days old) by autopsy. The other two were examined by ultrasonography at 56 and 95 days of gestation and found to be pregnant. These results suggest that porcine embryos derived from IVM oocytes have a potential to develop into live offspring after delipation and MVC vitrification. This study was supported by PROBRAIN.

DEGRADATION OF PENICILLIN G AND THREE SEMISYNTHETIC PENICILLINS BY BACILLUS CEREUS AND STAPHYLOCOCCUS AUREUS PENICILLINASE

1966 ◽  
Vol 12 (1) ◽  
pp. 35-42 ◽  
Author(s):  
J. A. Yurchenco ◽  
M. W. Hopper ◽  
G. H. Warren
Keyword(s):  
Staphylococcus Aureus ◽  
Bacillus Cereus ◽  
Enzymatic Degradation ◽  
Penicillin G ◽  
Aureus Infection ◽  
Penicillanic Acid ◽  
Potassium Penicillin ◽  
And Staphylococcus Aureus

An in vivo procedure is described for determining the relative sensitivities of potassium penicillin G and three semisynthetic penicillins to degradation by Bacillus cereus and Staphylococcus aureus penicillinases. The inactivating concentrations (IC50) of the penicillinases necessary to reduce the protective activity of each of the penicillins against an S. aureus infection in mice from PD95 to a PD50 level was determined. Conventional in vitro studies were carried out for purposes of comparison. After interaction with B. cereus penicillinase, Wy-3206 [6-(2-methoxy-1-naphthamido) penicillanic acid] had the greatest residual therapeutic activity, followed in order by nafcillin [6-(2-ethoxy-1-naphthamido)penicillanic acid], methicillin [sodium 6-(2, 6-dimethoxybenzamido)penicillinate monohydrate], and potassium penicillin G. Penicillin G proved to be the most sensitive to enzymatic degradation by S. aureus penicillinase, whereas nafcillin and methicillin were resistant to the highest concentration employed. These findings were, in general, supported by the in vitro results.

Penicillin Desensitization — A Case Review

1976 ◽  
Vol 10 (6) ◽  
pp. 333-337 ◽  
Author(s):  
Vincent Gotz
Keyword(s):  
New York ◽  
Renal Function ◽  
Serum Creatinine ◽  
Blood Urea Nitrogen ◽  
Bacterial Endocarditis ◽  
Blood Cultures ◽  
Penicillin Allergy ◽  
Penicillin G ◽  
Subacute Bacterial Endocarditis ◽  
Potassium Penicillin

The penicillin class of antibiotics are often life-saving and their use may be required despite prior evidence of allergic manifestations to these drugs. A case report of a patient with subacute bacterial endocarditis who was allergic to penicillin yet requiring treatment with it is presented. The mechanisms and manifestations of penicillin allergy, skin testing for penicillin allergy and the theory and procedures for penicillin desensitization are also discussed. and incontinence. On 8-28-75, edema and petechiae of the lower extremítíes and a new díastolíc heart murmur were noted. Blood cultures were positive for enterococcus sensitive to penicillin. The diagnosis of subacute bacterial endocarditis due to enterococcus was made at this time and intravenous potassium penicillin G 2,400,000 units every four hours and streptomycin 500 mg intramuscularly twice daily were initiated. On 9-1-75, the patient developed a florid rash with pruritus over his entire body. Despite the rash, penicillin was continued along with diphenhydramine. This therapy proved to be ineffective with the rash increasing in intensity and becoming unbearable to the patient. Penicillin was discontinued and cephalothin 2 g intravenously every four hours was initiated with continuation of streptomycin. Repeat blood cultures ten days later remained positive for enterococcus. The murmurs became louder progressing to Grade IV systolic and diastolic over both the mitral and aortic areas. Cephalothin was discontinued and vancomycin 500 mg intravenously every six hours was substituted. The patient's renal function began to slowly deteriorate as evidenced by a blood urea nitrogen of 60 mg% and serum creatinine of 2.5 mg%. One day prior to admission, the vancomycin dose was decreased to 500 mg every eight hours. It was decided to transfer the patient to The New York Hospital for re-evaluation and possible desensitization to penicillin. The patient was admitted to The New York Hospital on 10-2-75 with a blood urea nitrogen of 49 mg% and a serum creatinine of 2.5 mg%. Blood cultures on admission showed no growth. The initial decision was to continue with vancomycin 500 mg intravenously every eight hours and streptomycín 500 mg intramuscularly twice daily unless renal function further deteriorated or ototoxicity developed. The plan was to continue treatment for a duration of six weeks. On 10-6-75, the vancomycin dose was lowered to 250 mg every six hours. Desired therapeutic response was not being obtained on this regimen and renal function continued to worsen (creatinine clearance of 15 ml/minute and a serum creatinine of 3.4 mg%). In view of these findings, it was elected to desensitize the patient to penicillin. A penicillin desensitization schedule was outlined (see Table 1) and therapy was initiated on 10-7-75 without premedication with antihistamines or corticosteroids. Adequate precautions were observed. The patient had intravenous 5 percent dextrose running, along with a tourniquet and tracheostomy set at the bedside. Syringes of epinephrine (1 mg), aminophylline (250 mg), diphenhydramine (50 mg), and methylprednisolone (125 mg) were prepared and ready at the bedside for suppression of allergic and/or anaphylactic episodes. The patient received injections of potassium penicillin G at 20 minute intervals, first intradermally, then subcutaneously and intramuscularly, and finally intravenously. The patient was closely observed during the desensitization and was at no time left unattended. Before each injection, the patient was questioned and examined for manifestations of allergic reactions. Table 1 summarizes the penicillin desensitization schedule. The starting dose of 0.01 U. of penicillin was increased ten-fold at 20 minute intervals up until the drug was begun intravenously. The desensitization was performed over a period of approximately two and one-half hours without complication. After the desensitization procedure was completed, a continuous intravenous infusion of penicillin (20 million units daily) was begun. Due to the degree of renal impairment, the pharmacist recommended alternating 10 million units of sodium penicillin G with 10 million units of potassium penicillin G with careful monitoring of the electrolytes. The dose of streptomycin was decreased to 500 mg daily. The day following desensitization, the patient complained of pruritus without evidence of rash. Diphenhydramine 50 mg was administered orally every four hours as needed, providing relief. The patient continued to do well until 10-24-75 when his gait was noted to be increasingly unsteady. Caloric responses were noted to be hypoactive and the patient complained of nausea with occasional vomiting. Signs and symptoms were suggestive of vestibular toxicity. As a result, the dose of streptomycin was decreased to 250 mg daily. Shortly thereafter, the patient began to complain of dysuria, frequency, and occasional incontinence. A cystoendoscopy was performed showing bladder neck obstruction with bilateral reflux, indicating the necessity for urologic surgery. A cardiac catheterization was also performed that demonstrated marked aortic regurgitation and the necessity for valve replacement. Surgical intervention was deferred at this time at the patient's request. The patient completed his six-week course of antibiotics on 11-12-75 and was discharged on 11-22-75. He is to be readmitted in the near future for cardiac and urologie surgical procedures.

Sign in / Sign up

Export Citation Format

Share Document