Penicillin G acylases (PGAs) are industrially important enzymes primarily used for the synthesis of first- and second-generation cephalosporins or penicillins. This study aims to establish a high-efficiency biosynthetic system for cefradine on the purpose of significantly enhancing its catalytic efficiency in cefradine synthesis and developing its potentials for industrial application. In this study, we identified and engineered penicillin G acylase and obtained a highly active mutant KsPGA M7(M168F/F313G) for the synthesis of cefradine. The mutant achieved a conversion rate over 95% in the scaled-up reaction. To validate its industrial applicability, we immobilized both the wild-type and mutant enzymes and applied them in continuous flow reactions, which achieved a space-time yield of 2 800 g/(L·d). This study lays a foundation for the future applications of penicillin G acylases in the industrial synthesis of cefradine.
Penicillin Amidase/biosynthesis* ; Protein Engineering/methods* ; Cephradine/metabolism* ; Escherichia coli/metabolism* ; Enzymes, Immobilized/metabolism* ; Recombinant Proteins/biosynthesis*

Introduction: Chronic bullous disease of childhood (CBDC) is a rare immune-mediated subepidermal vesiculobullous eruption, characterized by linear IgA deposition along the basement membrane zone of the skin. Although mostly idiopathic, CBDC may be triggered by factors such as infection, and drugs. Clinical and immunohistopathological features of drug-induced cases are heterogeneous and indistinguishable from the idiopathic form. Case report: A two-year-old Filipino male presented with pruritic vesicles and bullae on the back several days after finishing a course of cefuroxime, and cefaclor. Examination revealed multiple tense vesicles and bullae, some coalescing into a rosette pattern with central crusts on the perioral, scalp, neck, back, perineal, and perianal areas. Histopathology showed a subepidermal split with neutrophilic and eosinophilic infiltrates. Direct immunofluorescence revealed strong linear deposition of IgA, and granular deposits of C3 and IgM at the basement membrane zone, thus confirming the di- agnosis of CBDC. Dapsone at 2mg/kg/day was started, with oral prednisolone (1.3mg/kg/day), and cloxacillin syrup (40mg/kg/day). Topical care with betamethasone dipropionate and mupirocin ointment was included. After eight weeks, patient showed significant im- provement with few vesicles and resolved lesions healing with post-inflammatory hyperpigmentation. Conclusion We report a case of a two-year-old male presenting with vesiculobullous lesions after a course of cefuroxime, and cefaclor. As both were given and withdrawn in a period of close proximity, it is difficult to determine the probable culprit drug. Spontaneous resolution upon withdrawal of the suspected drug is variable. Systemic therapy such as dapsone may be necessary for treatment.
Linear IgA Bullous Dermatosis ; Cefaclor ; Cefuroxime

PURPOSE: Cefaclor, a second-generation oral cephalosporin, is known to cause IgE-mediated hypersensitivity. Assays of serum-specific IgE (sIgE) to cefaclor are commercially available via the ImmunoCAP system (Thermo Fisher Scientific). While serum levels of sIgE >0.35 kU/L are considered indicative of an allergy, some patients with cefaclor allergy show low serum IgE levels. This study aimed to evaluate the proper cut-off levels of sIgE in the diagnosis of immediate hypersensitivity to cefaclor. MATERIALS AND METHODS: A total of 269 patients with drug allergy history, who underwent assays of sIgE to cefaclor at Ajou University hospital and Dong-A University Hospital, were reviewed retrospectively. Among them, 193 patients exhibited cefaclor-induced immediate hypersensitivity with certain or probable causality of an adverse drug reaction according to the WHO-UMC (the World Health Organization-the Uppsala Monitoring Centre) algorithm, and 76 controls showed delayed hypersensitivity reactions to non-antibiotics. RESULTS: In total, 126 of the 193 patients (65.3%) experienced anaphylaxis; they had higher serum sIgE levels than patients with immediate hypersensitivity who did not experience anaphylaxis (6.36±12.39 kU/L vs. 4.28±13.61 kU/L, p < 0.001). The best cut-off value for cefaclor-induced immediate hypersensitivity was 0.11 kU/L, with sensitivity of 80.2% and specificity of 81.6%. A cut-off value of 0.44 kU/L showed the best sensitivity (75.4%) and specificity (65.7%) for differentiating anaphylaxis from immediate hypersensitivity reactions. CONCLUSION: Patients with cefaclor anaphylaxis exhibit high serum IgE levels. A cut-off value of 0.11 kU/L of sIgE to cefaclor is proper for identifying patients with cefaclor allergy, and 0.44 kU/L may be useful to detect anaphylaxis.
Anaphylaxis ; Cefaclor* ; Diagnosis ; Drug Hypersensitivity ; Drug-Related Side Effects and Adverse Reactions ; Global Health ; Humans ; Hypersensitivity* ; Hypersensitivity, Delayed ; Hypersensitivity, Immediate ; Immunoglobulin E* ; Retrospective Studies ; Sensitivity and Specificity

Dental avulsion, defined as the complete displacement of a tooth from the alveolar bone with consequent loss of the blood and nerve supply, was reported as one of the most severe dental injuries. Avulsion can cause tissue ischemia, which leads to pulp necrosis.Apexification is a conventional treatment method that induces an apical calcified barrier in immature roots with pulp necrosis. However, root development characterized by an increase in the root thickness and length cannot be achieved by apexification.The purpose of this case report was to describe the radiographic and clinical outcomes of regenerative endodontic treatment for the avulsed and necrosed permanent tooth with an immature root after replantation in a 5-year-old girl; the treatment was performed using a mixture of ciprofloxacin, metronidazole and cefaclor, CollaTape and Biodentine.
Apexification ; Cefaclor ; Child, Preschool ; Ciprofloxacin ; Dental Pulp Necrosis ; Female ; Humans ; Ischemia ; Methods ; Metronidazole ; Replantation ; Tooth

BACKGROUND/AIMS: Several studies have reported on the clinical aspects of adverse drug reactions (ADRs). To date, no study has evaluated serious adverse drug reactions (SADRs) in Korea. The current study evaluates the clinical expression of SADRs in a Korean hospital. METHODS: We reviewed a total of 3,386 cases of SADR occurring between March 2012 and November 2015 in a single tertiary care institution (Regional Pharmacovigilance Center). RESULTS: When classified by organ system, the most common SADRs were white cell and reticuloendothelial system disorders (n = 511). Skin/appendage (n = 296) and gastrointestinal (n = 216) disorders were the fourth- and eighth-most common SADRs, respectively. The three most common single symptoms were leukopenia (n = 499 events), hypotension (n = 444) and anaphylaxis (n = 215). Leukopenia was mainly caused by anti-tumor drugs, followed by piperacilin/tazobactam (n = 28), vancomycin (n = 10) and methimazole (n = 6). Hypotension was most often caused by propacetamol injection (n = 145), while anaphylaxis was mainly caused by cefaclor (n = 19), ranitidine (n = 12), iopamidol (n = 10) and multi-vitamin infusion (n = 9). CONCLUSIONS: Significant differences were noted in the clinical aspects of ADRs and SADRs. Additional studies are warranted to further assess SADRs in response to frequently used causative drugs.
Anaphylaxis ; Cefaclor ; Drug Hypersensitivity ; Drug-Related Side Effects and Adverse Reactions* ; Hypotension ; Iopamidol ; Korea* ; Leukopenia ; Methimazole ; Mononuclear Phagocyte System ; Pharmacovigilance ; Ranitidine ; Tertiary Healthcare* ; Vancomycin

PURPOSE: To investigate the changes in urinary nerve growth factor (uNGF) levels after acute urinary tract infection (UTI) and to assess the role of uNGF in predicting UTI recurrence in women. METHODS: Women with uncomplicated, symptomatic UTIs were enrolled. Cephalexin 500 mg (every 6 hours) was administered for 7-14 days to treat acute UTIs. Subsequently, the patients were randomized to receive either sulfamethoxazole/trimethoprim 800 mg/160 mg daily at bedtime, or celecoxib 200 mg daily for 3 months and were monitored for up to 12 months. NGF levels in the urine were determined at baseline, 1, 4, and 12 weeks after the initiation of prophylactic therapy, and were compared between women with first-time UTIs and recurrent UTIs, sulfamethoxazole/trimethoprim and celecoxib-treated women, and no UTI recurrence and UTI recurrence that occurred during the follow-up period. Twenty women free of UTIs served as controls. RESULTS: A total of 139 women with UTI and 20 controls were enrolled in the study, which included 50 women with a first-time UTI and 89 women with recurrent UTIs. Thirty-seven women completed the study. Women with recurrent UTIs (n=23) had a trend of lower uNGF levels than women with first-time UTIs (n=14). During follow-up, 9 women had UTI recurrence. The serial uNGF levels in women with UTI recurrence were significantly lower than those in women who did not have UTI recurrence during the follow-up period. CONCLUSIONS: The lower levels of uNGF in women with recurrent UTI and the incidence of UTI recurrence during follow-up suggest that lower uNGF might reflect the defective innate immunity in women with recurrent UTI.
Biomarkers ; Celecoxib ; Cephalexin ; Female ; Follow-Up Studies ; Humans ; Immunity, Innate ; Incidence ; Inflammation ; Nerve Growth Factor* ; Recurrence ; Urinary Tract Infections* ; Urinary Tract*

Actinomycosis ; complications ; drug therapy ; pathology ; Aged, 80 and over ; Amoxicillin-Potassium Clavulanate Combination ; therapeutic use ; Anti-Bacterial Agents ; therapeutic use ; Cephalexin ; therapeutic use ; Ciprofloxacin ; therapeutic use ; Clindamycin ; therapeutic use ; Coinfection ; drug therapy ; Drug Resistance, Bacterial ; Escherichia coli Infections ; complications ; drug therapy ; Humans ; Male ; Pseudomonas Infections ; complications ; drug therapy ; Skin Diseases, Bacterial ; complications ; drug therapy ; pathology ; Staphylococcal Skin Infections ; complications ; drug therapy ; Thigh ; Trimethoprim, Sulfamethoxazole Drug Combination ; therapeutic use

PURPOSE: Cefaclor is widely prescribed for various infectious diseases. As its consumption increases, the number of hypersensitivity reactions to cefaclor has increased. This study aimed to evaluate the immunologic findings of immediate hypersensitivity to cefaclor. MATERIALS AND METHODS: We enrolled 47 patients with immediate hypersensitivity to cefaclor from Ajou University Hospital and Asan Medical Center. Serum specific IgE, IgG1, and IgG4 antibodies to cefaclor-human serum albumin (HSA) conjugate were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The most common phenotype was anaphylaxis (Group I, 78.7%), followed by urticaria (Group II, 21.3%). The detection of specific IgE, IgG1, and IgG4 to cefaclor-HSA conjugate by ELISA tended to be higher in Group I (40.5%, 41.7%, 21.6%) than in Group II (20.0%, 20.0%, 0%) with no statistical significance. Significant associations were found between specific IgE and IgG1 or IgG4 (p<0.001, p=0.019). ELISA inhibition tests showed significant inhibitions by both free cefaclor and cefaclor-HSA conjugate. For basophil activation tests in patients having no specific IgE antibody, the CD63 expression level on basophils increased with incubations of free cefaclor. CONCLUSION: The most common manifestation of immediate hypersensitivity to cefaclor was anaphylaxis, most of which was mediated by IgE; however, a non-IgE mediated direct basophil activation mechanism was suggested in a subset of anaphylaxis patients.
Adolescent ; Adult ; Aged ; Anaphylaxis/*chemically induced/immunology ; Anti-Bacterial Agents/adverse effects/*immunology ; Antigens, CD63 ; Basophils/metabolism ; Cefaclor/*adverse effects/immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Hypersensitivity, Immediate/chemically induced/diagnosis/*immunology ; Immunoglobulin E/*blood ; Immunoglobulin G/immunology ; Male ; Middle Aged ; Retrospective Studies ; Skin Tests ; Urticaria/chemically induced/diagnosis/immunology ; Young Adult

PURPOSE: To report a case of Pseudomonas fluorescens infection following endoscopic dacryocystorhinostomy and silicone tube intubation in a healthy patient who was using steroid nasal spray. In addition, a literature review is conducted. CASE SUMMARY: A 72-year-old female patient came to our clinic with tearing and hyperemia in the right eye. Ten months prior, she had undergone endoscopic dacryocystorhinostomy and silicone tube intubation due to nasolacrimal duct obstruction in the right eye. Six months after the first operation, dacryocystorhinostomy revision with silicone tube exchange was performed due to obstruction of the nasal bony orifice. In addition, the patient was using a steroid nasal spray. On slit lamp examination, conjunctival injection, marked inflammation and punctal edema around the tube were observed. The silicone tube was removed and the tube cultured. Pseudomonas fluorescens was isolated from the tube contents. The patients was treated with topical 0.3% gatifloxacin 4 times a day, methylol cephalexin lysinate 1000 mg 3 times a day and the nasal spray was discontinued. Two weeks later, all symptoms were resolved after treatment with antibiotic treatment. CONCLUSIONS: A case of Pseudomonas fluorescens canaliculitis which occurred in healthy patient who was using steroid nasal spray is presented with a literature review. Pseudomonas fluorescens canaliculitis can be treated by using proper antibiotics.
Anti-Bacterial Agents ; Cephalexin ; Corneal Ulcer ; Dacryocystitis ; Dacryocystorhinostomy ; Edema ; Eye ; Female ; Fluoroquinolones ; Humans ; Hyperemia ; Inflammation ; Intubation ; Nasolacrimal Duct ; Porphyrins ; Pseudomonas ; Pseudomonas fluorescens ; Silicones ; Canaliculitis

To explore the enzymatic route of cefatrizine synthesis, alpha-amino acid ester hydrolase (AEH) gene was cloned from the whole genome of Xanthomonas rubrillineans, and expressed heterologously in Escherichia coli BL21 (DE3). The effects of temperature, pH and substrates' molar ratio upon the transformation yield of cefatrizine by purified recombinant AEH were investigated. The monomer of AEH was determined as 70 kDa by SDS-PAGE. The optimal pH and temperature reaction were (6.0 +/- 0.1) and 36 degrees C for cefatrizine synthesis. The transformation yield was 64.3% under 36 degrees C, pH (6.0 +/- 0.1), when the concentrations of two substrates were about 30 mmol/L (7-ATTC) and 120 mmol/L (HPGM x HCl), respectively, and the enzyme consumption was 22 U/mL. The results pave the way for optimization of the industrial enzymatic synthesis of cefatrizine.
Carboxylic Ester Hydrolases ; biosynthesis ; genetics ; Catalysis ; Cefatrizine ; metabolism ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Kinetics ; Recombinant Proteins ; biosynthesis ; genetics ; Xanthomonas ; enzymology