Synthesis, identification and in vitro biological evaluation of some novel quinoline incorporated 1,3-thiazinan-4-one derivatives.

The present study describes the synthesis of two new series of 3-hydroxy-N-(4-oxo-2-phenyl-1,3-thiazinan-3-yl)-8-(trifluoromethyl)quinoline-2-carboxamide derivatives (4a-j) and 3-((7-chloroquinolin-4-ylamino)methyl)-2-phenyl-1,3-thiazinan-4-one derivatives (5a-7j). All the compounds were synthesized in moderate to good yield by one-pot three component cyclo-condensation reaction. The newly synthesized compounds were characterized by FT-IR, 1H, 13C NMR and elemental analysis. The compounds were screened for their in vitro antibacterial activity against a panel of pathogenic bacterial strains, antitubercular activity against Mycobacterium tuberculosis H37Rv and also for their in vitro antimalarial activity against Plasmodium falciparum. Among the synthesized compounds two of them (4f and 5f) showed excellent antibacterial activity against C. tetani at 15.6µg/mL. Some of them exhibited excellent antitubercular (4f &5f) and good antimalarial (4f, 5f &6f) activity compared with the first line drugs.

Adjuvants for Clostridium tetani and Clostridium diphtheriae vaccines updating.

It's known that diphtheria and tetanus are a contagious lethal diseases over the years, they caused by pathogenic microbes corynebacterium diphtheria and Clostridium tetani, respectively. The diseases result from the production of bacterial toxin. Vaccination with bacterial toxoid vaccines adsorbed on particulates adjuvants still are the best way to prevent this epidemic diseases from spread. The particulate vaccines have been shown to be more efficient than soluble one for the induction of the immune responses. Nanoparticles can be engineered to enhance the immune responses. As well known the immune response to inactivate killed and subunit vaccine enhances by alum adjuvants. The adjuvants examined and tested after reducing its size to particle size, thus mimic size of viruses which is considered smallest units can derive the immune system. The major issue is minimizing the adjuvant particles, to gain insight of resulting immunity types and impact on immune response. The adjuvant effect of micro/nanoparticles appears to largely be a consequence of their uptake into antigen presenting cells.

Isolation and Antibiogram of Clostridium tetani from Clinically Diagnosed Tetanus Patients.

Clostridium tetani, the etiologic agent of tetanus, produces a toxin that causes spastic paralysis in humans and other vertebrates. This study was aimed for isolation, identification, and determination of antimicrobial susceptibility of C. tetani from clinically diagnosed tetanus patients. Isolation was done from deep-punctured tissues of the foot and arm injuries of 80 clinically diagnosed tetanus patients from the Pakistan Institute of Medical Sciences hospital. We successfully screened out five C. tetani isolates out of 80 samples based on the strain-specific characteristics confirmed through biochemical testing and toxin production. A disc diffusion method was used for antimicrobial susceptibilities and C. tetani isolates showed susceptibility to cefoperazone, chloramphenicol, metronidazole, penicillin G, and tetracycline, but were found to be resistant to erythromycin and ofloxacin. During animal testing, all the infected mice developed symptoms of tetanus. The results showed that identification of C. tetani is possible using biochemical and molecular tools and that the strains of C. tetani isolated had not developed resistance against the antibiotics most often used for the treatment of tetanus.

[Bactericidal Activity of Constructed Recombinant Fusion Protein Pheromonicin-CT].

OBJECTIVE: To construct engineering peptide pheromonicin-Clostrzaum tretant krn-ui), and to test its bactericidal activity. METHODS: We amplified the gene of variable regions from hybridoma cells which secreted monoclonal antibody (mAb) against antigen in the membrane of Clostridium tetani and linked the small antibody mimetic to the channel-forming domain of colicin Ia to create Ph-CT. The Ph-CT was purified by CM sepharose ion-exchange column. Its in vitro antibacterial activity was evaluated by colony culture with different doses of Ph-CT (final concentration 2, 4, 8, and 16 microg/mL,respectively). Then we inoculated culture medium with CT strains and different doses of Ph-CT (final concentration of 4 and 16 microg/mL). The in vivo antibacterial activity of Ph-CT was evaluated by cumulative survival of mice. After 16 hours' anaerobic culture, the mice was treated with filtered CT medium or CT medium. RESULTS: We constructed Ph-CT successfully. CT colonies appeared in the CT medium treated with Ph-CT (2, 4 microg/mL), while no colony appeared in the CT medium treated with Ph-CT (8, 16 microg/mL). All mice survived when they were injected with filtered CT medium treated with Ph-CT (4, 16 microg/mL) and CT medium treated with Ph-CT (16 microg/mL). Three (50%) mice survived when they were injected with CT medium treated with Ph-CT (4 microg/mL). All mice in the control groups died after CT infections. CONCLUSION: Ph-CT may be of value as antibiotics against Clostridium tetani.

Disinfection methods for spores of Bacillus atrophaeus, B. anthracis, Clostridium tetani, C. botulinum and C. difficile.

To evaluate disinfection methods for environments contaminated with bioterrorism-associated microorganism (Bacillus anthracis), we performed the following experiments. First, the sporicidal effects of sodium hypochlorite on spores of five bacterial species were evaluated. Bacillus atrophaeus was the most resistant to hypochlorite, followed in order by B. anthracis, Clostridium botulinum and Clostridium tetani, and Clostridium difficile. Subsequently, using B. atrophaeus spores that were the most resistant to hypochlorite, the sporicidal effects of hypochlorite at lower pH by adding vinegar were evaluated. Hypochlorite containing vinegar had far more marked sporicidal effects than hypochlorite alone. Cleaning with 0.5% (5000 ppm) hypochlorite containing vinegar inactivated B. atrophaeus spores attached to vinyl chloride and plywood plates within 15 s, while that not containing vinegar did not inactivate spores attached to cement or plywood plates even after 1 h. Therefore, the surfaces of cement or plywood plates were covered with gauze soaked in 0.5% hypochlorite containing vinegar, and the sporicidal effects were evaluated. B. atrophaeus spores attached to plywood plates were not inactivated even after 6 h, but those attached to cement plates were inactivated within 5 min. On the other hand, covering the surfaces of plywood plates with gauze soaked in 0.3% peracetic acid and gauze soaked in 2% glutaral inactivated B. atrophaeus spores within 5 min and 6 h, respectively. These results suggest that hypochlorite containing vinegar is effective for disinfecting vinyl chloride, tile, and cement plates contaminated with B. anthracis, and peracetic acid is effective for disinfecting plywood plates contaminated with such microorganism.

Microbiologic characterization and antimicrobial susceptibility of Clostridium tetani isolated from wounds of patients with clinically diagnosed tetanus.

Clostridium tetani is the etiologic agent of the muscle-spasming disease tetanus. Despite an effective vaccine, tetanus is an ongoing problem in some developing countries. Diagnosis by bacterial culture is not done because it is generally unnecessary and the entry of route of the bacteria can be inapparent. We attempted to isolate and evaluate C. tetani from the wounds of 84 patients with tetanus. We effectively isolated C. tetani from 45 patients. All strains tested positive by polymerase chain reaction for the gene encoding tetanus neurotoxin. Antimicrobial susceptibilities were determined by disc diffusion and E-test. All C. tetani isolates were susceptible to penicillin and metronidazole but resistant to co-trimoxazole. Despite treatment with high doses of penicillin, C. tetani was isolated after 16 days of intravenous penicillin in two cases. These data show that the intravenous route for penicillin may be inadequate for clearing the infection and emphasizes wound debridement in the treatment of tetanus.

Tetanus in a subcutaneous drug abuser: ineffectiveness of intrathecal baclofen.

This report discusses the issues involved in the diagnosis and management of tetanus. A 29-year-old female with a history of illicit drug use presented with progressive tetany secondary to an abscess on her forearm from subcutaneous injections. She was managed in the intensive care unit for 29 days and was discharged to the ward. The main issue in her management was control of tetanic episodes interfering with ventilation and resistant to intrathecal baclofen. Other therapeutic strategies aimed at reducing spasms included magnesium, atracurium, pancuronium, midazolam, propofol and dantrolene. We conducted searches on Medline, PUBMED and the Cochrane Database of Systematic Reviews using the following terms: tetanus, treatment, therapy, drug abuse, magnesium, baclofen and human anti-tetanus immunoglobulin (hAIG). The current strategies involved in treating tetanus are discussed.

Menstrum for culture preservation and medium for seed preparation in a tetanus toxin production process containing no animal or dairy products.

AIMS: To completely eliminate animal and dairy products from the lyophilization menstrum and the seed medium used to produce tetanus toxin with Clostridium tetani. METHODS AND RESULTS: Tetanus toxin production in a recently developed fermentation medium lacking animal and dairy products was studied with different seed media. It was found that soy peptone could completely replace the beef heart infusion plus animal peptone previously used as seed medium. In addition, we found that cells lyophilized in soy milk could replace the usual type of cells lyophilized in cow's milk. CONCLUSIONS: We have now developed a complete tetanus toxin production process containing no animal and dairy products. SIGNIFICANCE AND IMPACT OF THE STUDY: Toxoid preparations made from toxin produced with animal and dairy products can contain undesirable contaminants such as the prion causing bovine spongiform encephalopathy (Mad Cow's Disease) or antigenic peptides that stimulate anaphylactic reactions and other undesirable immune reactions in immunized hosts. The new vegetable-based process described here avoids such unfortunate possibilities.

Analysis of the case fatality rate of tetanus among adults in a tertiary hospital in Turkey.

The aim of this study was to present the epidemiological and clinical characteristics of tetanus as related to case fatality in 43 adult patients hospitalized between 1990 and 2000 in Turkey's largest state tertiary hospital. All patients were > or =15 years of age, with the mean age being 45 years. Sixty-five percent of the patients were male. Most patients (77%) were from rural Anatolia. The most common clinical manifestations were trismus (100%), abdominal rigidity (93%), dysphagia (81%), and risus sardonicus (72%). The overall case fatality rate was 58%. In a multivariable logistic regression model an incubation period of < or =8 days versus >8 days was a significant predictor of case fatality (odds ratio, 4.8; P=0.044; 95%CI, 1.04-22.26), as was each day of delay between onset of symptoms and admission to the hospital (odds ratio, 7.4; P=0.035; 95%CI, 1.15-48.28). Although the case fatality rate of tetanus in our clinic has declined in the last 5 years, the overall rate was high. This suggests a need for adult vaccination programs and/or a plan to educate the public to recognize symptoms early and then rapidly seek medical care.

[Study on the effect of a biostimulant on the growth and toxigenic function of Clostridium tetani strain Copenhagen-471]. / Izuchenie vliianiia biostimuliatora na rost i toksinoobrazuiushchuiu funktsiiu Clostridium tetani "Kopenhagen-471."

Bakstim, a new biostimulating preparation obtained from the organs of the immune system of animals, was developed. The impact of Bakstim on the growth and toxigenic function of C. tetani production strain Copenhagen-471 was evaluated. The addition of the preparation to Gluzman commercial medium for obtaining tetanus toxoid led to an increase in the yield of bacterial biomass from 1.9 to 4-fold and an increase in the toxoid production from 2 to 2.8-fold. The optimum concentration of this biostimulant ensuring the maximum yield of tetanus toxin from the production culture was determined (1,000 mg/l). Bakstim will supposedly be used as additive to nutrient media for the production of tetanus toxoid.

Assessment of the efficacy of a low temperature hydrogen peroxide gas plasma sterilization system.

The STERRAD 100 sterilization system (Johnson & Johnson Medical Ltd) uses low temperature hydrogen peroxide gas plasma for sterilization of heat labile equipment. The efficacy of the machine was tested by contaminating a standard set of instruments with different organisms and using a filtration method to assess recovery of organisms from the wash fluids of instruments post-sterilization. Experiments were performed under clean (the organism only) and dirty (organism mixed with egg protein) conditions. A parallel study conducted using a 3M STERIVAC ethylene oxide sterilizer could not be completed owing to closure of the ethylene oxide plant. For sterilization of instruments with long and narrow lumens, hydrogen peroxide adaptors are necessary. The STERRAD 100 sterilizer can achieve effective sterilization of heat labile instruments with a reduction of 6 log10 cfu/mL of organisms tested. This method has the advantages over ethylene oxide sterilization of safety, ease of maintenance and no requirement for aeration time.

Prophylactic effect of ornidazole on experimental tetanus in mice.

Tetanus is still responsible for many deaths, especially in the developing parts of the world. In this study, the prophylactic effect of ornidazole on experimental tetanus in mice was investigated. The initial minimum lethal dose (MLD) of Clostridium tetani spores was determined in mice and 100 MLD was applied to mice in experiments. Ornidazole was then administered at the dose of 20 mg, 40 mg and 80 mg/kg. In addition, penicillin and horse tetanus serum were also administered to other groups. According to the present results, ornidazole decreased the number of deaths in mice significantly in a dose and drug administration time dependent manner. These results suggest that, together with essential wound care and active immunization, ornidazole (or another nitroimidazole) may be a useful and supportive therapeutic agent in tetanus prophylaxis.

La penicilina benzatina: previene el tétanos? / The benzatine penicillin: does it prevent tetanus?

El tétano es un problema de salud pública a nivel mundial, especialmente en los países en desarrollo. Chile no escapa a esta realidad, aunque la incidencia es mucho menor. La alta letalidad de esta enfermedad obliga a realizar un manejo adecuado de todos aquellos pacientes con heridas sospechosas, el que incluye aseo y debridación local junto al uso de la antitoxina tetánica. No existen evidencias de que la antibioterapia influya en su prevención

The role of cell population kinetics in the efficacy of penicillin--an experimental analysis and stochastic modeling of the tumour tetanus and wound tetanus of the mouse.

When investigating tetanus lethality summation curves of mice under comparable quantitative conditions following a temporarily limited administration of penicillin, the curves obtained can be calculated by the kinetics of tumour cells or wound fibroblasts. In particular, it has been shown that the optimal efficacy of penicillin, after short-time usage as compared with a long-time administration schedule, is determined by the generation time of the tetanus rods as a function of the mitotic cycle of the "pace-making" tumour cells or wound fibroblasts. Further variables of the mathematical model imply the pharmacokinetics of penicillin and the recovery process of the "hit" tetanus rods. From these results some basic experimental and clinical tetanus issues can be elucidated; thus, the mitosis theory of tetanus is being verified for the stage of incubation and of clinical manifestation, while the classical necrosis theory of the pathogenesis of tetanus infection should be valid only for the final stage.