The purpose of this study was to evaluate the viability of periodontal ligament cells in rat teeth using slow cryo-preservation method under pressure by means of MTT assay and WST-1 assay. Eighteen teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both sides of the first and second maxillary molars were extracted as atraumatically as possible under Tiletamine anesthesia. The experimental groups were group 1 (Immediate control), group 2 (Cold preservation at 4degrees C for 1 week), group 3 (Slow freezing), group 4 (Slow freezing under pressure of 3 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in 37degrees C water bath, then MTT assay and WST-1 assay were processed. One way ANOVA and Tukey method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 4 showed significantly higher viability of periodontal ligament cells than group 2 and 3 (p < 0.05), but showed lower viability than immediate control group. By the results of this study, slow cryo-preservation method under pressure suggests the possibility for long term cryo-preservation of the teeth.
Anesthesia ; Animals ; Baths ; Dimethyl Sulfoxide ; Eosine Yellowish-(YS) ; Female ; Freezing ; Humans ; Molar ; Periodontal Ligament ; Rats ; Tiletamine ; Tooth

The purpose of this study was to evaluate the viability of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure by means of MTT assay, WST-1 assay. 12 teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both side of the first and second maxillary molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group 1 (Immediate extraction), group 2 (Slow freezing under pressure of 3 MPa), group 3 (Slow freezing under pressure of 2 MPa), group 4 (Slow freezing under no additional pressure), group 5 (Rapid freezing in liquid nitrogen under pressure of 2 MPa), group 6 (Rapid freezing in liquid nitrogen under no additional pressure), group 7 (low-temperature preservation at 0degrees C under pressure of 2 MPa), group 8 (low-temperature preservation at 0degrees C under no additional pressure), group 9 (low-temperature preservation at -5degrees C under pressure of 90 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in 37degrees C water bath, then MTT assay, WST-1 assay were processed. One way ANOVA and Tukey HSD method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 7 (0degrees C/2 MPa) showed higher viability of periodontal ligament cells than other group (2-6, 8) and this was statistically significant (p < 0.05), but showed lower viability than group 1, immediate extraction group (no statistical significance). By the results of this study, low-temperature preservation at 0degrees C under pressure of 2 MPa suggest the possibility for long term preservation of teeth.
Anesthesia ; Animals ; Baths ; Cryopreservation ; Dimethyl Sulfoxide ; Eosine Yellowish-(YS) ; Female ; Freezing ; Humans ; Molar ; Nitrogen ; Periodontal Ligament ; Rats ; Tiletamine ; Tooth

The purpose of this study was to evaluate the viability of periodontal ligament cell in rat teeth using slow cryopreservation method with magnetic field through MTT assay and TUNEL test. For each group, 12 teeth of 4 weeks old white female Sprague-Dawley rat were used for MTT assay, and 6 teeth in TUNEL test. The Maxillary left and right, first and second molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group1 (immediately extraction), group 2 (cold preservation at 4degrees C for 1 week), group 3 (rapid cryopreservation in liquid nitrogen), group 4 (slow cryopreservation with magnetic field of 1 G), and group 5 (slow cryopreservation). F medium was used as preservation medium and 10% DMSO as cryoprotectant. After preservation and thawing, the MTT assay and TUNEL test were processed. One way ANOVA and Scheffe method were performed at the 95% level of confidence. The value of optical density obtained after MTT analysis was divided by the value of eosin staining for tissue volume standardization. In both MTT assay and TUNEL test, it had showed no significant difference among group 3, 4, and 5. And group 3 had showed higher viability of periodontal ligament cell than group 2. From this study, slow cryopreservation method with magnetic field can be used as one of cryopreservation methods.
Anesthesia ; Animals ; Cryopreservation ; Dimethyl Sulfoxide ; Eosine Yellowish-(YS) ; Female ; Humans ; In Situ Nick-End Labeling ; Magnetic Fields ; Magnetics ; Magnets ; Molar ; Periodontal Ligament ; Rats ; Tiletamine ; Tooth

Zoletil is a non-opioid, non-barbiturate animal anesthetic and proprietary combination of two drugs, a dissociative anesthetic drug, tiletamine, with the benzodiazepine anxiolytic drug, zolazepam. Zoletil has greater potency than ketamine. Zoletil is abused for recreational purposes, especially by people with easy access to medicine. However, in Korea, it is available over-the-counter. Here we report on a case of an 83-year-old woman who received injection of seven vials of "Zoletil 50" by her daughter and presented with an altered mental change. Her mental state was stupor and vital sign was hypotension, bradycardia. Her blood tests indicated metabolic and respiratory acidosis and hyperkalemia. She was treated with intravenous naloxone and flumazenil but was not responsive. She was admitted to the ICU and treated with supportive therapy. Her mental state showed transient recovery, however, her clinical manifestation worsened and she expired.
Acidosis, Respiratory ; Aged, 80 and over ; Animals ; Benzodiazepines ; Bradycardia ; Drug Combinations ; Female ; Flumazenil ; Hematologic Tests ; Humans ; Hyperkalemia ; Hypotension ; Ketamine ; Korea ; Naloxone ; Nuclear Family ; Stupor ; Tiletamine ; Vital Signs ; Zolazepam

Zoletil is a non-narcotic, nonbarbiturate, injectable veterinary anesthetic agent, which is routinely used as a veterinary anesthetic. Zoletil is an equal weight (1:1 ratio) combination of tiletamine hydrochloride and zolazepam hydrochloride. Tiletamine is a dissociative anesthetic agent with pharmacological similarity to ketamine, and zolazepam is a nonphenothiazine, diazepinone minor tranquillizer similar to diazepam. Zoletil is the term used in Europe and Telazol is used in the USA. Adverse effects of zoletil in animals include salivation, vomiting, tachycardia, seizures, central nervous system stimulation, apnea, and prolonged recovery time. A 30-year old male who was poisoned with zoletil visited our emergency center and presented with several symptoms. We report on a case of Zoletil poisoning.
Animals ; Apnea ; Central Nervous System ; Diazepam ; Drug Combinations ; Emergencies ; Europe ; Humans ; Ketamine ; Male ; Salivation ; Seizures ; Tachycardia ; Tiletamine ; Veterinary Medicine ; Vomiting ; Zolazepam

Objective To identify tiletamine, zolazepam and their metabolites in samples from drug facilitated sexual assault by gas chromatography-quadrupole time of flight mass spectrometry （GC-QTOF-MS）. Methods Urine samples of victims were collected, and detected by GC-QTOF-MS after liquid-liquid extraction and concentration. The molecular formula of fragments ions was identified by determination of accurate mass numbers, to detect related substances. Results Tiletamine, zolazepam, three metabolites of tiletamine and two metabolites of zolazepam were identified in urine samples from actual cases. Conclusion GC-QTOF-MS provides abundant and accurate information of fragment ions mass numbers, which can be used for qualitative identification of tiletamine, zolazepam and their metabolites in drug facilitated sexual assault.
Chromatography, High Pressure Liquid/methods* ; Forensic Toxicology/methods* ; Gas Chromatography-Mass Spectrometry/methods* ; Humans ; Sex Offenses ; Tandem Mass Spectrometry/methods* ; Tiletamine/blood* ; Zolazepam/blood*

BACKGROUND: The aim of this study was to investigate the neuroprotective effects of sevoflurane after severe forebrain ischemic injury. We also examined the relationship between the duration of ischemia and neuronal cell death. METHODS: Male Sprague-Dawley rats (300-380 g) were subjected to 6 (each n = 6) or 10 min (each n = 10) of near-complete forebrain ischemia while anesthetized with either 50 mg/kg of zoletil given intraperitoneally or inhaled sevoflurane (2.3%). Ischemia was induced by bilateral common carotid artery occlusion plus hemorrhagic hypotension (26-30 mmHg). Histologic outcomes were measured 7 days after ischemia in CA1 pyramidal cells of the rat hippocampus. RESULTS: The mean percentage of necrotic cells in the hippocampal CA1 area decreased in the sevoflurane group compared to the zoletil group (25% vs. 40% after 6 min ischemia, respectively: P = 0.004 and 44% vs. 54% after 10 min of ischemia, respectively P = 0.03). The percentage of apoptotic cells was similar in all groups. The percentage of necrotic cells in each anesthetic groups was significantly higher in the 10 min ischemia group compared to the 6 min ischemia group (P = 0.004 in the sevoflurane group, P = 0.03 in the zoletil group). CONCLUSIONS: The present data show that sevoflurane has neuroprotective effects in rats subjected to near-complete cerebral ischemia. Longer duration of ischemia is associated with more neuronal injury when compared to ischemia of shorter duration.
Anesthetics, Inhalation ; Animals ; Brain Ischemia ; Carotid Artery, Common ; Drug Combinations ; Hippocampus ; Humans ; Hypotension ; Ischemia ; Male ; Methyl Ethers ; Neurons ; Neuroprotective Agents ; Prosencephalon ; Pyramidal Cells ; Rats ; Rats, Sprague-Dawley ; Tiletamine ; Zolazepam

To compare the effects of ketamine and tiletaminezolazepam (TZ) drugs widely used for the chemical restraint and immobilization of primates, on various physiological parameters and blood gas values in cynomolgus monkeys (Macaca facicularis). Rectal temperature, heart rate, respiration rate and blood gas analysis were measured before treatment and at 1, 10, 20, 30, 40, 50 and 60 min after administration. Additionally, in both groups, induction and maintenance times were compared. Heart rate, respiration rate, rectal temperature, pH and pCO2 were not significant different in the two groups. However, pO2 in the ketamine-treated group was significantly lower at 30 and 40 min than in the TZ-treated group. The induction time was short in both groups, and the maintenance time was longer in the TZ-treated group (67.8+/-6.5 min) than in the ketamine-treated group (42.3+/-6.7 min). However, decreased rectal temperatures must be watched and prevented following TZ administration to cynomolgus monkeys. It was considered that ketamine may be useful for short duration anesthesia including handling, physical examination, blood sampling and TZ may be useful for prolonged anesthesia including minor surgery and other surgical procedure.
Animals ; Body Temperature/drug effects ; Carbon Dioxide/blood ; Excitatory Amino Acid Antagonists/*pharmacology ; Female ; Heart Rate/drug effects ; Hydrogen-Ion Concentration ; Immobilization/*physiology ; Ketamine/*pharmacology ; Macaca fascicularis ; Male ; Partial Pressure ; Respiratory Mechanics ; Restraint, Physical/*methods ; Tiletamine/*pharmacology ; Time Factors