A simultaneous detection and quantification method for determining the Phenylalkylamine derivatives, such as methamphetamine (MA), amphetamine (AM), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), ketamine (KT), norketamine (NKT), phentermine (PT), fenfluramine (FFA) and phenmetrazine (PM), in oral fluid was developed and validated according to international guidelines. The validated method was applied to actual oral fluid samples collected from drug abuse suspects. The recovery of phenylalkylamines from oral fluid collection devices was also assessed. Oral fluid specimens from 20 drug abuse suspects submitted by the police were collected using Salivette$^{TM}$, Quantisal$^{TM}$ or direct expectoration. The samples were screened using a biochip array analyzer. For confirmation, the samples were analyzed by GC-MS in selected-ion monitoring (SIM) mode after extraction using automated SPE with a mixed-mode cation exchange cartridge and derivatization with trifluoroacetic anhydride (TFAA). The results from the immunoassay were consistent with those from GC-MS. All the oral fluid samples gave positive results for MA, AM, PT and/or PM. The detection of phenylalkylamines in oral fluid can provide a better indication of recent use than urine or hair. Therefore, the oral fluid specimen was useful for demonstrating phenylalkylamines abuse in the driving under the influence of drug (DUID) as an alternative specimen for urine.
We investigated the effect of temperature and stock density on the collection efficiency of oral fluid in the pig farm in Korea. Three pig farms with similar breeding environmental conditions were selected and four pens of each farm (total 12 pens) were tested for the collection efficiency of oral fluid from pigs. Collection rate was considered as significant when oral fluid was collected from 70% of pigs within a pen. In the case of growing pigs, when internal temperature of pig barn increased by one designated degree (5℃), the oral fluid collection rate significantly decreased by 24.7% (P<0.05). The collection rate of oral fluid also decreased by 7.1% (P<0.05) as the density rate increase by one designated degree (12.5%). It was estimated that the collection efficiency of oral fluid decreased when the internal temperature of pig barn was 30℃ or higher, or barn density is higher 25% or high. On the other hand, in the case of stall-housing sows, unlike growing pigs, there was no significant differences according to the temperature, so oral fluid collection was considered to be efficient even in hot season.
The objective of this study was to evaluate the usefulness of detection of PRRSV and PRRSV-specific antibodies in oral fluids for monitoring of PRRSV infection in endemic farms. The level of PRRSV and anti-PRRSV antibodies in serum and oral fluids was evaluated in five age groups of pigs (6, 9, 12, 16 weeks of age and gilts). The samples (25 serums and 5 oral fluids/per a farm) were collected from 5 different farms endemically infected by PRRSV. Both serum and oral fluid samples were tested for PRRSV by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and for anti-PRRSV antibodies by two commercial PRRSV ELISA kits. ELISA mean s/p ratios (2.98 vs 1.63) and positive rate (84.0% vs 68.8%) of the oral fluid samples showed significantly higher levels but had similar patterns to the seroprofile of the blood samples. The PRRSV positive rate of oral fluid and serum samples was 40.0% and 44.0% respectively. In conclusion, the use of oral fluids for PRRS monitoring in endemic farms is strongly recommended.
Kim, Ju-Hyun;Kim, Soung-Min;Oh, Jin-Sil;Myoung, Hoon;Lee, Jong-Ho;Choi, Jin-Young
Korean Journal of Cleft Lip And Palate
/
v.14
no.1_2
/
pp.37-44
/
2011
Amniotic fluid is a complex and biological reservoir that provides mechanical cushioning and has many nutrients required for fetal growth and development. During our main research works about the fetal surgery of congenital facial defects, we reviewed several recent articles about the effectiveness and composition of amniotic fluid. Among these review processes, amniotic fluid, as the convenient medium to store sking grafts, was focused especially for its growth factors and rich nutrients, and we summarized some experimental investigations of skin grafts stored in amniotic fluid in rats. We reviewed mainly the article, "Turhan-haktanir N. et al. Histological assessment of skin grafts in amniotic fluid and saline. J Plast Surg Hand Surg 2010;44:226-30."
Oral fluid has become increasingly popular as an alternative specimen in the field of driving under the influence of drugs (DUID) and work place drug testing. In this study, an analytical method for the detection and quantification of ${\Delta}^9$-tetrahydrocannabinol (THC) and its metabolite, 11-nor-9-carboxy-${\Delta}^9$-tetrahydrocannabinol (THC-COOH) in oral fluid by SPE and GC-MS was established and fully validated. The stability of THC and THC-COOH in oral fluid during storage was also determined by examining the THC and THC-COOH concentration changes depending on time and container materials. Oral fluid samples were kept over 21 days at room temperature, $-4^{\circ}C$ and $-20^{\circ}C$ in two different specimen collection tubes; glass and polypropylene tubes. Three replicates for each condition with different temperature and types of a container were analyzed at five different time points over 21 days. When oral fluid samples were stored in glass tubes, the loss of both THC and THC-COOH was less than 10% at all room temperature, $-4^{\circ}C$ and $-20^{\circ}C$. However, in polypropylene tubes, the loss of both THC and THC-COOH increased significantly over the study period. In particular, the concentration of THC decreased more rapidly than that of THC-COOH at room temperature and the maximal percentage of THC lost was 90.3% after 21 days. The result indicates that it would be necessary to collect oral fluid samples in glass containers and cool the samples until analysis in order to prevent the degradation of analytes.
Few topics in operative and perioperative patient management generate more controversy than that of appropriate fluid and electrolyte therapy. especially, controversy has swirled around colloid vs crystalloid therapy and the composition of administered fluids, agreement among clinicians as to what fluid therapy is appropriate, and in what amount, is rare. This controversy likely will be enhanced by Arieff' s provocative article. He described 11 adults and 2 pediatric patients. All developed fatal postoperative pulmonary edema, seemingly caused solely by excessive postoperative fluid administration. From January 1999 to December 1999, we investigated 24 patients, which were operated by orthognathic surgery, about the intraoperative fluid therapy and the associated effect in orthognathic surgery, which is regarded as one of the major surgery of oral and maxillofacial surgery. First, They were devided into two groups, that is one-jaw surgery and two-jaw surgery, and each groups were devided by intraoperative fluid volume of 8ml/kg/hr. Subjective assesment was collected through use of a series of 3 questionnaries. In each questionnaire, a 5-point Liekert scale was used far assessment of following parameters of recovery from anesthesia: headache, dizziness, drowsiness, nausea/vomiting, thirst. The patient completed questionnaire 1 at 4 hour after surgery, questionnaire 2 was completed at 24 hours after surgery, and questionnaire 3 was completed at 48 hours after surgery. This study demonstrated that appropriated perioperative rehydration decreases postoperative adverse outcomes and improved the patient's perception of the postoperative period.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.34
no.2
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pp.113-118
/
2008
Temporomandibular joint disorder (TMD) can induce severe pain but, its pathogenic mechanisms remain poorly understood. In this study, we analyzed proteomes of human synovial fluid in the superior joint space in the patients with TMD, which is obtained during the treatment arthrocentesis. We've got this result that one of the spots was consistently down-regulated in synovial fluid of patients with TMD from analysis of protein pattern. Its molecular weight was estimated to be 33 kDa. Synoviolin was identified in our proteomics analysis of LC/MS/MS. This protein was recently reported as one of the proteins that might affect rheumatoid arthritis (RA). Synoviolin that might be associated with RA was detected in synovial fluid of patients with TMD. We can conclude that synoviolin might be involved not only in the pathogenesis of RA but also in TMD. In result, synoviolin might be involved in the pathogenesis of TMD and can be candidates as new therapeutic targets of TMD or early detection biomarkers.
Porcine circovirus-2 (PCV2) has been implicated in many clinical diseases/syndromes that are now referred to as PCV-associated diseases (PCVAD). Due to significant economic losses caused by PCVAD, many swine operations have launched extensive monitoring programs for PCV2. Traditional serum sampling is, however, rather expensive and laborious, hampering effective large scale pathogen surveillance. A field-based longitudinal study was conducted to assess the utility of pen-based oral fluid sample as an alternative to serum for herd PCV2 testing. Six pens (25 pigs/pen) at each of 3 different sites were used in the study. One oral fluid and 5 random serum samples per pen were collected at 3, 5, 8, 12, and 16 weeks of age, and the sera were pooled by pen for testing. All samples were tested for PCV2 by real-time PCR and for antibodies by indirect fluorescent antibody test (for both anti-PCV2 IgG and IgA) and 3 ELISA assays (blocking ELISA, indirect ELISA, and IgG/IgM sandwich ELISA). PCV2 DNA was detected in oral fluid samples sporadically until 8 weeks and in all pens at 16 weeks. PCV2-specific IgG was detected in oral fluid samples at 3 weeks and persisted until 5 to 8 weeks in all sites. Anti-PCV2 IgG and IgA were detectable in oral fluid samples collected at 16 weeks from all of the pens at 1 site. The detection of PCV2 and anti-PCV2 antibody in oral fluid samples correlated positively with results on pooled sera, suggesting that oral fluids can be a cost-effective alternative to serum for herd monitoring of PCV2 infection.
Kim, Eun-Mi;Lee, Ju-Seon;Choi, Hye-Young;Choi, Hwa-Kyung;Chung, Hee-Sun
YAKHAK HOEJI
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v.52
no.6
/
pp.419-425
/
2008
A qualitative and quantitative analytical method was developed for detection of methamphetamine (MA) and its main metabolite amphetamine (AM) in oral fluid. Oral fluids of eleven drug abusers were provided by Police, specimens were collected by stimulation with a cotton swab treated with 20 mg of citric acid ($Salivette^{(R)}$; Sarstedt, USA). As the preliminary test, oral fluid samples were screened for amphetamines by Fluorescence Polarization Immunoassay (TDxFLx, Abbott Co.). Extraction for MA was performed using solid-phase extraction (SPE) by $RapidTrace^{TM}$ (Zymark, USA) with mixed mode cation exchange cartridge, CLEAN $SCREEN^{(R)}$ (130 mg/3 ml, UCT) after dilution with phosphate buffer. Samples were evaporated and derivatized by pentafluoropropionic acid anhydride (PFPA). Quantitation of MA and AM was performed by gas chromatography-mass spectrometry (GC-MS) using selective ion monitoring (SIM), the quantitation ions were m/z 204 (MA), 208 (MA-$D_5$), 190 (AM) and 194 (AM-$D_5$). The selectivity, linearity of calibration, limit of detection (LOD) and quantification (LOQ) within- and between day precision, accuracy and recoveries were examined as parts of the method validation. All oral fluid samples gave positive results to immunoassay for MA (cut-off level, 50 ng/ml as d-amphetamine). Concentrations of MA and AM by GC-MS in eleven samples were ranged 104.2${\sim}$4603.3 ng/ml and 32.4${\sim}$268.6 ng/ml, respectively. Extracted calibration curves of MA and AM were linear over the two concentration range of 1${\sim}$100 and 50${\sim}$1000 ng/ml with correlation coefficient of above 0.999. LOQ of MA and AM was 1 and 3 ng/ml, respectively. The intraand inter-day run precisions (CV) for MA and AM were less than 10%, and the accuracies (bias) for MA and AM were also less than 10% at the two different concentrations 5 and 100 ng/ml at low calibration range, 50 and 1000 ng/ml at high calibration range. The absolute recoveries of MA and AM at low and high calibration ranges were more than 82% and 75%, respectively. In this study the qualitative and quantitative analytical method of MA in oral fluid was established. Oral fluid testing may detect drug use in past hours because of its shorter detection window than urine, and be useful in post-accident situations. So oral fluids will be most useful for testing drug abuse in the driving under the influence of drug (DUID) as the alternative specimens of urine.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.27
no.3
/
pp.204-208
/
2001
The purpose of this study is to examine the change of enzymeimmuno-assay for $prostaglandinE_2$ in the synovial fluid lavage specimen of patients with mandibular fracture patients without condylar fracture. For this study, fourteen patients (eight males, six females) with mandibular fractures without condylar fracture was investigated to analyse the synovial fluid from upper temporomandibular joint cavities. Synovial fluid was collected from TMJ cavities of mandibular fracture patients before open reduction and after one week of open reduction, and then stored in liquid nitrogen tank after centrifuge. Two synovial fluid lavage samples of TMJs of 2 asymptomatic served as normal controls referred from other data. The concentrations of $PGE_2$ were measured by use of $PGE_2$ EIA System ($Amersham^{(R)}$). The following results were obtained: 1) In nine patients, the concentrations of $PGE_2$ are lower after open reduction than before. 2) In three patients, the concentrations of $PGE_2$ are higher after open reduction than before. 3) There was no statistical significant between the preoperative group and postoperative 7 days (p>0.05), but there was some difference between the two groups. In conclusion, the results suggest that $PGE_2$ probably does not play as important role in the harm of TMJ.
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