Adolescents diagnosed with polycystic ovary syndrome under the Rotterdam criteria but not meeting the diagnosis under the updated guideline.

STUDY QUESTION: What are the characteristics of adolescents diagnosed with polycystic ovary syndrome (PCOS) based on the 2003 Rotterdam criteria, but who do not meet the diagnosis according to the international evidence-based guideline? SUMMARY ANSWER: Adolescents who had features of PCOS but did not meet the evidence-based guideline adolescent criteria exhibited unfavorable metabolic profiles compared to controls and shared considerable metabolic and hormonal features with adolescents who did meet the adolescent criteria. WHAT IS KNOWN ALREADY: The international evidence-based PCOS guideline recommended that ultrasound should not be used for the diagnosis of PCOS in girls with a gynecological age of <8 years. Thus far, few studies have evaluated the clinical characteristics of the girls diagnosed with PCOS based on the Rotterdam criteria but who do not meet the diagnosis according to the updated guideline. STUDY DESIGN, SIZE, DURATION: This is a retrospective study, and subjects attended for care from 2004 to 2022. PARTICIPANTS/MATERIALS, SETTING, METHODS: Adolescent girls with PCOS diagnosed according to the 2003 Rotterdam criteria and healthy controls. All participants were between 2 and 8 years since menarche. MAIN RESULTS AND THE ROLE OF CHANCE: Of the 315 girls diagnosed with PCOS according to the Rotterdam criteria, those with irregular menstruation (IM)/hyperandrogenism (HA)/polycystic ovary (PCO), IM/HA, HA/PCO, and IM/PCO phenotypes accounted for 206 (65.4%), 30 (9.5%), 12 (3.8%), and 67 (21.3%) participants, respectively. According to the evidence-based guideline, 79 girls (25.1%) with the HA/PCO or IM/PCO phenotypes were not diagnosed with PCOS, and aligned to the international guideline; they were designated as the 'at-risk' group. As expected, the girls meeting the evidence-based guideline adolescent criteria showed the worst metabolic profiles (degree of generalized or central obesity, frequency of insulin resistance, prediabetes or diabetes, and metabolic syndrome) and higher hirsutism scores than the at-risk group or controls. Approximately 90% of the at-risk group were not overweight or obese, which was similar to the controls. However, they showed worse metabolic profiles, with higher blood pressure, triglyceride, and insulin resistance parameters than controls; furthermore, these profiles were similar to those of the girls meeting the adolescent criteria. The at-risk group showed similarly elevated serum LH levels and LH/FSH ratio with the girls meeting adolescent criteria. LIMITATIONS, REASONS FOR CAUTION: We could not evaluate hormonal or ultrasound parameters in controls. WIDER IMPLICATIONS OF THE FINDINGS: Compared to the conventional Rotterdam criteria, the recent international evidence-based guideline-avoiding ultrasound in PCOS diagnosis in adolescents-still gives the opportunity to identify young girls at risk, aligned to the findings in this study. A practical approach to this adolescent population would involve establishing IM or HA (with ultrasound not indicated) and designating 'at-risk' PCOS status with regular check-ups for newly developed or worsening PCOS-related symptoms or metabolic abnormalities, with subsequent reassessment including ultrasound or anti-Müllerian hormone, once 8 years post-menarche. STUDY FUNDING/COMPETING INTEREST(S): No funding was received in support of this study. The authors have no conflicts of interest to disclose. TRIAL REGISTRATION NUMBER: N/A.

The Association between Metabolic Syndrome and Epithelial Cell Abnormalities Detected on Pap Smear: A Nationwide Population-Based Study.

Several epidemiologic studies have suggested the correlation between metabolic syndrome (MetS) and cervical cancer. The identification of epithelial cell abnormalities through cervical cytology implies lesions that may lead to cervical cancer in the long term, making screening a crucial measure for its prevention. We conducted a case-control study using data from the National Health Screening Programs under the Health Insurance System of South Korea between 2009 and 2017. Among women who underwent a Pap smear during this period, 8,606,394 tests reported negative results for epithelial cell abnormalities (controls, 93.7%), while 580,012 tests reported epithelial cell abnormalities (cases, 6.3%). Of these, the incidence of MetS was significantly higher in the case group, with 21.7% of cases and 18.4% of controls meeting the MetS criteria with p-Value of less than 0.0001; however, the effect size was small with odds ratio of 1.23. Logistic regression analysis revealed increased odds of epithelial cell abnormalities in women with MetS after adjusting for associated risk factors (AOR 1.202, 95% CI 1.195-1.210, p < 0.0001). These findings indicate that women with MetS have an elevated risk of developing epithelial cell abnormalities, reinforcing the importance of regular Pap smear screening to prevent cervical cancer progression in this population.

Primary ovarian high-grade endometrial stromal sarcoma: a case report.

BACKGROUND: Primary ovarian high-grade endometrial stromal sarcoma is a very rare disease. Even though it has poor prognosis, the gold standard treatment has not been established owing to its rarity. This report aimed to present therapeutic options for primary ovarian high-grade endometrial stromal sarcoma. CASE PRESENTATION: A 49-year-old Asian woman presented with disseminated intravascular coagulation due to ruptured primary high-grade ovarian endometrial stromal sarcoma with multiple intraperitoneal metastases. After the initial surgery, the patient underwent adjuvant chemotherapy with three courses of Adriamycin (75 mg/m2). We performed the secondary debulking operation including total hysterectomy, metastasectomy, omentectomy, peritonectomy, appendectomy, and hyperthermic intraperitoneal chemotherapy (paclitaxel 175 mg/m2). Currently she has been alive for 28 months under a new chemotherapy regimen. CONCLUSION: We suggest cytoreductive surgery with hyperthermic intraperitoneal chemotherapy could be a therapeutic option for primary high-grade ovarian endometrial stromal sarcoma with peritoneal dissemination.

Progression to prediabetes or diabetes in young Korean women with polycystic ovary syndrome: A longitudinal observational study.

OBJECTIVE: To investigate changes in glycaemic status in women with polycystic ovary syndrome (PCOS). DESIGN: Longitudinal observational study. PATIENTS: Women with PCOS who underwent baseline and follow-up screening tests for diabetes (n = 262). Four patients with type 2 diabetes (T2DM) at baseline and 6 patients who were taking drugs at the final follow-up were excluded. MEASUREMENTS: Changes in glycaemic classification based on fasting glucose, haemoglobin A1c and oral glucose tolerance test. RESULTS: The median length of follow-up was 2.9 years. The mean age and body mass index in the normoglycaemia group (n = 202) were 23.0 years and 21.6 kg/m2 , while it was 23.6 years and 22.9 kg/m2 in the prediabetes group (n = 50). In the normoglycaemia group, 38 (18.8%) and 2 (1.0%) developed prediabetes and T2DM, respectively. In the prediabetes group, 22 (44.0%) remained in the same category, 6 (12.0%) developed T2DM, while 22 (44.0%) achieved normoglycaemia. The incidence rate of T2DM was 9.3 per 1,000 person-years, which was significantly higher than that of the female population of similar age, and the incidence was higher in women with fasting glucose ≥ 5.6 mmol/L at baseline than in women with < 5.6 mmol/L. CONCLUSIONS: About 20% of normoglycaemic women had developed prediabetes or T2DM after a median time of 2.9 years. Meanwhile, nearly half of prediabetes women achieved normoglycaemia. Higher baseline fasting glucose levels were associated with an increased incidence of T2DM. Our results are the first to evaluate glycaemic status changes using all three parameters in patients with PCOS.

Coiled-coil domain containing 50-V2 protein positively regulates neurite outgrowth.

The coiled-coil domain containing 50 (CCDC50) protein is a phosphotyrosine-dependent signalling protein stimulated by epidermal growth factor. It is highly expressed in neuronal cells in the central nervous system; however, the roles of CCDC50 in neuronal development are largely unknown. In this study, we showed that the depletion of CCDC50-V2 impeded the neuronal development process, including arbor formation, spine density development, and axonal outgrowth, in primary neurons. Mechanistic studies revealed that CCDC50-V2 positively regulated the nerve growth factor receptor, while it downregulated the epidermal growth factor receptor pathway. Importantly, JNK/c-Jun activation was found to be induced by the CCDC50-V2 overexpression, in which the interaction between CCDC50-V2 and JNK2 was also observed. Overall, the present study demonstrates a novel mechanism of CCDC50 function in neuronal development and provides new insight into the link between CCDC50 function and the aetiology of neurological disorders.

Management of endometriosis-related infertility: Considerations and treatment options.

Endometriosis is a common inflammatory disease in women of reproductive age and is one of the major causes of infertility. Endometriosis causes a sustained reduction of ovarian reserve through both physical mechanisms and inflammatory reactions, which result in the production of reactive oxygen species and tissue fibrosis. The severity of endometriosis is related to ovarian reserve. With regard to infertility treatment, medical therapy as a neoadjuvant or adjuvant to surgical therapy has no definite beneficial effect. Surgical treatment of endometriosis can lead to ovarian injury during the resection of endometriotic tissue, which leads to the deterioration of ovarian reserve. To overcome this disadvantage, a multistep technique has been proposed to minimize the reduction of ovarian reserve. When considering surgical treatment of endometriosis in patients experiencing infertility, it should be kept in mind that ovarian reserve can be reduced both due to endometriosis itself and by the process of removing endometriosis. In cases of mild- to moderate-stage endometriosis, intrauterine insemination with ovarian stimulation after surgical treatment may increase the likelihood of pregnancy. In cases of severe endometriosis, the characteristics of the patient should be considered in a multidisciplinary manner to determine the prioritization of treatment modalities, including surgical treatment and assisted reproduction methods such as in vitro fertilization. The risk of cancer, complications after pregnancy, and infection during oocyte retrieval should also be considered when making treatment decisions.

Effects of cyclic adenosine monophosphate modulators on maturation and quality of vitrified-warmed germinal vesicle stage mouse oocytes.

BACKGROUND: It is still one of the unresolved issues if germinal vesicle stage (GV) oocytes can be successfully cryopreserved for fertility preservation and matured in vitro without damage after warming. Several studies have reported that the addition of cyclic adenosine monophosphate (cAMP) modulators to in vitro maturation (IVM) media improved the developmental potency of mature oocytes though vitrification itself provokes cAMP depletion. We evaluated whether the addition of cAMP modulators after GV oocytes retrieval before vitrification enhances maturation and developmental capability after warming of GV oocytes. METHODS: Retrieved GV oocytes of mice were divided into cumulus-oocyte complexes (COCs) and denuded oocytes (DOs). Then, GV oocytes were cultured with or without dibutyryl-cAMP (dbcAMP, cAMP analog) and 3-isobutyl-l-methylxanthine (phosphodiesterase inhibitor) during the pre-vitrification period for 30 min. RESULTS: One hour after warming, the ratio of oocytes that stayed in the intact GV stage was significantly higher in groups treated with cAMP modulators. After 18 h of IVM, the percentage of maturation was significantly higher in the COC group treated with dbcAMP. The expression of F-actin, which is involved in meiotic spindle migration and chromosomal translocation, is likewise increased in this group. However, there was no difference in chromosome and spindle organization integrity or developmental competence between the MII oocytes of all groups. CONCLUSIONS: Increasing the intracellular cAMP level before vitrification of the GV oocytes maintained the cell cycle arrest, and this process may facilitate oocyte maturation after IVM by preventing cryodamage and synchronizing maturation between nuclear and cytoplasmic components. The role of cumulus cells seems to be essential for this mechanism.

Implementation of a Blind Quality Control Program in Blood Alcohol Analysis.

Declared proficiency tests are limited in their use for testing the performance of the entire system, because analysts are aware that they are being tested. A blind quality control (BQC) is intended to appear as a real case to the analyst to remove any intentional or subconscious bias. A BQC program allows a real-time assessment of the laboratory's policies and procedures and monitors reliability of casework. In September 2015, the Houston Forensic Science Center (HFSC) began a BQC program in blood alcohol analysis. Between September 2015 and July 2018, HFSC submitted 317 blind cases: 89 negative samples and 228 positive samples at five target concentrations (0.08, 0.15, 0.16, 0.20 and 0.25 g/100 mL; theoretical targets). These blood samples were analyzed by a headspace gas chromatograph interfaced with dual-flame ionization detectors (HS-GC-FID). All negative samples produced `no ethanol detected' results. The mean (range) of reported blood alcohol concentrations (BACs) for the aforementioned target concentrations was 0.075 (0.073-0.078), 0.144 (0.140-0.148), 0.157 (0.155-0.160), 0.195 (0.192-0.200) and 0.249 (0.242-0.258) g/100 mL, respectively. The average BAC percent differences from the target for the positive blind cases ranged from -0.4 to -6.3%, within our uncertainty of measurement (8.95-9.18%). The rate of alcohol evaporation/degradation was determined negligible. A multiple linear regression analysis was performed to compare the % difference in BAC among five target concentrations, eight analysts, three HS-GC-FID instruments and two pipettes. The variables other than target concentrations showed no significant difference (P > 0.2). While the 0.08 g/100 mL target showed a significantly larger % difference than higher target concentrations (0.15-0.25 g/100 mL), the % differences among the higher targets were not concentration-dependent. Despite difficulties like gaining buy-in from stakeholders and mimicking evidence samples, the implementation of a BQC program has improved processes, shown methods are reliable and added confidence to staff's testimony in court.

A systematic review and meta-analysis comparing single port laparoscopic myomectomy with conventional laparoscopic myomectomy.

OBJECTIVE: We systematically evaluated if there are any clinical differences in the safety and feasibility of single port laparoscopic myomectomy when compared to conventional laparoscopic myomectomy. STUDY DESIGN: A meta-analysis and systematic review was conducted in accordance with the PRISMA statement. We searched different databases including Medline, Embase, Cochrane, and Google Scholar up to April 2018. Comparative studies as randomized controlled trials and prospective and retrospective cohort studies were included. RESULTS: A meta-analysis was performed on the safety, feasibility, and potential benefits. Two randomized controlled trials and six cohort studies (2 prospective and 4 retrospective) were meta-analyzed to compare the surgical outcomes of 907 patients. Rates of minor complications were not different between the two groups [OR 1.33 (0.67, 2.63), I2 = 0%]. Major complications requiring re-operation occurred in two cases in the single port group and in the conventional group respectively. There was no significant statistical difference between the groups. Conversion to laparotomy did not occur in either group, and the conversion rate from single port to the multi-port operation was 2.1% (6/288). The pain score one hour after operation was significantly lower in the single port group [MD -0.41 point (-0.63, -0.18), I2 = 3.7%]. However, this difference was not observed at six and 24 h postoperatively. There was no difference in the operation time, estimated blood loss, or hemoglobin decrease between the two groups. CONCLUSION: A meta-analysis showed that single port laparoscopic myomectomy is comparable to conventional laparoscopic myomectomy in terms of safety and feasibility and more advantageous in terms of immediate postoperative pain. If performed based on the appropriate patient selection criteria including size and number of myoma, single port laparoscopic myomectomy can have similar surgical outcomes to those of conventional laparoscopic myomectomy. Further research and resources are required to identify whether single port laparoscopic myomectomy is more beneficial in terms of cosmetic results, patient satisfaction, and pregnancy outcomes.

Adverse symptoms during short-term use of ulipristal acetate in women with uterine myomas and/or adenomyosis.

AIM: To evaluate various adverse symptoms during short-term use of ulipristal acetate in women with uterine myomas (n = 90), adenomyosis (n = 3) or both (n = 7). METHODS: One hundred premenopausal women who received ulipristal acetate for 4-12 weeks during 2016 to 2017 were selected. The medical records were reviewed and the following information was collected; adverse symptoms during medication, presence of menorrhagia or menstrual cramps, blood hemoglobin and liver function test. Adverse symptoms were recorded in the medical records as a checklist form including 76 specific progestin-related symptoms. RESULTS: Overall, the most frequent adverse symptom was amenorrhea (43%), followed by weight gain (29%), fatigue (27%), abdominal discomfort (21%), decreased menstrual flow (19%) and dizziness (18%). In 89 symptomatic women (with heavy menstrual bleeding and/or menstrual cramping pain and/or anemia), the most frequent adverse symptom was weight gain (27%) and fatigue (27%), followed by abdominal discomfort (21%), dry eye (18%), facial flushing (17%), dizziness (17%), headache (17%) and increased vaginal discharge (15%). Fourteen women stopped the medication due to unwanted adverse symptoms. Of this discontinuation group, major complaint was fatigue (50%), followed by weight gain (36%) and breast discomfort (35.7%). CONCLUSION: Adverse symptoms were common and discontinuation rate was somewhat higher during short-term course of ulipristal acetate. Information about incidence of various adverse symptoms should be given to women who willing to take ulipristal acetate.

Successful in Vitro Fertilization and Embryo Transfer after Transplantation of Cryopreserved Ovarian Tissue: Report of the First Korean Case.

For patients at risk of premature ovarian failure with cancer treatment, it is an important option to re-implant the ovarian tissue (OT) after cryopreservation to preserve endocrine function and fertility. With this technique, about 30% of pregnancy success rate and about 90 live births have been reported to date. However, there has been no case report of successful in vitro fertilization (IVF) and embryo transfer (ET) with oocytes collected from transplanted cryopreserved OT in Korea. We report a 30-year old woman with rectal cancer who underwent IVF and ET after cryopreserved OT thawing and re-implantation. She has been diagnosed with stage IIIC rectal cancer after surgery, and right ovary was removed and cryopreserved between cycles of chemotherapy. After completion of chemotherapy and radiotherapy, the patient underwent orthotopic transplantation of cryopreserved OTs. Three months after transplantation, the serum follicle-stimulating hormone level decreased from 91.11 mIU/mL to 43.69 mIU/mL. Thereafter, the patient underwent 11 ovarian stimulation cycles, and in 7 cycles, follicle growth was observed at the OT graft site. In one of these cycles, the oocyte was successfully retrieved and one embryo was transplanted after IVF. The patient was not pregnant, but the cryopreservation of OT can save the fertility after anticancer chemotherapy.

Symptom-alleviating effect and adverse effect of dienogest in Korean women with endometriosis.

The purpose was to evaluate the effects of dienogest on Korean women with endometriosis. A cross-sectional questionnaire-based survey was conducted for 100 premenopausal women. They had taken or were taking 2 mg of dienogest daily. We assessed the pelvic pain score and quality-of-life (QOL) score before and after taking dienogest as well as the prevalence of short-term (≤12 weeks) and long-term adverse effects (>52 weeks). Patients were classified into three groups: dienogest treatment immediately following surgery (A), dienogest treatment for a recurrence of endometriosis after surgery (B), or dienogest treatment without any surgery (C). In groups A and C, the median pain score (from 5 to 0, p <.001; from 7 to 1.5, p <.001) and median QOL score (from 10 to 5, p = .002; from 7.5 to 6.5, p = .008) were significantly decreased. Irregular bleeding and decreased menstrual flow were more prevalent in patients with dienogest intake of fewer than 12 weeks, while amenorrhea, weight gain, hair loss, and dorsal pain were more prevalent in patients with dienogest treatment of over 52 weeks. Accordingly, proper counseling is necessary when prescribing dienogest.

The serum estradiol/oocyte ratio in patients with breast cancer undergoing ovarian stimulation with letrozole and gonadotropins.

OBJECTIVE: To investigate the patterns of estradiol-oocyte ratio (EOR) and estradiol-mature oocyte ratio (EMOR) in patients with breast cancer undergoing controlled ovarian stimulation (COS) using letrozole and gonadotropins for fertility preservation. METHODS: This retrospective study included 32 breast cancer patients underwent COS for fertility preservation and no patients had previously undergone gonadotoxic treatments. Basal characteristics and in vitro fertilization (IVF) outcomes were compared according to the age of women (age <35 vs. age ≥35) and the starting phase of ovarian stimulation (early follicular phase vs. late follicular phase vs. luteal phase). RESULTS: Patients who were administered with a letrozole-combined regimen, the peak serum estradiol (E2) was maintained at a low level (386.3±346.9 pg/mL). EOR was 52.4±78.8 pg/mL, and EMOR was 71.0±41.2 pg/mL. When the 2 groups were compared according to the age of women (age <35 vs. age ≥35), EOR was 34.5 (22.2-46.5) pg/mL and 40.7 (19.3-65.3) pg/mL, respectively; EMOR was 79.8 (40.8-90.6) pg/mL and 68.8 (44.5-85.9) pg/mL, respectively. There was no significant difference in the IVF outcomes. When the 2 groups were compared according to the starting phase of ovarian stimulation, there were no significant differences in IVF outcomes, EOR and EMOR among the groups. CONCLUSION: Measuring the peak E2 concentration in breast cancer patients undergoing IVF for fertility preservation with a co-treatment of letrozole allows for the prediction of the numbers of oocytes and mature oocytes.

Advantages of Single-Port Laparoscopic Myomectomy Compared with Conventional Laparoscopic Myomectomy: A Randomized Controlled Study.

STUDY OBJECTIVE: To compare operative outcomes of single-port laparoscopic myomectomy (SP-LM) vs conventional laparoscopic myomectomy (CLM), including subjective and objective cosmetic aspects. DESIGN: Prospective randomized controlled trial (Canadian Task Force classification I). SETTING: University hospital. PATIENTS: Women with uterine myoma scheduled for laparoscopic myomectomy. INTERVENTIONS: Sixty-six women were assigned at random to either the SP-LM or CLM group. Surgical outcomes, including patient and observer scar assessments, were evaluated between the groups according to the intention-to-treat principle. MEASUREMENTS AND MAIN RESULTS: There were no significant differences in demographic characteristics and properties of myomectomy between the groups. There also were no differences in surgical outcomes, such as operation time, estimated blood loss, and complications, between the 2 groups. The mean total score of the Observer Scar Assessment Scale was lower in the SP-LM group at 1 week (13.0 ± 3.2 vs 18.3 ± 4.8; p < .001) and 8 weeks (9.9 ± 3.2 vs 14.3 ± 3.8; p < .001) after discharge. Similar results were obtained for the Patient Scar Assessment Scale at 1 week (11.6 ± 7.2 vs 18.5 ± 12.8; p = .024) and 8 weeks (9.5 ± 6.0 vs 18.8 ± 9.1; p < .001) after discharge. Postoperative pain and analgesic consumption did not differ between the groups, except in patient-controlled analgesia consumption at 6 hours after operation, which was lower in the SP-LM group (12.7 ± 6.3 mL vs 16.4 ± 6.2 mL; p = .039). Operative outcomes were similar in the 2 groups. CONCLUSION: SP-LM is associated with more favorable cosmetic outcomes and better patient satisfaction compared with CLM. There were no differences in operative outcomes and complications between the 2 modalities.

Plasma Cannabinoid Pharmacokinetics After Controlled Smoking and Ad libitum Cannabis Smoking in Chronic Frequent Users.

More Americans are dependent on cannabis than any other illicit drug. The main analytes for cannabis testing include the primary psychoactive constituent, Δ(9)-tetrahydrocannabinol (THC), equipotent 11-hydroxy-THC (11-OH-THC) and inactive 11-nor-9-carboxy-THC (THCCOOH). Eleven adult chronic frequent cannabis smokers resided on a closed research unit with unlimited access to 5.9% THC cannabis cigarettes from 12:00 to 23:00 during two ad libitum smoking phases, followed by a 5-day abstinence period in seven participants. A single cigarette was smoked under controlled topography on the last day of the smoking and abstinence phases. Plasma cannabinoids were quantified by two-dimensional gas chromatography-mass spectrometry. Median plasma maximum concentrations (Cmax) were 28.3 (THC), 3.9 (11-OH-THC) and 47.0 µg/L (THCCOOH) 0.5 h after controlled single cannabis smoking. Median Cmax 0.2-0.5 h after ad libitum smoking was higher for all analytes: 83.5 (THC), 14.2 (11-OH-THC) and 155 µg/L (THCCOOH). All 11 participants' plasma samples were THC and THCCOOH-positive, 58.3% had THC ≥5 µg/L and 79.2% were 11-OH-THC-positive 8.1-14 h after last cannabis smoking. Cannabinoid detection rates in seven participants 106-112 h (4-5 days) after last smoking were 92.9 (THC), 35.7 (11-OH-THC) and 100% (THCCOOH), with limits of quantification of 0.5 µg/L for THC and THCCOOH, and 1.0 µg/L for 11-OH-THC. These data greatly expand prior research findings on cannabinoid excretion profiles in chronic frequent cannabis smokers during ad libitum smoking. Smoking multiple cannabis cigarettes led to higher Cmax and AUC compared with smoking a single cigarette. The chronic frequent cannabis smokers exhibited extended detection windows for plasma cannabinoids, reflecting a large cannabinoid body burden.

Oral fluid cannabinoids in chronic frequent cannabis smokers during ad libitum cannabis smoking.

Oral fluid (OF) offers a simple, non-invasive, directly observable sample collection for clinical and forensic drug testing. Given that chronic cannabis smokers often engage in drug administration multiple times daily, evaluating OF cannabinoid pharmacokinetics during ad libitum smoking is important for practical development of analytical methods and informed interpretation of test results. Eleven cannabis smokers resided in a closed research unit for 51 days, and underwent four, 5-day oral delta-9-tetrahydrocannabinol (THC) treatments. Each medication period was separated by 9 days of ad libitum cannabis smoking from 12:00 to 23:00 h daily. Ten OF samples were collected from 9:00-22:00 h on each of the last ad libitum smoking days (Study Days 4, 18, 32, and 46). As the number of cannabis cigarettes smoked increased over the study days, OF THC, cannabinol (CBN), and 11-nor-9-carboxy-THC (THCCOOH) also increased with a significant effect of time since last smoking (Δtime; range, 0.0-17.4 h) and ≥88% detection rates; concentrations on Day 4 were significantly lower than those on Days 32 and 46 but not Day 18. Within 30 min of smoking, median THC, CBN, and THCCOOH concentrations were 689 µg/L, 116 µg/L, and 147 ng/L, respectively, decreasing to 19.4 µg/L, 2.4 µg/L, and 87.6 ng/L after 10 h. Cannabidiol and 11-hydroxy-THC showed overall lower detection rates of 29 and 8.6%, respectively. Cannabinoid disposition in OF was highly influenced by Δtime and composition of smoked cannabis. Furthermore, cannabinoid OF concentrations increased over ad libitum smoking days, in parallel with increased cannabis self-administration, possibly reflecting development of increased cannabis tolerance.

Impact of oral fluid collection device on cannabinoid stability following smoked cannabis.

Evaluation of cannabinoid stability in authentic oral fluid (OF) is critical, as most OF stability studies employed fortified or synthetic OF. Participants (n = 16) smoked a 6.8% delta-9-tetrahydrocannabinol (THC) cigarette, and baseline concentrations of THC, 11-nor-9-carboxy-THC (THCCOOH), cannabidiol (CBD), and cannabinol (CBN) were determined within 24 h in 16 separate pooled samples (collected 1 h before to 10.5 or 13 h after smoking). OF was collected with the StatSure Saliva Sampler™ and Oral-Eze® devices. Oral-Eze samples were re-analyzed after room temperature (RT) storage for 1 week, and for both devices after 4 °C for 1 and 4 weeks, and -20 °C for 4 and 24 weeks. Concentrations ±20% from initial concentrations were considered stable. With the StatSure device, all cannabinoids were within 80-120% median %baseline for all storage conditions. Individual THC, CBD, CBN and THCCOOH pool concentrations were stable in 100%, 100%, 80-94% and >85%, respectively, across storage conditions. With the Oral-Eze device, at RT or refrigerated storage (for 1 and 4 weeks), THC, CBD and THCCOOH were stable in 94-100%, 78-89%, and 93-100% of samples, respectively, while CBN concentrations were 53-79% stable. However, after 24 weeks at -20 °C, stability decreased, especially for CBD, with a median of 56% stability. Overall, the collection devices' elution/stabilizing buffers provided good stability for OF cannabinoids, with the exception of the more labile CBN. To ensure OF cannabinoid concentration accuracy, these data suggest analysis within 4 weeks at 4 °C storage for Oral-Eze collection and within 4 weeks at 4 °C or 24 weeks at -20 °C for StatSure collection. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.

Cannabinoids in oral fluid by on-site immunoassay and by GC-MS using two different oral fluid collection devices.

Oral fluid (OF) enables non-invasive sample collection for on-site drug testing, but performance of on-site tests with occasional and frequent smokers' OF to identify cannabinoid intake requires further evaluation. Furthermore, as far as we are aware, no studies have evaluated differences between cannabinoid disposition among OF collection devices with authentic OF samples after controlled cannabis administration. Fourteen frequent (≥4 times per week) and 10 occasional (less than twice a week) adult cannabis smokers smoked one 6.8% ∆(9)-tetrahydrocannabinol (THC) cigarette ad libitum over 10 min. OF was collected with the StatSure Saliva Sampler, Oral-Eze, and Draeger DrugTest 5000 test cassette before and up to 30 h after cannabis smoking. Test cassettes were analyzed within 15 min and gas chromatography-mass spectrometry cannabinoid results were obtained within 24 h. Cannabinoid concentrations with the StatSure and Oral-Eze devices were compared and times of last cannabinoid detection (t(last)) and DrugTest 5000 test performance were assessed for different cannabinoid cutoffs. 11-nor-9-Carboxy-THC (THCCOOH) and cannabinol concentrations were significantly higher in Oral-Eze samples than in Stat-Sure samples. DrugTest 5000 t(last) for a positive cannabinoid test were median (range) 12 h (4-24 h) and 21 h (1- ≥ 30 h) for occasional and frequent smokers, respectively. Detection windows in screening and confirmatory tests were usually shorter for occasional than for frequent smokers, especially when including THCCOOH ≥20 ng L(-1) in confirmation criteria. No differences in t(last) were observed between collection devices, except for THC ≥2 µg L(-1). We thus report significantly different THCCOOH and cannabinol, but not THC, concentrations between OF collection devices, which may affect OF data interpretation. The DrugTest 5000 on-site device had high diagnostic sensitivity, specificity, and efficiency for cannabinoids.