OBJECTIVE Pituitary adenomas account for approximately 10% of intracranial tumors and have an estimated prevalence of 15%-20% in the general US population. Resection is the primary treatment for pituitary adenomas, and the transsphenoidal approach remains the most common. The greatest challenge with pituitary adenomas is that 20% of patients develop tumor recurrence. Current approaches to reduce recurrence, such as intraoperative MRI, are costly, associated with high false-positive rates, and not recommended. Pituitary adenomas are known to overexpress folate receptor alpha (FRα), and it was hypothesized that OTL38, a folate analog conjugated to a near-infrared (NIR) fluorescent dye, could provide real-time intraoperative visual contrast of the tumor versus the surrounding nonneoplastic tissues. The preliminary results of this novel clinical trial are presented. METHODS Nineteen adult patients who presented with pituitary adenoma were enrolled. Patients were infused with OTL38 2-4 hours prior to surgery. A 4-mm endoscope with both visible and NIR light capabilities was used to visualize the pituitary adenoma and its margins in real time during surgery. The signal-to-background ratio (SBR) was recorded for each tumor and surrounding tissues at various endoscope-to-sella distances. Immunohistochemical analysis was performed to assess the FRα expression levels in all specimens and classify patients as having either high or low FRα expression. RESULTS Data from 15 patients (4 with null cell adenomas, 1 clinically silent gonadotroph, 1 totally silent somatotroph, 5 with a corticotroph, 3 with somatotrophs, and 1 somatocorticotroph) were analyzed in this preliminary analysis. Four patients were excluded for technical considerations. Intraoperative NIR imaging delineated the main tumors in all 15 patients with an average SBR of 1.9 ± 0.70. The FRα expression level of the adenomas and endoscope-to-sella distance had statistically significant impacts on the fluorescent SBRs. Additional considerations included adenoma functional status and time from OTL38 injection. SBRs were 3.0 ± 0.29 for tumors with high FRα expression (n = 3) and 1.6 ± 0.43 for tumors with low FRα expression (n = 12; p < 0.05). In 3 patients with immunohistochemistry-confirmed FRα overexpression (2 patients with null cell adenoma and 1 patient with clinically silent gonadotroph), intraoperative NIR imaging demonstrated perfect classification of the tumor margins with 100% sensitivity and 100% specificity. In addition, for these 3 patients, intraoperative residual fluorescence predicted postoperative MRI results with perfect concordance. CONCLUSIONS Pituitary adenomas and their margins can be intraoperatively visualized with the preoperative injection of OTL38, a folate analog conjugated to NIR dye. Tumor-to-background contrast is most pronounced in adenomas that overexpress FRα. Intraoperative SBR at the appropriate endoscope-to-sella distance can predict adenoma FRα expression status in real time. This work suggests that for adenomas with high FRα exp...

Pheromaxein, the 16-androstene steroid-binding protein with a relative molecular mass of 15,000 was isolated in sub-milligram quantities from the submaxillary gland and saliva of the Gottingen miniature boar, after a fourfold purification involving the following methods: ultrafiltration for submaxillary gland cytosols and ethanol precipitation for saliva, Concanavalin-A-Sepharose affinity chromatography, sodium dodecyl sulphate polyacrylamide gel electrophoresis, 'Extractigel-D' affinity chromatography (to remove sodium dodecyl sulphate) and fast protein-liquid chromatography. Yields of purified pheromaxein obtained after fast protein-liquid chromatography represented 10-20% of total protein present in an ultrafiltrate of a submaxillary gland cytosol. Fast protein-liquid chromatography separated the alpha- and beta-charge isomers of pheromaxein which were shown to have isoelectric points of 4.78 and 5.35 respectively on flat-bed isoelectric focusing. Some data are provided for the variable occurrence of the isomeric forms of pheromaxein in relation to different breeds of pig. Five 16-unsaturated steroids showed the highest binding to pheromaxein. Other steroids of the 5 alpha- and 5 beta-androstane series also showed some binding to pheromaxein, i.e. 17 beta-hydroxy-5 alpha-androstan-3-one (19.2%), with 5 alpha-androstan-3-one, which has a similar urinous odour to 5 alpha-androst-16-en-3-one, showing the greatest binding (42.6%) relative to 5 alpha-androst-16-en-3-one (100%).

Preoperative injection of folate-tagged NIR dye provides strong signal and visualization of NF pituitary adenomas. It is 100% sensitive and specific for detecting margin neoplasm and can predict postoperative MRI findings. Our results suggest that NIR fluorescence may be superior to white-light visualization alone and may improve resection rates in NF pituitary adenomas.

bAbilities to detect heterogeneity of ospC genotypes of the Lyme disease spirochete Borrelia burgdorferi in the tick vector by in vitro culture (IVC) and direct PCR (dPCR) were compared. IVC failed to detect one-third of the ospC genotypes detected by dPCR. Among IVC results, common ospC genotypes were overrepresented while occurrence of rare genotypes was underestimated.A bility to culture Borrelia burgdorferi has been essential for clinical, epidemiological, and pathogenicity studies of B. burgdorferi (1). Various culture medium formulations for Borrelia growth have been widely used to isolate B. burgdorferi from ticks, hosts, and clinical samples (2-4). More recently, culture-independent means of Borrelia detection, usually involving PCR, have become the gold standard (5). One widely used genetic marker for detection of B. burgdorferi is the gene encoding outer surface protein C (ospC). The ospC gene encodes an important major surface protein that has proven useful for discriminating several genotypes (6-9).A comparison of B. burgdorferi strains cultured from ticks with those cultured from clinical samples has led to conclusions that a limited set of ospC genotypes (A, B, I, and K) are invasive and cause systemic disease while the others are limited to ticks or persist as localized infections (8, 10). While several investigators have reported a cultivation effect of B. burgdorferi, all results are based on clinical samples and/or conserved genetic markers. In this study, we compared performances of direct PCR (dPCR) and in vitro cultivation (IVC) to detect 17 distinct Borrelia burgdorferi ospC genotypes from tick specimens.A total of 926 adult deer ticks were surface sterilized in providone-iodine and then rinsed in 70% EtOH. Ticks in one treatment group (n ϭ 420) were bisected by a dorsoventral medial cut, and one half of the ticks were placed directly into BSK-H medium (Sigma-Aldrich, St. Louis, MO) while the other half were used for DNA extraction. Ticks in a second treatment group (n ϭ 506) were dissected, and gut contents were placed in 1.5 ml BSK-H medium; the remainder of the tick body was used in DNA extraction as described above. All cultures were incubated at 34°C and checked biweekly for 4 weeks by dark-field microscopy.After 4 weeks, 25 l of each culture was transferred to a 96-well plate containing 75 l H 2 O, heated to 95°C for 10 min, and centrifuged at 6,000 ϫ g for 10 min, and the resulting supernatant was used as a template for PCR. DNA extraction was performed using the Epicentre Master Complete DNA and RNA purification kits (Epicentre Technologies, Madison, WI). PCR conditions for ospC and the reverse line blotting technique were as described previously (7,11). In addition to the 15 published probes, two new ospC probes were used in this study. We named these type-V (5=-GAG CCG CTT GAG CAG TTA AAC CAT TTG CAC C-3=) and type-W (5=-TCG TTT CGA TTT GCT TCT ACA CCC-3=).dPCR of ticks detected a higher overall infection rate (61.9%) than IVC (34.3%) ( 2 ϭ 140.64; degrees of freedom [df] ϭ 1; P Ͻ...

Aim: Two integrated headspace Early Intervention Teams (hEITs) were established in 2017 to bridge gaps between headspace, the national primary care youth mental health programme in Australia, and the state funded secondary and tertiary mental health services. This study aims to describe functioning and outcomes of patients referred to hEIT over a 6-month period.Methods: A retrospective file audit was conducted for all patients accepted into the service over a 6-month period in 2018. Measures of distress, functioning and client satisfaction were collected and analysed. Exemplar vignettes were created to construct hypothetical examples and illustrate research findings.Results: At admission, the three most common presentations were depression/anxiety, trauma and stress related, and psychotic disorders. During their time in the service, young people displayed a statistically significant improvement in functioning, reduction in self-harm in those 18 years and under, and a trend to reduction in distress scores. hEIT delivered a broad range of services covering social, occupational, educational, medical and mental health care, and the service was experienced positively by the patient cohort.Conclusions: hEIT appears to meet the needs of young people requiring greater care than primary care services can deliver. The integrated, wrap-around care coordination facilitates treatments across social, educational and health domains. Further exploration of young people who disengage from care, improved outcome data reporting and economic evaluation are indicated.