A Functional Agonist of Insect Olfactory Receptors: Behavior, Physiology and Structure.

Chemical signaling is ubiquitous and employs a variety of receptor types to detect the cacophony of molecules relevant for each living organism. Insects, our most diverse taxon, have evolved unique olfactory receptors with as little as 10% sequence identity between receptor types. We have identified a promiscuous volatile, 2-methyltetrahydro-3-furanone (coffee furanone), that elicits chemosensory and behavioral activity across multiple insect orders and receptors. In vivo and in vitro physiology showed that coffee furanone was detected by roughly 80% of the recorded neurons expressing the insect-specific olfactory receptor complex in the antenna of Drosophila melanogaster, at concentrations similar to other known, and less promiscuous, ligands. Neurons expressing specialized receptors, other chemoreceptor types, or mutants lacking the complex entirely did not respond to this compound. This indicates that coffee furanone is a promiscuous ligand for the insect olfactory receptor complex itself and did not induce non-specific cellular responses. In addition, we present homology modeling and docking studies with selected olfactory receptors that suggest conserved interaction regions for both coffee furanone and known ligands. Apart from its physiological activity, this known food additive elicits a behavioral response for several insects, including mosquitoes, flies, and cockroaches. A broad-scale behaviorally active molecule non-toxic to humans thus has significant implications for health and agriculture. Coffee furanone serves as a unique tool to unlock molecular, physiological, and behavioral relationships across this diverse receptor family and animal taxa.

Granulosa cell tumor of the ovary: cytologic findings.

OBJECTIVE: To describe the cytologic findings in 6 adult variants of granuloma cell tumors (AGCTs) and 1 juvenile variant (JGCT), emphasizing differences between recurrent/metastatic (REC AGCT) and nonrecurrent tumors (NED AGC7). STUDY DESIGN: Imprints and fluids were evaluated for: hypercellularity, Call-Exner bodies (CEB), sheets, single cells/naked nuclei, nuclear grooves, single cell necrosis and established histologic criteria of atypia in AGCT (>3 mitoses, pleomorphism, hyperchromasia, prominent nucleoli). RESULTS: All AGCTs and JGCT showed hypercellularity, clusters, sheets, single cells and naked nuclei. CEBs and grooves were seen only in AGCTs. Fluids had less cellularity than imprints, fewer clusters, grooves, single cells/naked nuclei and no sheets, CEBs, necrosis or vacuoles. Hyperchromasia and nucleoli were more striking in JGCT than AGCTs. All REC AGCTs had cytoplasmic vacuoles, while NED AGCTs did not. Prominent nucleoli were 3 times more common in REC AGCTs than NED AGCTs. Increased mitoses and necrosis were seen in 1 REC AGCT. CONCLUSION: CEBs and grooves are not seen in JGCT. JGCT shows more striking cellular atypia than AGCT (REC AGCTs and NED AGCTs). When evaluating pelvic washes/ascitic fluid a high index of suspicion is necessary, as tumor cells can be overlooked. AGCTs showing cytoplasmic vacuoles, prominent nucleoli, mitoses and necrosis are suggestive of aggressive behavior, and that information should be conveyed in cytology reports.

Interobserver and intraobserver variability in the calculation of the lipid-laden macrophage index: implications for its use in the evaluation of aspiration in children.

The lipid-laden macrophage index (LLMI) is a semiquantitative test used to evaluate aspiration in children. We assessed the reliability and reproducibility of LLMI by calculating interobserver and intraobserver variability among pathologists, with and without expertise in cytopathology. Forty-nine bronchoalveolar washes/lavages were blindly reviewed by four reviewers and assigned an LLMI. Three pathologists (two cytopathologists, one pathology fellow) reviewed slides twice and one cytotechnologist reviewed them once. Intraclass correlation coefficient (ICC) with 95% confidence interval (C.I.) was used to measure overall intraobserver and interobserver agreement. Interobserver agreement was also calculated separately for each pair of reviewers. ICC values did not indicate an acceptable level of interobserver agreement among pathologists, with (ICC = 0.67, 95% C.I.: 0.56-0.77) and without (ICC = 0.77, 95% C.I.: 0.61-0.84) the cytotechnologist included in the analysis. An ICC of 0.84 (95% C.I.: 0.78-0.89) indicated an acceptable level of intraobserver agreement among pathologists. When calculated separately for each pair of reviewers, all but two ICC values for interobserver agreement were less than 0.75 (the minimally acceptable value for a reliable clinical measurement), and the lower confidence limit of each of the 95% C.I. was far below the 0.75 cutoff. Using Lin's coefficient, intraobserver variability was only acceptable for two pathologists. Our study highlights the lack of precision and subjectivity of the LLMI, as well as the significant inter and intraobserver bias that may occur among experienced and inexperienced pathologists, and cytotechnologists. Clinicians and cytopathologists alike should be mindful of this potential pitfall and interpret LLMI scores with caution.

Microsatellite instability and DNA methylation of endometrial tumors and clinical features in young women compared with older women.

INTRODUCTION: Molecular genetic changes in endometrial cancers are important to identify possible family cancer syndromes and thus, to facilitate appropriate screening. Most studies in this regard have focused primarily on young women. We have assayed cancers for microsatellite instability (MSI) and DNA methylation from a large group of patients younger than 50 years and a comparable group of older women. We obtained personal and medical histories of the patients and their family cancer histories. METHODS: The Bethesda panel of markers was used for the detection of MSI. Methylation status of mismatch repair genes was ascertained using the methylation-specific DNA detection kit SALSA MS-MLPA No. ME011. RESULTS: There were 101 patients younger than 50 years and 112 older women. The 2 age groups did not differ in the percentage of patients who were obese, carried a diagnosis of diabetes, or previously had another cancer. The younger patients were more likely to be nulliparous, whereas the older patients were more likely to have hypertension. Among the younger group, 21 (20.8%) tumors revealed MSI, and 13 (61.9%) of these were unmethylated. For the older women, 35 (31.2%) had MSI tumors, and only 6 (17.1%) of these were unmethylated. Young women with a family history of a hereditary nonpolyposis colorectal cancer-related cancer were more likely to have a tumor revealing MSI and no methylation, but family history was less helpful in older women in this regard. CONCLUSION: We did not find personal risk factors or a history of an additional cancer to be different between the 2 age groups. The combination of MSI testing and DNA methylation studies resulted in the identification of presumptive hereditary nonpolyposis colorectal cancer syndrome in approximately 13% of women with endometrial cancer presenting at age younger than 50 years and in approximately 5% of older women. Family history was more helpful with younger women than with older women.

A comparison of the tolerability of two dilution volumes (0.5 mL and 1.0 mL) of a purified chick embryo cell rabies vaccine administered intramuscularly to healthy adult volunteers: A randomized, intraindividual, assessor-blind study.

BACKGROUND: The current recommendation of the manufacturer for administering purified chick embryo cell rabies vaccine (PCECV) is to reconstitute the contents with 1.0 mL of water for injection (WFI). However, it has been debated whether a lower volume of WFI (0.5 mL) is likely to cause less pain. OBJECTIVES: The aims of this study were to compare the tolerability of PCECV administered IM at a volume of 0.5 mL versus 1.0 mL of diluent and to determine the immunogenicity of the vaccine when administered according to the World Health Organization-recommended preexposure prophylaxis regimen for rabies immunization. METHODS: This comparative, intraindividual, assessor-blind study was conducted at the Department of Clinical Pharmacology, Topiwala National Medical College and Bai Yamunabai Laxman Nair Charitable Hospital Mumbai, India). Healthy volunteers aged 18 to 50 years received, by randomized sequence, 3 IM injections of PCECV, diluted in 0.5 mL or 1.0 mL of WFI, on study days 0, 7, and 28. Tolerability was assessed at 30 minutes and 24 hours after injection and included assessments for local and systemic reactions. For immunogenicity assessment, rabies virus-neutralizing antibody 0RVNA) titers were assayed at baseline and on day 49 (ie, 3 weeks after the third injection). RESULTS: Twenty-six subjects (24 men, 2 women; mean [SD] age, 22.4 [2.4] years; mean [SD] body weight, 59.0 [11.3] kg) entered the study. Twenty-five subjects were included in the tolerability assessment; 24 in the immunogenicity assessment. No statistically significant differences were found between dilutions in the frequency of local and systemic reactions. Most reactions were mild. All subjects developed RVNA titers >0.5 IU/mL (indicative of protection) by day 49. CONCLUSIONS: In this population of healthy volunteers, a full antigenic dose of PCECV in a dilution of 0.5 mL WFI is as well tolerated locally and systemically as in a dilution of 1.0 mL. All subjects developed levels of RVNA far exceeding 0.5 IU/mL, which is indicative of protection against rabies.