Melatonin interactions with cultured murine B16 melanoma cells.

Both in vitro and in vivo observations have suggested that melatonin modulates malignant cell growth. The present studies aimed to characterize the interactions of melatonin with cultured murine B16 melanoma cells. Time- and temperature-dependent specific melatonin accumulation by B16 murine melanoma cells was observed. B16 cells possessed a high affinity binding site (KD = 1.4 nM) which exhibited structural specificity in its affinity for analogues of melatonin (melatonin > 6-hydroxymelatonin = N-acetyl-5-hydroxytryptamine > 5-methoxytryptamine >> 5-hydroxytryptamine). Evidence for a lower affinity uptake system without structural specificity was also observed. Ninety-five per cent of the specific cell-associated melatonin in B16 cells was present in the soluble subcellular fraction of lysed cells; more than 97% of the cell-associated radioactivity was authentic melatonin. When the solubilized cell extracts from the binding assay were analysed by gel filtration immediately, all of the bound counts eluted at the void volume. Continuous exposure to melatonin for 48-120 h did not affect B16 cell proliferation as determined by cell counts, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay or [3H]thymidine incorporation. After 8-h pulse exposures to melatonin daily for 3 days, a 15% stimulation of B16 cell proliferation (p < 0.02) was observed at melatonin concentrations of 0.1 and 1 nM. The anti-oestrogen, tamoxifen, inhibited B16 cell growth and increased specific melatonin accumulation by B16 cells at 1 x 10(-6) M (p < 0.02). Cultured B16 murine melanoma cells possessed a specific, high affinity uptake system for melatonin which appeared to be altered by anti-oestrogen exposure.

Voluntary contraction of middle ear muscles: effects on input impedance, energy reflectance and spontaneous otoacoustic emissions.

Two types of measurements were performed on a subject able to voluntarily contract her middle ear muscles (MEM). First, wideband measurements (0-11 kHz) of middle ear input impedance and energy reflectance were obtained when the subject was relaxed and when she contracted her MEM. The changes in impedance observed with voluntary MEM contraction were similar to those reported in the literature for acoustically-elicited MEM contractions. The energy reflectance increased for frequencies below about 4 kHz. Second, the effects of voluntary MEM contraction on the frequencies and levels of spontaneous otoacoustic emissions (SOAEs) were measured and compared to effects evoked by contralateral acoustic stimulation. Effects on SOAEs appear to be a more sensitive indicator of MEM activity than changes in impedance, and the effects due to voluntary MEM contraction were qualitatively similar to those evoked by contralateral acoustic stimulation. These results suggest that in subjects with normally-functioning middle ears, only some effects on otoacoustic emissions caused by contralateral stimuli whose levels are below the contralateral acoustic reflex threshold can be unequivocally attributed to the action of cochlear efferents. The temporal aspects of SOAE frequency shifts caused by voluntary contraction of MEM show that voluntary contraction fatigues rapidly over a time period of tens of seconds.

The occupations of drink drivers: using occupational information to identify targetable characteristics of offenders.

Data collected by the Victoria Police at the time an alleged drink-driving offender undergoes an evidential breath test for the presence of alcohol were analyzed to investigate the possibility that occupational information could be used to define groups of drink-drivers with similar characteristics. Such groups could then be utilized in the development of targeted public education campaigns in the Victorian (Australia) context where there are already high levels of enforcement and mass-media publicity. It is argued that drink-drivers in this context are likely to share some characteristics which might reduce the impact of current programs. Analysis of the data relied on the application of a theory of the relationship between career choice and other behavioral characteristics (Holland, 1973, Making Vocational Choices, Prentice Hall, Englewood Cliffs, N.J.; Holland, 1975, Manual for the Vocational Preference Inventory, Consulting Psychologists Press, Palo Alto, CA). The analysis identified two potential groups which accounted for 58% of the male drink-drivers. These groups differed from other male drink drivers in a number of ways, underlining the potential for their use as targets in future campaigns. One occupational category accounted for 42% of the male drink drivers, including occupations such as carpenter, electrician, chef, mechanic, gardener, and laborer. The behavioral characteristics associated with these occupational codes in the Holland model included asocial, conforming, reserved, introspective, unpopular, orderly, careful, unimaginative, and defensive. The other occupational category accounted for 16% of the male drink drivers and included occupations such as business manager, company director, public servant, and sales representative. Behavioral characteristics associated with this combination included acquisitive, adventurous, ambitious, energetic, extroverted, friendly, and generous.

Ferromagnetism in the Mott insulator Ba2NaOsO6.

Results are presented of single crystal structural, thermodynamic, and reflectivity measurements of the double-perovskite Ba2NaOsO6. These characterize the material as a 5d1 ferromagnetic Mott insulator with an ordered moment of approximately 0.2microB per formula unit and TC=6.8(3) K. The magnetic entropy associated with this phase transition is close to Rln2, indicating that the quartet ground state anticipated from consideration of the crystal structure is split, consistent with a scenario in which the ferromagnetism is associated with orbital ordering.

Effects of contralateral acoustic stimulation on spontaneous otoacoustic emissions.

Spontaneous otoacoustic emissions (SOAEs) were measured in the ear canal of adult humans prior to, during, and following presentation of tonal and broadband stimuli to the contralateral ear. Tones were presented at a fixed level at ten frequencies relative to the SOAE. Broadband noise was presented at eight levels, from 6 to 76 dB SPL. Shifts in SOAE frequency and amplitude were observed for some subjects, for some tone conditions. Frequency shifts were always positive, whereas amplitude shifts were variable. No apparent pattern of tuning was seen, such that tones with a particular frequency relationship to the SOAEs induced greater changes in the SOAEs. Systematic changes in frequency and amplitude of SOAEs were observed for increasing level of broadband noise for all subjects. Results are discussed with respect to possible mechanism(s) responsible for the alterations in SOAEs: Transcranial conduction; the olivocochlear system; and/or the middle-ear reflex arc.

Characteristics of transient evoked otoacoustic emissions in normal-hearing and hearing-impaired children.

OBJECTIVE: Transient evoked otoacoustic emissions (TEOAEs) were measured in children with normal hearing and in children with hearing loss to investigate the characteristics of TEOAEs as they relate to overall amplitude and amplitude spectra of evoking stimuli, and to audiometric status. DESIGN: Three parameters of response measure (signal to noise ratio, amplitude and reproducibility) were assessed to determine accuracy of identification of varying degrees of hearing loss for broadband clicks, frequency-specific click bands and tone bursts. Forty-four children (66 ears) between 4 and 13 yr of age were evaluated for participation in the study. Fifty-nine ears with intact tympanic membranes were included in the final analyses: 14 ears with normal hearing and 45 ears with hearing loss. RESULTS: Children with normal hearing had robust responses that displayed nonlinear growth functions for broadband clicks and for tone bursts. Children with hearing loss had responses that decreased rapidly with decreasing stimulus level, if emissions were present at all. Data were analyzed using clinical decision analysis and receiver operator characteristic curves. Broadband clicks presented at 80 and 86 dB peSPL identified a hearing loss > or = 30 dB HL with a high degree of accuracy. Click responses filtered into octave bands centered at 500 and 1000 Hz did not improve classification of hearing loss, in fact, the 500 Hz band was particularly inaccurate. Results for click responses filtered into half-octave bands centered at 2000 and 4000 Hz were comparable with those for the broadband click, although the 2000 Hz band was superior for identification of hearing loss > or = 20 dB HL for an 80 dB peSPL click, and > or = 30 dB HL for an 86 dB peSPL click. Results for tone bursts centered at 500, 1000, 2000 and 4000 Hz, presented at 80 dB peSPL, were similar to results of the filtered click bands. Accuracy for identifying hearing loss increased with increasing center frequency. The 2000 and 4000 Hz tone bursts provided the best separation between normal-hearing and hearing-impaired ears, with 4000 Hz being slightly better. CONCLUSIONS: Data from this study suggest that TEOAEs in children can separate ears with normal hearing from those with hearing loss using a variety of stimulus and response conditions. Moreover, by using multiple stimulus levels it may be possible to distinguish between mild and moderate hearing losses.

Growth of Listeria monocytogenes and Yersinia enterocolitica colonies under modified atmospheres at 4 and 8 degrees C using a model food system.

The growth of Listeria monocytogenes and Yersinia enterocolitica colonies was studied on solid media at 4 and 8 degrees C under modified atmospheres (MAs) of 5% O2: 10% CO2: 85% N2 (MA1), 30% CO2: 70% N2 (MA2) and air (control). Colony radius, determined using computer image analysis, allowed specific growth rates (mu) and the time taken to detect bacterial colonies to be estimated, after colonies became visible. At 4 degrees C both MAs decreased the growth rates of L. monocytogenes by 1.5- and 3.0-fold under MA1 (mu = 0.02 h(-1)) and MA2 (mu = 0.01 h(-1)), respectively, as compared with the control (mu = 0.03 h(-1)). The time to detection of bacterial colonies was increased from 15 d (control) to 24 (MA1) and 29 d (MA2). At 8 degrees C MA2 decreased the growth rate by 1.5-fold (mu = 0.04 h(-1)) as compared with the control (mu = 0.06 h(-1)) and detection of colonies increased from 7 (control) to 9 d (MA2). At 4 degrees C both MAs decreased the growth rates of Y. enterocolitica by 1.5- and 2.5-fold under MA1 (mu = 0.03 h(-1)) and MA2 (mu = 0.02 h(-1)), respectively, as compared with the control (mu = 0.05 h(-1)). At 8 degrees C identical growth rates were obtained under MA1 and the control (mu = 0.07 h(-1)) whilst a decrease in the growth rate was obtained under MA2 (mu = 0.04 h(-1)). The detection of colonies varied from 6 (8 degrees C, aerobic) to 19 d (4 degrees C, MA2). Refrigerated modified atmosphere packaged foods should be maintained at 4 degrees C and below to ensure product safety.

Incidence of Campylobacter and Salmonella isolated from retail chicken and associated packaging in South Wales.

AIMS: To investigate the incidence of Campylobacter and Salmonella contamination associated with supermarket and butchers' shop chicken and related packaging. METHOD AND RESULTS: Three hundred raw samples (whole chicken, chicken breast with skin or chicken pieces) were purchased on a monthly basis for seven months. Packaging associated with the chicken was also sampled to provide isolation data for external and whole packaging. Campylobacter and Salmonella were isolated from 68% and 29% of retail chicken, respectively. Campylobacter was isolated from 3% of external and 34% of whole packaging overall. Salmonella was absent from external packaging but was isolated from 11% of whole packaging. No significant trends in isolation rates of the organisms were obtained during the period of sampling. CONCLUSIONS: The food industry and consumers should be made aware of the potential risk of Campylobacter and Salmonella on both the external and internal surfaces of packaging in addition to chicken itself. SIGNIFICANCE AND IMPACT OF THE STUDY: Chicken and chicken packaging is a potential vehicle for the introduction of pathogens in retail and domestic kitchens and in particular for the cross-contamination of Campylobacter and Salmonella.

Oxathiin carboxanilide, a potent inhibitor of human immunodeficiency virus reproduction.

Oxathiin carboxanilide (OC), NSC 615985, a compound originally synthesized as a potential fungicide, was demonstrated to be highly active in preventing human immunodeficiency virus (HIV)-induced cell killing and in inhibiting HIV reproduction. Virus-infected CD4+ lymphocytes were completely protected by 0.5 microM OC, whereas no toxicity was observed at concentrations below 50 microM OC. Production of infectious virus, viral p24 antigen, and virion reverse transcriptase were reduced by OC at concentrations that prevented viral cell killing. A variety of CD4+ T-cell lines were protected by OC from HIV cytopathicity, and OC inhibited two distinct strains of HIV-1. However, HIV-2 infections were unaffected by OC. OC had no direct effect on virions of HIV or on the enzymatic activities of HIV reverse transcriptase or HIV protease. Time-limited treatments of cells with OC before, during, or after exposure of cells to virus failed to protect cells from the eventual cytopathic effects of HIV, and OC failed to inhibit the production of virus from cells in which infection was established or from chronically infected cells. We conclude that the highly active OC has a reversible effect on some early stage of HIV-1 reproduction and cytopathicity. Pilot in vivo experiments showed that circulating concentrations of OC exceeding 1 microM could be achieved and sustained in hamsters for at least a week with no remarkable toxicological sequelae. OC represents a new class of anti-HIV agents that are promising candidates for drug development.

Identification of neonatal hearing impairment: experimental protocol and database management.

OBJECTIVE: The purposes of this article are to describe the overall protocol for the Identification of Neonatal Hearing Impairment (INHI) project and to describe the management of the data collected as part of this project. A well-defined protocol and database management techniques were needed to ensure that data were 1) collected accurately and in the same way across sites; 2) maintained in a database that could be used to provide feedback to individual sites regarding enrollment and the extent to which the protocol was complete on individual subjects; and 3) available to answer project questions. This article describes techniques that were used to meet these needs. DESIGN: This study was a prospective, randomized study that was designed to evaluate auditory brain stem responses, transient evoked otoacoustic emissions, and distortion product otoacoustic emissions as hearing-screening tools, and to relate neonatal test findings to hearing status, defined by visual reinforcement audiometry at 8 to 12 mo of age. Measures of middle-ear function also were obtained at some sites as part of the neonatal test battery. In addition, other clinical and demographic data were gathered to determine the extent to which factors, other than auditory status, influenced test behavior. Three groups were evaluated: neonatal intensive care unit (NICU) infants (those who spent 3 or more days in a NICU), well babies with risk factors for hearing loss, and well babies without risk factors. Six centers participated in the trial. The testers for the project included audiologists, technicians, audiology graduate students, and medical research staff. The same computerized neonatal test program was applied at each center. This program generated the neonatal test database automatically. Clinical and demographic data were collected by means of concise data collection forms and were entered into a database at each site. After the neonatal test, subjects from the NICU and at-risk well babies were evaluated with visual reinforcement audiometry starting at 8 to 12 mo of age. All data were electronically transmitted to the core site where they were merged into one overall database. This database was exercised to provide feedback and to identify discrepancies throughout the course of the study. In its final form, it served as the database on which all analyses were performed. RESULTS AND CONCLUSION: The protocol was a departure from typical hearing screening procedures in terms of 1) its regimented application of three screening measures; 2) the detailed information that was obtained regarding subject clinical and demographic factors; and 3) its application of the same procedures across six centers having diverse geographic location and subject demographics. A learning curve for successfully executing the study protocols was observed. Throughout the study, monthly reports were generated to monitor subject enrollment, check for data completeness, and to perform data integrity checks. In combination with monthly data reports and checks that occurred throughout the progression of the study, miscellaneous data audits were performed to check accuracy of neonatal testing programs and to cross-check information entered in the clinical and demographic database. The data management techniques used in this project helped to ensure the quality of the data collection process and also allowed for detailed analyses once data were collected. This was particularly important because it enabled us to evaluate not only the performance of individual measures as screening tools, but also permitted an evaluation of the influence of other variables on screening test results.

Structure-activity and cross-resistance evaluations of a series of human immunodeficiency virus type-1-specific compounds related to oxathiin carboxanilide.

A series of compounds related to the nonnucleoside reverse transcriptase (RT) inhibitor (NNRTI) oxathiin carboxanilide (UC84) were evaluated for activity against the human immunodeficiency virus (HIV) to determine structural requirements for anti-HIV activity. Twenty-seven compounds representative of the more than 400 Uniroyal Chemical Company (UC) compounds were evaluated for structure-activity relationships. Several of the compounds evaluated were highly active, with 50% effective concentrations in the nanomolar range and therapeutic indices of > 1,000. Highly synergistic anti-HIV activity was observed for each compound when used in combination with 3'-azido-3'-deoxythymidine; additive to slightly synergistic interactions were observed with the compounds used in combination with dideoxycytidine. In combination with the NNRTI costatolide, only UC38 synergistically inhibited HIV type 1. Residues in the RT which, when mutated, impart resistance to the virus isolates selected in cell culture, against virus variants with site-directed mutations, and against RTs containing defined single amino acid changes. The mutations included changes in RT amino acids 100, 101, 103, 106, 108, and 181. The results with isolates selected in cell culture indicate that the carboxanilide compounds interact with the RT at two vulnerable sites, selecting UC-resistant virus isolates with the Y-to-C mutation at position 181 (Y181C) or the L100I substitution. A resistant virus isolate containing both Y181C combination with calanolide A, an NNRTI which retains activity against virus with the single Y181C mutation, UC10 rapidly selected a virus isolate with the K103N mutation. The merits of selecting potential candidate anti-HIV agents to be used in rational combination drugs design as part of an armamentarium of highly active anti-HIV compounds are discussed.